Protective effect of gastrodin on A β 25-35-induced HT22 cell injury by improving glycolytic function through activating PI3K/Akt/BACH1 signaling axis / 中国药理学与毒理学杂志

OBJECTIVE To investigate whether gas-trodin(GAS)plays a neuroprotective role by activating PI3K/Akt/BACH1 signaling axis to improve glycolytic func-tion.METHODS HT22 cells were treated with Aβ25-35 for 24 h to establish cell damage model.GAS pretreated HT22 cells for 2 h,and Akt agonist SC79,Akt inhibitor MK2206,PI3K inhibitor LY294002 were added 0.5 h before GAS treatment to detect their protective mecha-nisms.Pharmacodynamic research of GAS in this model were divided into six groups:control group,GAS group(GAS 10 μmol·L-1),model group(Aβ25-35 20 μmol·L-1),model +GAS 2.5,5 and 10 μ mol·L-1 group).Mecha-nism research of GAS in this model was divided into 6 groups:control group,Aβ25-35 20 μmol·L-1 group,Aβ25-35 20 μmol·L-1 + GAS 10 μmol·L-1 group,Aβ25-35 + SC79 group(Aβ25-35 20 μmol·L-1 +SC79 10 μmol·L-1),Aβ25-35+MK2206+GAS group(A β 25-35 20 μ mol·L-1 +MK2206 10 μmol·L-1+GAS 10 μmol·L-1),Aβ25-35+LY294002+GAS group(Aβ25-35 20 μmol·L-1+LY294002 10 μmol·L-1+GAS 10 μmol·L-1).Cell viability was detected by MTT,mor-phological changes of cells were observed by micro-scope,ATP content was detected by chemilumines-cence,and pyruvate(PA)content was detected by colo-rimetry.Western blotting was used to detect the protein levels of transcription factor BACH1,key glycolysis enzyme hexokinase(HK1)and PI3K/Akt signaling path-way related proteins PI3K,p-PI3K,Akt and p-Akt.RESULTS The results showed that compared with the control group,the cell morphology of HT22 cells damaged by Aβ25-35 was damaged,the number of cells decreased,the cell body became smaller,the number of dead cells increased,the cell survival rate,ATP and PA contents decreased significantly,and the protein expressions of p-PI3K,p-Akt,BACH1 and HK1 were significantly down-regulated.GAS treatmentcansignificantlyimprovethemor-phology of HT22 cells damaged by Aβ25-35,increase cell survival rate,ATP and PA contents,and up-regulate the expression of p-PI3K,p-Akt,BACH1 and HK1 proteins.SC79 also significantly increased cell survival rate,ATP content,protein expression of BACH1 and HK1.However,the above ameliorative effect of GAS on HT22 cell dam-age induced by Aβ25-35 was antagonized by LY294002 and MK2206.CONCLUSION GAS exerts a neuroprotec-tive effect on Aβ25-35-induced HT22 cell injury by improv-ing glycolytic function through activating PI3K/Akt/BACH1 signaling axis.

Gastrodin injection alleviates lung injury caused by focal cerebral ischemia in rats through NGF/TrkA pathway-mediated activation of the anti-inflammatory pathway / 南方医科大学学报

OBJECTIVE@#To investigate the therapeutic mechanism of gastrodin injection for alleviating lung injury caused by focal cerebral ischemia in rats and the role of the NGF-TrkA pathway in mediating this effect.@*METHODS@#Forty SD rats were equally randomized into normal group, sham-operated group, model group and gastrodin group, and in the latter two groups, rat models of focal cerebral ischemia were established by embolization of the right middle cerebral artery. After successful modeling, the rats were treated with intraperitoneal injection of gastrodin injection at the daily dose of 10 mg/kg for 14 days. After the treatment, the wet/dry weight ratio of the lung tissue was determined, the pathological changes in the lung tissue were observed using HE staining, and the levels of IL-10 and TNF-α in the arterial blood were detected with ELISA. The expressions of NF-κB p65 and TNF-α in the lung tissue were detected with Western blotting, and the expressions of NGF and TrkA were detected using immunohistochemical staining and Western blotting.@*RESULTS@#Compared with the normal control and sham-operated groups, the rats in the model group showed obvious inflammatory lung injury, significantly increased wet/ dry weight ratio of the lungs (P < 0.01), increased TNF-α level in arterial blood (P < 0.01), and significantly up-regulated protein expressions of NF-κB p65 (P < 0.01), TNF-α (P < 0.01), NGF (P < 0.05) and TrkA(P < 0.05) in the lung tissue. Treatment with gastrodin injection obviously alleviated lung inflammation, decreased the wet/dry weight ratio of the lungs (P < 0.05), and significantly lowered TNF-α level (P < 0.01) and increased IL-10 level in the arterial blood in the rat models (P < 0.01); gastrodin injection also significantly decreased the protein expressions of NF-κB p65 and TNF-α (P < 0.05) and up-regulated the expressions of NGF and TrkA in the lung tissue of the rats (P < 0.05).@*CONCLUSION@#The NGF/TrkA pathway may participate in cerebral ischemia-induced inflammatory lung injury, which can be obviously alleviated by gastrodin through the activation of the anti-inflammatory pathway mediated by the NGF/TrkA pathway.

