CN106176780A - A kind of new opplication of gastrodine - Google Patents
A kind of new opplication of gastrodine Download PDFInfo
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Abstract
The invention discloses the new opplication of a kind of gastrodine；The application in preparing preventing/treating hepatic ischemia-reperfusion injury medicine of the described gastrodine.The present invention has carried out internal pharmacodynamic study to gastrodine, and experimental result shows, gastrodine can reduce rat model serum lipid overoxidation product assay, has stronger antioxidant activity；Release to inflammatory factor has obvious inhibiting effect；There is suppression liver inner cell apoptosis activity；Significantly raised NO concentration can improve liver microcirculation, hepatic ischemia-reperfusion injury is had obvious protective function.Gastrodine of the present invention can directly be used alone, it is possible to is mutually combined application, can include that plant extract composition compound recipe form uses with other drug, it is also possible to use in a usual manner.
Description
Technical field
The invention belongs to pharmaceutical technology field, be specifically related to the new opplication of a kind of gastrodine.
Background technology
Tissue or organ can cause damage after certain time ischemia, not only can not make its merit after recovered blood perfusion
Can recover with structure, make damage increase the weight of referred to as postischemic reperfusion damage (ischemia-reperfusion further on the contrary
injury ,I/RI).There is stream in a large amount of Ca2+ after Reperfu-sion, and generate a large amount of oxygen-derived free radicals, be extensive tissue cell insult
Main pathogenesis.The most multiple disease such as Dalayed neuronal necrosis, irreversible shock, myocardial infarction, acute internal organs
The generation of nonfunction and organ transplant rejection etc., development are all relevant with ischemia-reperfusion.
Hepatic ischemia-reperfusion injury (HIRI) is the pathological state that surgery of liver is common.Owing to the anatomical structure of liver is multiple
Miscellaneous, blood supply is enriched, and often need to block hepatic blood supply for minimizing intraoperative hemorrhage in liver surgery.After but liver blood flow blocks
HIRI is also resulted in while minimizing is hemorrhage.This damage often betides hepatocarcinoma excision, liver transplantation, serious wound and infection etc.
Pathological process.HIRI may result in liver dysfunction, and severe patient can cause liver failure, even dead.Current research shows HIRI's
Pathogenesis is the most fully aware of, following factor may be had to participate in: oxidativestress damage, inflammatory cell infiltration, calcium overload, enzyme
Effect, hepatocellular apoptosis etc..Along with being embedded in of research is improved HIRI aspect and had remarkable progress, but effect is the most undesirable.
HIRI is to cause postoperative Primary graft failure, cause the major reason that liver transplantation is failed.The most how alleviating HIRI is
Surgery of liver particularly spare-part surgery is badly in need of the thorny problem solved at present!
At present it has been recognized that HIRI is the result that much different mechanism interacts, therefore its preventing and treating the most just has different sides
Method.Conventional method mainly has Ischemic reperfusion (IP) and medical preconditioning.(Gong NQ, Ye QF, Jang HY, et
a.l P rotection of grafts by m ed icin e p recondi tioning and Ischem ia p
recondit ion ing in liver transp lan tation [ J]. C hin JH epa Su rg, 2000,
6:282-284.) but, (S chu lz R, the W alz such as this protective effect of liver IP is the most limited, Schu lz
MK, Beh rends M, et a.l M in im al p rotection of the liver by ischem ic
precond ition ing in pigs [ J] . Am J Phys iol Heart C irc Physiol, 2001,
280:H 198-H207.) find that IP protective effect can maintain 1～2 h, IP effect after experience 200min long period ischemia
Disappear, therefore limit it in clinical application.For how improving the self supporting capacity of liver in liver surgery,
Block the problem that liver blood flow exists HIRI, transfer its endogenous protection mechanism, alleviate liver I/RI, be facing of surgery of liver
Bed demand.In liver transplantation, Ischemic reperfusion is implemented more limited, therefore uses medical preconditioning to induce IP and mould
Intend the Delayed Protection of IPC or directly protect the liver after I/RI, there is wide potential applicability in clinical practice, be worth into one
Step research.
