CN104257877A - Applications of Xinnaoxin capsule in preparation of medicines treating cerebral ischemia-reperfusion injury - Google Patents
Applications of Xinnaoxin capsule in preparation of medicines treating cerebral ischemia-reperfusion injury Download PDFInfo
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- CN104257877A CN104257877A CN201410276400.5A CN201410276400A CN104257877A CN 104257877 A CN104257877 A CN 104257877A CN 201410276400 A CN201410276400 A CN 201410276400A CN 104257877 A CN104257877 A CN 104257877A
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- 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)
- A61K36/41—Crassulaceae (Stonecrop family)
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- A—HUMAN NECESSITIES
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- 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)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/81—Solanaceae (Potato family), e.g. tobacco, nightshade, tomato, belladonna, capsicum or jimsonweed
- A61K36/815—Lycium (desert-thorn)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
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Abstract
Applications of a Xinnaoxin capsule in preparation of medicines treating cerebral ischemia-reperfusion injury are disclosed. The applications prove that: the capsule significantly reduces behavioral scores, a cerebral infarction rate, a NO content and NOS activity of rats, improves blood rheological dynamic indexes and relaxes NE-precontracted vessels in a concentration-dependent manner.
Description
Technical field
The invention belongs to field of medicaments, be specifically related to the application of XINNAOXIN JIAONANG in preparation treatment cerebral ischemia reperfusion injury medicine.
Background technology
At present, ischemic cerebrovascular has a strong impact on human survival quality.Cerebral ischemia, also known as cerebral infarction, is that a class causes ischemia, anoxia due to cerebral blood flow supply obstacle, cause the disease of the necrosis of limitation brain tissue ischemia or cerebral malacia, and cerebral ischemia reperfusion injury is the important pathophysiological mechanism causing multiple cerebrovascular disease.Cerebral ischemia reperfusion injury refers to that ischemic tissue of brain is after recovery hemoperfusion, and brain tissue impairment increases the weight of further, and its mechanism is not yet thoroughly illustrated.In recent years, along with the progress of shock treatment on clinical medicine and applying of artery bypass art and thrombolytic therapy etc., make the research of ischemical reperfusion injury become one of project of current concern, wherein Drug protective effect also more and more comes into one's own.
XINNAOXIN JIAONANG is S&P Pharmaceutical Co., Ltd.'s product, and the accurate word Z20025866 of traditional Chinese medicines, cures mainly supplementing QI and nourishing YIN, blood circulation promoting and blood stasis dispelling.But act in treatment cerebral ischemia reperfusion injury and have not been reported.
Summary of the invention
Goal of the invention: the new opplication that the object of the present invention is to provide XINNAOXIN JIAONANG.
The invention discloses the application of XINNAOXIN JIAONANG in preparation treatment cerebral ischemia reperfusion injury medicine.
Above-mentioned XINNAOXIN JIAONANG, is preferably made up of following parts by weight component: Radix Rhodiolae 3 ~ 6 parts, Fructus Lycii 5 ~ 15 parts, Fructus Hippophae 3 ~ 9 parts.
Beneficial effect: present invention demonstrates XINNAOXIN JIAONANG and obviously can reduce the neurological deficit score of rat, cerebral infarction rate, NO content and NOS vigor, improve hemorheology dynamics index, and can the pre-shrunk blood vessel of concentration dependent ground diastole NE.
Accompanying drawing explanation
Fig. 1 is the impact of XINNAOXIN JIAONANG on MCAO rat model cerebral infarction;
Fig. 2 is the impact (HE dye, × 100) of XINNAOXIN JIAONANG on MCAO rat model neuron morphology;
Fig. 3 is the impact (HE dye, × 200) of XINNAOXIN JIAONANG on MCAO rat model neuron morphology;
Fig. 4 XINNAOXIN JIAONANG is on the impact of the diastolic rate of NE preshrinking blood vessel
note: compare with sham operated rats,
#p<0.05,
##p<0.01; Compare with model group,
*p<0.05,
*p<0.01.
