CN113521059A - New application of pueraria lobata small molecules in regulating alcohol dependence behavior - Google Patents
New application of pueraria lobata small molecules in regulating alcohol dependence behavior Download PDFInfo
- Publication number
- CN113521059A CN113521059A CN202111036980.7A CN202111036980A CN113521059A CN 113521059 A CN113521059 A CN 113521059A CN 202111036980 A CN202111036980 A CN 202111036980A CN 113521059 A CN113521059 A CN 113521059A
- Authority
- CN
- China
- Prior art keywords
- alcohol
- tolerance
- daidzin
- dependence
- puerarin
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 208000007848 Alcoholism Diseases 0.000 title claims abstract description 48
- 201000007930 alcohol dependence Diseases 0.000 title claims abstract description 44
- 235000010575 Pueraria lobata Nutrition 0.000 title claims abstract description 19
- 150000003384 small molecules Chemical class 0.000 title claims abstract description 18
- 230000001105 regulatory effect Effects 0.000 title claims abstract description 13
- 241000219781 Pueraria montana var. lobata Species 0.000 title claims abstract 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 187
- ZQSIJRDFPHDXIC-UHFFFAOYSA-N Daidzein Natural products C1=CC(O)=CC=C1C1=COC2=CC(O)=CC=C2C1=O ZQSIJRDFPHDXIC-UHFFFAOYSA-N 0.000 claims abstract description 149
- RXUWDKBZZLIASQ-UHFFFAOYSA-N Puerarin Natural products OCC1OC(Oc2c(O)cc(O)c3C(=O)C(=COc23)c4ccc(O)cc4)C(O)C(O)C1O RXUWDKBZZLIASQ-UHFFFAOYSA-N 0.000 claims abstract description 50
- 235000007240 daidzein Nutrition 0.000 claims abstract description 50
- HKEAFJYKMMKDOR-VPRICQMDSA-N puerarin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1C1=C(O)C=CC(C2=O)=C1OC=C2C1=CC=C(O)C=C1 HKEAFJYKMMKDOR-VPRICQMDSA-N 0.000 claims abstract description 50
- GMTUGPYJRUMVTC-UHFFFAOYSA-N Daidzin Natural products OC(COc1ccc2C(=O)C(=COc2c1)c3ccc(O)cc3)C(O)C(O)C(O)C=O GMTUGPYJRUMVTC-UHFFFAOYSA-N 0.000 claims abstract description 49
- KYQZWONCHDNPDP-UHFFFAOYSA-N Daidzoside Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=C2C(=O)C(C=3C=CC(O)=CC=3)=COC2=C1 KYQZWONCHDNPDP-UHFFFAOYSA-N 0.000 claims abstract description 49
- KYQZWONCHDNPDP-QNDFHXLGSA-N daidzein 7-O-beta-D-glucoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=C2C(=O)C(C=3C=CC(O)=CC=3)=COC2=C1 KYQZWONCHDNPDP-QNDFHXLGSA-N 0.000 claims abstract description 49
- 230000007774 longterm Effects 0.000 claims abstract description 28
- 230000001154 acute effect Effects 0.000 claims abstract description 27
- 230000035945 sensitivity Effects 0.000 claims abstract description 27
- 230000006399 behavior Effects 0.000 claims abstract description 22
- 241000219780 Pueraria Species 0.000 claims abstract description 14
- 206010048010 Withdrawal syndrome Diseases 0.000 claims abstract description 11
- 230000035622 drinking Effects 0.000 claims description 20
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 6
- 241000628997 Flos Species 0.000 claims description 5
- 201000010099 disease Diseases 0.000 claims description 5
- 230000000694 effects Effects 0.000 abstract description 34
- 241000255588 Tephritidae Species 0.000 description 47
- 241000255581 Drosophila <fruit fly, genus> Species 0.000 description 46
- 244000046146 Pueraria lobata Species 0.000 description 10
- 238000012549 training Methods 0.000 description 9
- 238000003556 assay Methods 0.000 description 6
- 239000003814 drug Substances 0.000 description 6
- 238000000034 method Methods 0.000 description 5
- IKHGUXGNUITLKF-UHFFFAOYSA-N Acetaldehyde Chemical compound CC=O IKHGUXGNUITLKF-UHFFFAOYSA-N 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 230000004044 response Effects 0.000 description 4
- 238000011282 treatment Methods 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 3
- 241000255925 Diptera Species 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 239000007640 basal medium Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000012546 transfer Methods 0.000 description 3
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 208000025746 alcohol use disease Diseases 0.000 description 2
- 239000001045 blue dye Substances 0.000 description 2
- 230000006735 deficit Effects 0.000 description 2
- AUZONCFQVSMFAP-UHFFFAOYSA-N disulfiram Chemical compound CCN(CC)C(=S)SSC(=S)N(CC)CC AUZONCFQVSMFAP-UHFFFAOYSA-N 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- 201000009032 substance abuse Diseases 0.000 description 2
- 101710088194 Dehydrogenase Proteins 0.000 description 1
- 102000017934 GABA-B receptor Human genes 0.000 description 1