Medical preconditioning is that the pharmacological action utilizing some active substance direct or indirect reaches similar Ischemic reperfusion
Protective effect, strengthen tissue or the cell toleration to ischemical reperfusion injury, thus alleviate damage.HIRI is had pre-
The drug main calcium channel blocker to be had for the treatment of effect, free radical scavenger, improve the medicine of microcirculation or cellular energy metabolism
Deng.But many of which medicine has bigger toxic and side effects, have impact on its application clinically, it is difficult to meet clinical needs.
Therefore, develop a kind of medicine that can solve an above-mentioned difficult problem to be very important.
Summary of the invention
It is an object of the invention to provide the new opplication of a kind of gastrodine.
The object of the present invention is achieved like this, and described gastrodine is preparing preventing/treating hepatic ischemia reperfusion damage
Application in vulnerary thing.
Rhizoma Gastrodiae (Gastrodia elata.BL) is the dry tuber of orchid Rhizoma Gastrodiae, is rare Chinese medicine, property is sweet,
Flat, return Liver Channel.The traditional Chinese medical science thinks that its primary efficacy has relieving spasm by subduing liver-wind, suppressing liver-YANG, dispelling wind and removing obstruction in the collateral.The chemistry extracted from Rhizoma Gastrodiae
Composition has Gastrodine, gastrodia elata genin, vanillyl alcohol, vanillin, β-steroid paddy alcohol, daucosterol etc., and wherein active component content is
High effective monomer is Gastrodine, the entitled p-methylol benzene-β-D-pyranglucoside semihydrate (β-D-Gluc of chemistry
Opyranoside), also known as gastrodine.Modern pharmacology research display, gastrodine has increase central authorities and peripheral arterial blood vessel
Compliance, reduces peripheral vascular resistance, increases cardiovascular and cerebrovascular vessel blood flow, produces gentle hypotensive effect, and to myocardial cell, brain
Organize the most protected effect, have simultaneously calmness, hypnosis, ease pain, strengthen immunity etc. effect, be widely used in the treatment heart clinically
Cerebrovascular, microcirculqtory system disease, evident in efficacy, and non-evident effect.Common dosage forms dosage form has gastrodine the most on the market
Injection, gastrodini, gastrodine capsule.
Gastrodine (i.e. to methylol benzene-β-D-pyranglucoside) is one of effective monomer component of Rhizoma Gastrodiae.Modern medicine
Reason research shows, gastrodine has calmness, analgesia, a soporific function, also can convulsion, anxiety, cerebral blood flow can be increased, subtract
Lack vascular resistance, improve vertebral-basilar artery insufficiency, there is neuroprotective, (Ju Guichun. Rhizoma Gastrodiae and the medicine of preparation thereof
Reason effect and clinical application research progress (J). China's Pharmaceutical, 2008,17 (1): 64-66.) animal experiment study display gastrodine
The apoptosis that cerebral ischemia can be suppressed to induce, has neuroprotective, thin to the rat brain microvascular endothelial of In vitro culture
Born of the same parents (BMEC) ischemic injuries has certain protective effect.(Yang Jie, Zhao Chaohua, Zong Changhong, etc. transience MCAO rat
Injection gastrodine has neuroprotective (J). Journal of the Fourth Military Medical University, and 2008,29 (4): 295-297.；Hu Jinghong,
Si Yinchu, Hong Qingtao. the protective effect (J) of Gastrodine on Cultivated Rat Brain Microvessel Endothelial Cells by Mimic Cerebral Ischemia. in
Central China medical magazine (former Chinese Medicine journal), 2007,22 (2): 124-126.； Zeng X H.Zhang Y , Zhang
S M , etal A microdialysis is study of effect s of gast rodin on
neurochemical changes in the ischemic/ reperfused ret cerebral hippocampus
(J) .Biol Pham Bull, 2007,30 (4): 801.) research shows that gastrodine also has nootropics anti-aging effects.(king
Clear monarch, practises Yang Yanbin, Liu Jia. the gastrodine impact (J) on senescence-acceleratad mice cerebral tissue Aging-associated gene expression. solve
Cut open scientific advance, 2007,13 (4): 353-357.) foreign study shows that gastrodine is effective antianxiety drugs.(Jung
JW, Yoon BH , Oh HR.et al.Anxiolvtie-like ef fects of Gast rodia elata and it
S phenolie const ituents in mice (J) .Biol Pharm Bull, 2006,29 (2): 261-265.)
