CN105104755A - Compound phyllanthus urinaria preparation and application thereof in piglet diet - Google Patents
Compound phyllanthus urinaria preparation and application thereof in piglet diet Download PDFInfo
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Abstract
The invention provides a compound phyllanthus ruinaria preparation and an application thereof in piglet diet. The compound phyllanthus urinaria preparation is prepared by the steps that phyllanthus urinaria herbs and uraria crinita roots are mixed according to the ratio of 1:1 after impurities are removed and then processed through ultramicro-pulverization (the pulverization degree is 200-mesh). By adding the compound phyllanthus urinaria preparation in the basic piglet diet, environmental stress caused by weaning to piglets can be obviously reduced, the digestive ability of the gastrointestinal tract is improved, piglet growth is promoted, the antioxidant activity of the livers is improved, liver damage is reduced, the effects are related to the preparation adding doses, and the effect is most obvious by adding a high dose of the preparation. Compared with other health care products of the piglets, the compound phyllanthus ruinaria preparation has the advantages that the toxic and side effects are few, no reverse reaction exists, the functions of resisting oxidation, improving the organism immunity and the like are further achieved, the preparation can be used for a long time, the raw materials are easy to obtain and low in cost, the extraction separation technology is simple, production is convenient, and the development value is high.
Description
Technical field
The invention belongs to animal husbandry field, be specifically related to the application on a kind of compound Underleaf pearl formulation and piglet diet thereof.
Background technology
At present, along with China to livestock breeding industry attention, livestock breeding industry has had significant progress, particularly large scale of pig farm having qualitative leap especially in recent years.But the continuous expansion of Compact Develop, cultivation density greatly improves, drug abuse, the use of excessive or some the unconventional raw materials of the excessive pursuit speed of growth, feed nutrition, the mycotoxin contamination in feedstuff is day by day serious, and pig farm mycotoxin increasing pressure is large, further infringement is caused to swinery, cause swinery body condition decline swinery, occurred the disease taking liver diseases as principal character, affected growing and growth performance of swinery.
Chinese herbal medicine is the comparatively ideal additive of a class, not only has certain inhibitory action to mycotoxin, can also directly anti-bacteria and virus breeding, treatment swinery disease, thus minimizing antibiotic residue, the situation of the generation of drug-fast bacteria, is more and more taken seriously in Compact Develop.In Compact Develop, the effect of Chinese herbal medicine mainly contains the effect of following components.
Chinese herbal medicine can improve local flavor and the palatability of feed, Appetizing spleen-tonifying, and promoting the circulation of qi treatment for relieving indigestion and constipation, improves a poor appetite, and improves feed intake, obviously improves gastrointestinal function, improves digestive ferment secretion and vigor, and then promotes swinery growth.Former sea is forever waited and is added Chinese herbal medicine nursing growing and fattening pigs at basal diet, and test finds: growing and fattening pigs average weight weightening finish more than the control group 9.0kg(of test group
p< 0.01), feed conversion rate increases by 7.8%, remarkable in economical benefits.The beautiful brightness of official etc. utilizes the compound of the natural plants compositions such as honeysuckle, Radix Codonopsis, the Radix Astragali, garlic, adds in child care pig basal diet, omnidistance contrast.Found that: the average daily gain of each test group extremely significantly (
p< 0.01) higher than blank group, wherein the feedstuff-meat ratio of 1% Chinese herbal medicine group decreases compared with blank group, and significant difference (
p< 0.05), this compound Chinese herb can promote the growth of swinery as seen, reduces feedstuff-meat ratio.
Chinese herbal medicine can be dispelled heat, spleen benefiting and stimulating the appetite, the promoting flow of qi and blood circulation by heat clearly, strengthens metabolism, strengthens swinery physique and anti-stress ability.According to Yin Fugui etc., the neutrophil leucocyte of the weanling pig after Chinese herbal medicine of feeding, lymphocyte and the obvious increase compared with blank group of monocyte number (
p< 0.05), and the obviously obvious minimizing compared with blank group of MDA with NO content (
p< 0.05), compared with control group significant difference (
p< 0.05), illustrate this group Chinese herb compound can improve pigling immunity, alleviate wean cause to piglet stress injure.