- 108060003377 GABA-B receptor Proteins 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- WJBLNOPPDWQMCH-MBPVOVBZSA-N Nalmefene Chemical compound N1([C@@H]2CC3=CC=C(C=4O[C@@H]5[C@](C3=4)([C@]2(CCC5=C)O)CC1)O)CC1CC1 WJBLNOPPDWQMCH-MBPVOVBZSA-N 0.000 description 1
- 206010028813 Nausea Diseases 0.000 description 1
- 208000007107 Stomach Ulcer Diseases 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 208000029650 alcohol withdrawal Diseases 0.000 description 1
- 108010081577 aldehyde dehydrogenase (NAD(P)+) Proteins 0.000 description 1
- VWYANPOOORUCFJ-UHFFFAOYSA-N alpha-Fernenol Chemical compound CC1(C)C(O)CCC2(C)C3=CCC4(C)C5CCC(C(C)C)C5(C)CCC4(C)C3CCC21 VWYANPOOORUCFJ-UHFFFAOYSA-N 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000003542 behavioural effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- UDSAIICHUKSCKT-UHFFFAOYSA-N bromophenol blue Chemical compound C1=C(Br)C(O)=C(Br)C=C1C1(C=2C=C(Br)C(O)=C(Br)C=2)C2=CC=CC=C2S(=O)(=O)O1 UDSAIICHUKSCKT-UHFFFAOYSA-N 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000001627 detrimental effect Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 229960002563 disulfiram Drugs 0.000 description 1
- 229940000406 drug candidate Drugs 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 230000002996 emotional effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000003777 experimental drug Substances 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 201000005917 gastric ulcer Diseases 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000010197 meta-analysis Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229960005297 nalmefene Drugs 0.000 description 1
- 229960003086 naltrexone Drugs 0.000 description 1
- DQCKKXVULJGBQN-XFWGSAIBSA-N naltrexone Chemical compound N1([C@@H]2CC3=CC=C(C=4O[C@@H]5[C@](C3=4)([C@]2(CCC5=O)O)CC1)O)CC1CC1 DQCKKXVULJGBQN-XFWGSAIBSA-N 0.000 description 1
- 239000003887 narcotic antagonist Substances 0.000 description 1
- 230000008693 nausea Effects 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 238000001543 one-way ANOVA Methods 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 206010036067 polydipsia Diseases 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 208000020016 psychiatric disease Diseases 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229940126586 small molecule drug Drugs 0.000 description 1
- 229960003928 sodium oxybate Drugs 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/488—Pueraria (kudzu)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/30—Drugs for disorders of the nervous system for treating abuse or dependence
- A61P25/32—Alcohol-abuse
Abstract
The invention discloses a new application of a pueraria flower micromolecule for regulating alcohol dependence behaviors, provides a basis for the pueraria flower micromolecule to treat alcohol dependence, discloses an application of the pueraria flower micromolecule to reducing relative alcohol consumption and alcohol preference in a withdrawal reaction period, discloses an application of the pueraria flower micromolecule to reducing alcohol sensitivity and acute alcohol tolerance, and discloses an application of the pueraria flower micromolecule to improving long-term alcohol tolerance. The invention determines that the pueraria flower micromolecules can obviously regulate alcohol dependence behaviors and have a protection effect on body damage caused by alcohol; the function of treating alcohol dependence of the small molecules of the pueraria lobata through multi-component, multi-path and multi-target point is determined; it is determined that puerarin, daidzin and daidzein have effects of regulating alcohol dependence behavior and treating alcohol dependence.
Description
Technical Field
The invention belongs to the biomedical technology, and particularly relates to a new application of pueraria lobata flower micromolecules in regulating alcohol dependence behaviors.
Background
The WHO published a Global Drinking and health report "Global status report on alcohol and health 2018" in 2018. Reports indicate that nearly 300 million people die in 2016 due to alcohol drinking, accounting for 5.3% of the total death worldwide; the prevalence of alcohol use disability and alcohol dependence in the population of Chinese in 2016 has reached worldwide levels, with 6% of men and 1% of women in China dying from alcohol-related illness; alcohol dependence seriously impairs physical health of drinkers, such as gastric ulcer, liver cirrhosis, irreversible disorders of the nervous system and the like, and a trend toward younger drinking has also been shown in recent years. Therefore, the research on alcohol dependence has important medical and social values.