Gastrodine is widely used clinically, treatment dizziness, vertebro-basilar artery insufficiency, cerebral infarction, the headache of blood vessel character
Etc. disease, evident in efficacy, the auxiliary for hypertension, diabetic peripheral neuropathy is treated, effect affirmative, clinic weight
Depending on.
There are some researches show that gastrodine can strengthen the compliance of blood vessel, improved internal organs anaerobic condition, some are supplied by brain
The central nervous system disease that insufficiency of blood causes has preferable therapeutical effect.There are some researches show at present, gastrodine to brain and
Heart ischemia reperfusion damage has a protective effect, but there is not been reported to the effect of hepatic ischemia-reperfusion injury.
The present invention has carried out internal pharmacodynamic study to gastrodine.Experimental result shows, gastrodine can reduce rat model
Serum lipid overoxidation product assay, has stronger antioxidant activity；Release to inflammatory factor has obvious inhibiting effect；
There is suppression liver inner cell apoptosis activity；Significantly raised NO concentration can improve liver microcirculation, to hepatic ischemia-reperfusion injury
There is obvious protective function.
Gastrodine of the present invention can directly be used alone, it is possible to is mutually combined application, can include plant with other drug
Extract composition compound recipe form uses, it is also possible to use in a usual manner.
Detailed description of the invention
Below in conjunction with embodiment, the present invention is further illustrated, but is any limitation as the present invention never in any form,
Based on present invention teach that any conversion or replacement made, belong to protection scope of the present invention.
The new opplication of gastrodine of the present invention is that described gastrodine is preparing preventing/treating hepatic ischemia reperfusion
Application in damage medicine.
Gastrodine preparation in the new opplication of described gastrodine be described gastrodine adds pharmaceutically acceptable
Adjuvant makes tablet, granule, capsule, pill, suspensoid or injection.
With specific embodiment, the present invention will be further described below:
Embodiment 1
The gastrodine impact on hepatic ischemia-reperfusion injury in rats:
1, reagent and animal
Gastrodine, content 98.8%, lot number: JT20161014, Kun Yao group produces.
Male SD rat 40,250-300g, Kunming Medical University's Experimental Animal Center provide, production licence:
SCXK(Yunnan) K2015-0002, licence issuing authority: Kunming Office of Science and Technology.20 ~ 25 DEG C of (temperature difference per day≤3 of laboratory animal room temperature
DEG C), humidity 40% ~ 70%, illumination 12h:12h light and shade replaces, illumination 150 ~ 300lx, noise≤60 Db；Laboratory animal uses to be permitted
Can demonstrate,prove: SYXK(Yunnan) K2014-0001, issuing unit: Kunming Office of Science and Technology.
Malonaldehyde (MDA) testing cassete, superoxide dismutase (SOD) testing cassete, glutamic oxaloacetic transaminase, GOT (AST) testing cassete, paddy
Pyruvic transaminase (ALT), glutathion (GSH) testing cassete, superoxide dismutase (SOD), glutathion (GSH), malonaldehyde
(MDA) test kit builds up biological reagent company purchased from Nanjing；TUNEL test kit: source: Roche, lot number: 10348000；Anti-
Cleaved Caspase-3 antibody, Cell Signaling Technology, lot number: 19；Alexa Fluor568 donkey anti-rabbit
IgG (H+L), Molecular Probes, lot number: 1134929；Alexa Fluor568 goat anti rat IgG (H+L),
Molecular Probes, lot number: 1259374；Triton X-100, Sigma, lot number: 031M0301V；BCA protein quantification tries
Agent box, Wei Ao bio tech ltd, Shanghai, lot number: WB0123；Interleukin-11 β (IL-1 β): lot number DZE30419；Interleukin
6(IL-6): lot number DZE30646；Tumor necrosis factor α (TNF-α): lot number DZE30635；Above reagent is purchased from Shanghai and breathes out spirit
Bio tech ltd.