Common Leafflower Herb (
phyllanthusurinariaL.) for Euphorbiaceae (
euphorbiaceae) Leafflower (
phyllanthusL.) plant, also known as Sagina subulata, spire pearl, Herba Phyllanthi Urinariae etc.Supplementary Amplifications of the Compendium of Materia Medica claims: this grass have clear liver and improve vision, the effect such as inducing diuresis for removing edema, eliminating inflammation and expelling toxin.Main component includes the materials such as lignanoids, organic acid, alkaloid, flavones, tannin class.Since Thyagarajan Late Cambrian has been delivered since Chinese herbal medicine Common Leafflower Herb has good effect to hepatitis type B virus, a lot of scholar starts the research to Common Leafflower Herb, the effect especially on protection liver.Tiger tail wheel (
urariacrinita) penetrate the herb of plant for long fringe cat tail, penetrate also known as timothy grass, iron gold mace, system sky grass, dog tail.Crinite uraria root main root is sturdy, cylindrical, and side root is many and thin, and coring quantity can reach tens; Main root diameter is about 2cm, color and luster cadmium yellow, and likeness in form ginseng, fragrant odour, therefore also referred to as " stone ginseng ".It is higher that it dries rate, can reach about 50%, relevant to old tender degree.The effects such as " Guangxi Chinese medicinal herbal " is middle claims it to have promoting qi circulation and relieving pain, clearing and antitussive, promoting blood circulation and removing obstruction in channels.Tiger tail wheel chemical composition is the material such as flavones, glycoside, amino acid mainly.
In recent years, the pharmacological research of Common Leafflower Herb and brave tail wheel is mainly concentrated on to anti-oxidation function, anti-liver injury and improves immunity of organisms aspect.In Common Leafflower Herb and brave tail wheel, all there is higher active material: flavone compound.Most of flavone compound has the function of obvious resisting oxidation free radical, and flavones belongs to phenolic acid class, and when there being some metal ion, this structure is easily oxidized, causes the unrighted acid on cell membrane protected, avoids oxidized.Also containing gallic acid material in Common Leafflower Herb, play obvious anti-oxidation function.The research such as Ma Xie shows: the alcohol extract Common Leafflower Herb with 70% 2 times, its general flavone content can reach 1.84%.Zhu Yuanping also finds in the research to brave tail wheel determination of total flavonoids, and containing more rich general flavone compound in crinite uraria root, content reaches 1.10mg/g.Blue or green according to Zheng Xiu, Common Leafflower Herb water extract hydroxy radical, superoxide anion have very strong Scavenging activity, illustrate that Common Leafflower Herb water extract has certain antioxidation.Find according to researchs such as Xiao Shuwen, crinite uraria root portion extract can remove Superoxide anion free radical, and effect is better than vitamin C, has good in vitro anti-oxidation.
Common Leafflower Herb element in Common Leafflower Herb and hypophyllanthin all have liver-protective effect, and flavonoid also has significantly protection liver effect, and this may be relevant with these material anti peroxidation of lipid.Zhou Shiwen etc. use Common Leafflower Herb and Effect of Phyllanthus Amarus Co. to mouse stomach administration, use CCl afterwards
4gavage, makes mouse liver injury model, finds, in test group serum, AST, ALT are active significantly reduces, and each dosage group hepatic injury degree has certain recovery, and visible Common Leafflower Herb and Effect of Phyllanthus Amarus Co. are to CCl
4the hepatic injury caused can have more significant repairing effect.Zhou Jun etc., by after phyllanthol extract gavage mouse, use CCl respectively
4, TAA and D-GN carry out lumbar injection to mouse, cause hepatic injury, found that, test group obviously reduces by CCl
4, TAA and D-GN destroy liver and cause the rising of glutamic-pyruvic transaminase (ALT), this and Common Leafflower Herb induce liver enzyme activity to have certain relation.According to Syamasunder etc., Cacumen Securinegae Suffruticosae extract can to CCl
4galactosamine causes the damage of primary cultured hepatocyte to play a protective role.Chen Qiu bravely waits and uses crinite uraria root flavone compound gavage to raise mouse, and continuous 21d, uses CCl afterwards
4intraperitoneal injection of mice, cause hepatic injury, the visible test group of result can obviously reduce AST, ALT, MDA level, and improves SOD, T-AOC, ALB, TP level, the hepatic injury pathologic structure change of each test group significantly alleviates, and illustrates that brave tail wheel chromocor compound is to CCl
4the chemical damage caused has obvious protected effect.
Summary of the invention
The object of the present invention is to provide a kind of compound Underleaf pearl formulation and the application on piglet diet thereof.
