| ObjectiveIn this report,we observed the cytoprotection effect of Huangqi injection and its molecular mechanism on the cell injury caused by nicotine in isolated human umbilical vein endothelial cell in votro.And further research the relationship of nicotine on the human umbilical vein endothelial cell injury with the nicotinic acetylcholine receptor a7.Methods1 Human umbilical vein endothelial cells culture in vitro.2 MTr method was used to investigate inhibition rate of Nicotine injury in Human umbilical vein endothelial cells generation.3 The Nitrate reductase assay,Chemical colorimetry assay,Enzyme linked immunosorbent assay were used to investigate the level of total NO,total NOS,ET-1.4 RT-PCR PCR was used to detect the expression of nicotinic acetylcholine receptor a7 mRNA.Results1 The affect of Nicotine on the the growth of HUVEC and the intervention of astragalusMTT method was used to test the proliferation rate.The results showed,compared with the normol HUVEC,the proliferation rates of cells treated by the concentration of 10-3 mol/l 10-4mol/l,10-5mol/l of nicotine were reduced to varying degrees,and the cell proliferation rate was decreased significantly for 10-4mol/l(P＜0.01).D-tubocurarine,as a blocker for nicotine's receptor,was found to block nicotine'affect on the HUVEC.Compare with the nicotine treated group,10-8mol/l d-tubocurarine- treated group the cell proliferation rate increased significantly (P＜0.01).It shows that,the effect of nicotine on the HUVEC is closely related to the concentration.,d-tubocurarine can block this effect on the nicotinic acetylcholine receptor.The result of MTT also show that,compared separately with the nicotine treated group and the normol group,the cell vegetation of the Astragalus treated group is raised(two comparisons(P＜0.01).compared with the nicotine treated group,the cell vegetation of the astragalus and nicotine treated group is also raised(P＜0.05).2 The affect of Nicotine on the the cell'function of HUVEC and the intervention of Radix astragaliWe tested separately with nitrate reductase assay of total NO levels,chemical colorimetric assay of total NOS levels,enzyme-linked immunosorbent assay of ET-1 level.The results showed that the nicotine injury HUVEC cells after the total NO level and the total NOS were significantly lower(P＜0.01),but significantly increased for ET-1 level(P＜0.01).Total NO levels and total NOS in Radix astragali-treated group were significantly raised(P＜0.01),but obviously decreased for ET-1(P＜0.01).Compared with the nicotine treated group,astragalus and nicotine combined treatment group,the total NO level and the level of total NOS significantly increased(P＜0.05), decreased for ET-1(P＜0.05).3 The effect of nicotine on the expression of nicotinic acetylcholine receptor a7 on HUVECThe result of PCR showed that the levels of nicotinic acetylcholine receptor a7 mRNA was up-regulated in Nicotine treated group than the other groups,Imaging analysis of electrophoresis gel band was brighter than the other groups also。ConclusionThis study suggests that nicotine can affect HUVEC ceils,the cellular levels of total NO and total NOS significantly increased(P＜0.01),ET-1 levels decreased significantly(P＜0.01).The 1 concentration of Chinese medicine Astragalus(200ug/l) could significantly promote the proliferation of HUVEC cells,at the same time,the Radix astragali treated group,the total cellular levels of NO and total NOS significantly increased,ET-1 levels decreased significantly,Chinese medicine Astragalus can promote HUVEC cell proliferation and reverse the affect of HUVEC injuryed by nicotine.The expression of nicotinic acetylcholine receptor a7 mRNA in nicotine -injured HUVEC is upregulaed,so we can conclude that nicotine can induce the the expression of nicotinic acetylcholine receptor a7 on HUVEC.According to result of this experiment,we also know d-tubocurarine can block nicotine's effect on nicotinic acetylcholine receptor.
|
PDF Full Text Request