Cardiovascular protective properties of gastrodin

Cardiovascular diseases cause significant morbidity and mortality worldwide, incurring a major public health burden. Gastrodia elata Blume is a traditional Chinese herbal medicine that has been widely used to treat central nervous system and cardiovascular diseases. Gastrodin, as the major active component in Gastrodia elata Blume, can confer protection against cardiovascular diseases. In this review, we summarize the anti-inflammatory actions, anti-cardiac hypertrophy, anti-hypertension, anti-atherosclerosis, and angiogenic effects of gastrodin, as well as its protective effects on vascular cells and against myocardial ischemia-reperfusion injury. The medical potential of gastrodin in diabetes-related cardiovascular diseases is also discussed.

Systematic review and Meta-analysis of efficacy and safety of gastrodin in treatment of tension-type headache / 中国中药杂志

The efficacy of gastrodin as a Chinese herbal medicine extract in the treatment of tension-type headache has been confirmed. This paper systematically reviewed the efficacy and safety of gastrodin in the treatment of tension-type headache, aiming to provide a new choice for the treatment of this disease. In this study, four Chinese databases, four English databases and two trial registries were searched from the date of establishment to September 2020. The related randomized controlled trials(RCTs) were screened out according to the predetermined criteria. The bias risk assessment tool developed by Cochrane collaboration was used to evaluate the quality of the reports. RevMan 5.4.1 was used for Meta-analysis, and GRADE system for the evidence-based evaluation on the quality of outcome indicators. A total of 177 articles were retrieved and 8 articles were finally included for analysis, with a total sample size of 1 091 cases, which included 565 cases in the treatment group and 526 cases in the control group. The overall quality of included stu-dies was not high. The results of Meta-analysis are as follows:(1)In terms of headache frequency, gastrodin group was better than wes-tern medicine group(MD=-2.90, 95%CI[-3.76,-2.03], P<0.000 01).(2)In terms of number of abnormal blood vessels in TCD, gastrodin group was better than western medicine group(MD=-88.96, 95%CI[-102.36,-75.55], P<0.000 01).(3)In terms of effective rate, gastrodin group was better than western medicine group(RR=1.47, 95%CI[1.29, 1.68], P<0.000 01). The results of subgroup analysis are as follows:(1)Effective rate based on age, for the patients upper age limit 40-46 years old, gastro-din group was better than western medicine group(RR=1.69, 95%CI[1.50, 1.90], P<0.000 01); for the patients upper age limit 55-60 years old, gastrodin group was better than western medicine group(RR=1.27, 95%CI[1.16, 1.38], P<0.000 01).(2)Effective rate based on dosage form, both the gastrodin capsules and injection groups were better than western medicine group(RR_(capsules)=1.42, 95%CI[1.08, 1.88], P=0.01; RR_(injection)=1.50, 95%CI[1.26, 1.77], P<0.000 01). GRADE evaluation showed that the above outcomes had low quality of evidence. Only one article detailed the occurrence of adverse reactions and thus the present study cannot make a positive conclusion on the safety of gastrodin in the treatment of tension-type headache. The small number and low quality of the included reports affected the reliability of the results. In the future, more high-quality randomized controlled trails are needed to improve the evaluation on the efficacy and safety of gastrodin in the treatment of tension-type headache.

Antioxidant mechanism of gastrodin combined with isorhynchophylline in inhibiting MPP~+-induced apoptosis of PC12 cells / 中国中药杂志

Gastrodiae Rhizoma-Uncariae Ramulus cum Uncis is the most frequently used herbal pair in the treatment of Parkinson's disease(PD). Gastrodin and isorhynchophylline are important components of Gastrodiae Rhizoma-Uncariae Ramulus cum Uncis herb pair with anti-Parkinson mechanism. This study aimed to investigate the effect of gastrodin combined with isorhynchophylline on 1-methyl-4-phenylpyridinium(MPP~+)-induced apoptosis of PC12 cells and their antioxidant mechanism. The leakage of lactate dehydrogenase(LDH) from cells to media was analyzed by spectrophotometry. Apoptotic cells were labeled with Annexin V-fluorescein isothiocyanate(FITC) and propidium iodide(PI) and analyzed by flow cytometry. The cell cycle was analyzed using propidium iodide(PI) staining. Lipid peroxidation(LPO) level was analyzed by spectrophotometry. The mRNA expression of caspase-3 was examined by Real-time RT-PCR. The protein expressions of heme oxygenase 1(HO-1) and NADPH: quinoneoxidore-ductase 1(NQO-1) were determined by Western blot. Gastrodin combined with isorhynchophylline reduced the percentage of Annexin V-positive cells and cell cycle arrest in MPP~+-induced PC12 cells. Gastrodin combined with isorhynchophylline down-regulated the mRNA expression of caspase-3, up-regulated the protein expressions of HO-1 and NQO-1, and reduced LPO content in MPP~+-induced PC12 cells. PD98059, LY294002 or LiCl could partially reverse these changes pretreated with gastrodin combined with isorhynchophylline, suggesting that gastrodin combined with isorhynchophylline inhibited MPP~+-induced apoptosis of PC12 cells and oxidative stress through ERK1/2 and PI3 K/GSK-3β signal pathways. Our experiments showed that gastrodin combined with isorhynchophylline could down-re-gulate the mRNA expression of caspase-3 and up-regulate the protein expressions of HO-1 and NQO-1, so as to reduce oxidative stress and inhibit apoptosis.