For achieving the above object, the present invention adopts following technical scheme:
A kind of compound Underleaf pearl formulation, after removing impurity by strain herb under leaf and crinite uraria root, 1:1 ratio mixes in mass ratio, forms through ultramicro grinding process (degree of grinding is at 200 orders).
The application of a kind of compound Underleaf pearl formulation in piglet diet.
The application of a kind of compound Underleaf pearl formulation in weanling pig basal diet.
The content of described compound Underleaf pearl formulation in daily ration is 0.025-0.035wt%.
Have no it and have promotion piglet growth, improve liver antioxidation activity, what alleviate hepatic injury effect has bibliographical information.
The invention has the advantages that:
1, compound Underleaf pearl formulation does not have report to have promotion piglet growth at present, improves liver antioxidation activity, alleviates hepatic injury effect.The present invention finds that compound Underleaf pearl formulation can promote piglet growth, improves and improves liver antioxidation activity, alleviate hepatic injury, and compared with other health products of existing pig, poison is secondary little, has no adverse reaction, can Long-Time Service; 2, compound Underleaf pearl formulation raw material is easy to get, inexpensive, and extraction and separation process is simple, and convenient for production, Development volue is high.
Accompanying drawing explanation
Fig. 1 blank group liver organization HE coloration result.
Fig. 2 model group liver organization HE coloration result.
Fig. 3 low dose group liver organization HE coloration result.
Dosage group liver organization HE coloration result in Fig. 4.
Fig. 5 high dose group liver organization HE coloration result.
Fig. 6 DDB control group liver organization HE coloration result.
Detailed description of the invention
Below in conjunction with embodiment, the present invention is described in detail:
The preparation of embodiment 1 compound Underleaf pearl formulation
Under every 5kg leaf, the 1:1 ratio mixing in mass ratio of strain herb and 5kg crinite uraria root, forms through ultramicro grinding process (degree of grinding is at 200 orders).
Embodiment 2 compound Underleaf pearl formulation is to the shadow of piglet growth performance
1 materials and methods
1.1 material
Male earner pig is castrated in the wean of (28 ± 1) d, raises on pig farm, Nan Ping.
Compound Underleaf pearl formulation: under every 5kg leaf, strain herb and 5kg crinite uraria root are in the mixing of 1:1 ratio, form through ultramicro grinding process (degree of grinding is at 200 orders).
experimental animal and design
Choose 30 ages in days and castrate male earner pig in the wean of (28 ± 1) d, average weight, at 8.87 ± 0.49kg, chooses 5 nursing cotes at same nursery house, cleans and hurdle of doing experiment after sterilizing.Test hurdle is set as 5 groups, and often organize 6 pigs, every hurdle repeats 1 pig, by the principle that body weight is close, is respectively dosage group in compound Common Leafflower Herb high dose group, compound Common Leafflower Herb, compound Common Leafflower Herb low dose group, DDB control group, blank group.
Wean castrates male earner swine rearing in same nursery house, and temperature, humidity are suitable, well-ventilated, free choice feeding and drinking-water, feeding cycle 28d.Duration of test, vaccine immunity and expelling parasite are by pig farm conventional program process.Experimental design scheme is in table 1.
Table 1 experimental design
1.3 assay method
Test breeding cycle 28d, before on-test, weanling pig was weighed, every 3d once weighs afterwards, the consumption of each experimental group feed every day is recorded in process of the test, the inspection registration every day spirit of each test group swinery, ight soil and hair color, register the death toll of each test group breathing problem, disease of digestive tract morbidity number and treatment situation, each test group simultaneously.
Average daily gain=average daily gain every day summation/test number of days
Average daily ingestion amount=every day feed consumption summation/(test number of days × each test group head number)
Feedstuff-meat ratio=every day feed consumption summation/(average daily gain every day summation × each test group head number)
The incidence of disease=often group morbidity number/each test group head number × 100%
1.4 data statistic analysis
All adopt Office2013Excell and SPSS19.0 statistical software to carry out statistical analysis to test data, data result with " mean ± standard deviation (
± SD) " represent, carry out Multiple range test by one-way analysis of variance and LSD method.
