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.

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.

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.

Research progress on Gastrodia elata prevention and treatment neuropsychiatric diseases / 中草药

Gastrodia elata is a kind of precious Chinese materia medica, which showed good effect in the treatment of headache, improve learning and memory, antidepressant, and other neuropharmacology effects. In this paper, the neuropharmacology effects and corresponding chemical constituents of G. elata in the last ten years were summarized and sorted by referring the literature of CNKI and PubMed, as well as inquired the development of G. elata related drugs and health care products in the official website of the Chinese Food and Drug Administration, which provides reference to develop nervous and mental diseases products of G. elata.

Comparison on main chemical composition contents in three variants of Gastrodia elata from southwest China / 中草药

Objective To compare and principal component analyze the main chemical composition contents in three variants of Gastrodia elata from southwest of China in order to provide reference for further researches and evaluations on the quality of G. elata. Method Sulfuric acid-phenol method was used to measure the contents of polysaccharides of three variants of G. elata, the contents of free amino acid of three variants of G. elata were estimated by ninhydrin colorimetric method, and the total phenol content of three variants of G. elata were measured by Folin-Ciocaileu colorimetry. HPLC method was applied to determine the contents of gastrodin and hydroxyphenyl methanol of three variants of G. elata from southwest China, and the data was analyzed by principal component analysis. Results The results indicated that the contents of main chemical composition had great difference in different variants of G. elata from southwest China, and the coefficient of variation (CV) of polysaccharide contents was the smallest with 18.025%, and CV of hydroxyphenyl methanol was the biggest with 48.978%. Among the components in this study, the contents of free amino acid, total polyphenol, and gastrodine of G. elata f. glauca from Beichuan in Sichuan province were 2.873%, 0.805%, and 0.862%, respectively, which were all extremely different from the other materials. G. elata f. viridis from Dafang in Guizhou province had the highest content of polysaccharide with the value of 29.225%, which had significantly higher content of polysaccharide than that of G. elata f. glauca from Beichuan and had extremely significantly higher content of polysaccharides than that of the rest. G. elata f. glauca from Zhaotong in Yunnan province had the highest content of hydroxyphenyl methanol, then G. elata f. elata from Guangyuan in Sichaun province, with no significant difference between them. Conclusion There was no significant difference among the regions or variants in main chemical composition contents. The quality of G. elata f. glauca from Beichuan in Sichuan province was the best from the perspective of main chemical composition.

Transcriptome analysis on symbiotic molecular mechanism of Armillaria mellea and Gastrodia elata / 中草药

Objective In this study, Armillaria mellea and Zhaotong Gastrodia elata vegetative stems were used as the experimental material to reveal the symbiosis molecular mechanism of A. mellea infecting G. elata through comparative transcriptome sequencing analysis. Methods Trizor reason (invitrogen) was used to extract RNA from vegetative propagation stem samples, and the sequencing library was established and sequenced. Results The results showed that 38 838 sequences were annotated when G. elata symbiosis with A. mellea. Under false discovery rate (FDR) 1 screening conditions, there were significant differences in expression levels among the 23 333 genes, in which 1 595 genes up-regulated and 21 738 down-regulated expression. Based on the transcriptome analysis, unigenes associated with the extracellular enzyme gene cellulase, xylanase, laccase, and polygalacturonase genes were 21, 6, 39, and 6 genes, respectively, in which the number of corresponding differentially expressed genes were 9, 3, 23, and 4, respectively. The expression of all these unigenes were down-regulated except for one cellulase genes. The number of unigenes related to anti-oxidant enzymes superoxide dismutase, peroxidase, and catalase were 13, 7, and 17, respectively. The differentially expressed genes respectively were 6, 7, and 7, all of which were down-regulated. In addition, these genes were selectively confirmed by real-time PCR. Conclusion The life activites of A. mellea which infect and symbiosis with G. elata declined, meanwhile G. elata did not produce biological stress for A. mellea. This study provides a lot of valuable genetic resources for further studying the symbiosis molecular mechanism of A. mellea and G. elata.