Study on Absorption Characteristics of Gastrodia elata Powder with Different Particle Sizes Based on Rat Everted Intestinal Sac Model in vitro / 中国药房

OBJECTIVE：To compar e the absorpt ion characteristics of gastrodin ，parishin A ，parishin B and parishin C of Gastrodia elata powder，and to explore the effect of particle size on intestinal absorption of above components. METHODS ：Based on everted intestinal sac model ，using accumulative absorption amount （Q）and absorption rate constant （Ka）as indexes ，UPLC-MS/MS method was used to determine the absorption of gastrodin ，parishin A ，parishin B and parishin C from different doses （2.5，5，10 g/L） of G. elata powder with different particle sizes （fine powder 146 μm，superfine powder 52 μm，ultrafine powder 37 μm）in different segments（duodenum，jejunum，ileum and colon ）. RESULTS ：Q and Ka of gastrodin and parishin B （intestinal segment ），Q（colon） and Ka（ileum and colon ）of parishin C in 2.5 g/L G. elata superfine powder ；Q and Ka of gastrodin （intestinal segment ），Q and Ka of parishin B （duodenum，jejunum，ileum）and Ka of parishin C （colon）in 2.5 g/L G. elata ultrafine powder ；Q of gastrodin （duodenum），Q of parishin A and parishin B （intestinal segment ）and Q of parishin C （duodenum，jejunum）in 5 g/L G. elata superfine powder ；Q（duodenum jejunum ，colon）and Ka（intestinal segment ）of gastrodin ，Q of parishin B （duodenum，ileum and colon）and Q of parishin C （duodenum，ileum）in 5 g/L G. elata ultrafine powder ；Q and Ka of parishin B （jejunum，ileum），Q of parishin C （jejunum，ileum）in 10 g/L G. elata superfine powder as well as Q（colon）and Ka（duodenum）of gastrodin ，Q （duodenum，ileum，colon）and Ka（duodenum，colon）of parishin B ，Q（duodenum，ileum）and Ka（duodenum）of parishin C in 10 g/L G. elata ultrafine powder were all increased significantly ，compared with the same dose of G. elata fine powder （P＜0.05 or P＜0.01）. Ka of parishin A （jejunum）and Q of parishin C （duodenum）in 2.5 g/L G. elata superfine powder ；Ka of parishin A （jejunum，ileum）， Q and Ka of parishin C （duodenum，jejunum）in 2.5 g/L G. elata ultrafine powder ；Ka of gastrodin （jejunum，ileum and colon ），Ka of parishin A （colon），Ka of parishin B （ileum）and Ka of parishin C （jejunum，ileum）in 5 g/L G. elata superfine powder ；Ka of gastrodin and parishin C （jejunum，ileum and colon ），Q（jejunum，colon）and Ka（colon）of parishin A ，Ka of parishin B （jejunum，ileum）in 5 g/L G. elata ultrafine powder ；Q and Ka of parishin A （ileum）in 10 g/L G. elata superfine powder ；Q（duodenum）and Ka（jejunum） of parishin A ，Ka of parishin C （jejunum）in 10 g/L G. elata ultrafine powder were decreased significantly ，compared with the same dose of G. elata fine powder （P＜0.05 or P＜0.01）. Q of gastrodin （colon），Q（colon）and Ka（ileum，colon）of parishin A ，Q and Ka of parishin B （jejunum，colon），Q and Ka of parishin C （ileum，colon）in 2.5 g/L G. elata fine powder ；Q and Ka of gastrodin （colon），Q（ileum，colon）and Ka（jejunum，ileum，colon）of parishin A ，Ka of parishin C （colon）in 2.5 g/L G. elata superfine powder；Q（colon）and Ka（jejunum，ileum，colon）of parishin A and C ，Q and Ka（ileum，colon）of parishin B in 2.5 g/L G. elata ultrafine powder ；Q and Ka of gastrodin ，parishin A and C （colon），Ka of parishin B （colon）in 5 g/L G. elata fine powder ；Q and Ka of gastrodin and parishin A （colon），Q and Ka of parishin C （jejunum，ileum，colon）in 5 g/L G. elata superfine powder ；Q and Ka of gastrodin（ileum，colon），Q of parishin A （jejunum，ileum，colon），Q and Ka of parishin B （jejunum，colon），Q（jejunum，colon） and Ka（jejunum，ileum，colon）of parishin C in 5 g/L G. elata ultrafine powder ；Q of gastrodin （colon），Q and Ka of parishin A ，B and C （jejunum，ileum，colon）in 10 g/L G. el ata fine powder ；Q of gastrodin （colon），Q and Ka of parishin A and C （jejunum， ileum，colon），Q and Ka of parishin B （colon）in 10 g/L G. elata superfine powder ；Q（colon）and Ka（jejunum，ileum，colon）of gastrodin，Q and Ka of parishin A and C （jejunum，ileum，colon），Q（jejunum，ileum，colon）and Ka（ileum，colon）of parishin B in 10 g/L G. elata ultrafine powder were decreased significantly ，compared with duodeum of the same group （P＜0.05）. Q and Ka of gastrodin（jejunum）in 2.5 g/L G. elata superfine pow