Alcohol Dependence (AD) has become a worldwide medical and sociological challenge since the twentieth century. Internationally, it is commonly recognized as an Alcohol Use Disorder (AUD). The features of alcohol use disorders include uncontrolled excessive drinking, compulsive drinking, and negative emotional states when not drinking, which may lead to relapse of alcohol dependence. Alcohol use disorders are defined by the operating criteria of the handbook of diagnosis and statistics of mental Disorders (DSM) and the International Classification of Diseases (ICD): despite negative psychological, physical, behavioral and social consequences, alcohol consumption continues, which must be brought to diagnostic compliance (minimum 12 months) during the course of treatment. Alcohol use impairment is diagnosed according to DSM-5 and the severity is measured by the number of eligible doses. In ICD-11, this disease is diagnosed as either "alcohol dependence", which is a more severe manifestation of alcohol use impairment, or "detrimental alcohol use patterns".
There are many drugs available worldwide for the treatment of alcohol dependence including disulfiram, dehydrogenase inhibitors, opioid antagonists naltrexone, nalmefene, and the like. All of these treatments were validated with different indications supported by Meta analysis. The reason why the patient feels discomfort such as nausea, headache, accelerated heartbeat and the like when abstinence from alcohol and sulfur is caused is that the patient can generate acute toxic reaction in vivo after taking alcohol, and acetaldehyde oxidation is blocked through the activity of acetaldehyde dehydrogenase, so that the concentration of acetaldehyde in the body is obviously increased. Sodium oxybate, mainly by interacting with GABA-B receptors, has a high risk of abuse. Although the curative effect of the medicaments is confirmed in the long-term clinical application process, the side effects are also prominent, such as possible medicament abuse, adverse effects on the spirit and body of patients and poor medicament metabolism, and another prominent side effect is that the patients who have abstinence from alcohol can drink the medicaments quickly and repeatedly.
Disclosure of Invention
One of the purposes of the invention is to provide a new application of the pueraria lobata flower micromolecules for regulating alcohol dependence behaviors and provide a basis for the pueraria lobata flower micromolecules to treat alcohol dependence.
The other purpose is to provide the application of the pueraria flower micromolecules in reducing the relative drinking amount and alcohol preference in the withdrawal reaction period.
The third purpose is to provide the application of the pueraria lobata small molecules in reducing alcohol sensitivity and acute alcohol tolerance.
The fourth purpose is to provide the application of the pueraria flower micromolecules in improving long-term alcohol tolerance.
In order to further realize the invention, the pueraria flower micromolecules adopt puerarin, daidzin or daidzein.
The puerarin which can realize the purpose of the invention can reduce the relative drinking amount, alcohol preference, alcohol sensitivity and acute alcohol tolerance in the withdrawal reaction period when being 1-5 mmol/L; puerarin can improve long-term alcohol tolerance at 2.5-5 mmol/L.
The content of daidzin in the extract can be reduced to 0.25-5mmol/L, so that the relative drinking capacity and alcohol preference in the withdrawal reaction period can be reduced; the daidzin can reduce alcohol sensitivity and acute alcohol tolerance at 1-5 mmol/L; the daidzin can improve long-term alcohol tolerance at 0.25-2.5 mmol/L.
The daidzein which can realize the purpose of the invention can reduce the relative drinking capacity and alcohol preference in the withdrawal reaction period when the daidzein is 1-2.5 mmol/L; the daidzin can reduce alcohol sensitivity and acute alcohol tolerance at 1-5mmol/L, and can improve long-term alcohol tolerance.
The fifth purpose of the invention is to provide the application of the pueraria flower micromolecule in treating alcohol dependence and diseases caused by the alcohol dependence.
Compared with the prior art, the invention has the beneficial effects that:
the fruit fly alcohol dependence behavior model is constructed, and on the basis, the small pueraria lobata molecules are found and determined to be capable of remarkably adjusting alcohol dependence behavior and have a protection effect on body damage caused by alcohol; the method finds and determines that the small molecules of the pueraria lobata flower play a role in treating alcohol dependence through 'multi-component-multi-path-multi-target point'; it is found and determined that puerarin, daidzin and daidzein have the effects of regulating alcohol dependence behavior and treating alcohol dependence.
Drawings
Figure 1 is a graph of the effect of puerarin on alcohol preference during the withdrawal reaction phase of drosophila alcohol (n = 150);
fig. 2 is the effect of puerarin on the relative alcohol consumption during the withdrawal reaction period of drosophila alcohol (n = 150);
figure 3 is the effect of puerarin on drosophila alcohol sensitivity (n = 150);
fig. 4 is the effect of puerarin on drosophila acute alcohol tolerance (n = 150);
fig. 5 is the effect of puerarin on long-term alcohol tolerance of drosophila (n = 150);
figure 6 is the effect of daidzin on alcohol preference during the withdrawal response period of drosophila alcohol (n = 150);
fig. 7 is a graph of the effect of daidzin on the relative alcohol consumption during the withdrawal response period for drosophila alcohol (n = 150);
figure 8 is the effect of daidzin on drosophila alcohol sensitivity (n = 150);
fig. 9 is the effect of daidzin on acute alcohol tolerance of drosophila (n = 150);
fig. 10 is the effect of daidzin on long term alcohol tolerance of drosophila (n = 150);
figure 11 is a graph of the effect of daidzein on alcohol preference during the drosophila alcohol withdrawal response (n = 150);
fig. 12 is a graph showing the effect of daidzein on the relative alcohol consumption during the withdrawal response period of drosophila alcohol (n = 150);
figure 13 is the effect of daidzein on drosophila alcohol sensitivity (n = 150);
figure 14 is the effect of daidzein on acute alcohol tolerance of drosophila (n = 150);
fig. 15 is the effect of daidzein on long-term alcohol tolerance of drosophila (n = 150).