Detailed description of the invention
Embodiment 1:
XINNAOXIN JIAONANG by parts by weight be the Radix Rhodiolae of 3 parts, the Fructus Hippophae of the Fructus Lycii of 5 parts and 3 parts mixes.
Embodiment 2:
XINNAOXIN JIAONANG by parts by weight be the Radix Rhodiolae of 4 parts, the Fructus Hippophae of the Fructus Lycii of 10 parts and 6 parts mixes.
Embodiment 3:
XINNAOXIN JIAONANG by parts by weight be the Radix Rhodiolae of 5 parts, the Fructus Hippophae of the Fructus Lycii of 12 parts and 8 parts mixes.
Embodiment 4:
XINNAOXIN JIAONANG by parts by weight be the Radix Rhodiolae of 6 parts, the Fructus Hippophae of the Fructus Lycii of 15 parts and 9 parts mixes.
Embodiment 5:
The zoopery of XINNAOXIN JIAONANG treatment cerebral ischemia reperfusion injury
1 experiment material
1.1 medicines and reagent
XINNAOXIN JIAONANG, content is chocolate brown powder, 0.5g/ grain, lot number: 120803, S&P Pharmaceutical Co., Ltd.; During experiment, remove capsule shell, grinding content, is configured to desired concn with distilled water.Hydergine, specification: 1mg, lot number: 2L850T, Tianjin Huajin Pharmaceutical Co., Ltd.; During experiment, be ground to powder, be configured to desired concn with distilled water.
Nitric oxide (NO) test kit, nitricoxide synthase (NOS) test kit, the bright test kit of coomassie all build up Science and Technology Ltd. purchased from Nanjing; TTC is purchased from Nanjing You Sibo company limited.Other reagent are analytical pure.
1.2 instrument
Electronic balance (Beijing Sai Duolisi balance company limited); Microplate reader (Thermo, the U.S.); 96 well culture plates (Costar, the U.S.); Electric homogenate machine; Beijing Sai Kexide SA-6000 blood rheological instrument; Beijing Sai Kexide SC-2000 platelet aggregation instrument; Desk centrifuge Kubota5800 (Kubo field company, Japan); The super calorstat of numerical control; Muscular tension sensor, model JH-2; Constant temperature smooth muscle groove, model HW-400SE; BL-410 biological functional system, is more than Chengdu TME Technology Co., Ltd.'s product.
1.3 laboratory animal
Adult male SD rats, body weight 200 ~ 250g, is provided by Yangzhou University's comparative medicine center, laboratory animal production licence number: SCXK (Soviet Union) 2012-0004.Animal sub-cage rearing, keep 12h circadian rhythm, room temperature 22 ± 1 DEG C, freely drinks water and ingests.
2 experimental techniques
2.1 administrations, grouping and modeling
Rat is divided into 6 groups at random: sham operated rats, model group, positive drug group, XINNAOXIN JIAONANG low dose group, middle dosage group and high dose group.Gastric infusion, every day 1 time, successive administration 7d; Dosage: hydergine group is 0.8mg/kg/d, the glad basic, normal, high dosage group of heart and brain is 100,200,400mg/kg/d, perform the operation after last administration 1h.Model group and sham operated rats gavage same volume water.Corresponding index is detected respectively at after Reperfu-sion 1h, 24h.
The preparation of rat medium-sized artery thromboembolism model (middle cerebral artery occlusion, MCAO): operation adopts improvement Longa line brush to copy middle cerebral artery cerebral ischemia re-pouring model on the right side of rat.After rat 10% chloral hydrate lumbar injection (1g/kg) anesthesia, lain on the back fixing.Be separated right common carotid artery (CCA), internal carotid artery (ICA) and external carotid artery (ECA) hanging wire is for subsequent use, ligation ECA and CCA, after closing ICA distal end with bulldog clamp folder, a kerf is made rapidly in ECA and ICA crotch, insert heating one end from incision and become smooth, spherical fishing line (diameter is 0.25mm, marks apart from 2cm place, pommel).After line inserts ICA, ligation fishing line and porch ICA section slightly in porch, then unclamp the bulldog clamp that folder closes ICA, slightly withdraws after continuing to insert fishing line to slightly resistance, be (18.5 ± 0.5) about mm to line insertion depth, realize middle cerebral artery occlusion and cause cerebral ischemia.Ligation porch again, stays about 1cm, skin suture outside fishing line.The end of a thread that stays of lift gently after 2h is to there being resistance, and realizing middle cerebral artery fills with again, then modeling completes.A sham operated rats ligation ECA and ICA.In whole modeling process, keep animal anus temperature at about 37 DEG C.