Study on the Mechanism of Improvement Effects of Gastrodin Injection on Methamphetamine Induced Neurotoxic Damage in Rats via nNOS Pathway / 中国药房

OBJECTIVE：To stu dy the mechanism of improvement effects of Gastrodin injection on methamphetamine induced neurotoxic damage in rats via nNOS pathway. METHODS ：SD rats were randomly divided into control group ，methamphetamine group，regular-dose of gastrodin group ，double-dose of gastrodin group ，negative control （NC）adenovirus group ，NC adenovirus+ methamphetamine group ，NC adenovirus+gastrodin group and nNOS adenovirus+gastrodin group ，with 10 rats in each group. Control group was given normal saline intraperitoneally ，twice a day. Methamphetamine group was given methamphetamine intraperitoneally（7.5 mg/kg），twice a day. Regular-dose and double-dose of gastrodin groups were respectively given different doses of Gastrodin injection （10，20 mg/kg）intraperitoneally 30 min earlier ，once a day ，and then given methamphetamine intraperitoneally by the same way as methamphetamine group. NC adenovirus group was given NC adenovirus （4.8×107 PFU）3 μL once in the striatum and normal saline intraperitoneally ，twice a day. NC adenovirus+methamphetamine group was given NC adenovirus by the same way and methamphetamine （7.5 mg/kg）intraperitoneally，twice a day. NC adenovirus+gastrodin group was given NC adenovirus+methamphetamine by the same way ，meanwhile given Gastrodin injection intraperitoneally （20 mg/kg）30 min before methamphetamine ，once a day. nNOS adenovirus+gastrodin group was given nNOS adenovirus and methamphetamine by the same way ，meanwhile given Gastrodin injection intraperitoneally （20 mg/kg）30 min before methamphetamine ，once a day. Each group was given relevant medicine intraperitoneally 1 mL/100 g，for consecutive 3 days. The stereotyped behavior of rats were observed and scored ；the apoptotic rate ，the protein expression of apoptotic factors （Bcl-2，Bax，Cleaved caspase- 3），the levels of oxidative stress factors （MDA，SOD，GPx） and NO ，the protein expression of nNOS were detected. RESULTS ： Compared with control group ，stereotyped behavior score ，cell apoptosis rate of striatum ，protein expression of Bax ，Cleaved caspase-3 and nNOS ，the levels of MDA and NO were increased significantly in methamphetamine group ；while the protein expression of Bcl- 2 and the levels of SOD and GPx were decreased significantly （P＜0.05 or P＜0.01）. Compared with methamphetamine group ，stereotyped behavior score ，cell apoptosis rate of striatum ，protein expression of Bax ，Cleaved caspase- 3 and nNOS ，the levels of MDA and NO were decreased significantly in regular-dose and double-dose of gastrodin groups ；while the protein expression of Bcl- 2，the levels of SOD and GPx were increased significantly ，and most above indexes in double-dose of gastradin group were better than regular-dose of gastrodin group （P＜0.05 or P＜0.01）. Compared with NC adenovirus group ，cell apoptosis rate of striatum ，protein expression of Bax ，Cleaved caspase- 3 and nNOS ，the levels of MDA and NO were increased significantly in NC adenovirus+methamphetamine group ；while the protein expression of Bcl- 2，the levels of SOD and GPx were decreased significantly （P＜0.01）. Compared with NC adenovirus+methamphetamine group ，cell apoptosis rate of striatum ，protein expression of Bax ，Cleaved caspase- 3 and nNOS ，the levels of MDA and NO were decreased significantly in NC adenovirus+ gastrodin group ；while the protein expression of Bcl- 2，the levels of SOD and GPx were increased significantly （P＜0.01）. Compared with NC adenovirus+gastrodin group ，cell apoptosis rate of striatum ，protein expression of Bax ，Cleaved caspase- 3 and nNOS，the levels of MDA and NO were increased significantly in nNOS adenovirus+gastrodin group ；while the protein expression of Bcl- 2，the levels of SOD and GPx were decreased significantly （P＜0.01）. CONCLUSIONS ：Gastrodin injection can protect rats against neurotoxic damage induced by methamphetamine ，and the effect is related to the inhibition of nNOS-mediated apoptosis and oxidative stress.