2, experimental technique
2.1 animal models and packet
Laboratory animal is divided into sham operated rats, liver I/RI model group (model group) and gastrodine pretreatment low dosage and height at random
Dosage group (60,120mg/kg)+liver I/RI model group (gastrodine pretreatment low, high dose group).Preoperative 12 h fasting, freely
Drinking-water.Pentobarbital sodium (30 mg/kg) solution intraperitoneal injection of anesthesia rat with 1%, takes median abdominal incision about 2 cm and enters abdomen,
Free hepatoduodenal ligament, presss from both sides with not damaged bulldog clamp and closes Reperfu-sion 6 h after portal vein, Hepatic artery and common bile duct 30 min
Making hepatic ischemia reperfusion animal model (Zhao Haifeng, Kong Rui, Sun Bei, etc. Sodium butyrate ischemia hot to rat liver is again
The research [J] of perfusion injury protective effect. China's general surgery magazine, 2008,17 (1): 91-93.).Sham operated rats is swum
Do not block from hepatoduodenal ligament, preoperative 20 min of gastrodine pretreated group by tail vein injection gastrodine (60,
120mg/kg), model group gives normal saline.Respectively organize rat after postoperative 6 h from postcava blood sampling about 5 mL；Cut
Take leftlobe of liver, remainder split-80 DEG C frozen, hepatic tissue biochemical indicator to be measured and caspase-3 level.
2.2 observation index and detection method
(1) liver functional testing:
Take serum after rat vein blood is centrifugal, detect glutamic oxaloacetic transaminase, GOT (AST) with respective kit method, glutamate pyruvate transaminase (
ALT) level.
(2) hepatic tissue biochemical indicator detection:
Hepatic tissue SOD, GSH and MDA level is detected with respective kit method.
(3) hepatic tissue caspase-3 horizontal detection:
Rat liver is inserted in the paraformaldehyde solution of 4% and fix 12-16h.The liver fixed is carried out frozen section, puts
In frozen protection liquid ,-20 C save backup, and to carry out Cleaved Caspase-3 immunohistochemical staining, TUNEL contaminates
Color.Frozen section, after 0.01M PBS rinses, is placed in 0.3%Triton X-100 30min, and rear directly resisting with corresponding one hatches
(4 C are overnight).Cut into slices and rinse 3 times with PBS, every all over 5min, then with corresponding fluorescence two anti-room temperature reaction 1-2h, rear continue with
PBS rinses, and carries out mounting by mountant.Coloration result is taken pictures through Nikon fluorescence microscope, and divides with Adobe Photoshop
Analysis；In situ cell apoptosis detection description as appended by test kit is carried out.Specifically, frozen section first rinses with PBS, then
Hatch with 0.1%TritonX-100, then directly hatch with TUNEL reactant liquor (being made up of marking fluid and enzyme reaction solution),
Lucifuge reaction 60min under the conditions of 37 degree.Coloration result is directly observed under fluorescence microscope and takes pictures.
(4) inflammatory factor detection:
Take serum after rat vein blood is centrifugal, operate in strict accordance with test kit description, measure IL-1, IL-6, TNF-α.
2.3 statistical procedures
Data with meansigma methods and standard deviation (± SD) represent.Statistical analysis uses one factor analysis of variance, compares application between group
Student Newman-Keul ' s test tests analysis.P < 0.05 thinks have significant difference.
3, result
The horizontal detection result of 3.1 Serum ALT and AST
Compare with sham operated rats, model group rats Serum ALT and AST level significantly raised (equal P < 0.05), and and model
Group compares, and Serum ALT and the AST level of gastrodine pretreated group substantially lower (equal P < 0.05), are shown in Table 1.
Table 1 respectively organize rat blood serum transaminase level compare (± SD), n=10
| Group | ALT(U/L) | AST(U/L) |
| Sham operated rats | 44.5±4.6 | 90.1±9.1 |
| Model group | 85.7±6.1* | 234.3±16.3* |
| Gastrodine pretreatment low dose group | 69.3±5.3▲ | 163.1±11.4▲ |
| Gastrodine pretreatment high dose group | 61.2±4.0▲ | 140.5±13.8▲▲ |
Note: compare with sham operated rats, * P ＜ 0.05, * * P ＜ 0.01；Compare with model group, ▲ P ＜ 0.05, ▲ ▲ P ＜
0.01。
3.2 biochemical indicator testing results
Comparing with sham operated rats, in model group hepatic tissue, SOD and GSH level substantially reduces (P < 0.05), and MDA level
Significantly raised (P < 0.01).Comparing with model group, in gastrodine pretreated group hepatic tissue, SOD and GSH level significantly raises,
MDA level significantly reduces, (P < 0.05 or P < 0.01), is shown in Table 2.