2.2 rat functional defect scorings
Whole scoring adopts mono blind method, and namely scoring person does not know laboratory animal grouping situation.Adopt 5 points of standards of grading of longa.0 be divided into asymptomatic; 1 be divided into can not full extension offside fore paw; 2 are divided into and turn-take to offside; 3 are divided into and topple over to offside; 4 are divided into and can not walk voluntarily, loss of consciousness.Reject 0 point, 4 points and postoperative death animal, the animal reaching 1,2,3 point is modeling success.The scoring of rat functional defect is measured respectively after recovering blood supply 1h and 24h.
The preparation of 2.3 brain tissue homogenates
After rat femoral gets blood, put to death by its de-cervical vertebra immediately, take out cerebral tissue, rinsing in the normal saline of pre-cooling on ice bath, removing blood, filter paper is wiped dry, weighs.Volume ratio adds the tissue homogenate that normal saline makes 10% by weight, 2500r/min, and centrifugal 15min, gets supernatant for subsequent use.
The mensuration of 2.4 cerebral tissue indices
In cerebral tissue, the mensuration of NO content, NOS vigor illustrates by test kit and operates, and protein content adopts Coomassie Brilliant Blue to measure.
The mensuration of 2.5 cerebral infarction rates
Brain is got after rat sacrificed by decapitation, brain removes olfactory bulb, cerebellum and low brain stem, get left hemisphere, be cut into 5 along coronalplane, be placed in 2%TTC phosphate buffer, carry out TTC dyeing, strict lucifuge in dyeing course, hatches 30min in 37 DEG C of water baths, and then dislocation is containing fixing in the PBS solution of 4% paraformaldehyde, white portion is infarction tissue, and RED sector is normal structure.Computing formula: cerebral infarction rate (%)=infarction tissue's weight/full brain weight × 100%.
The observation of 2.6 neuron morphologies
Brain is taken out rapidly, decerebellation and medulla oblongata after rat sacrificed by decapitation.Specimen to be fixed in 10% neutral formalin buffer 1 week, again fixes 12h, organize with ethanol dehydration after drawing materials, dimethylbenzene is transparent, paraffin embedding, and paraffin slicing machine is cut into slices, carry out haematoxylin-Yihong (Haematoxylin-eosinStaining, HE) dyeing.
Concrete steps are as follows:
1. wrong microtome continuous coronal section, slice thickness 5 μm;
2. sheet is conventional with dimethylbenzene dewaxing, through ethanol at different levels to washing: the ethanol 1min-80% ethanol 1min-75% ethanol 1min-steaming shop washing 2min of dimethylbenzene (I) 5min-dimethylbenzene (II) 5min-dehydrated alcohol 2min-95%;
3. lignin dyeing soaks 15min, tap water twice;
4. sour ethanol differentiation 30s; Carry out water soaking l0min;
5. 0.5% eosin stain soaks 2min, tap water;
6. dehydration is advised, transparent, mounting: 95% ethanol (I) 1min-95% ethanol (II) 1min-dehydrated alcohol (I) lmin-anhydrous second liquor-saturated (II) 1min-dimethylbenzene (I) 1min-dimethylbenzene (II) 1min-resinene is closed.