Fresh Gastrodia elata ameliorates learning and memory impairments in sleep interruption induced mice / 中草药

Objective: To study the learning and memory enhancement effect of fresh Gastrodia elata (FG) on deficits induced by sleep interruption (SI) in mice. Methods: The contents of gastrodin and p-hydroxybenzyl alcohol in FG were determined by HPLC, and the content of polysaccharide were determined by phenol-sulfuric acid method. Then the learning and memory enhancement effects of FG on sleep deficits induced mice were studied. A total of 60 ICR male mice were randomly divided into the control group, the SI model group, the modafinil group, and the FG groups (3 and 9 g/kg). The mice were sleep interrupted continuously for 14 d, behavioral tests were performed by using open field test, novel object recognition (NOR) experiment, Morris water maze (MWM) task, and passive avoidance method. The levels of SOD and MDA in the serum and the hippocampus, Ach, Glu and NE in the hippocampus were measured. Results: There were no significant changes in locomotor activities among all groups. Compared with the control group, the discrimination index (DI) of SI model group in NOR was decreased significantly, the longer escape latency in MWM was observed in SI mice group, in passive avoidance test the errors times increased and the latent period decreased. In addition, the levels of MDA in the serum and the hippocampus were increased, while the contents of SOD, Ach, Glu and NE in the serum and the hippocampus were decreased significantly. In comparison with the SI group, FG (3 and 9 g/kg) treatment markedly enhanced the discriminative ability by elevating DI in NOR, improved the acquisition and retention of spatial memory by decreasing escape latency, decreased the errors times, and prolonged the latency time in passive avoidance test. Administration of FG significantly reduced the elevated MDA level in the serum and the hippocampus and raised the reduced SOD, Ach, Glu and NE levels in the hippocampus. Conclusion: The results reveal that FG treatment can improve SI-induced learning and memory impairments, and ameliorate oxidative stress damage and raise neurotransmitter content. FG is a potential traditional Chinese medicine for improving learning and memory impairments.

Research progress on antidepressant substances and their mechanisms in Gastrodia elata / 中草药

Gastrodia elata is a valuable traditional Chinese medicine with many pharmacological activities such as calming, anti-depression, anti-anxiety, anti-oxidation, anti-virus, and anti-tumor. In this paper, the research progress of the pharmacodynamic basis and antidepressant mechanism of anti-depression effect of G. elata is summarized. And combined with the current clinical application of G. elata anti-depression, a reference is provided for the further research and development of G. elata.

Research progress on biosynthesis of gastrodin / 中草药

Gastrodin is a major active ingredient in Gastrodia elata, which is one of the traditional rare medicinal herbs in China. It has lots of pharmacological activities, such as reducing blood pressure, anti-epileptic, anti-tumor, and protecting nerve, etc. With the increasing demand for gastrodin in the market and the inherent problems of traditional methods on obtaining gastrodin, new methods are urgently needed to solve various difficulties in the actual production of gastrodin. Biosynthesis is a new method instead of the traditional acquisition method, and has made great progress and achievements in gastrodin acquisition. Therefore, it is necessary to systematically review the biosynthesis of gastrodin from three aspects of biosynthetic pathway, plant transformation and microbial transformation in this paper, hoping to provide valuable reference for further improvement and perfection of gastrodin production method in the future to meet public increasing demand for gastrodin.

The protective effects of gastrodin on methamphetamine-induced neurotoxicity / 中国药理学通报

Aim To compare the protective effects of gastrodin and melatonin on methamphetamine ( MA )-induced neurotoxicity of cortical neurons. Methods Cortical neuron cells were treated with different concentrations of methamphetamine for 24 h, then the cell viability was detected by CCK-8 kit to choose the optimal concentration of MA. The cells were treated with different concentrations of gastrodin or melatonin 2 h before the treatment of cells with optimal concentration of MA for 24 h. The optimal concentrations of gastrodin and melatonin were determined by CCK-8 kit as well. Cortical neuron cells were randomly divided into con trol group, MA group, gastrodin plus MA group, and melatonin plus MA group. Then the apoptotic rate of cells was detected by TUNEL method . The expression of Caspase-3 in cells was assessed by immunofluorescence. Results The optimal concentration of MA was 0. 5 mmol • L"1. After treating with MA, the numbers of cortical neurons decreased, the length of synapses became shorter, and the cell vacuolation, the number of dead cells increased. The optimal concentration of gastrodin was 25 mg • L'1, and the optimal concentration of melatonin was 10 jimol • L"1. Gastrodin treatment could reduce the apoptosis of cortical neurons induced by MA, and the expression of Caspase-3 decreased as well, and there was no significant difference compared with the intervention effect of gastrodin with melatonin. Conclusions MA has neurotoxic damage to cortical neurons. Gastrodin could attenuate MA-in-duced neurotoxicity via alleviating the apoptosis induced by MA.

Effect of acupuncture plus medication on expression of Bcl-2 and Bax in hippocampus in rats with Alzheimer's disease / 中国针灸

