Protective Effect of Total Ginsenoside Ginseng Root on Learning and Memory Impairment and Anxiety in Rats Induced by Hindlimb Suspension / 中国实验方剂学杂志

Objective:To investigate the effect of total ginsenoside ginseng root on the learning and memory impairment and anxiety of hindlimb suspension rats by detecting the performance of rats in the water maze， elevated plus maze， and the expression of hypothalamic-pituitary-adrenal （HPA） axis， inflammatory factors and tryptophan pathway related factors through the intervention of ginsenosides in hindlimb suspension rats. Method:The Wistar male rats were divided into normal group， hindlimb suspension model group， Huperzine A group （0.1 mg·kg^-1）， and total ginsenoside ginseng root low and high dose groups （100， 200 mg·kg^-1）， with 8 rats in each group. Except for the normal group， the rats in the other groups maintained a -30° hindlimb suspension state for 24 h. The normal group and the model group received intragastric administration of 10 mL·kg^-1 pure water . After 28 days of continuous administration， the water maze and elevated plus maze behavioral tests were performed. After the tests， blood was taken from the abdominal aorta， and the rat brain cortex was peeled off on ice， quenched with liquid nitrogen， and stored at -80 ℃ for later use. LC-MS/MS was used to detect neurotransmitter levels of dopamine， acetylcholine， glutamate， <italic>γ</italic>-aminobutyric acid and tryptophan pathway metabolites （tryptophan， 5-hydroxytryptamine， 5-hydroxyindoleacetic acid， kynurenine， 3-hydroxykynurenine， and kynurenine） in rat brain cortex. An enzyme-linked immunosorbent assay （ELISA） kit was used to detect the levels of inflammatory factors interleukin-6 （IL-6）， interleukin-10 （IL-10， the HPA axis-related hormone corticotropin （ACTH）， and the level of corticosterone （CORT）. Result:Compared with the normal group， the escape latency in the water maze significantly increased， the number of crossings was significantly reduced， and the number of open-arm entry and the percentage of open-arm entry were significantly reduced in the elevated plus maze in model group （<italic>P</italic><0.05，<italic> P</italic><0.01）， the content of dopamine， acetylcholine， glutamic acid， and <italic>γ</italic>-aminobutyric acid in the cortex decreased， kynurenine and kynurenic acid showed an upward trend， 3-hydroxykynurenine， 5-hydroxytryptamine， 5-hydroxyindole acetic acid showed a downward trend， and the levels of IL-6， IL-10， ACTH， and CORT in the serum significantly increased （<italic>P</italic><0.05， <italic>P</italic><0.01）. Compared with the model group of rats， total ginsenoside ginseng root low and high dose groups group reduced the avoidance latency in the water maze， and increased the number of crossings and the number of open arms of the elevated plus maze， dopamine， acetylcholine， glutamate， and <italic>γ</italic>-aminobutyl content increased， while kynurenine and kynurenic acid showed a downward trend， 3-hydroxykynurenine， serotonin， and 5-hydroxyindole acetic acid showed an upward trend， and IL-6， IL-10， ACTH， and CORT factor levels were down-regulated（<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:Hindlimb suspension for 28 days in simulated microgravity can impair the learning and memory ability of rats and cause anxiety-like behaviors. Total ginsenoside ginseng root can improve their learning and memory impairment and anxiety-like behaviors. The mechanism may be mainly related to inhibiting body inflammation and regulating HPA axis imbalance.

Anti-fatigue Analysis of Common Mechanisms of Interaction of Ginseng Radix et Rhizoma "Tonifying Qi" and Notoginseng Radix et Rhizoma "Enriching Blood" / 中国实验方剂学杂志

Objective:To analyze the common active ingredients， potential target genes and pathways of Ginseng Radix et Rhizoma "Tonifying Qi" and Notoginseng Radix et Rhizoma "Enriching blood" in alleviating fatigue based on the network pharmacology technology. And the compound ingredients of total Ginsenoside Ginseng Root and Notoginseng total Saponins were selected to verify the core target genes in vitro. Method:The main active ingredients and related targets of Ginseng Radix et Rhizoma and Notoginseng Radix et Rhizoma were screened by traditional Chinese medicine systems pharmacology （TCMSP）. The data of fatigue genes were established by GeneCards comprehensive database and Human Mendelian Genetic Integrated Database（OMIM）. Depending， The data sets of fatigue-related genes are established based on the data bank of GeneCards and OMIM. The intersecting genes of drugs and disease were obtained by R software. Cytoscape software was used to establish the regulatory network among the active ingredients， drug targets and fatigue-related genes. PPI network of intersecting genes was constructed by STRING 11.0 software， and the core genes were screened by CytoHubba software and Matthews correlation coefficient （MCC） algorithm. Based on the results of network analysis， 24 male SPF ACR mice were randomly divided into control group， total Ginsenoside Ginseng Root group （0.08 g·kg-1） and Notoginseng total Saponins group （0.08 g·kg-1）. The corresponding drugs were given for 3 weeks. The expressions of core genes in muscle tissue were detected by real-time fluorescence quantitative PCR. Result:The 20 active components and 181 drug targets were screened from TCMSP. 33 intersecting genes of diseases and drugs were obtained when compared with GeneCards and OMIM comprehensive database using R software. 10 core genes including aryl hydrocarbon receptor （AHR）， androgen receptor （AR）， glutathione S-transferase P1 （GSTP1）， cysteine proteinase-3（Caspase-3）， cytochrome p450 enzyme 3A4 （CYP3A4）， intercellular adhesion molecule 1 （ICAM1） and nuclear factor kappa B inhibitor alpha （NFKBIA） were screened out by the algorithm of MCC. Total Ginsenoside Ginseng Root and Notoginseng total Saponins had no significant effect on GSTP1 and ICAM1 genes， but they could significantly inhibit the expressions of AHR， CYP3A4， Caspase-3， NFKBIA and AR （P<0.05，P<0.01）， and there were no significant difference in anti-fatigue effect between total Ginsenoside Ginseng Root and Notoginseng total Saponins groups. Conclusion:The mechanism of anti-fatigue of Ginseng Radix et Rhizoma and Notoginseng Radix et Rhizoma may be related to the regulation of AHR， CYP3A4 and Caspase-3 genes， and there is no significant difference in their anti-fatigue effects， through the analysis of network and experimental verification.