2.7 hemorheology of rat index determinings
The mensuration of whole blood viscosity and plasma viscosity: after rat behavior experiment terminates, femoral artery gets blood, liquor sodii citratis (3.8%) anticoagulant.From the 6mL whole blood being added with anticoagulant, take out 2mL add blood rheological instrument, measure low cutting (10s) respectively, whole blood viscosity under high (200s) 2 shear rates.Get 4mL whole blood again with the centrifugal 10min of 2500r/min, get upper plasma 2mL and measure plasma viscosity.
Platelet aggregation test: adopt Beijing Sai Kexide SC-2000 platelet aggregation instrument, under room temperature by the whole blood of anticoagulant with the centrifugal 8min of 1000r/min, prepare platelet rich plasma (PRP), place in 2mL Ep pipe and close lid preservation, put in constant temperature hole for subsequent use, remaining anticoagulation 3000r/min centrifugal 30min gained supernatant is platelet poor plasma (PPP).Get 200 μ L PPP and put into circular cup in order to zeroing, getting 200 μ L PRP adds in the circular cup filling stirrer unshakable in one's determination, PPP circular cup is extracted after PPP zeroing, put into PRP circular cup, being that 3.5 μm of ol/LADP20 μ L press mensuration key adding final concentration simultaneously, obtaining 1,3min platelet aggregation rate (PAG) and max platelet rate (MPAG) numerical value.
The preparation of 2.8 isolated rat thorax artery rings and drug treating
After rat femoral gets blood, open breast, rapid taking-up thoracic aorta, be soaked in oxygen saturation K-H balance liquid, clean residual blood, divest epicardial fat and connective tissue, the blood vessel handled well is cut into the vascular ring of 4 ~ 5mm, vascular ring one end is connected tonotransducer, and the other end is fixed in bath, with BL-410 biological functional system record aortic annulus tension force.Mounted vascular ring is placed in 37 DEG C of thermostatic bath temperature bath 60min, continues to pass into 95%O
2and 5%CO
2mist, aortic annulus adds tranquillization load 1.5 ~ 2g, changes K-H liquid 1 time, repeatedly rinse, add the K-H liquid of high K+, hatch 15min after specimen 60min every 20min, blood vessel is reached and shrinks balance.Add K-H liquid temperature bath 30min, every 10min changes a not good liquor, makes tension force return to baseline values.Add 3 μ L10
-6the norepinephrine of mol/L, treats tension stability after 15min, changes liquid, rejoins K-H liquid, adds 10 of 2 μ L afterwards
-9the Ach of mol/L, treats tension stability after 10min, is discharged by K-H liquid.10 are added successively according to above method
-8mol/L, 10
-7mol/L, 10
-6mol/L, 10
-5the Ach of mol/L, sequentially determining aortic annulus tension variation.
2.9 statistical procedures
All data are with mean ± standard deviation
represent, adopt one factor analysis of variance method (one-way ANOVA) to experimental data statistical analysis, P<0.05 is for there being statistical significance.
3 experimental results
3.1 XINNAOXIN JIAONANG are on the impact of cerebral ischemia/reperfusion injury of rats 1h and 24h neurological deficits score
During Reperfu-sion 1h, ischemia-reperfusion neurologic impairment symptom is the most obvious, recovers to some extent after 24h.After Reperfu-sion 1h, there was no significant difference between the neurological deficits score three groups of model group, XINNAOXIN JIAONANG group, hydergine group and rats in sham-operated group.Reperfu-sion 24h XINNAOXIN JIAONANG group and hydergine group neurological deficits score are starkly lower than model group.In table 1.
Table 1 XINNAOXIN JIAONANG is on the impact of cerebral ischemia/reperfusion injury of rats 1h and 24h neurological deficits score
Note: compare with sham operated rats,
#p<0.05,
##p<0.01; Compare with model group,
*p<0.05,
*p<0.01
3.2 XINNAOXIN JIAONANG are on the impact of NO content and NOS vigor in MCAO rat model brain
Experimental result shows, compares with model group, and the basic, normal, high dosage group of XINNAOXIN JIAONANG and hydergine group obviously can reduce the content of MCAO rat model NO, reduces the vigor of NOS.In table 2.