OBJECTIVE@#To explore the mechanism of acupuncture plus medication on treatment of Alzheimer's disease (AD).@*METHODS@#Sixty adult SD rats were randomly divided into a normal group, a sham operation group, a model group, an electroacupuncture (EA) group, a gastrodin group and an EA+gastrodin group, 10 rats in each one. The rat model of AD was established by intraperitoneal injection of D-galactose and bilateral hippocampal injection of Aβ1-40. Two weeks after modeling, the rats in the EA group and EA+gastrodin group were treated with EA at "Baihui" (GV 20) "Dazhui" (GV 14) and bilateral "Zusanli" (ST 36), 30 min per treatment, once a day for consecutive 4 weeks. The rats in the gastrodin group and EA+gastrodin group were treated with intraperitoneal injection of gastrodin, once a day for consecutive 4 weeks. The rats in the normal group, model group and sham operation group were not treated. The morphology of hippocampal neurons was observed by using HE staining. The expression of Bcl-2 and Bax in the hippocampal CA1 area was detected by using immunohistochemical method. The expression of Bcl-2 and Bax protein in hippocampus was detected by using Western blot.@*RESULTS@#The HE staining results showed the arrangement of neurons in the hippocampal CA1 area was regular in the normal group and the sham operation group, and the cytoplasm and nucleus were clearly visible. The neurons in the model group were severely damaged; the cell arrangement was not close, and the cell morphology was incomplete. Compared with the model group, the cell morphology of each intervention group was significantly improved. The immunohistochemistry results showed that, compared with the normal group and the sham operation group, the expression of Bcl-2 in the hippocampal CA1 region in the model group was decreased (<0.05), but the expression of Bax was enhanced (<0.05); compared with the model group, the expression of Bcl-2 was increased (all <0.05) and the expression of Bax was decreased (all <0.05) in all intervention group; compared with the EA group or the gastrodin group, the expression of Bcl-2 was enhanced (<0.05) and the expression of Bax was decreased (<0.05) in the EA+gastrodin group. The result of Western blot method was consistent with that of immunohistochemistry method.@*CONCLUSION@#EA and gastrodin could significantly inhibit the expression of Bax and up-regulate the expression of Bcl-2, and the combination of EA and gastrodin has the most significant effect. This indicates that EA combined with gastrodin has synergistic effect on inhibiting the apoptosis of neurons in hippocampus in AD rats, which may be one of the mechanisms of EA plus medication on AD lesions.

Effect of Gastrodin on Early Brain Injury and Neurological Outcome After Subarachnoid Hemorrhage in Rats / 神经科学通报·英文版

Gastrodin is a phenolic glycoside that has been demonstrated to provide neuroprotection in preclinical models of central nervous system disease, but its effect in subarachnoid hemorrhage (SAH) remains unclear. In this study, we showed that intraperitoneal administration of gastrodin (100 mg/kg per day) significantly attenuated the SAH-induced neurological deficit, brain edema, and increased blood-brain barrier permeability in rats. Meanwhile, gastrodin treatment significantly reduced the SAH-induced elevation of glutamate concentration in the cerebrospinal fluid and the intracellular Ca overload. Moreover, gastrodin suppressed the SAH-induced microglial activation, astrocyte activation, and neuronal apoptosis. Mechanistically, gastrodin significantly reduced the oxidative stress and inflammatory response, up-regulated the expression of nuclear factor erythroid 2-related factor 2, heme oxygenase-1, phospho-Akt and B-cell lymphoma 2, and down-regulated the expression of BCL2-associated X protein and cleaved caspase-3. Our results suggested that the administration of gastrodin provides neuroprotection against early brain injury after experimental SAH.

Comparison of in Vitro Dissolution of Active Ingredients in Gastrodiae Rhizoma Powder with Different Particle Size / 中国实验方剂学杂志

Objective: To compare in vitro dissolution behaviors of active ingredients (gastrodin,parishin A,p-hydroxybenzyl alcohol,parishin B and parishin C) in Gastrodiae Rhizoma powder with different particle size.Method: In vitro dissolution of Gastrodiae Rhizoma powder in different dissolution media (water,artificial gastric juice and artificial intestinal juice) were detected by stirring paddle method.Dissolution of these five components in Gastrodiae Rhizoma powder with different particle size was determined by HPLC,mobile phase was acetonitrile-0.1% phosphoric acid solution for gradient elution,column temperature was set at 40℃ and detection wavelength was 220 nm.Result: In water and artificial intestinal juice,the dissolution rates of five active components in three kinds of Gastrodiae Rhizoma ultrafine powders were higher than that of the fine powder and the finest powder;in artificial gastric juice,the dissolution rates of gastrodin and p-hydroxybenzyl alcohol in Gastrodiae Rhizoma ultrafine powders were higher than that of the other powders,and the dissolution rate of parishin A in Gastrodiae Rhizoma ultrafine powders was lower than that of the other powders.Conclusion: An appropriate degree of superfine grinding can promote the dissolution of active ingredients in Gastrodiae Rhizoma,but not as fine as possible.The dissolution medium has an obvious influence on the dissolution behaviors of active components,which provides a reference for screening optical particle size of Gastrodiae Rhizoma powder in clinical application.

Effect of Gastrodia elata on improving sleep in mice and its mechanism / 中草药

Objective: To investigate the sleep promoting effect of Gastrodia elata on mice and its effect on the central dopamine (DA) system, and study the mechanisms of gastrodin, the active constituent of G. elata. Methods: Five groups of mice were treated by gavage with saline, gastrodia, olive oil, gastrodia mixed with olive oil and positive control Estazolam, respectively, for 20 d (10 mice per group). Hypnosis induced by suprathreshold and subthreshold doses of Pentobarbital sodium were used to evaluate the effect of G. elata on sleep in mice. Sleep latency, occurrence rate and duration were recorded at the 7th and 20th day. The DA content in the brain tested by ELISA and the expression of DA receptor subtypes detected by qRT-PCR were used to determine the effect of G. elata on central DA system. Western blotting was further used to detect the expression levels of ERK pathway related protein. Results: After 20 d of gavage, the sleep latency of mice was significantly shortened, the sleep occurrence rate was increased, the sleep duration was prolonged, and the content of brain DA was significantly increased. At the same time, the expression levels of all the dopamine receptor subtypes were significantly up-regulated. Furthermore, the gastrodin, the active constituent of G. elata, could activate the dopamine receptor D2 rather than the D1-mediated signaling pathway. Conclusion: G. elata might regulate sleep by up-regulating the activity of central DA system. Gastrodin, the active constituent of G. elata, could play a regulating role through D2-mediated signaling pathway.

