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Objective: To investigate the chemical metabolites from Cercospora lagenariae MT-45, an endophytic fungus isolated from Huperzia serrata (Thunb.) Trev. Methods: The compounds were isolated and purified by using silica gel column chromatography and semi-preparative liquid chromatography. The structures were established using physicochemical properties and MS and NMR. The anti-inflammatory activities of all the isolates were also preliminarily investigated by using in vitro model. Results: Nine polyketide derivatives including cercolagenlic acid A (1), alternariol (2), alternariol 9-methyl ether (3), (+)-nigrosporaol A (4), alternarienonic acid B (5), 2-methyl-5-carboxymethyl-7-hydroxychromone (6), 2,5-dimethyl-7-hydroxychromone (7), 1-deoxy- rubralactone (8), and (-)-alternarlactam (9) were isolated from C. lagenariae MT-45 fermented on brown rice solids. Conclusion: Compound 1 is a new compound, and compound 6 can exhibit a certain inhibition on the nitric oxide production in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells with an IC50 value of (57.5 ± 1.2) μmol/L.
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Objective: To analyze the differences in protein of thallus of Huperzia serrata (in vitro cultures, strains A, B, and C, respectively) with different Hup A content, and to explore related enzymes that may be synthesized with Hup A accumulation. Methods: Quantitative proteomics was performed in vitro cultures of Huperzia serrata with different Hup A content using quantitative proteomics tandem mass tag (TMT) techniques, followed by differential protein analysis: GO, KEGG and other biological information analysis. Results: Strain B hadthe lowest Hup A content and strain C had the highest Hup A content, which was twice than that of B. There were 78 differential proteins between the strain B and C. Analysis of differential protein GO enrichment showed that MF accounted for 28.75%; Analysis of differential protein expression showed that three strains shared two differential proteins (P93541, Q8RXU4) in the alkaloid metabolic pathway starting from amino acids. P93541 protein was down-regulated in the low-yield strain B and up-regulated in the relatively high-yield strains A and C. The Q8RXU4 protein was up-regulated in the low-yield strain B and down-regulated in the relatively high-yield strains A and C. Conclusion: This study found that the difference in Hup A content was positively correlated with the protein expression. Two enzymes P93541 and Q8RXU4 that may be related to Hup A accumulation were analyzed, providing a basis for bioinformatics analysis of Hup A biosynthesis.
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Objective: To investigate the non-alkaloids in Huperzia serrata. Methods: The constituents were mainly isolated and purified by means of silica gel, RP18, and Sephadex LH-20 column chromatographies. The structures were identified based on analyses of the spectral data of NMR and MS. Results: Totally 11 constituents were obtained from dichloromethane fraction of the plant and they were 3β-hydroxyserrat-14-en-21β-yl-p-dihydrocoumarate (1), seratenediol-3,21-diacetate (2), seratenediol-3-acetate (3), 21-epi-serratenediol-3-acetate (4), 3α,21β,24-trihydroxyserrat-14-en (5), 21-epi-serratenediol (6), serratenediol (7), 3α,21β-dihydroxy- serrat-14-en-16-one (8), 3β,21β,24-trihydroxyserrat-14-en (9), 3α,21β,24-trihydroxyserrat-14-en-16-one (10), and 1-dibenzofuranol (11). Conclusion: Compounds 1-10 are serratene-type triterpenoids and compound 11 is a dibenzofuranol. Compounds 1 is a new compound named as serratcoumarate and compounds 2 and 11 are isolated from the plant for the first time.
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Objective: Huperzia serrata, whose growth is limited by high temperature, is a rare medicinal plant with the treatment function for Alzheimer's disease (AD). To research the effect of high temperature on the structure and function of cell membrane and chloroplast, and to provide the evidence for production practices. Methods: H. serrata was processed at 25, 30, 35, and 40°C, respectively, then the content changes of malondial dehyde (MDA) and conductivity rate, and the content changes of total chlorophyll, chlorophyll a, chlorophyll b, and chlorophyll a/b values were measured. The changes of the chloroplast ultra microstructure were observed under the transmission electron microscope (TEM). Results: The changes of MDA and conductivity rate in the process at 35 and 40°C were significantly higher than those of the control group; After processed at 40°C for 4 d, the total chlorophyll was decreased significantly, and became the lowest on the day 6, just was 58% compared to the control group; the change trends to the contents of chlorophyll a, chlorophyll b, and total chlorophyll were similar; TEM observation revealed that after processed at 35°C for 4 d, the chloroplast structure appeared deformation, and after processed at 40°C for 4 d, the chloroplast structure subjected obvious destruction capsule fuzzy, fracture in different degrees, thylakoid in disorder, matrix lamellar irregular, and so on. Conclusion: According to the changes of physiological index, ultramicroscopic structure, and external morphology of chloroplast, the suitable temperature for H. serrata is 25-30°C, 40°C is the limited temperature, causing death after 4 d stress, and 35°C has obvious impact on the growth, long-time stress in 35°C could also cause plant deaths.
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Object To compare the hupzine A (Hup A) in Huperzia serrata (Thunb.) Trev. obtained by different extracting methods and investigate the amount of alkaloids and the content of Hup A from different parts of the plants and from different places. Methods Using HPLC for the determination of Hup A. Results The content of Hup A in the stem and leaf is richer than that in the root. The content of Hup A from Guizhou, Guangdong and Anhui Provinces is 0.018%, 0.021% and 0.020% repectively; The difference of extract method of Hup A is no prominence. Conclusion The content of Hup A in the ground is richer than that of underground, and there are some difference in the content of Hup A obtained from different places.