Determination of the Contents of Gastrodin, 4-Hydroxybenzyl Alcohol and Gastrodia Polysaccharides in Different Processed Products of Gastrodia elata Bl. f. glauca S. Chow / 中国药学杂志

OBJECTIVE: To evaluate the qualities of different processed products of Gastrodia elata Bl. f. glauca S. Chow, such as steamed, wine-processed, ginger-processed and tribulus-processed products, and determine the optimum processing method of Gastrodia elata. METHODS: HPLC was employed to determine gastrodin and 4-hydroxybenzyl alcohol, and phenol-sulfuric acid method was used to detect the content of gastrodia polysaccharide. RESULTS: Gastrodin and gastrodia polysaccharides had the highest contents(1.901 and 3.246 mg·g-1, respectively) in the tribulus-processed products; and the highest content of 4-hydroxybenzyl alcohol in ginger-processed Gastrodia elata was 0.80 mg·g-1; gastrodin in two kinds of wine-processed products was lower than the others, while the content of gastrodia polysaccharide in Gastrodia elata processed by wine-soaking for three days was obviously less than those in the other products. CONCLUSION: The traditional tribulus-processed method is better than the modern steaming method. This study can provide scientific basis for the mechanism of traditional processing method of Gastrodia elata.

Research on variable-temperature drying process of Zhaotong Gastrodia elata Bl. f. glauca S. Chow / 中草药

Objective: To put an end to events of sulfur content over-standard in Zhaotong Gastrodia elata Bl. f. glauca S. Chow (ZGE) and to improve the drying efficiency. Methods: Effects of steaming time and drying parameters on phenomenal and quality characteristics of ZGE were investigated. Drying process of ZGE adapt to modern process equipment was explored. Results: Optimal steaming time for second grade fresh ZGE was 4 min under 95℃. Suitable drying process parameters were as follows: ZGE was natural cooling at room temperature for 12 h after steamed. ZGE was drying at 30℃ for 1 d, and then the temperature was raised with the speed of 2℃/12 h to 40℃. Then with the speed of 5℃/12 h until to 60℃, all the work can be stopped until the ZGE was dried completely. It totally took about 6 d. ZGE appeared light brown and less deformation, and the fold was shallow, without hollow. The content of gastrodin was 0.93%, soluble extract content was 20.1%, and the content of total ash was 2.65%. All quality indexes were corresponded to the standard of Chinese Pharmacopoeia 2010. Conclusion: Drying process of ZGE was applied to modern processing equipment. It can not only guarantee the phenomenal but also the quality of ZGE. Meanwhile, it can improve the production efficiency and was suited to demonstrate and extend in Zhaotong region.

Genetic diversity of Gastrodia elata based on SRAP analysis / 中草药

Objective: In order to correctly identify the different germplasm resources of Gastrodia elata and gain the excellent germplasm resources, SRAP molecular marker was used to analyze the genetic diversity of G. elata. Methods: G. elata was collected from seven different areas, which included 24 smaples of G. elata f. elata, G. elata f. g1auca and G. elata f. viridis, the DNA figerprint of G. elata was constructed with SRAP molecular marker, and the genetic diversity was analyzed. Results: Six huandred Thirty-seven belts were amplified by 33 primers pairs, and the polymorphic percentage was 73.16%. The range of genetic similarity coefficient was 0.404 0-0.908 0, the genetic similarity coefficient among G. elata f. elata was in the range of 0.906 6-0.996 4, and those of G. elata f. g1auca, G. elata f. viridis, and hybrid was in the range of 0.410 4-0.999 6, 0.541 0-0.950 4 and 0.578 2, respectively. They showed small genetic differences. The analysis of molecular variance showed higher percentages of genetic variation within population than those among populations in all species. Populations showed higher genetic structure (FST = 0.33, P < 0.05). In addition, artificial cultivation had influence to the genetic differentiation, but not significantly. So in terms of different cultivation conditions so far had no significant impact on the genetic differentiation of G. elata. Mantel's test has been used to detect the correlation between genetic distance and geographic distance of all sorts of germplasm resources, the result had no significant correlation, and due to the limited number of samples, the result is not representative. Conclusion: SRAP molecular marker method can more effectively reflect the genetic polymorphisms of G. elata. Compared with other two phenotypes, G. elata f. elata is more conservative and has lower genetic diversity. The other two variants have higher genetic diversity.

