| PurposeInvestigate the roles of SDF-1/CXCR4-R7biological axis in the proliferation ofvascular smooth cells induced by ox-LDL.MethedThe human VSMC were cultured in vitro, cultured by tissue explant method. Andthey were divided into normal control group (Control), ox-LDL group (model group),SDF-1group, AMD3100group, AMD3100control group, to observe the role ofSDF-1/CXCR4axis in the proliferation of VSMC. Observation of the role ofSDF-1/CXCR7axis in the proliferation of VSMC by RNAi silencing of the CXCR7receptor, they were randomly divided into normal control group (Control), ox-LDLgroup (model group), SDF-1group, Lip2000control group, negative control group,positive control group, siRNA1group, siRNA2group, siRNA3group. The experimentswere divided into normal control group (Control), ox-LDL group (model group),SDF-1group, AMD3100group, RNAi CXCR7group, to analysing the role of SDF-1/CXCR4-R7axis in the proliferation of VSMC. The rate of cell proliferation wasdetected using MTT assay, while the Cell cycle percentage was detected by flowcytometry analysis, RNA expression was analyzed using Realtime PCR.Result(1) Experiments on the role of the SDF-1/CXCR4axis in VSMC.①MTTmethod was used to measure the proliferation rate. The proliferation rate in the modelgroup was significantly higher than control group, but lower than the SDF-1group(P<0.05), and there was no significance difference when compared with the AMD3100group (P>0.05); there was no significance difference between the control group and the AMD3100control group (P>0.05).②we use flow cytometry to test the cell cycle.Compared with normal group, in the model group the number of cells in S phaseincreased significantly (P<0.05), in G2/M phase decreased significantly (P<0.05);compared with the model group, in the SDF-1group, S increased (P<0.05), the G2/Mphase was decreased (P<0.05).After the addition of the CXCR4inhibitor AMD3100,compared with SDF-1group, S phase cells decreased significantly (P<0.05), but G2/Mphase cells increased significantly;compared with the model group, there was nosignificant difference in all cell phase(>0.05); there was no significant differencebetween the cell cycle of the AMD3100control group and the normal group (P>0.05).③Compared with normal group, CXCR4RNA expression in the model group is anupward trend, but no significant difference (P>0.05); the expression level of RNA inthe SDF-1group increased significantly (P<0.05). After the addition of CXCR4specific inhibitor, CXCR4RNA expression had no significant difference,comparedwith the SDF-1group; CXCR4RNA expression was higher than that in the normalgroup significantly (P<0.05); AMD3100control group, CXCR4RNA expression wassignificantly lower than that in SDF-1group, AMD3100group (P<0.05); and had nosignificant difference, compared with the control group,the model group. Theexpression of CXCR7RNA in model group was significantly increased than thenormal group; in the SDF-1group, the expression level of RNA increased significantlythan the model group (P<0.05). After the addition of the CXCR4inhibitor AMD3100,CXCR7RNA expression, compared with the SDF-1group, had no significantdifference; compared with the normal group,model group increased significantly(P<0.05); CXCR7RNA expression in the AMD3100control group, compared withnormal control group, had no significant difference.(2) Experiments on the role of the SDF-1/CXCR7axis in VSMC.①MTTmethod was used to measure the proliferation rate. The proliferation rate in the modelgroup was significantly higher compared with control group, positive controlgroup,but lower than the SDF-1group,negative control group, Lip2000group(P<0.05), and there was no significance difference when compared with siRNA1group, siRNA2group and siRNA3group; there was no significant difference in siRNA1group, siRNA2group and siRNA3group (P>0.05).②Measurement of cell cycle byflow cytometry.Compared with normal group,in the model group S phrase increasedsignificantly (P<0.05), G2/M decreased significantly (P<0.05); compared with theSDF-1group, in the model group, S phrase decreased significantly (P<0.05)(P<0.05),G2/M phase increased significantly (P<0.05); gene silencing of CXCR7receptor, Sphase cells decreased significantly compared with group SDF-1(P<0.05), but G2/Mphase cells increased significantly;there was no significant difference compared withthe model group.③Compared with normal group,the CXCR4RNA expression in themodel group had an upward trend, but the difference was not significant; theexpression level of CXCR4RNA in the SDF-group increased significantly comparedwith the normal group (P<0.05); and compared with the Lip2000control group,negative control group, positive control group, siRNA1group, group siRNA2, groupsiRNA3had no significant difference. The expression of CXCR7RNA in model groupwas significantly increased, compared with the normal group (P<0.05); the expressionlevel of CXCR7RNA in the SDF-1group increased significantly than the model group(P<0.05), and and had no significant difference compared with the Lip2000controlgroup, negative control group; gene silencing of CXCR7receptor, CXCR7RNAexpression,compared with SDF-1group, Lip2000control group, negative controlgroup, the model group decreased significantly (P<0.05); compared with the normalgroup, the positive control group, increased significantly (P<0.05);CXCR7RNAexpression in the positive control group,compared with normal control group, had nosignificant difference.(3) Experiments on the role of the SDF-1/CXCR4-R7axis in VSMC.①MTTmethod was used to measure the proliferation rate. The proliferation rate in the modelgroup was significantly higher compared with control group, but lower than the SDF-1group (P<0.05), and there was no significance difference when compared with theAMD3100, RNAiCXCR7group (P>0.05); there was no significance differencebetween the AMD3100group and the RNAiCXCR7group (P>0.05).②to test the cell cycle by flow cytometry, the model group compared with normal group, Sincreased significantly (P<0.05); the SDF-1group compared with the model group,Sincreased (P<0.05), G2/M decreased significantly (P<0.05). After the addition of theCXCR4inhibitor AMD3100, compared with SDF-1group, S phase cells decreasedsignificantly (P<0.05), G2/M phase cells increased significantly; compared with themodel group, there was no significant difference in all phase (P>0.05). Gene silencingof CXCR7receptor, and there is no significant difference in cell cycle compared withmodel group (P>0.05), compared with the SDF-1group of cells in S phase decreasedsignificantly (P<0.05).③the CXCR4RNA expression between the model group andthe normal group, there is an upward trend, but the difference was not significant; theexpression level of CXCR4RNA in the SDF-1group, increased significantly comparedwith the normal group (P<0.05). After the addition of the CXCR4inhibitor AMD3100,CXCR7gene silencing, CXCR4RNA expression, compared with SDF-1group, had nosignificant difference (P>0.05); compared with the normal group significantlyincreased (P<0.05). The expression of CXCR7RNA in model group was significantlyincreased compared with normal group (P<0.05), the CXCR7RNA expression in theSDF-1, increased significantly than the model group (P<0.05). After the addition of theCXCR4inhibitor AMD3100, CXCR7RNA expression, compared with the SDF-1group had no significant difference; compared with the normal group, the model groupobviously increased (P<0.05); after CXCR7gene silencing, CXCR7expression ofRNA, compared with SDF-1group, model group, AMD3100group decreasedsignificantly (P<0.05), but significantly higher than that in the normal group (P<0.05).Conclusion(1) Stromal cell derived factor-1can promote the proliferation of vascularsmooth muscle cells induced by ox-LDL.(2) The SDF-1/CXCR4-R7axis participates in the value-added role of stromalcell derived factor-1promotes ox-LDL induced vascular smooth muscle cells. |
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