Glycoside constituents from Bletilla striata / 中草药

Objective: To study the glycoside chemical constituents of Bletilla striata grown in Guizhou Province. Methods: The glycoside chemical constituents were separated and purified by silica gel column chromatography, MCI, gel column chromatography, medium pressure preparation, highperformance liquid chromatography and other modern separation methods. The structures were identified based on the spectral data. Results: Thirteen glycosides were isolated from the 95% ethanol extracts: 1-methyl-3-phenylpropyl-β-D-glucopyranoside (1), 1-(4-β-D-glucopyranosyl oxybenzyl) 4-ethyl (2R)-2-isobutylmalate (2), 3’,5- dimethoxy-bibenzyl-3-O-β-D-glucopyranoside (3), syringaresinol mono-β-D-glucoside (4), 4-O-(6’-O-glucosyl-p-coumaroyl)-4- hydroxybenzyl alcohol (5), (4-hydroxyphenyl) methyl-β-D-glucopyranoside (6), 4-methylphenyl-β-D-glucopyranoside (7), benzyl-β-D- glucopyranoside (8), phenyl-β-D-glucopyranoside (9), 4-[(acetyloxy) methyl] phenyl-β-D-glucopyranoside (10), batatasin III-3-O- glucoside (11), gastrodin (12) and 4-(4-β-D-glucopyranosyl-oxybenzyl)-(2R)-2-isobutylmalate (13). Conclusion: Compound 1 is a new natural product. Compounds 3-6 are isolated from this plant for the first time; Compounds 2, 7-13 are obtained from this genus for the first time.

Gastrodin induced HO-1 and Nrf2 up-regulation to alleviate H2O2-induced oxidative stress in mouse liver sinusoidal endothelial cells through p38 MAPK phosphorylation

Nuclear factor erythroid-related factor 2 (Nrf2) has been implicated in several detoxifying and antioxidant defense processes. Nrf2-mediated heme oxygenase-1 (HO-1) expression was demonstrated to play a key role against oxidative stress. Gastrodin (GSTD) is a well-known active compound isolated from the roots of Rhizoma gastrodiae, a plant used in ancient Chinese traditional medicine. The aim of this work was to investigate whether GSTD could alleviate H2O2-induced oxidative stress in mouse liver sinusoidal endothelial cells (LSECs). In LSECs exposed to 1 mM H2O2, treatment with GSTD (1, 10, or 50 µM) resulted in higher cell viability than the untreated control. Treated cells maintained a higher Bcl2/Bax ratio and suppressed caspase-9 expression compared with untreated cells, reducing cell apoptosis. GSTD was protective for H2O2-induced oxidative injury by reducing the generation of intracellular reactive oxygen species and malondialdehyde. HO-1 and Nrf2 expressions were synergistically upregulated by GSTD. Inhibition of HO-1 by 10 µM zinc protoporphyrin resulted in less protective effects on cell viability and malondialdehyde reduction by GSTD treatment in H2O2-exposed LSECs. Additionally, phosphorylated p38 in LSECs exposed to H2O2 was elevated by GSTD. Inhibition of p38 phosphorylation by SB203580 did not induce Nrf2 and HO-1 expression after 1 or 10 µM GSTD treatment and the protective effect on cell viability and malondialdehyde reduction in H2O2-exposed LSECs was reduced. The data conclusively demonstrated that GSTD-induced HO-1 and Nrf2 expression is involved in protection of LSECs from H2O2-induced oxidative injury, which may be regulated by p38 phosphorylation.

Electroacupuncture plus Gastrodin Improves Learning-memory Ability Possibly by Up-regulating Expression of SIRT 1 and PGC-1 ɑ in Hippocampal CA 1 Region of Alzheimer's Disease Rats / 针刺研究

