| Objective:To study the genetic variation of Tetrastigma hemsleyanum Diels et Gilg in different places,the content of major chemical constituents,the antioxidant activity in vitro,the anti-inflammatory,analgesic and anti-hepatic injury effects.Choose the place where the quality of Tetrastigma hemsleyanum Diels et Gilg is good,which can be planted in the forest to alleviate the need for the Tetrastigma hemsleyanum Diels et Gilg.Method:1.The genomic DNA was extracted by using the modified CTAB method,and the ISSR-PCR reaction system was established by ISSR molecular marker technique.Genetic diversity analysis of 16 different places of Tetrastigma hemsleyanum Diels et Gilg was carried out.2.The determination of water loss rate was carried out by measuring the quality of fresh roots,stems,leaves and dry weight after drying.The UV and HPLC were used to determine the total Flavonoids,total saponins,total phenol,polysaccharides and kaempferol,quercetin,isoquercitrin 7 indicators of the composition of the content.3.The used of response surface method optimized extraction process of Tetrastigma hemsleyanum Diels et Gilg,the antioxidant capacity of the clover was determined by the ABTS method and DPPH method.4.Pharmacological models were established by xylene-induced mouse ear swelling,acetic acid writhing and CCl4-induced liver injury,were used to carry out anti-inflammatory,analgesic and anti-hepatic injury effects.Result:1.The 16 different places of Tetrastigma hemsleyanum Diels et Gilg has a large genetic difference,can be divided into A,B,C three categories,Class A includes S1 FJ-SC,S2 FJ-SW,S4 FJ-SX,S5 FJ-SY,S6 FJ-ZP,S7 FJ-LC,S8 FJ-DT,S12 FJ-S,S14 ZJ-LQ,S15 FJ-LA;Class B includes S3 FJ-JY,S9 FJ-YA,S10 FJ-TN,S11 FJ-YX,S13 FJ-YC;Class C includes S16 ZJ-KH.2.In the aspect of water loss rate,the lowest water loss rate from the root of S2 FJ-SW,from the stem of S8 FJ-DT stem,from the leaves of S7 FJ-LC,was 62.01%,66.84%,80.70%.In the aspect of total flavonoids,total saponins,total phenolics,polysaccharides and kaempferol,quercetin,isoquercitrin content.According to parts considerations,The highest contents of total flavonoids and kaempferol from the stem of S12 FJ-S,which respectively were 113.07 mg·g-1,7830.61 μg·g-1,The highest contents of total saponins and total phenolics from the stem of S8 FJ-DT,which respectively were 114.17 mg·g-1,22.36 mg·g-1;The highest content of polysaccharide from the root of S5 FJ-SY was 96.35 mg·g-1;The highest contents of quercetin and isoquercitrin in the leaves of S11 FJ-YX,which respectively were 803.60μg·g-1,9188.72 μg·g-1;From the whole plant,The highest contents of total flavonoids,total saponins and total phenolics from S12 FJ-S,which respectively were 66.97 mg·g-1,49.28 mg·g-1,15.16 mg·g-1;The highest content of polysaccharide from S5 FJ-S was 52.48 mg·g-1;The highest content of kaempferol from S13 FJ-YC was 5197.23 μg·g-1;The highest contents of quercetin and isoquercitrin from S11 FJ-YX,which respectively were 317.52 μg·g-1,3105.86 μg·g-1.3.The optimal extraction process was as follows:extraction time 40 min,Liquid ratio of 26:1,ethanol concentration of 50%,extraction temperature of 50 ℃.In this process conditions to extract of Tetrastigma hemsleyanum Diels et Gilg from 16 different places,The results showed that the Tetrastigma hemsleyanum Diels et Gilg from S5 FJ-SY,S8 FJ-DT,S9 FJ-YA,S12 FJ-S eliminated DPPH and ABTS free radicals,which were stronger than those 12 places.4.The different places of Tetrastigma hemsleyanum Diels et Gilg were with anti-inflammatory and analgesic effects,but there were significant differences.Among them,the anti-inflammatory and analgesic effect of S5 FJ-SY was better than of S11 FJ-YX,S1 FJ-SC.Polysaccharide from S5 FJ-SY of Tetrastigma hemsleyanum Diels et Gilg has the effect of anti-liver injury.Conclusion:The 16 different places of-were genetic differences,Through the study of chemical composition,antioxidant,anti-inflammatory and analgesic effects,it was the best for the S5 FJ-SY,S12 FJ-S. |
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