吴晶魁+杨乔 李洋洋

[摘要]為了探究中药水蛭干预作用下p38MAPK信号转导通路对早期动脉粥样硬化（AS）大鼠血管平滑肌细胞（VSMCs）增殖与凋亡的影响及其可能机制。实验通过生化分析仪检测中药水蛭对大鼠血脂（TG，TC，LDLC，HDLC）水平的调控；ELISA检测血清中TGFβ1的水平；免疫组织化学法（IHC）检测VSMCs的PCNA，Caspase3表达水平；Western blot检测中药水蛭对VSMCs中MKK3，p38及Cmyc蛋白表达的影响；HE染色观察大鼠胸主动脉形态变化。结果显示，中药水蛭可使TC，LDLC明显降低，HDLC升高，TGFβ1，PCNA的表达水平明显下降，Caspase3表达上调，MKK3，p38及Cmyc蛋白表达水平下降，以及抑制内膜增厚、减少斑块形成。以上结果表明中药水蛭可能通过调降血脂，降低TGFβ1水平，影响p38MAPK信号通路蛋白表达，从而抑制VSMCs的增殖、促进其凋亡，抑制内膜增厚、减少斑块形成等环节，进而干预早期AS进程。

[关键词]水蛭；动脉粥样硬化；p38MAPK信号通路；血管平滑肌细胞；增殖与凋亡

Effect of leech on VSMCs in early atherosclerosis rats via

p38MAPK signaling pathway

WU Jingkui1， YANG Qiao1*， LI Yangyang2， XIE Dongjie1

（1Tianjin Medical University General Hospital， Tianjin 300052， China；

2The First Affiliated Hospital of Henan University of Science and Technology， Luoyang 471003， China）

[Abstract]To explore the effect of leech on proliferation and apoptosis of vascular smooth muscle cells（VSMCs） in early atherosclerosis rats via p38MAPK signaling pathway and investigate its possible mechanism Biochemical analyzer was used to examine the regulation of leech on levels of triglycerides（TG）， total cholesterol（TC）， lowdensity lipoprotein（LDLC）， and highdensity lipoprotein（HDLC） in blood lipid of rats The expression of transforming growth factorbeta 1（TGFβ1） in serum was detected by ELISA Immunological histological chemistry （IHC） was taken to measure the expression levels of proliferating cell nucleus antigen（PCNA） and cell apoptosis proteinase3（Caspase3）， while the protein expression levels of MKK3， p38 and Cmyc were detected by Western blot In addition， hematoxylin and eosin（HE） staining was used to observe the morphological change of thoracic aortas The results showed that leech decreased the levels of TC， LDLC obviously and increased HDLC， suppressed the expression levels of TGFβ1 and PCNA， upregulated Caspase3， downregulated the expression levels of MKK3， p38， and Cmyc protein HE staining indicated that it could inhibit intimal thickening and reduce plaque formation The above results indicated that leech may affect the protein expression of the p38MAPK signaling pathway to inhibit proliferation and promote the apoptosis of VSMCs via reducing blood lipid levels and suppressing TGFβ1， aiming at inhibiting intimal thickening and reducing plaque formation， tand then slowing down the process of early atherosclerosis

[Key words]leech； atherosclerosis； p38MAPK signaling pathway； vascular smooth muscle cells； proliferation and apoptosisendprint

血管平滑肌细胞（vascular smooth muscle cells，VSMCs）的激活、迁移与增殖是动脉粥样硬化（atherosclerosis，AS）发生的重要病理过程[1]。一般情况下，VSMCs保持静止，即处于不增殖的状态[2]，但是在血管损伤后VSMCs将会异常增殖，同时引起早期AS病理改变[3]。有研究指出，VSMCs增殖与凋亡的失衡是AS发生过程中斑块形成的的重要环节[4]。转化生长因子β1（transforming growth factorbeta1，TGFβ1）具有激活细胞内信号转导通路以及促进细胞增殖、分化的作用[5]。p38丝裂原活化蛋白激酶（p38 mitogenactivated protein kinase，p38MAPK）是调控细胞增殖与死亡的重要通路，可参与细胞增殖、分化和凋亡等多种生理过程[6]，TGFβ1作为外在刺激因子，对p38MAPK信号通路的激活具有重要作用[7]。血管壁的组成细胞主要包括VSMCs和内皮细胞，成熟的内皮细胞不具有增殖活性，因此在早期AS的发生主要为VSMCs增殖所致；……