interpretation of result
Table 2 compound Underleaf pearl formulation is on the impact of piglet growth performance
Note: there is in same row different lowercase person, expression significant difference (
p<0.05), have different capitalization person represent difference extremely significantly (
p<0.01)
As known from Table 2, the average end of compound Common Leafflower Herb high dose group weighs and average daily gain obviously overweights blank group, and significant difference (
p< 0.05).In compound Common Leafflower Herb dosage group, compound Common Leafflower Herb low dosage, DDB control group and blank group difference not significantly (
p>0.05), illustrate that compound Underleaf pearl formulation has certain growth promoting function to pig, and have certain relation with additive capacity, DDB is not remarkable to the effect of promotion pig growth performance.The average daily ingestion amount difference of each test group not significantly (
p>0.05), compound Common Leafflower Herb is to raising swinery feed intake DeGrain.Between each test group feedstuff-meat ratio difference not significantly (
p>0.05).
discuss
3.1 compound Common Leafflower Herbs are on the impact of piglet growth performance
The wean of piglet enter nursery house be to piglet once greatly stress, comprise that feed changes stress, new growing environment stress, new colony formed stress, stress the waiting of vaccine immunity.Piggy after wean is also in growth and development process, during each viscera tissue organ all increases rapidly, and each function is all not yet perfect, particularly mycotoxin is being had to deposit in case, very easily cause weanling pig metabolic disorder, immune system is easy to suffer damage, and causes immunity degradation, and is vulnerable to the infringement of pathogenic microorganism.Further, ablactation stress can cause piglet micro ecology of gastrointestinal tract to be lacked of proper care, and affects piglet digesting and assimilating and growing feed.Research shows, is rich in active material in some Chinese herbal medicine, can effectively anti-bacteria breeding increase and anti-virus ability, improve gastrointestinal bacterial flora balance, improve pipe intestinal digesting absorbability, thus improve wean pig feed intake, strengthen anti-stress ability, improve the overall physique level of wean pig.Miao Xu etc. are added with the basal diet of Chinese herbal medicine by feeding to small weaning pig, and observe the influence of this formula to its growth performance, test finds, Chinese herb feed additive group end is heavy, daily gain increases all to some extent compared with blank group, and significant difference (
p< 0.01).This test, the average end of compound Common Leafflower Herb high dose is heavy, average daily gain is compared than blank and increased weight to some extent, and significant difference (
p< 0.05), the average end of dosage group in compound Common Leafflower Herb, low dose group, DDB control group is heavy, average daily gain and blank group difference remarkable (
p> 0.05), illustrate that compound Underleaf pearl formulation can improve wean pig growth performance, and have certain relation with additive capacity.
4 brief summaries
Compound Underleaf pearl formulation can reduce wean to piglet cause stress, improve GI digestion power, so promote piglet growth, improve food conversion ratio.
Embodiment 3 compound Underleaf pearl formulation is on the impact of the anti-carbon tetrachloride hepatic injury of piglet
1 materials and methods
1.1 material
Male earner pig is castrated in the wean of (28 ± 1) d, raises on pig farm, Nan Ping.
Peanut oil: Luhua Group Co., Ltd., Shandong;
Bifendate: Community in Baiyunshan, Guangzhou group of stars medicine company limited company;
Carbon tetrachloride (CCl
4): Chemical Reagent Co., Ltd., Sinopharm Group;
AST, ALT, ADA, LDH kit: be all purchased from Nanjing and build up Bioengineering Research Institute;
Digital display electronic thermostat water bath: Changzhou Guohua Electric Appliance Co., Ltd.;
18 series ultraviolet visible spectrophotometer: Beijing Puxi General Instrument Co., Ltd;
Peace section AnkeTGL-16G-A high speed desktop refrigerated centrifuge: Anting Scientific Instrument Factory, Shanghai;
1.2 experimental animals and design
Method, with example 2, adds a group model group.
Table 3 experimental design
1.3 assay method
1.3.1 the preparation of liver injury model
Preparation 5%CCl
4peanut oil: first measure 5ml/CCl with graduated cylinder
4, pour in beaker, then measure 95mL peanut oil, fully stir, make it mix, matching while using before test.Each test group is fed after 28d, fasting 12h, presses 1.5ml/kg body weight lumbar injection 5%CCl respectively
4peanut oil solvent in compound Common Leafflower Herb high dose group, dosage group in compound Common Leafflower Herb, compound Common Leafflower Herb low dose group, DDB control group.In model group, by 1.5ml/kg body weight lumbar injection peanut oil solvent in blank group control group.After 24h, model is set up.
