丹蒌片抗动脉粥样硬化模型小鼠主动脉平滑肌细胞增殖的机制研究  

Mechanism of Danlou Tablet on the Proliferation of Vascular Smooth Muscle Cells in Mice Model with Atherosclerosis

作　　者：缪静 周鑫斌[2] 毛威[2] 陈洁[2] 潘孔寒[1] MIAO Jing;ZHOU Xin-bin;MAO Wei;CHEN Jie;PAN Kong-han(Department of Critical Care Medicine,Sir Run Run Shaw Hospital,Zhejiang University School of Medicine,Hangzhou 310006;Department of Cardiology,First Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou 310006)

机构地区：[1]浙江大学医学院附属邵逸夫医院重症医学科,杭州310016 [2]浙江中医药大学附属第一医院心内科,杭州310006

出　　处：《中国中西医结合杂志》2020年第7期811-816,共6页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：浙江省教育厅科研项目(No.Y201327471);浙江省中医药科技计划项目(No.2014ZB043);浙江省自然科学基金(No.LY16H150004)。

摘　　要：目的研究丹蒌片对动脉粥样硬化(AS)模型ApoE-/-小鼠血清抵抗素水平、血管平滑肌细胞(VSMC)增殖的影响,并探讨其可能机制。方法32只ApoE-/-小鼠按随机数字表法分为正常组、模型组、他汀组及丹蒌组,每组8只。正常组ApoE-/-小鼠喂饲基础饲料12周,余各组均采用高糖高脂饲料连续喂饲12周制备AS模型。正常组不予灌胃干预,模型组、他汀组及丹蒌组于第5周时分别给予生理盐水、阿托伐他汀片混悬液(1.5 mg/mL)、丹蒌片混悬液(0.34 g/mL)连续灌胃干预8周。干预结束后,ELISA检测血清抵抗素水平,免疫组化染色观察主动脉细胞增殖核抗原(PCNA)阳性表达,透射电镜观察VSMC表型改变,Real-time PCR、Western Blot检测ApoE-/-小鼠主动脉ERK1、ERK2 mRNA和ERK1/2、pERK1/2蛋白表达。结果与正常组比较,模型组血清抵抗素水平升高(P<0.05),PCNA阳性表达增加(P<0.01),血管内ERK1、ERK2 mRNA和ERK1/2、pERK1/2蛋白表达上升(P<0.01,P<0.05),透射电镜下可见主动脉VSMC胞浆内肌丝减少,粗面内质网、线粒体等细胞器增多,有较多的吞噬大量脂质形成的空泡,呈典型合成型改变。与模型组比较,他汀组与丹蒌组血清抵抗素水平降低(P<0.05),PCNA阳性表达减少(P<0.01),血管内ERK1、ERK2 mRNA和ERK1/2、pERK1/2蛋白表达下降(P<0.05,P<0.01),透射电镜下主动脉合成型VSMC明显减少。结论丹蒌片可明显抑制VSMC异常增殖,其作用机制可能是通过降低血清抵抗素水平,抑制抵抗素对ERK信号通路的激活,减缓VSMC的增殖效应,从而达到抗VSMC增殖作用。Objective To study the effect of Danlou Tablet on the serum resistin level,vascular smooth muscle cells(VSMC)proliferation in atherosclerosis(AS)model ApoE-gene knockout mice,and to explore its mechanism.Methods Totally 32 ApoE-gene knockout mice were assigned to the control group,the model group,the Atorvastatin group and the Danlou group by random digit table,8 in each group.Mice in the control group were given basic forage for 12 weeks,while mice in the other three groups were given with high fat forage to establish AS model.Then mice in the normal group did not receive gavage intervention,the model group,the Atorvastatin group and the Danlou group were administered with normal saline,Atorvastatin suspension(1.5 mg/mL),and Danlou Tablet suspension(0.34 g/mL)by gastrogavage for 8 successive weeks,respectively.After intervention serum resistin level was detected by ELISA.Proliferating cell nuclear antigen(PCNA)positive expression in aorta were observed by Immunohistochemical staining.The VSMC phenotypic modulation was detected by transmission electron microscope(TEM).mRNA and protein expression levels of ERK1,ERK2 and ERK1/2,pERK1/2 in thoracic aorta were measured by Real-Time PCR and Western Blot.Results Compared with the control group,serum resistin level,PCNA positive expression,and expression levels of ERK1,ERK2 mRNA and ERK1/2,pERK1/2 protein significantly increased in the model group(P<0.01,P<0.05).TEM demonstrated decreased intracytoplasmic filaments,increased organelles such as rough endoplasmic reticulum and mitochondria,and vacuoles formed by phagocytosis of a large amount of lipids in aortic VSMC,synthetic transformation were showed.Compared with the model group,serum resistin level,PCNA positive expression,expression levels of ERK1,ERK2 mRNA,ERK1/2 and pERK1/2 protein significantly decreased in Atorvastatin group and Danlou group(P<0.05,P<0.01).The transform VSMC in aortic was fewer in the medicated groups than that in the model group.Conclusions Danlou Tablet could significantly inhibit the prolifer

关 键 词：丹蒌片 动脉粥样硬化 ERK信号通路 血管平滑肌细胞 

分 类 号：R285.5[医药卫生—中药学] R-332[医药卫生—中医学]