Table 2 respectively organize each Biochemical Indices In Serum of rat comparison (± SD), n=10
| Group | SOD(U/mg albumen) | GSH(mg/mg albumen) | MDA(nmol/mg albumen) |
| Sham operated rats | 305.1±16.2 | 3.13±0.20 | 1.98±0.57 |
| Model group | 204.2±34.5* | 1.88±0.75* | 4.67±0.89** |
| Gastrodine pretreatment is low | 286.5±15.2▲ | 2.64±0.13▲ | 3.56±0.72▲ |
| Gastrodine pretreatment is high | 377±12.4▲▲ | 3.76±0.65▲▲ | 2.09±0.47▲▲ |
Note: compare with sham operated rats, * P ＜ 0.05, * * P ＜ 0.01；Compare with model group, ▲ P ＜ 0.05, ▲ ▲ P ＜
0.01。
And the mechanism of hepatic ischemia-reperfusion injury is multiple with the formation of oxygen-derived free radicals, inflammatory cell infiltration, hepatocellular apoptosis etc.
Factor is relevant.(Kang KJ. Mechanism of hepatic ischemia/reperfusion injury and
protection against reperfusion injuryl[J]. Transplant Proc, 2002, 4(7):2659-
2661.) display of this result of study, hepatic ischemia/reperfusion injury 30 min, in the Ischemia and Reperfusion in vivo in Rats model of Reperfu-sion 6 h, give Rhizoma Gastrodiae
Element can substantially reduce Serum ALT and AST level (P < 0.05), alleviates hepatic pathology damage.These results prompting gastrodine is pre-
Process and liver I/RI is had protective effect.Research is had to also confirm that in liver I/RI animal model process LAN SOD or give
GSH, (He SQ, Zhang YH, Venugopal SK, et al. Delivery of antioxidative enzyme
genes protects against ischemia/reperfusion-induced liver injury in mice[J].
Liver Transpl, 2006, 12(12):1869-1879.； Schauer RJ, Gerbes AL, Vonier D, et
al. Glutathione protects the rat liver against reperfusion injury after
Prolonged warm ischemia [J]. Ann Surg, 2004,239 (2): 220-231.) or the base removing that freers
Agent, as Edaravone can alleviate liver I/RI.(Shi Chenlei, Qin Huadong, Shi Tiefeng. Edaravone is to rats'liver I/RI
The protective effect [J] of damage. China's general surgery magazine, 2010,19 (1): 28-31.)
This experimental result shows, after giving gastrodine, in the hepatic tissue of ischemia-reperfusion, SOD and GSH level is significantly raised；With
The MDA level of Shi Zuowei products of oxidative stress is significantly reduced, and shows that gastrodine can improve ischemia-reperfusion liver organization
Oxidation resistance.
3.3 on apoptotic impact
In our current research, we have detected apoptotic cell (TUNE method) and the signaling molecule relevant to apoptosis (such as Caspase-
3)。
As shown in table 3, hepatic ischemia/reperfusion injury 30 min, in the Ischemia and Reperfusion in vivo in Rats model of Reperfu-sion 6 h, TUNEL method in brain
Apoptotic cell and Cleaved Caspase-3 immuno positive (Cleaved Caspase-3-ir) cell of dye marker all go out
Now increase (comparing with sham operated rats, P < 0.01), after giving gastrodine, then can significantly reduce TUNEL method dye marker in brain
Apoptotic cell and Cleaved Caspase-3 immuno positive (Cleaved Caspase-3-ir) cell (compare with model group,
P < 0.05 or P < 0.01).