sample collection
After modeling 24h, carry out vena cava anterior blood sampling respectively to each test group hemostix, whole blood sample is placed in centrifuge tube, at room temperature slant setting 2h after gathering, and until whole blood coagulation and after separating out serum, then puts into centrifuge, with 3000rmin
-1centrifugal 15min, gets serum and is placed in vitro ,-40 DEG C of preservations.
sample determination
AST, ALT, ADA, LDH press the UL of kit
-1description method of operating, measures the content of each index respectively.
interpretation of result
2.1 compound Underleaf pearl formulations are on the impact of piglet serum alt and AST activity
The measurement result of table 4AST and ALT
As known from Table 4, in model group, the active and AST activity of ALT is all apparently higher than each test group, and difference extremely remarkable (
p< 0.01), visible lumbar injection CCl
4cause hepatocellular injury, and model group hepatocellular injury is the most serious.The ALT of compound Common Leafflower Herb high dose group is active and AST activity is all extremely remarkable (
p< 0.01) lower than dosage group, compound Common Leafflower Herb low dose group, DDB control group in compound Common Leafflower Herb, illustrate that these four groups of test group hepatic injury degree have certain difference, compound Common Leafflower Herb high dose group degree of injury is minimum, may be relevant with addition, embody the dependence effect of doses.In compound Common Leafflower Herb, low dose group, DDB contrast group difference not significantly (
p> 0.05), illustrate that these three groups of hepatocellular injury degree are similar.
compound Underleaf pearl formulation is on the impact of ADA and LDH activity in piglet serum
The measurement result of table 5ADA and LDH
As known from Table 5, in model group ADA active and LDH activity is all extremely remarkable (
p< 0.01) higher than each test group, illustrate that liver cell sustains damage, and model group hepatocellular injury is the most serious.Active and the LDH activity of the ADA of compound Common Leafflower Herb high dose group all lower than dosage group, compound Common Leafflower Herb low dose group, DDB control group in compound Common Leafflower Herb, and difference extremely remarkable (
p< 0.01).Illustrate, these four groups of test group hepatic injury degree have certain difference, and compound Common Leafflower Herb high dose group degree of injury is minimum, may be relevant with additive capacity, embodies the dependence effect of doses.In compound Common Leafflower Herb, pearl low dose group, DDB contrast group difference not significantly (
p> 0.05), illustrate that these three groups of hepatocellular injury degree are similar.
3 discuss
Common chemically hepatotoxicant matter has CCl
4, ethanol, D-Gal amine etc.CCl
4acute hepatic injury model is drugs to the common model one of of liver with or without toxic action and the effect of medicine anti-liver injury.CCl
4there is liver property, after being absorbed by body, by the metabolic activation of liver cell pigment P450, CCl
4c-Cl key homolysis in molecule, thus generate trichloromethyl free radical (CCl
3), then under aerobic conditions, be oxidized to and there is highly active trichloromethyl peroxylradicals (CCl
3oO), start the phospholipid molecule attacked on liver plasma membrane afterwards, cause liver lipid peroxidation, there is a large amount of degraded in membrane phospholipid, finally causes the destroy integrity of liver plasma membrane, affect its 26S Proteasome Structure and Function.Liang Daoming etc. are by the CCl of various dose
4intraperitoneal injection of mice, sets up acute liver, and result shows lumbar injection CCl
4the hepatic injury that mouse is stable can be set up out, the CCl of various dose
4determine the degree difference to some extent of hepatic injury.Long-tail BUGY CRAXONE etc. is being inquired into experimental liver injury model, injects CCl by the mode in gavage, abdominal cavity
4, cause acute liver injury of rats, found that lumbar injection CCl
4the liver injury model effect set up is better; This experiment adopts 5%CCl
4peanut oil solvent lumbar injection piggy, attacks poison to liver, ALT, the AST of model group, ADA, LDH activity all higher than blank group, and difference extremely significantly (
p< 0.01), visible 5%CCl
4peanut oil solvent lumbar injection causes obvious damage to piglet liver, and the modeling of acute chemical liver injury model is set up.