Genetic Polymorphism of Gastrodia elata Bl.Populations from Guizhou Based on ISSR Analysis / 中华中医药杂志

Objective:To construct the fingerprinting and analyze the genetic relationship in Gastrodia elata Bl.Methods:Inter-simple sequence repeat(ISSR) markers were used to investigate 23 populations of wild and cultivated Gastrodia elata Bl.in Guizhou Province.4 effective primers which were screened from 24 ISSR primers detected 32 loci in 23 samples,29 loci were polymorphic,amounting to 90.63% of the total loci..The average number of bands was 8 and the percentage of polymorphic loci(PPB)was from 80% to 100%.UPGMA dendrogram was analyzed by NTSYSpc,ver.2.2.Results:Similarity coefficients of the 23 populations were from 0.13 to 1.00.Individuals of the same population did not strictly cluster together and the genetic diversity of individuals in different populations was considerably high.Conclusion:Geographical distribution,variant growing conditions and artificial selection were significant factors for the rich genetic polymorphism of Gastrodia elata Bl.

Distinction between Gastrodia elata and its four falsified specimens and determination of gastrodin by capillary electrophoresis / 中草药

To develop a method for the distinction of genuine Gastrodia elata B1. from its four falsified plants of common occurrence and determination of its active principle, gastrodin by capillary electrophoresis. Methods　The electrophoretic conditions were a buffer solution containing 24 mmol/L borate (pH=9.4), at a voltage of 16 kV. Results　The electrophorograms obtained under the above conditions showed easily distinguishable differences. Baseline separation of gastrodin was achieved and its quantitative analysis can be accomplished within 15 min. The linear correlation equation was Y=-0.048+7.79X (r=0.999). Conclusion　This method could serve as an easy and precise method for the quality control of G. elata.

Influences of wild,degradative,and rejuvenative Armillearia mellea on yield of Gastrodia elata and content of gastrodin / 中草药

Objective By rejuvenation of degradative strain,the effects of Armillearia mellea to promote the growth of Gastrodia elata and to accumulate the gastrodin in G.elata could be recovered.Methods G.elata was inoculated with same variety of G.elata with A.mellea of wild strain,degradative strain for continuously asexual reproduction and rejuvenative strain,and the yield of G.elata and the content of gastrodin of G.elata were determined.Results The significant differences(P

HPLC Fingerprint of Gastrodia elata / 中草药

Objective To establish HPLC fingerprint for evaluating and controlling the quality of Gastrodia elata. Methods HPLC Analysis and similarity calculation were used for establishing fingerprint chromatogram and classifying 18 different original samples; LC-MS and pure compound comparisons were employed for the fingerprint peaks identification. Results HPLC Fingerprint chromatogram was cons- tructed with 27 fingerprint peaks, 17 of which were common fingerpring peaks and 12 main compounds were identified. According to the similarity to standardize herbal medicine, 18 samples could be classified three groups, Anhui-Henan, Shanxi-Sichuan, and Yunnan habitats. The two critical points were 0.80 and 0.58 for correlation coefficient and 0.88 and 0.73 for cosine values. Conclusion The selected fingerprint peaks are diagnostic for G. elata and the constituted HPLC fingerprint chromatogram could be used for identifying different habitats and served as a powerful tool for further quality control of G. elata.

Studies on cultivation of Gastrodia elata to lower expense and increase yield / 中草药

Object As stated in the subject. Methods By field cultivation.Results In sexual reproduction, (1) short tree branches can be used instead of the conventional tree trunks. Sow four layers of seed on to two timber layers. The peretration rate of Amillaria mellea (Vahl. ex Fr.) Karst into the germinated Gastrodia elata Bl.protocorm can attain well over 50%. After half year, the yield of both small and medium sized gastrodia tubers (2) Sow four layers of seed onto every two layers of long tree trunks may double the yield. In asexual propagation, the use of short tree branches not only can save timber material but also maintain the yield.Conclusion This new cultivation method can be used to save timber while maintaining the quality and quantity in the production of G. elata.

Distinction between Gastrodia elata and its four falsified specimens and determination of gastrodin by capillary electrophoresis / 中草药

Object To develop a method for the distinction of genuine Gastrodia elata B1 from its four falsified plants of common occurrence and determination of its active principle, gastrodin by capillary electrophoresis Methods The electrophoretic conditions were a buffer solution containing 24 mmol/L borate (pH=9 4), at a voltage of 16 kV Results The electrophorograms obtained under the above conditions showed easily distinguishable differences Baseline separation of gastrodin was achieved and its quantitative analysis can be accomplished within 15 min The linear correlation equation was Y=-0 048+7 79X (r=0 999) Conclusion This method could serve as an easy and precise method for the quality control of G elata