Objective:To explore the effect and related mechanism on inflammatory response,proliferation and apoptosis of oxLDL-induced vascular smooth muscle cell of matrine.Methods:Theatherosclerotic model was conducted through treating human aort-icvascular smooth muscle cell with oxidized low density lipoprotein(oxLDL).Cell viability and proliferation was detected by CCK-8 as-say.The mRNA level of interleukin 1 beta(IL-1β),tumor necrosis factor alpha(TNF-α),IL-10 and IL-13 was tested by quantitative real-time reverse transcription PCR(qRT-PCR).Cell apoptosis was measured by flow cytometry.The expression of proliferation marker proteins antigen identified by monoclonal antibody(Ki-67) and proliferating cell nuclear antigen(PCNA),apoptosis marker proteins B cell lymphoma 2(Bcl-2) and Bcl-2-associated X protein(Bax),signal transducer and activator of transcription 3(STAT3) and signal transducer and activator of transcription 5(STAT5) was detected by Western blot.Results: Compared with control group,the mRNA level of IL-1β and TNF-α in model group was largely increased with decreased mRNA level of IL-10 and IL-13(P<0.01).Compared with model group,the mRNA level of IL-1β and TNF-α in treatment group was attenuated with enhancive mRNA level of IL-10 and IL-13(P<0.05).Cell proliferation and apoptosis in model group was higher than control group.Cell proliferation and apoptosis in treatment group was lower than model group(P<0.05).Compared with control group,the expression of Ki-67,PCNA and Bax in model group was augmented with decreased expression of Bcl-2(P<0.01).Compared with model group,the expression of Ki-67,PCNA and Bax in treatment group declined with elevated expression of Bcl-2(P<0.05).The expression of p-STAT3 and p-STAT5 in model group was higher than control group(P<0.01).The expression of p-STAT3 and p-STAT5 in treatment group was lower than model group(P<0.05).Compared with model group,the expression of Ki-67,PCNA and Bax in treatment group and model+Ruxolitinib group was decreased with enhancive expression of Bcl-2(P<0.05).The expression of Ki-67,PCNA and Bax in model+matrine+Ruxolitinib group was observably lower than model group and the expression of Bcl-2 in model+matrine+Ruxolitinib group was observably higher than model group(P<0.01).Compared with model group,the mRNA level of IL-1β and TNF-α in treatment group,model+Ruxolitinib group and model+matrine+Ruxolitinib group was attenuated with enhancive mRNA level of IL-10 and IL-13(P<0.05).Conclusion:Matrine represses inflammation,proliferation and apoptosis of vascular smooth muscle cell by inhibiting activation of JAK/STAT3 signal pathway in atherosclerosis.%目的:探讨苦参碱对氧化型低密度脂蛋白(oxLDL)诱导的血管平滑肌细胞炎症反应及增殖凋亡的影响及分子机制.方法:oxLDL处理人主动脉血管平滑肌细胞建立动脉粥样硬化模型.CCK-8实验分析细胞活力和增殖.实时定量PCR(qRT-PCR)检测炎症因子白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、IL-10和IL-13的mRNA水平.流式细胞术分析细胞凋亡.蛋白印迹检测增殖标记蛋白细胞增殖核抗原-67(Ki-67)和增殖细胞核抗原(PCNA),细胞凋亡标记蛋白 B细胞淋巴瘤-2(Bcl-2) 和Bcl-2相关蛋白X(Bax),信号转导及转录激活子3(STAT3)和信号转导及转录激活子5(STAT5)的表达.结果:与对照组相比,造模组促炎因子IL-1β和TNF-α的mRNA水平大大提高,抗炎因子IL-10和IL-13的mRNA水平则显著降低(P<0.01).与造模组相比,模型加药组促炎因子IL-1β和TNF-α的mRNA水平显著降低,抗炎因子IL-10和IL-13的mRNA水平明显升高(P<0.05).造模组细胞增殖倍数和细胞凋亡率明显高于对照组,模型加药组细胞增殖倍数和细胞凋亡率明显低于造模组(P<0.05).与对照组相比,造模组Ki-67、PCNA和Bax的表达显著升高,Bcl-2显著降低(P<0.01).与造模组相比,模型加药组Ki-67、PCNA和Bax的表达明显减少,Bcl-2表达明显增多(P<0.05).造模组p-STAT3和p-STAT5相对蛋白表达量显著高于对照组(P<0.01).模型加药组p-STAT3和p-STAT5相对蛋白表达量明显低于造模组(P<0.05).与造模组相比,模型加药组和模型+Ruxolitinib组Ki-67、PCNA和Bax的表达明显减少,Bcl-2表达明显增多(P<0.05);模型加药+Ruxolitinib组Ki-67、PCNA和Bax的表达显著减少,Bcl-2表达显著增多(P<0.01).与造模组相比,模型加药组和模型+Ruxolitinib组及模型加药+Ruxolitinib组IL-1β和TNF-α的mRNA水平都明显降低,IL-10和IL-13的mRNA水平都明显升高(P<0.05).结论:苦参碱可通过抑制JAK/STAT3通路的活化起到抗炎和抑制血管平滑肌细胞增殖和凋亡的作用.
展开▼