Detailed Description
The invention is further described with reference to the following figures and detailed description.
The application of the pueraria lobata small molecule for regulating alcohol dependence behaviors:
1. principal material
1.1 Primary reagents
Bromophenol blue dye (Sigma-alorich, MKCD 9610), agar powder (Cali-Bio, CB 300005), sucrose (Sigma-alorich, WXBC 9911V), yeast extract (OXOID, 4255382-02), sterile, enzyme-free water (Beijing Soilebao Tech Co., Ltd., 20201116).
1.2 Experimental animals
Male drosophila melanogasterw 1118 (5905) From the Blumeton Drosophila resource center. Setting a fruit fly constant-temperature illumination incubator: the fruit flies are raised in the incubator at the temperature of 25 +/-1 ℃, the relative humidity of 45-65% and the illumination time of 12 h/darkness of 12 h.
1.3 Experimental drugs
Puerarin (Puerarin, Shanghai-sourced leaf Biotechnology Co., Ltd., LOT: SO2M 91354875), Daidzein (Daidzin, Shanghai-sourced leaf Biotechnology Co., Ltd., LOT: P11J11FII 5623), Daidzein (Daidzein, Shanghai-sourced leaf Biotechnology Co., Ltd., LOT: CO6NY 5504)
2. Experimental methods
2.1 preparation of Small-molecule drug-containing culture Medium for flos Puerariae Lobatae
Firstly, preparing 1% agar, subpackaging culture tubes with 4-5mL per bottle, solidifying the agar, putting into a small test tube cover, and cooling for later use; then, 50mL of dye food solution was prepared. According to a molar concentration calculation method, the micromolecular solution and the blue dye food solution are mixed, so that the final concentration of each micromolecular drug is 0.25mmol/L, 1mmol/L and 5 mmol/L.
2.2 Experimental method for influence of pueraria flower micromolecules on fruit fly alcohol dependence behaviors
2.2.1 relative alcohol consumption and alcohol preference determination
(1) The puerarin determines the relative drinking capacity and alcohol preference after the fruit fly alcohol is abstined: after 2 days of recovery, the flies were randomly divided into 2 groups and transferred to 2 groups of bottles containing different solutions in gaseous atmosphere for 3 days. Wherein, the solution on the filter paper of the control group is 100 microliter of pure water; the experimental group was alcohol training group and the solution on the filter paper was 100 μ L75% alcohol. After 30min of each training, transfer back to basal medium. Then, the fruit flies in the alcohol training group are divided into a blank group, 0.25mmol/L puerarin, 1mmol/L puerarin and 5mmol/L puerarin for intervention during the withdrawal period, and the drinking capacity and the alcohol preference of the fruit flies are measured after the withdrawal period.
(2) The relative drinking capacity and alcohol preference of the daidzin to the drosophila alcohol after withdrawal are determined: after 2 days of recovery, the flies were randomly divided into 2 groups and transferred into 2 groups of children containing different solutions in gaseous atmosphere for 3 days. Wherein, the solution on the filter paper of the control group is 100 microliter of pure water; the experimental group was alcohol training group and the solution on the filter paper was 100 μ L75% alcohol. After 30min of each training, transfer back to basal medium. Then, the fruit flies in the alcohol training group are divided into a blank group, 0.25mmol/L daidzin, 1mmol/L daidzin and 5mmol/L daidzin for intervention during the withdrawal period, and the alcohol drinking amount and the alcohol preference of the fruit flies are measured after the withdrawal period.
(3) Determination of relative alcohol consumption and alcohol preference of daidzein to drosophila alcohol: after 2 days of recovery, the flies were randomly divided into 2 groups and transferred to 2 groups of bottles containing different solutions in gaseous atmosphere for 3 days. Wherein, the solution on the filter paper of the control group is 100 microliter of pure water; the experimental group was alcohol training group and the solution on the filter paper was 100 μ L75% alcohol. After 30min of each training, transfer back to basal medium. Then, the fruit flies in the alcohol training group are divided into a blank group, 0.25mmol/L daidzein, 1mmol/L daidzein and 5mmol/L daidzein for intervention during the withdrawal period, and the relative alcohol drinking amount and alcohol preference of the fruit flies are measured after the withdrawal period.