Table 2 XINNAOXIN JIAONANG is on the impact of NO content and NOS vigor in MCAO rat model brain
Note: compare with sham operated rats,
#p<0.05,
##p<0.01; Compare with model group,
*p<0.05,
*p<0.01
3.3 XINNAOXIN JIAONANG are on the impact of MCAO rat model cerebral infarction volume
Experimental result shows, compares with model group, and XINNAOXIN JIAONANG and hydergine group obviously can reduce MCAO rat model cerebral infarction volume.In table 3 and Fig. 1.
Table 3 XINNAOXIN JIAONANG is on the impact of MCAO rat model cerebral infarction rate
Note: compare with sham operated rats,
#p<0.05,
##p<0.01; Compare with model group,
*p<0.05,
*p<0.01
3.4 XINNAOXIN JIAONANG are on the impact of MCAO rat model Determination of Hemorheological Indexes
Experimental result shows, the basic, normal, high dosage group of XINNAOXIN JIAONANG and hydergine group can significantly reduce MCAO rat model whole blood viscosity and plasma viscosity, simultaneously can remarkable anticoagulant.In table 4 and table 5.
Table 4 XINNAOXIN JIAONANG is on the impact of MCAO rat model blood plasma and whole blood viscosity thereof
Note: compare with sham operated rats,
#p<0.05,
##p<0.01; Compare with model group,
*p<0.05,
*p<0.01
Table 5 XINNAOXIN JIAONANG is on the impact of MCAO rat model platelet aggregation rate
Note: compare with sham operated rats,
#p<0.05,
##p<0.01; Compare with model group,
*p<0.05,
*p<0.01
3.5 XINNAOXIN JIAONANG are on the impact of MCAO rat model cerebral cortex neurons pathology damage
As shown in Figure 2,3, rats in sham-operated group cortical area cerebral tissue structural integrity, neurocyte is intensive.Neuron plasma enriches, and nucleus is placed in the middle, and nuclear staining is clear, and form is normal; Vascular endothelial cell is complete, has no hemorrhage and erythrocyte caulked, Guan Zhou dense structure.When ischemia 1 day, there is the damage of bright product and the inflammatory reaction of local in MCAO model group rats cerebral tissue, rat brain lesions position interstitial is loose in sieve-like, paintedly obviously shoals, visible neuronal arrangement is at random, a large amount of wall regeneration, nuclear targeting is unclear, karyopycnosis engrain, the shrinkage of part cell space, N euron loss is obvious, and glial cells hyperplasia is obvious, the gap increase of blood vessel pipe week.Compared with model group, the complete variable neuronal cell showed increased of XINNAOXIN JIAONANG group, degeneration necrosis scope of organization relative decrease, degree alleviate.
3.6 XINNAOXIN JIAONANG are to the diastole effect of MCAO rat model NE preshrinking blood vessel
Experimental result shows, compares with model group, and hydergine group and XINNAOXIN JIAONANG middle and high dosage group can the obvious pre-shrunk blood vessels of diastole MCAO rat model NE.See Fig. 4.
Claims (2)
1. the application of XINNAOXIN JIAONANG in preparation treatment cerebral ischemia reperfusion injury medicine.
2. XINNAOXIN JIAONANG according to claim 1, is characterized in that, is made up of following parts by weight component: Radix Rhodiolae 3 ~ 6 parts, Fructus Lycii 5 ~ 15 parts, Fructus Hippophae 3 ~ 9 parts.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105147866A (en) * | 2015-10-13 | 2015-12-16 | 北京普瑞博思投资有限公司 | Traditional Chinese medicine composition for improving cerebral circulation and atherosclerosis and application thereof |
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| CN103316151A (en) * | 2013-07-08 | 2013-09-25 | 三普药业股份有限公司 | Combined drug of rhodiola rosea extract, medlar extract and sea-buckthorn fresh pulp powder extract as well al preparation and application thereof |
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| CN105147866A (en) * | 2015-10-13 | 2015-12-16 | 北京普瑞博思投资有限公司 | Traditional Chinese medicine composition for improving cerebral circulation and atherosclerosis and application thereof |
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