ANO1抑制剂对自发性高血压大鼠血管平滑肌细胞增殖的影响

EFFECTS OF AN ANO1 INHIBITOR ON PROLIFERATION OF VASCULAR SMOOTH MUSCLE CELLS IN SPONTANEOUSLY HYPERTENSIVE RATS

目的探讨钙激活氯通道ANO1抑制剂T16A(inh)-A01(A01)对自发性高血压大鼠血管平滑肌细胞(SHR-VSMC)增殖的影响。方法取SHR-VSMC及其对照WKY大鼠的血管平滑肌细胞(WKY-VSMC),均分为对照组(加入体积分数为0.001的DMSO)和A01处理组(分别加入浓度为1、5、10、20μmol/L的A01)。采用MTT法检测各组细胞存活率,Western blot法检测各组增殖细胞核抗原(PCNA)表达,观察A01对细胞增殖能力的影响。结果 MTT结果显示,与WKY-VSMC对照组比较,SHR-VSMC对照组的生长速度明显加快(t=3.702,P<0.01)。与SHR-VSMC对照组比较,A01处理组的细胞存活率呈剂量依赖性下降,其中10、20μmol/L A01的抑制作用比较差异有统计学意义(F=9.916,q=4.468~7.483,P<0.01)。Western blot结果显示,与WKY-VSMC对照组相比,SHR-VSMC对照组的PCNA表达水平显著升高(t=2.871,P<0.01),A01(20μmol/L)处理24h后,SHR-VSMC的PCNA表达部分被抑制(t=2.064,P<0.01)。结论 ANO1抑制剂能明显抑制SHR-VSMC的异常增殖。

Objective To investigate the effects of a calcium-activated chloride channel ANO1 inhibitor,T16 A（inh）-A01（A01）,on the proliferation of vascular smooth muscle cells in spontaneously hypertensive rats（SHR-VSMC）. Methods Both SHR-VSMC and vascular smooth muscle cells in control WKYrats（WKY-VSMC）were divided into control group（volume fraction 0.001 DMSO）and A01-treated groups（1,5,10,and 20μmol/L A01）.The cell survival rate was determined by MTT assay.The expression of proliferating cell nuclear antigen（PCNA）was determined by Western blot.The impact of A01 on cell proliferation was evaluated. Results According to the results of the MTT assay,cell growth in the control group of SHR-VSMC was significantly accelerated than that in the control group of WKY-VSMC（t=3.702,P〈0.01）.Compared with the control group of SHR-VSMC,the A01-treated group of SHR-VSMC had cell viability reduced in a dose-dependent manner;particularly,cells treated with 10 or 20μmol/L A01 had significantly reduced cell viability compared with the control group（F=9.916,q=4.468-7.483,P〈0.01）.The results of Western blot showed that the expression of PCNA was significantly higher in the control group of SHR-VSMC than in the control group of WKY-VSMC（t=2.871,P〈0.01）.The expression of PCNA in SHR-VSMC was partially suppressed after 24 htreatment with A01（20μmol/L）（t=2.064,P〈0.01）. Conclusion ANO1 inhibitors can significantly inhibit the abnormal proliferation of SHR-VSMC.