2.2.2 sensitivity, acute alcohol tolerance assay
(1) Determination of puerarin sensitivity and acute alcohol tolerance: after the fruit flies recover for 2 days, the fruit flies are randomly divided into 4 groups, a blank group, 0.25mmol/L puerarin, 1mmol/L puerarin and 5mmol/L puerarin, and after puerarin intervenes for 3 days, ST50 of the first alcohol exposure of each group of fruit flies is measured by using a sensitive device, namely the first alcohol exposure of each group of fruit flies to alcoholSensitivity of (2), 4h Re-alcohol Exposure determination of Drosophila ST501Acute alcohol tolerance of each group of drosophila was calculated.
(2) Daidzin sensitivity, acute alcohol tolerance assay: after the fruit flies recover for 2 days, the fruit flies are randomly divided into 4 groups, a blank group, 0.25mmol/L daidzin, 1mmol/L daidzin and 5mmol/L daidzin, after 3 days of daidzin intervention, ST50 of the first alcohol exposure of each group of fruit flies is measured by using a sensitive device, namely the sensitivity of each group of fruit flies to alcohol, and ST50 of the fruit flies is measured by 4 hours of alcohol exposure again1Acute alcohol tolerance of each group of drosophila was calculated.
(3) Daidzein sensitivity, acute alcohol tolerance assay: after 2 days of fruit fly recovery, the fruit flies are randomly divided into 5 groups, a blank group, 0.25mmol/L daidzein, 1mmol/L daidzein and 5mmol/L daidzein, after 3 days of daidzein intervention, ST50 of first alcohol exposure of each group of fruit flies is measured by using a sensitive device, namely the sensitivity of each group of fruit flies to alcohol, and ST50 of the fruit flies is measured by 4 hours of second alcohol exposure1Acute alcohol tolerance of each group of drosophila was calculated.
2.2.3 Long term alcohol tolerance assay
(1) Determination of puerarin long-term alcohol tolerance: after the fruit flies recover for 2 days, the fruit flies are randomly divided into 4 groups, a blank group, 0.25mmol/L puerarin, 1mmol/L puerarin and 5mmol/L puerarin, the puerarin intervenes for 4 days, ST50 of the fruit flies is measured by a sensitive device every 24 hours, and the long-term alcohol tolerance of the fruit flies is calculated.
(2) Daidzin long term alcohol tolerance assay: after the fruit flies recovered for 2 days, the fruit flies were randomly divided into 4 groups, a blank group, 0.25mmol/L daidzin, 1mmol/L daidzin and 5mmol/L daidzin, the daidzin intervention was carried out for 4 days, ST50 of the fruit flies was measured every 24 hours by using a sensitive device, and the long-term alcohol tolerance of the fruit flies was calculated.
(3) Daidzein long-term alcohol tolerance assay: after the fruit flies recover for 2 days, the fruit flies are randomly divided into 4 groups, a blank group, 0.25mmol/L daidzein, 1mmol/L daidzein and 5mmol/L daidzein, the daidzein is intervened for 4 days, ST50 of the fruit flies is measured by a sensitive device every 24 hours, and the long-term alcohol tolerance of the fruit flies is calculated.
2.3 statistical analysis
Experimental data are expressed as mean ± s.e.m. In the experimental process, 10 male fruit flies are used in each bottle, 3-5 bottles are used in each batch, and the total number of batches is more than or equal to 3. All statistical treatments were performed using GraphpadPrism8 statistical analysis software. And analyzing and comparing the difference of each group of Drosophila flos puerariae flower after each small molecule is dried on the relative alcohol consumption, alcohol preference and the like by adopting a Two-tailedTTest method. And comparing the variation of the sensitivity, acute alcohol tolerance and long-term alcohol tolerance behavior of each group after each group of Drosophila and Pueraria lobata micromolecular stem prognosis by adopting an One-wayANOVA method.PThe difference is significant if < 0.05,Pthe more remarkable is that < 0.01,P< 0.001 is extremely significant.
3. Results of the experiment
3.1 Effect of Puerarin on Drosophila alcohol preference and relative alcohol consumption
As can be seen from FIG. 1, the alcohol preference of the model group is increased relative to the blank group (P<0.001); (1 mmol/L, 2.5mmol/L, 5mmol/L puerarin reduced alcohol preference after withdrawal relative to model group: (P<0.001，P<0.01）。
As can be seen from FIG. 2, the relative alcohol consumption of the model group was significantly increased relative to the blank group (P<0.001); relative to the model group, puerarin 1mmol/L, 2.5mmol/L, and 5mmol/L can reduce the relative alcohol consumption of Drosophila ()P<0.001，P<0.01）。
Therefore, the puerarin can reduce the alcohol preference and the relative drinking amount after the fruit flies are abstinent.
3.2 Effect of Puerarin on Drosophila alcohol sensitivity and acute alcohol tolerance
As can be seen from FIG. 3, it was found that different concentrations of puerarin did not significantly affect ST50 (relative to the blank group) (ST 50)P>0.05), which indicates that puerarin does not affect the sensitivity of fruit flies to alcohol.