The effect that 3.1 compound Common Leafflower Herbs are active to ALT in Swine serum and AST is active
ALT and AST is non-specific cell internal strength energy enzyme, and ALT major part exists liver, and be secondly in the tissues such as kidney, cardiac muscle, skeletal muscle, AST mainly exists in the tissues such as cardiac muscle, liver, skeletal muscle, kidney.In normal swine serum, comparision contents is low, but when injuries of tissues and organs, cytoplasma membrane permeability increases, and intracellular ALT and AST just will enter blood, causes enzymatic activity in serum to raise.So serum alt height that is active and AST activity can as the indicateing arm observing hepatic injury.Li Lanlan etc., by the intervention of Cacumen Securinegae Suffruticosae extract to L-02 cell, put into CCl afterwards
4cultivate in cultivation, carry out every Biochemical Indexes such as ALT, AST afterwards, found that nutrient solution supernatant, in the nutrient solution of the L-02 cell of Cacumen Securinegae Suffruticosae extract intervention, ALT and AST is active obviously reduces, and illustrates that L-02 cellular damage obviously alleviates; After Su Jiehan etc. give mouse stomach one week compound Cacumen Securinegae Suffruticosae extract, disposable to mouse peritoneal injection CCl
4peanut oil solvent, causes acute liver, finds CCl
4in model group, ALT and AST is active significantly raises, ALT and the AST activity of compound Common Leafflower Herb group is starkly lower than and model group, and difference extremely significantly (
p<0.01), visible lumbar injection CCl
4mouse liver can be caused to damage, and serum alt and AST activity raise.This result of the test shows, 5%CCl
4after peanut oil solvent lumbar injection, in each test group all there is rising in various degree in AST and ALT activity, and wherein in model group, AST and ALT activity is the highest; Compared with model group, each test group AST of compound Underleaf pearl formulation and ALT activity all decrease, and difference extremely significantly (
p<0.01), after visible compound Underleaf pearl formulation can reduce hepatic injury, AST and ALT activity raises, and alleviates hepar damnification Be very effective.
3.2 compound Underleaf pearl formulations are to the effect of LDH and ADA activity in Swine serum
Lactic dehydrogenase (LDH) is glycolytic ferment, mainly exist in histiocytic kytoplasm, wherein content is the highest in kidney, can catalysis lactic acid be pyruvic acid, enzyme activity is in the tissue significantly higher than the vigor in serum, as long as so tissue is subject to slight damage or necrosis, in tissue, lactic dehydrogenase (LDH) enters into blood, and the lactic dehydrogenase (LDH) in blood will there will be significant rising.So when liver organization sustains damage, in blood, lactic dehydrogenase (LDH) vigor will there will be obvious rising.Adenosine deamination acid (ADA) is relevant nucleic acid metabolism enzyme active in Cellular Immunity, adenosine deamination acid (ADA) activity is the sensitive indicator of reflection hepatic injury, serum adenosine deamination acid (ADA) major part is all discharge by liver, when liver sustains damage, cause the permeability of liver plasma membrane to strengthen, the activity of the ADA enzyme in serum will raise.The bright grade of Chen Du is experimental animal with Tilapia mossambica, and the Common Leafflower Herb of various dose of feeding, uses CuSO
4poisoningly cause Tilapia mossambica hepatic injury, found that LDH and the ADA activity in model group significantly improves, the test group LDH of Common Leafflower Herb of feeding and ADA activity extremely significantly (
p<0.01) lower than model group, illustrate that hepatic injury can cause LDH and ADA activity in blood to raise.This result of the test shows, and compares with model group, compound Underleaf pearl formulation each test group serum ADA and LDH activity extremely significantly (
p<0.01) reduce, visible compound Underleaf pearl formulation can protect liver successful.
4 brief summaries
In this test, the compound Underleaf pearl formulation adding various dose in wean pig basal diet is fed after 28d, lumbar injection CCl
4, cause acute liver damage, by the mensuration of serum alt, AST, ADA, LDH, compound Common Leafflower Herb high dose group, middle dosage group, low dose group group compared with model group, inequality heteropole significantly (
p<0.01), and compound Common Leafflower Herb high dose group and middle dosage, low dose group difference extremely significantly (
p<0.01).Visible, compound Underleaf pearl formulation has significantly protection and repair to piglet hepatic injury.
