The Effect Of HSP22and Atorvastatin In The Injury Of HUVECs Induced By Ox-LDL

Objective:Research of the function of the heat shock protein22(HSP22) in ox-LDL leadingthe human umbilical vein endothelial cell(HUVECs) damage and the statin’sintervention effect, and then discuss it’s mechanism.Method:The experiment had three groups: The normal control group, the empty plasmidtransfection control group(con group) and the shRNA plasmid transfectiongroup(shRNAgroup), adding Lipo and empty plasmid（pGcsi-U6/Neo/GFp/shRNA）in HUVECs of the con group, adding Lipo and shRNA plasmid（pGcsi-U6/Neo/GFp/shRNA/HSP22） in HUVECs of the shRNA group. Aftersuccessful transfection, each group had four subgroups: The control group, theox-LDL group, the statin group and the statin+ox-LDL group. Using the MTT,Westernblot to select appropriate ox-LDL and atorvastatin intervene concentrationand time. Observing the ox-LDL inducing environment’s HSP22expressing situation,using ELISA to detect TNF-α expressing situation, using the Western blot to detectthe eNOS, p38MAPK, erk and p-erk’s expressing situation.Result:1. ox-LDL and atorvastatin’s appropriate intervention time and concentration:(1)160μg/ml ox-LDL intervene for48h, the expression of HSP22reach to thehighest(P<0.01)，and ox-LDL has obviously inhibitory effect to the cell(P<0.05)..(2)40μg/ml atorvastatin intervene for48h, the expression of HSP22is thelowest(P<0.05)，and atorvastatin has obvious promoting effect to the cell(P<0.05)..2. The expression of HSP22protein: Under the same intervene condition, theHSP22protein expression in the shRNA group was obviously lower than the normalcontrol group and the con group(P<0.01). In each group, the HSP22expressing of theox-LDL subgroups were significantly higher than the control subgroups and theox-LDL+statin subgroups (P<0.01). 3. The expression of the p38MAPK: Under the same intervene condition, theexpression of the p38MAPK in the shRNA group was obviously higher than thenormal control group and the con group(P<0.01). In each group, the expression of thep38MAPK in the ox-LDL subgroups were higher than the control subgroups and theox-LDL+statin subgroups (P<0.05).4. The expression of the erk and p-erk: Under the same intervene condition, theexpression of the erk and p-erk in the shRNA group were lower than the normalcontrol group and the con group(P<0.05). In each group, the expression of the erk andp-erk in the ox-LDL subgroups were higher than the control subgroups and theox-LDL+statin subgroups (P<0.05).5. The expression of TNF-α: Under the same intervene condition, the expressionof the TNF-α in the shRNA group was obviously higher than the normal controlgroup and the con group(P<0.01). In each group, the expression of the TNF-α in theox-LDL subgroups were obviously higher than the control subgroups and theox-LDL+statin subgroups (P<0.01).6. The expression of eNOS: Under the same intervene condition, the expressionof the eNOS in the shRNAgroup was obviously lower than the control group and thecon group(P<0.01). In each group, the eNOS expression of the ox-LDL subgroupswere obviously lower than the control subgroups and the ox-LDL+statin subgroups(P<0.01).Conclusion:1. The ox-LDL damaging HUVECs can induce the expression of HSP22.2. In the HUVECs damage caused by ox-LDL, HSP22can express the functionof inhibiting of cell apoptosis and protecting the endothelial cells, it’s mechanismmay be related to the down regulation of p38MAPK, promote erk, p-erk and inhibitthe expression of TNF-α; and it can promote the expression of eNOS.3. The statin’s protection to the endothelial cells may be related to the downregulation of p38MAPK, erk, p-erk and inhibit the expression of TNF-α; and it canpromote the expression of eNOS.4. The statin’s protectionto the endothelial cells may have no relation to HSP22.