As can be seen from FIG. 4, 1mmol/L, 2.5mmol/L, 5mmol/L puerarin reduced acute alcohol tolerance of Drosophila (R) ((R))P<0.05）。
Therefore, puerarin can reduce acute alcohol tolerance of drosophila to alcohol.
3.3 Effect of Puerarin on Long-term alcohol tolerance of Drosophila
As can be seen in FIG. 5, 5mmol/L puerarin significantly increased alcohol tolerance relative to the blank group 24h after puerarin administration (first alcohol exposure) ((S))P<0.05); 48h after puerarin administration (third alcohol exposure), 2.5mmol/L and 5mmol/L puerarin significantly increased alcohol tolerance relative to the blank group (P<0.01，P<0.01); after puerarin is administrated for 72 hours (fourth alcohol exposure), 1mmol/L, 2.5mmol/L and 5mmol/L puerarin significantly increase the alcohol tolerance relative to the blank group ((P<0.01); 96h after puerarin administration (fifth alcohol exposure); 2.5mmol/L and 5mmol/L puerarin significantly increased alcohol tolerance relative to the blank group: (P<0.05，P<0.01）。
Thus, it is shown that puerarin can increase long-term alcohol tolerance with time, i.e., puerarin can improve long-term alcohol tolerance of drosophila.
3.4 Effect of Glycine on Drosophila alcohol preference and relative alcohol consumption
As can be seen from FIG. 6, the alcohol preference of the model group is increased relative to the blank group (P<0.001); relative to the model group, 0.25mmol/L, 1mmol/L, 2.5mmol/L, 5mmol/L daidzin reduced alcohol preference after the withdrawal of Drosophila: (P<0.01，P<0.0001）。
As can be seen from FIG. 7, the relative alcohol consumption of the model group was significantly increased relative to the blank group (P<0.001); relative to the model group, the relative alcohol consumption of the fruit flies can be reduced by 0.25mmol/L, 1mmol/L, 2.5mmol/L and 5mmol/L daidzeinP<0.001，P<0.01）。
Thus, it was shown that daidzin can reduce alcohol preference and relative alcohol consumption after withdrawal from drosophila.
3.5 Effect of daidzin on Drosophila alcohol sensitivity and acute alcohol tolerance
As can be seen from FIG. 8, 1mmol/L, 2.5mmol/L, and 5mmol/L of daidzin ST50 were all increased by (relative to the blank group) ((P<0.001，P<0.05), which shows that 1mmol/L, 2.5mmol/L and 5mmol/L daidzin can reduce the sensitivity of fruit flies to alcohol.
As can be seen from FIG. 9, 1mmol/L, 2.5mmol/L, 5mmol/L daidzin significantly reduced acute alcohol tolerance of Drosophila (R) ((R))P<0.05）。
Therefore, the daidzin can reduce the sensitivity and acute alcohol tolerance of the fruit flies.
3.6 Effect of daidzin on Long-term alcohol tolerance of Drosophila
As can be seen from FIG. 10, 1mmol/L daidzin significantly increased alcohol tolerance relative to the blank group 24h after daidzin administration (r) ((r))P<0.05); (Soyaside significantly increased alcohol tolerance at 0.25mmol/L, 1mmol/L, 2.5mmol/L relative to blank group 48h after daidzin administration (third alcohol exposure) (P<0.01，P<0.01); after 72h of daidzin administration (fourth alcohol exposure), both 0.25mmol/L and 1mmol/L of daidzin significantly increased alcohol tolerance relative to the blank group (C) ((C))P<0.01); after 96h of daidzin administration (fifth alcohol exposure); (5 mmol/L daidzin significantly reduced alcohol tolerance relative to the blankP<0.01）。
Thus, low concentrations of daidzin were shown to improve long-term alcohol tolerance in drosophila.
3.7 Effect of daidzein on Drosophila alcohol preference and relative alcohol consumption
As can be seen from FIG. 11, the model group's preference for alcohol increases over the blank group (P<0.001); (1 mmol/L and 2.5mmol/L daidzein reduced alcohol preference of drosophila after withdrawal relative to model group: (P<0.05）。
As can be seen from FIG. 12, the relative alcohol consumption of the model group was significantly increased relative to the blank group (P<0.001); 1mmol/L and 2.5mmol/L daidzein can reduce the relative alcohol intake of Drosophila relative to model group: (P<0.05）。
Therefore, the daidzein can reduce the alcohol preference and the relative drinking amount after the fruit fly is abstinent.
3.8 Effect of daidzein on Drosophila alcohol sensitivity and acute alcohol tolerance
As can be seen from FIG. 13, 1mmol/L, 2.5mmol/L relative to the blank group(5 mmol/L daidzein ST50 has different degrees of significant increaseP<0.0001), which shows that daidzein can reduce the sensitivity of fruit fly to alcohol.
As can be seen from FIG. 14, 1mmol/L, 2.5mmol/L, 5mmol/L daidzein reduced acute alcohol tolerance of Drosophila (R) ((R))P<0.05，P<0.0001）。
Therefore, the daidzein can reduce the sensitivity and acute alcohol tolerance of the fruit flies.