Table 3 gastrodine on apoptotic impact (± SD), n=10
Note: compare with sham operated rats, * P ＜ 0.05, * * P ＜ 0.01；Compare with model group, ▲ P ＜ 0.05, ▲ ▲ P ＜
0.01。
In liver I/RI, hepatocellular apoptosis is a major reason of hepar damnification, and during apoptosis
The activation of caspase-3 is most important link, and wherein caspase-3 is most important apoptosis executor.Caspase-3's
Activation can directly or indirectly act on enzyme action (Porter AG, the J nicke RU. of key protein during apoptosis performs
Emerging roles of caspase-3 in apoptosis[J]. Cell Death Differ, 1999, 6(2):
99-104.).This experiment shows, in the animal model of liver I/RI, gastrodine can substantially suppress TUNEL method in rats'liver
The apoptotic cell of dye marker and Cleaved Caspase-3 immuno positive (Cleaved Caspase-3-ir) cell, can
Alleviate liver I/RI, hepatocellular apoptosis after pointing out gastrodine can suppress ischemia-reperfusion.
3.4 impacts on inflammatory factor
After Reperfu-sion 6h, compare with sham operated rats, model group TNF-α, IL-1 β and NO level the most significantly raised (P <
0.05 or P < 0.01), NO level significantly raised (P < 0.05)；Compare with model group, gastrodine pretreated group TNF-α and
IL-1 β level significantly reduces, and NO level significantly raises (P < 0.05 or P < 0.01), is shown in Table 4.
Table 4 on the impact of inflammatory factor (± SD, n=10)
Note: compare with sham operated rats, * P ＜ 0.05, * * P ＜ 0.01；Compare with model group, ▲ P ＜ 0.05, ▲ ▲ P ＜
0.01。
This laboratory observation arrives, after Reperfu-sion: the Serum ALT of gastrodine pretreated group and AST level are significantly lower than model
Group；SOD and GSH level is significantly higher than model group, and serum MDA level substantially relatively model group reduces.Also find liver Reperfu-sion
Rear TNF-α and IL-1 inflammatory factor level raise, and the release of gastrodine pretreated group TNF-α and IL-1 is pressed down
System, the index of reperfusion injury declines the most accordingly, alleviates liver reperfusion injury by reducing TNF-α and IL-1 release.Ba
Jt etc. [3] find that the TNF-α of Kupffer Cell secretion can be combined formation TNFR-TRADD-FADD by the TNFR1 on liver plasma membrane
Complex also raises Caspase-3 gene expression, activates the cascade reaction of C aspase family, active cell apoptosis, I
Further by the situation of liver cell apoptosis after TUNEL method detection Reperfu-sion, found that the cell of gastrodine pretreated group
Apoptosis is significantly inhibited, and shows that gastrodine can suppress liver cell apoptosis, and this effect may be withered no by suppression
Cell-stimulating, the release of minimizing TNF-α realize.Have been reported that and think after liver Reperfu-sion that microcirculation disorders in early days is main and NO
Producing relevant, the concentration of NO is unbalance is a major reason of microcirculation disturbance after hepatic ischemia reperfusion.We also observe
NO level change after liver Reperfu-sion, after finding Reperfu-sion, the NO level in blood significantly raises, gastrodine pretreated group NO water
Flat elevation amplitude is big compared with model group, illustrate that gastrodine can improve the micro-of liver after Reperfu-sion by adjustment NO concentration ratio
Circulation, thus play protective effect.
To sum up, gastrodine can alleviate hepatic ischemia-reperfusion injury in rats, and possible mechanism of action includes reducing oxygen freely
The generation of base, suppression inflammatory cytokine release, reduce liver cell apoptosis and improve Reperfu-sion after microcirculation etc..
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For Yuan, under the premise without departing from the principles of the invention, it is also possible to make some improvements and modifications, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (2)
1. the new opplication of a gastrodine, it is characterised in that described gastrodine is preparing preventing/treating hepatic ischemia reperfusion
Application in damage medicine.
2. the gastrodine preparation in the new opplication of the gastrodine described in a claim 1, it is characterised in that described gastrodine
The pharmaceutically acceptable adjuvant of middle addition makes tablet, granule, capsule, pill, suspensoid or injection.
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| CN111150737A (en) * | 2020-01-17 | 2020-05-15 | 昆明医科大学 | New application of gastrodin |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103566198A (en) * | 2012-08-06 | 2014-02-12 | 中国医学科学院医药生物技术研究所 | Application of gastrodia elata and gastrodin in preparation of product used for treating non-alcoholic fatty liver disease |
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| 刘凌钊等: ""天麻在肾缺血再灌注损伤中的作用机制"", 《广东医学》 * |
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