Embodiment 4 compound Common Leafflower Herb is on the impact of piglet liver organization structure
1 materials and methods
1.1 material
The concentrated sulfuric acid, potassium bichromate, formaldehyde (being mixed with the solution of 10%), ethanol (being mixed with 50% respectively, 70%, 80%, 90%, 95%, 100% solution), dimethylbenzene, have more than needed Chemical Co., Ltd. in Tianjin
Fusing point 50-52 DEG C paraffin I, 52-54 DEG C paraffin II, 56-58 DEG C paraffin III, beeswax, neutral gum, traditional Chinese medicines Group Co., Ltd
Asepsis environment-protecting H-E dye liquor kit: Bioengineering Research Institute is built up in Nanjing
AE31 is inverted electron microscope: Motic company
Paraffin slicing machine: Leica company
202-2A digital display Constant Temp. Oven: Shanghai and minister instrument manufacturing Co., Ltd.
method
1.2.1 the collection of liver organization
Each test group lumbar injection 5wt%CCl
4after peanut oil solvent 24h, respectively get two for each group, bloodletting takes liver, and the liver of taking-up puts into ice physiological saline, then liver is cut into the tissue block of size 1cm × 1cm × 0.5cm, fixes more than 48h in 10wt% formalin.
liver histopathology checks
Hepatic tissue section forming steps
Hepatic tissue blocking is taken out in 1 washing, rinses 24h with water.
The flushed hepatic tissue blocking of 2 dehydration,---70% alcohol (60min)---80% alcohol (60min)---95% alcohol (60min)---100% alcohol (40min)---100% alcohol (40min) of putting into 50% alcohol (60min) in order.
3 transparent dimethylbenzene: alcohol=1:1(45min)---dimethylbenzene (8min)---dimethylbenzene: paraffin=1:1(30min).
4 waxdip 50-52 DEG C paraffin I (45min)---52-54 DEG C of paraffin II (45min)---56-58 DEG C of paraffin III (45min).
Hepatic tissue blocking is swung in embedded box, FFPE by 5 embeddings.
After embedded hepatic tissue blocking is carried out repairing block by 6 sections, be fixed on slicer and cut into slices, slice thickness is 5 μm.
The hepatic tissue sheet cut is put into 40 DEG C of water-baths and is extended piece by 7 exhibition sheets.
Albumen glycerine spreads upon on slide by 8 bonding dies, and pick up from water-bath with the section of slide by expansion afterwards, keep flat and dry into thermostatic drying chamber, bake out temperature is 50 DEG C, and the time is 24h.
The dried paraffin section dimethylbenzene of 9 dewaxing rehydration carries out dewaxing twice, each 10min, then puts into concentration 100%, 95%, 90%, 80%, 70% alcoholic solution 10min at different levels, uses water rinse 3min afterwards.
Liver tissue slices after 10 dyeing rehydrations carries out HE dyeing.
11 mounting neutral gums carry out mounting.
12 microscopic examinations carry out pathologic examination under section being placed in light microscopic.
result
2.1 blank group liver microstructures
Microscopic observation blank group liver organization, visible lobuli hepatis clear in structure, liver cell is without obvious sex change, necrosis, and liver rope is normal radial arrangement, and without inflammatory cell infiltration around lobuli hepatis central area, liver cell nuclear obviously (as Fig. 1).
model group liver microstructure
Microscopic observation model group liver organization, visible model group pathological change is obvious, and extent of disease involves whole lobuli hepatis, liver cell arrangement disorder, downright bad, swelling, the visible a large amount of necrosis region of lobuli hepatis central area, have more inflammatory cell, liver cell nuclear be concentrated, disintegration or disappearance (as Fig. 2).
compound Common Leafflower Herb low dose group liver microstructure
Microscopic observation compound Common Leafflower Herb low dose group liver organization, visible lobuli hepatis profile is more clear, and lobuli hepatis central area has necrosis region, visible have inflammatory cell, moderate steatosis, central liver rope fracture, misaligned, there are liver cell nuclear disappearance, disintegration (as Fig. 3).
dosage group liver microstructure in compound Common Leafflower Herb
Dosage group liver organization in Microscopic observation compound Common Leafflower Herb, visible lobuli hepatis profile is substantially clear, and lobuli hepatis central necrosis district reduces, the steatosis of slight hepatic cell, liver rope presents the neat radioactivity arrangement of a part, there is the nucleus (as Fig. 4) of a small amount of regeneration.
compound Common Leafflower Herb high dose group liver microstructure
Microscopic observation compound Common Leafflower Herb high dose group liver organization, visible lobuli hepatis is clear, and lobuli hepatis mesozone necrosis region is less, and liver cell is swelling a little, and liver rope becomes radioactivity marshalling substantially, visible part Binucleate Hepatocytes (as Fig. 5).
dDB control group liver microstructure
Microscopic observation DDB control group liver organization, visible lobuli hepatis structural deterioration is not obvious, and there is necrotic area lobuli hepatis mesozone, has inflammatory cell around.Liver rope ruptures on a small quantity, and nucleus has a small amount of disappearance (as Fig. 6).