3.9 Effect of daidzein on Long-term alcohol tolerance of Drosophila
As can be seen from FIG. 15, the ST50 of the fruit flies is a general increasing trend as time increases overall. After the daidzein is administrated for 24 hours (first alcohol exposure), compared with the blank group at the same period, 1mmol/L daidzein can obviously prolong the alcohol tolerance (P<0.05); after 48h of daidzein administration (second alcohol exposure), 1mmol/L, 2.5mmol/L and 5mmol/L daidzein significantly extended alcohol tolerance relative to the same blank group; (daidzein, 1mmol/L, 2.5mmol/L, and 5mmol/L of daidzein significantly extended alcohol tolerance relative to the same blank after 72h of daidzein administration (third alcohol exposure) (P<0.01); also after daidzein administration for 96h (fourth alcohol exposure), 1mmol/L and 2.5mmol/L daidzein significantly extended alcohol tolerance relative to the same blank group (ii) ((iii))P<0.01）。
Thus, daidzein can affect long-term alcohol tolerance in drosophila.
In conclusion, the pueraria lobata small molecules can be used for regulating alcohol dependence behaviors and can be applied to treating alcohol dependence. Wherein puerarin, daidzin and daidzein all have effects of regulating alcohol dependence behavior and treating alcohol dependence, so can be used for treating alcohol dependence and diseases caused by alcohol dependence.
Claims (8)
1. New application of pueraria lobata small molecules in regulating alcohol dependence behaviors is characterized in that: the flos Puerariae Lobatae small molecule can be used for reducing relative drinking amount and alcohol preference in withdrawal reaction period.
2. New application of pueraria lobata small molecules in regulating alcohol dependence behaviors is characterized in that: the flos Puerariae Lobatae small molecule can be used for reducing alcohol sensitivity and acute alcohol tolerance.
3. New application of pueraria lobata small molecules in regulating alcohol dependence behaviors is characterized in that: the application of the pueraria flower micromolecule in improving long-term alcohol tolerance.
4. Novel use of a small pueraria flower molecule according to any one of claims 1 to 3 for modulating alcohol dependence behaviour, characterized in that: the flos Puerariae Lobatae small molecule is puerarin, daidzin or daidzein.
5. The novel use of small molecules of pueraria lobata as claimed in claim 4 for modulating alcohol dependence behavior, characterized in that: when the puerarin is 1-5mmol/L, the relative drinking capacity and alcohol preference, alcohol sensitivity and acute alcohol tolerance in the withdrawal reaction period are reduced; the puerarin can improve long-term alcohol tolerance when concentration is 2.5-5 mmol/L.
6. The novel use of small molecules of pueraria lobata as claimed in claim 4 for modulating alcohol dependence behavior, characterized in that: when the daidzin is 0.25-5mmol/L, the relative drinking capacity and alcohol preference in the withdrawal reaction period are reduced; the daidzin is 1-5mmol/L to reduce alcohol sensitivity and acute alcohol tolerance; the daidzin content of 0.25-2.5mmol/L can improve long-term alcohol tolerance.
7. The novel use of small molecules of pueraria lobata as claimed in claim 4 for modulating alcohol dependence behavior, characterized in that: when the daidzein is 1-2.5mmol/L, the relative drinking capacity and alcohol preference in the withdrawal reaction period are reduced; the daidzin content of 1-5mmol/L can reduce alcohol sensitivity and acute alcohol tolerance, and improve long-term alcohol tolerance.