OBJECTIVE: To observe the effect of electroacupuncture (EA) combined with Gastrodin on learning-memory ability and expression of silent information regulator 2 homologous protein 1(SIRT 1) and peroxisome proliferator activated receptor γ coactivator (PGC-1 ɑ) of hippocampal CA 1 region in Alzheimer's disease(AD) rats, so as to explore its mechanism under-lying improvement of AD. METHODS: Sixty male SD rats were randomly divided into normal control (normal), sham operation (sham), model, EA, Gastrodin and EA＋ Gastrodin groups (n＝10 in each). The AD model was established by intraperitoneal injection of D-Galactose (120 mg•kg－1•d－1) combined with bilateral hippocampal injection of β amyloid 1-40(Aβ 1-40). EA was applied at "Baihui"(GV 20), "Dazhui"(GV 14) and "Zusanli"(ST 36) for 30 min, once daily for 4 weeks. For rats of the Gastro-din group and EA＋ Gastrodin group, intraperitoneal injection of gastrodin(10 mg/kg) was conducted once daily for 4 weeks. Morris water maze tests were used to assess the rat's learning-memory ability. Nissl staining was used to assess the morphological changes of neurons in the hippocampal CA 1 area. The expression of SIRT 1 and PGC-1 ɑ of hippocampal CA 1 region was mea-sured by immunohistochemical staining. RESULTS: 1) Morris water maze tests showed that, compared with the normal and sham group, the escape latency was significantly prolonged (P<0.05), and the percentage of platform quadrant residence duration and the platform crossing times were considerably decreased in the model group (P<0.05). After the intervention, the escape latency was obviously shortened (P<0.05), and the percentage of platform quadrant residence duration and the platform crossing times were markedly increased in the EA, Gastrodin and EA＋Gastrodin groups relevant to the model group (P<0.05). 2) Nissl staining showed that, in comparison with the normal group or sham group, the number of cells in the hippocampal CA 1 area was decreased and the arrangement was disorganized in the model group. The number of cells in CA 1 area was relatively higher in the 3 treatment groups than in the model group. 3) The expression levels of SIRT 1 and PGC-1 ɑ proteins in the hippocampal CA 1 area were significantly down-regulated in the model group than in the normal and sham groups (P<0.05). After the intervention, the expression levels of SIRT 1 and PGC-1 ɑ in the EA, Gastrodin and EA＋Gastrodin groups were significantly up-regulated compared with the model group (P<0.05). The effects of EA＋Gastrodin were significantly superior to those of simple EA and simple Gastrodin in shortening the escape latency, up-regulating the expression levels of SIRT 1 and PGC-1 ɑ as well as in increasing the percentage of platform quadrant residence time and platform crossing times (P<0.05). CONCLUSION: Both EA and Gastrodin can improve the learning-memory ability of AD rats, which may be related to their effects in up-regulating the expression of SIRT 1 and PGC-1 ɑ and reducing neuronal injury in the CA 1 region of hippocampus, suggesting a protective role of EA on hippocampal neurons. The effect of EA combined with Gastrodin is markedly better than that of EA and Gastrodin alone.

Role of mitochondrial permeability transition pore in mediating the inhibitory effect of gastrodin on oxidative stress in cardiac myocytes / 南方医科大学学报

OBJECTIVE@#To explore the role of mitochondrial permeability transition pore (mPTP) in mediating the protective effect of gastrodin against oxidative stress damage in H9c2 cardiac myocytes.@*METHODS@#H9c2 cardiac myocytes were treated with HO, gastrodin, gastrodin+HO, cyclosporin A (CsA), or CsA+gas+HO group. MTT assay was used to detect the survival ratio of H9c2 cells, and flow cytometry with Annexin V-FITC/PI double staining was used to analyze the early apoptosis rate after the treatments. The concentration of ATP and level of reactive oxygen species (ROS) in the cells were detected using commercial kits. The mitochondrial membrane potential of the cells was detected with laser confocal microscopy. The expression of cytochrome C was detected with Western blotting, and the activity of caspase-3 was also assessed in the cells.@*RESULTS@#Gastrodin pretreatment could prevent oxidative stress-induced reduction of mitochondrial membrane potential, and this effect was inhibited by the application of CsA. Gastrodin significantly lowered the levels of ROS and apoptosis-related factors in HO-exposed cells, and such effects were reversed by CsA. CsA significantly antagonized the protective effect of gastrodin against apoptosis in HO-exposed cells.@*CONCLUSIONS@#Gastrodin prevents oxidative stress-induced injury in H9c2 cells by inhibiting mPTP opening to reduce the cell apoptosis.

Determination of six ingredients of gastrodin from gastrodia elata with quantitative analysis of multi-components by single marker / 中草药

Objective To establish a quantitative analysis of multi-components by single marker (QAMS) for determining contents of six gastrodin compositions in Gastrodia elata, gastrodin, p-hydroxybenzyl alcohol, parishin A, parishin B, parishin C and parishin E. Methods Separation was carried out on Agilent Zorbax SB-C18 Column (250 mm × 4.6 mm, 5 μm) by using acetonitrile (A)-0.05% Phosphoric acid aqueous solution (B) as the mobile phase, and a linear gradient elution was employed (0-5 min, 2% A, 5-20 min, 2%-30% A, 20-25 min, 30%-2% A) with detection wavelength at 220 nm, flow velocity at 1.0 mL/minand column temperature at 25 ℃. Five relative correction factors (RCFs) of the five compositions using gastrodin as the internal standard were applied to determine their contents in all samples It was established to determine the contents of gastrodin, p-hydroxybenzyl alcohol, parishin A, parishin B, parishin C, parishin E by the external standard method (ESM) at the same time. The feasibility and accuracy of QAMS method were verified by comparing the content results determined by ESM and QAMS. Results Within the linear range, the RCFs of gastrodin, p-hydroxybenzyl alcohol, parishin A, parishin B, parishin C, parishin E were 0.663, 1.278, 1.435 and 1.591, respectively, with good repeatability in different experimental conditions. There was no significant difference between the QAMS method and ESM method. Conclusion The QAMS method is feasible and accurate for the determination of the six chemical compositions, and which can be used for quality control of G. elata.