discuss
3.1CCl
4
the Histopathologic change of caused acute chemical liver injury piggy hepatocellular injury
Liver is removing toxic substances organ important in body, primarily of liver cell composition, comprises nucleus, cytoplasm, mitochondria, endoplasmic reticulum, golgiosome etc. in liver cell.Lobuli hepatis is made up of a large amount of liver cells, and liver cell nuclear is rounded, and liver cell nuclear is impaired, just liver cell disintegration is described.Liver cell is the arrangement in strand radioactivity centered by central vein, when liver sustains damage, liver cell morphosis will change, and occurs sex change and necrosis, liver rope arrangement disorder, the metabolism of body and various functions all can be affected, and body will morbid state.Common chemically hepatotoxicant matter has CCl
4, ethanol, D-Gal amine, cortex hormone of aadrenaline thioacetamide etc.CCl
4acute hepatic injury model is drugs to the classical animal model one of of liver with or without toxic action and medicine anti-liver injury.The Puerarin gavage such as Zhao Chunjing raises 5d, uses CCl afterwards
4hypodermic injection rat, causes acute hepatic injury model, and result is visible, and model group liver obviously damages, and has occurred massive inflammatory cells infiltrated, nuclear hyperchromatism, liver rope irregular arrangement, and the injury of liver of Puerarin of feeding obviously alleviates.This test adopts lumbar injection 5%CCl
4peanut oil solvent causes the acute chemical liver injury of child care pig, and in visible model group, the pathology of hepatic injury has appearred in liver organization under microscopic visualization.Extent of disease involves whole lobuli hepatis, liver rope arrangement disorder, downright bad, swelling, and the visible a large amount of necrosis region of lobuli hepatis central area, have more inflammatory cell, liver cell nuclear disappears.CCl
4being chemically poisonous substance, is cause the most classical conventional material of chemical damage, from the hepatocellular a series of pathology of model group, adopts lumbar injection CCl
4the acute chemical liver injury making piglet is feasible.
the mechanism of hepatic injury and compound Underleaf pearl formulation are to the repair of hepatic injury
CCl
4there is liver property, after being absorbed by body, by the metabolic activation of liver cell pigment P450, CCl
4c-Cl key homolysis in molecule, thus generate trichloromethyl free radical (CCl
3), then under aerobic conditions, be oxidized to and there is highly active trichloromethyl peroxylradicals (CCl
3oO), start the phospholipid molecule attacked on liver plasma membrane afterwards, cause liver lipid peroxidation, there is a large amount of degraded in membrane phospholipid, finally causes the destroy integrity of liver plasma membrane, affect its 26S Proteasome Structure and Function.This peroxidating and liver cell inflammatory degenerative close relation.Hepatocellular inflammatory degenerative shows as cell granulations sex change, bubble sex change, inflammatory cell infiltration, steatosis and meronecrosis etc.This test lumbar injection CCl
4after modeling, the hepatic injury situation of each dosage group of compound Common Leafflower Herb is compared with model group, there is alleviating to a certain degree, gently the most obvious with compound Common Leafflower Herb high dose group degree of injury, visible hepatocellular structure is recovered normally substantially, and lobuli hepatis is clear, lobuli hepatis mesozone necrosis region is less, liver cell is swelling a little, and liver rope becomes radioactivity marshalling substantially, visible part Binucleate Hepatocytes.Illustrate that compound Underleaf pearl formulation is to CCl
4the acute liver damage caused improves significantly, and repairing effect is remarkable.
brief summary
By observing the Histopathology of liver, compound Underleaf pearl formulation all serves repair in various degree to liver cell, and hepatocellular sex change and necrosis alleviate all to some extent, and visible compound Underleaf pearl formulation serves obvious anti-liver injury effect.
Claims (3)
1. a compound Underleaf pearl formulation, is characterized in that: described compound Underleaf pearl formulation is in mass ratio for 1:1 ratio mixes after removing impurity by strain herb under leaf and crinite uraria root, and form through ultramicro grinding process, degree of grinding is at 200 orders.
2. the application of an a kind of compound Underleaf pearl formulation as claimed in claim 1 in piglet diet.
3. application according to claim 2, is characterized in that: the content of described compound Underleaf pearl formulation in daily ration is 0.025-0.035wt%.
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