Study on the Effects of Ginseng Root Extract on Autophagy ，Proliferation and Cytokine Secretion of Mice Splenic Lymphocytes and Its Mechanism / 中国药房

OBJECTIVE：To study the effects of gin seng root extract on autophagy ，proliferation and cytokine of splenic lymphocyte of mice ，and to study its mechanism. METHODS ：The splenic lymphocyte of mice were divided into blank control group，positive control group （concanavalin A ，5 μg/mL），different concentration of ginseng root extract groups （32，125，500 μg/mL，by lyophilized powder ）. After 48 h of culture with the corresponding medicine ，acridine orange staining method was used to detect autophagy of splenic lymphocyte ；CCK-8 assay was used to detect the cell proliferation ；ELISA assay was used to determine the levels of IL- 4，IL-6 and TNF-α in cell culture；Western blotting method was used to detect the expression of LC 3B and Beclin- 1，as well as the phosphorylation of AMPK ，AKT and mTOR. RESULTS ：Compared with blank control group ，32， 125，500 μg/mL ginseng root extract could increase the number of acidic autophagosomes in splenic lymphocytes of mice（P＜0.05 or P＜0.01）；125，500 μg/mL Ginseng root extract could significantly enhance the survival rate of splenic lymphocytes，the levels of IL- 4，IL-6 and TNF-α and the transformation of LC3BⅠ to LC 3BⅡ，significantly increased the protein expression of Beclin- 1， quinacrine cationic liposomes for treating non-small cell @163.com lung cancer[J]. J Drug Target ，2015，23（3）：232-243. the phosphorylation of AMPK protein ，and significantly reduced the phosphorylation level of AKT and mTOR proteins ，with statistical significance （P＜0.05 or P＜0.01）. CONCLUSIONS ：Ginseng root extract can induce autophagy of mice splenic lymphocytes，activate the activity of splenic lymphocytes ，regulate the secretion of cytokine by activating AMPK and inhibiting the activation of AKT which can inhibit the activity of mTOR ，so as to exert immune enhancement effect.

Microbial transformation of ginsenosides extracted from Panax ginseng adventitious roots in an airlift bioreactor

Background: Ginsenoside is the most important secondary metabolite in ginseng. Natural sources of wild ginseng have been overexploited. Although root culture can reduce the length of the growth cycle of ginseng, the number of species of ginsenosides is reduced and their contents are lower in the adventitious roots of ginseng than in the roots of ginseng cultivated in the field. Results: In this study, 147 strains of ß-glucosidase-producing microorganisms were isolated from soil. Of these, strain K35 showed excellent activity for converting major ginsenosides into rare ginsenosides, and a NCBI BLAST of its 16S rDNA gene sequence showed that it was most closely related to Penicillium sp. (HQ608083.1). Strain K35 was used to ferment the adventitious root extract, and the fermentation products were analyzed by high-performance liquid chromatography. The results showed that the content of the rare ginsenoside CK was 0.253 mg mL-1 under the optimal converting conditions of 9 d of fermentation at pH 7.0 in LL medium, which was significantly higher than that in the adventitious roots of ginseng. Conclusion: These findings may not only solve the problem of low productivity of metabolite in ginseng root culture but may also result in the development of a new valuable method of manufacturing ginsenoside CK.

