Study On The Material Basis And Blood Activating Effect Of Tricyrtis Maculata From Shaanxi Province

Objective To explain the material basis and effect of T.maculata of activating blood circulation and removing blood stasis,and to lay the theoretical foundation and experimental basis for the research and development of new drugs.Methods 1.The natural product chemical separation method（semi-preparation HPLC,silica gel column,sephadex column,etc.）was used to separate and purify the ethyl acetate extract from T.maculata,and modern spectroscopic analysis techniques(¹H-NMR,¹³C-NMR,IR,HR-ESI-MS,DEPT,¹H,¹H-COSY,etc.)were used to identify the structure of the compounds;2.Antioxidant activity of the monomer compounds isolated from the ethyl acetate extract of T.maculata was determined by DPPH and ABTS free radical scavenging experiments;3.Using human umbilical vein endothelial cells as the research object,the anti-inflammatory ability of monomer compounds was evaluated by using lipopolysaccharide-induced inflammation model,and the monomer compounds with significant anti-inflammatory ability were screened;4.The survival environment of blood vessels in vitro was simulated,and different vasoactive substances were given to stimulate the mesenteric vessels ring of SD rats for the detection of vascular function.The changes of vascular tension were recorded by the vascular tension detection system,and the effects of each monomer compound on the ring were evaluated;5.FeCl₃ was used to induce carotid artery thrombosis in SD rats.The thrombus weight and formation rate were used as indexes to evaluate the inhibitory effect of the active components in T.maculata on thrombus formation.Results 1.11 compounds were isolated and identified from the ethyl acetate fraction of T.maculata,including one new phenylpropanoid compound with high molecular weight.All other known compounds,except compound 8,were isolated from this plant species for the first time;2.Compound 7 exhibited the most antiradical activity for DPPH with IC₅₀ value of 0.028±0.001 m M（p<0.001）,which was nearly 3-fold higher than that of Trolox(IC₅₀=0.073±0.001 m M).To the best of our knowledge,the antioxidant activities of compounds1,2,4,6,and 10 were firstly reported;3.Compounds 2,5,8,9 and 10 all have protective effects on inflammatory cells.In compound 2 and 9 of them on inflammation model of protection is the most significant（p<0.05）,the cell vitality is almost equal with the blank control group;4.Compounds 1,2,6 and 10 were selected,which can inhibit PE contraction of vascular function.Compound 2 can make blood vessels shrinkage rate from 90.15%±10.70%to 23.10%±5.03%（p<（15）（13）（15）（20））;Compounds 2,3,and 8 can inhibit KCl contraction of vascular function for:including compound 8 can make blood vessels shrinkage rate from93.06%±12.50%to 49.39%±9.46%（p<0.05）;Compounds 4,5,7 and 8 had vasodilating effects,and these four compounds were all lignans,among which compounds 7 and 8 had vasodilating effects nearly equal to that of the positive control;5.Based on the results of in vitro activity screening,compound 8,which has a more comprehensive effect,was selected as the active compound to study the mechanism of inhibition of thrombosis of T.maculata.The final results showed that compound 8 could reduce the weight of thrombus from 2.30±0.05 mg to 1.70±0.03 mg in the blank control group,and the inhibition rate of thrombus formation could reach 76.92%±3.9%（p<0.001）（13）Conclusion Phenylpropanoid compounds were the main material basis of ethyl acetate fraction of T.maculate;Both in vitro and in vivo experiments were used as the experimental design basis to study the blood-activating effect of this paper The results of in vitro activity experiments showed that compound 8 had significant antioxidant activity,stronger anti-inflammatory activity at 1μM,and inhibitor of carotid artery thrombosis in rats showed that compound 8 had high inhibitory rate of thrombosis.