8. Novel use of a small pueraria flower molecule according to any one of claims 1 to 7 for modulating alcohol dependence behaviour, characterized in that: the application of the pueraria lobata small molecule in treating alcohol dependence and diseases caused by the alcohol dependence.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111036980.7A CN113521059A (en) | 2021-09-06 | 2021-09-06 | New application of pueraria lobata small molecules in regulating alcohol dependence behavior |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111036980.7A CN113521059A (en) | 2021-09-06 | 2021-09-06 | New application of pueraria lobata small molecules in regulating alcohol dependence behavior |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113521059A true CN113521059A (en) | 2021-10-22 |
Family
ID=78123011
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111036980.7A Pending CN113521059A (en) | 2021-09-06 | 2021-09-06 | New application of pueraria lobata small molecules in regulating alcohol dependence behavior |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113521059A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2112703A1 (en) * | 1991-07-01 | 1993-01-02 | Bert L. Vallee | Method for the inhibition of aldh-i useful in the treatment of alcohol dependence or alcohol abuse |
| CN1671373A (en) * | 2002-06-27 | 2005-09-21 | 人类生物研究基金公司 | Compounds useful for the inhibition of ALDH |
| CN102846771A (en) * | 2012-07-04 | 2013-01-02 | 贵州省生物研究所 | Composition with antialcoholism and hepatoprotective effect |
| CN104430890A (en) * | 2014-12-12 | 2015-03-25 | 长春科技学院 | Pueraria flower juice and milk beverage and preparation method thereof |
-
2021
- 2021-09-06 CN CN202111036980.7A patent/CN113521059A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2112703A1 (en) * | 1991-07-01 | 1993-01-02 | Bert L. Vallee | Method for the inhibition of aldh-i useful in the treatment of alcohol dependence or alcohol abuse |
| US5204369A (en) * | 1991-07-01 | 1993-04-20 | The Endowment For Research In Human Biology | Method for the inhibition of aldh-i useful in the treatment of alcohol dependence or alcohol abuse |
| CN1671373A (en) * | 2002-06-27 | 2005-09-21 | 人类生物研究基金公司 | Compounds useful for the inhibition of ALDH |
| CN102846771A (en) * | 2012-07-04 | 2013-01-02 | 贵州省生物研究所 | Composition with antialcoholism and hepatoprotective effect |
| CN104430890A (en) * | 2014-12-12 | 2015-03-25 | 长春科技学院 | Pueraria flower juice and milk beverage and preparation method thereof |
Non-Patent Citations (6)
| Title |
|---|
| DAVID M.PENETAR，等: "A single dose of kudzu extract reduces alcohol consumption in a binge drinking paradigm", 《DRUG ALCOHOL DEPEND》 * |
| RENEE C. LIN，等: "Isoflavonoid Compounds Extracted from Pueraria lobata Suppress Alcohol Preference in a Pharmacogenetic Rat Model of Alcoholism", 《ALCOHOLISM: CLINICAL AND EXPERIMENTAL RESEARCH》 * |
| 周吉银，等: "葛根总黄酮及葛根素解酒的药理研究进展", 《中国医院药学杂志》 * |
| 彭景华，等: "葛根及其主要异黄酮成分解酒药理研究进展", 《中草药》 * |
| 杨馥嘉，等: "中医药治疗酒精依赖的研究进展", 《中国药物依赖性杂志》 * |
| 陈荣富，等: "酒依赖治疗药物研究进展", 《中国药物滥用防治杂志》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104450590B (en) | One strain has bacillus subtilis and the application thereof of bacteriostatic activity | |
| CN113755409B (en) | Bifidobacterium longum for relieving insulin resistance and application thereof | |
| CN110777098A (en) | Lactobacillus with effect of preventing and treating diabetic encephalopathy and separation method and application thereof | |
| CN101120950A (en) | Application of realgar microorganism extracting liquid in preparing anti-tumor medicine | |
| KR102623984B1 (en) | Cannabinoid compositions and their use in the manufacture of drugs for treating neurodegenerative diseases, including Parkinson's disease and Alzheimer's disease. | |
| CN111329982B (en) | Novel application of bupleurum tenue granules in combination with chloroquine phosphate | |
| CN113521059A (en) | New application of pueraria lobata small molecules in regulating alcohol dependence behavior | |
| EP2116252A1 (en) | The use of epimedium flavones and effective components thereof for the preparation of medicaments of promoting proliferations and differentiations of nerve cells | |
| CN115569156B (en) | Ginseng and ganoderma lucidum fungus extract for remarkably improving bronchial asthma inflammation and application thereof | |
| Cannon et al. | A rapid technique for producing ethanol dependence in the rat | |
| CN109381473A (en) | Scutelloside is in the application for preparing gestational diabetes Fetal neurotubules malformation drug | |
| CN114081893A (en) | Application of combined anthocyanin and pectin in preparation of medicine for preventing and/or treating non-alcoholic fatty liver disease | |
| CN1268382C (en) | Blood lead content reducing medicine for treating plumbism | |
| CN103099779A (en) | Prescription and preparation process for irinotecan hydrochloride injection | |
| CN102600210B (en) | A kind of compound DM Cough agent and preparation method thereof | |
| CN108743697A (en) | A kind of application of littleleaf indianmulberry root and stem extract in treating acute viral pharyngitis | |
| CN109248193A (en) | A kind of pharmaceutical composition and preparation method thereof for snoring | |
| CN110974878B (en) | Traditional Chinese medicine spray for treating avian infectious rhinotracheitis of green-shell laying hens and preparation method thereof | |
| TWI837529B (en) | Use of lactobacillus fermentum gkf3 for preparing oral composition of reducing tobacco addiction | |
| US20060024327A1 (en) | Composition for controlling blood sugar | |
| CN106902175A (en) | A kind of Chinese medicine ointment for treating perianal eczema | |
| CN1965837A (en) | Ofloxacin solution containing clustered water used for animal | |
| TW202317161A (en) | Use of lactobacillus fermentum gkf3 for preparing oral composition of reducing tobacco addiction | |
| CN105997962B (en) | Application of coenzyme Q10 in treating periodic paralysis | |
| CN115386523A (en) | Lactococcus lactis and application thereof in resisting helicobacter pylori infection |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20211022 |
|
| RJ01 | Rejection of invention patent application after publication |