Antagonistic Evaluation of Chromobacterium sp. JH7 for Biological Control of Ginseng Root Rot Caused by Cylindrocarpon destructans

Cylindrocarpon destructans is an ascomycete soil-borne pathogen that causes ginseng root rot. To identify effective biocontrol agents, we isolated several bacteria from ginseng cultivation soil and evaluated their antifungal activity. Among the isolated bacteria, one isolate (named JH7) was selected for its high antibiotic activity and was further examined for antagonism against fungal pathogens. Strain JH7 was identified as a Chromobacterium sp. using phylogenetic analysis based on 16S rRNA gene sequences. This strain was shown to produce antimicrobial molecules, including chitinases and proteases, but not cellulases. Additionally, the ability of JH7 to produce siderophore and solubilize insoluble phosphate supports its antagonistic and beneficial traits for plant growth. The JH7 strain suppressed the conidiation, conidial germination, and chlamydospore formation of C. destructans. Furthermore, the JH7 strain inhibited other plant pathogenic fungi. Thus, it provides a basis for developing a biocontrol agent for ginseng cultivation.

Chlamydospore Induction from Conidia of Cylindrocarpon destructans Isolated from Ginseng in Korea

Cylindrocarpon destructans causes root rot disease in ginseng and can survive for a long time, producing chlamydospores. We optimized conditions to induce chlamydospore production from the conidia of C. destructans, isolated from Korean ginseng. This will provide the basis for testing the efficacy of control agents targeting these chlamydospores.

Antagonistic Effect of Streptomyces sp. BS062 against Botrytis Diseases

The use of microorganisms and their secreted molecules to prevent plant diseases is considered an attractive alternative and way to supplement synthetic fungicides for the management of plant diseases. Strain BS062 was selected based on its ability to inhibit the mycelial growth of Botrytis cinerea, a major causal fungus of postharvest root rot of ginseng and strawberry gray mold disease. Strain BS062 was found to be closely related to Streptomyces hygroscopicus (99% similarity) on the basis of 16S ribosomal DNA sequence analysis. Postharvest root rot of ginseng and strawberry gray mold disease caused by B. cinerea were controlled up to 73.9% and 58%, respectively, upon treatment with culture broth of Streptomyces sp. BS062. These results suggest that strain BS062 may be a potential agent for controlling ginseng postharvest root rot and strawberry gray mold disease.

Genetic Diversity and Pathogenicity of Cylindrocarpon destructans Isolates Obtained from Korean Panax ginseng

We analyzed the genetic diversity of Cylindrocarpon destructans isolates obtained from Korean ginseng (i.e., Panax ginseng) roots by performing virulence tests and nuclear ribosomal gene internal transcribed spacer (ITS) and mitochondrial small subunit (mt SSU) rDNA sequence analysis. The phylogenetic relationship analysis performed using ITS DNA sequences and isolates from other hosts helped confirm that all the Korean C. destructans isolates belonged to Nectria/Neonectria radicicola complex. The results of in vivo and ex vivo virulence tests showed that the C. destructans isolates could be divided into two groups according to their distinctive difference in virulence and the genetic diversity. The highly virulent Korean isolates in pathogenicity group II (PG II), together with foreign isolates from P. ginseng and P. quinquefolius, formed a single group. The weakly virulent isolates in pathogenicity group I, together with the foreign isolates from other host plants, formed another group and exhibited a greater genetic diversity than the isolates of PG II, as confirmed by the mt SSU rDNA sequence analysis. In addition, as the weakly virulent Korean isolates were genetically very similar to the foreign isolates from other hosts, they were likely to originate from hosts other than the ginseng plants.

The History of Dai-kenchu-to and the Component Crude Drugs / 日本東洋医学雑誌

A Kampo medicine, Dai-kenchu-to, has been used clinically for treatments of various ailments such as vomiting, stomachache, and abnormal intestinal peristalsis caused by abdominal chill. Recently, it is often used to prevent intestinal obstruction after abdominal operations. We searched ancient Chinese and Japanese medical texts for the indications and pharmacological characteristics of Dai-kenchu-to and its constituent herbs (<i>Zanthoxylum</i> fruit, dried Ginger rhizome, Ginseng root, and Malt sugar). We clarified the applications and the cautions of Dai-kenchu-to in this paper. Dai-kenchu-to has rarely been used in China. However, it was often used for the remedy of severe abdominal pain caused by chilling, worm-ileus and hernia in the medieval period of the Edo era in Japan. For these reasons, evidence is considered as described below. i) Japanese people did not have the habit of eating meat in those days. ii) Japanese people used to drink a lot of water. iii) Severe abdominal pain occurred frequently due to wearing traditional Japanese clothing, which does not protect well against cold. iv) Abdominal diagnosis was advanced in Kampo medical methods. We found two precautions in the ancient Japanese medical texts. One is that a purgative should be avoided when used in an applicable case of Dai-kenchu-to. The other is that Dai-kenchu-to should not be given in the case of high fever. It is supposed that the botanical origins and processing of the four herbs used in the medieval period of the Edo era are the same as those of today. Our findings suggest that the pharmacological contribution of the four herbs in Dai-kenchu-to is mainly due to <i>Zanthoxylum</i> fruit and dried Ginger rhizome, and that Ginseng root and Malt sugar harmonize between the condition of patient and the pharmacological action of <i>Zanthoxylum</i> fruit and dried Ginger rhizome.