鼠妇提取物可抑制ILK调节骨桥蛋白诱导的血管平滑肌细胞增殖  

Pillbug extracts inhibit osteopontin-induced vascular smooth muscle cell proliferation by modulation of integrin kinase protein pathway

作　　者：段哲萍[1] 于新江[2] 李菁菁[3] 赵娟[3] Duan Zhe-ping Yu Xin-jiang Li Jing-jing Zhao Juan(Second Department of Oncology Department of Cardiac Surgery, Hebei General Hospital, Shijiazhuang 050051, Hebei Province, China Department of Cytobiology, Hebei Medical University, Shijiazhuang 050017, Hebei Province, China)

机构地区：[1]河北省人民医院肿瘤二科,河北省石家庄市050051 [2]河北省人民医院心脏外科,河北省石家庄市050051 [3]河北医科大学细胞生物教研室,河北省石家庄市050017

出　　处：《中国组织工程研究》2017年第16期2582-2586,共5页Chinese Journal of Tissue Engineering Research

基　　金：河北省中医药管理局课题(2015138);河北省高等学校科学技术研究青年基金(2011177)~~

摘　　要：背景:前期实验通过MTT法及细胞划痕实验检测出鼠妇可影响血管平滑肌细胞的增殖和迁移过程,但其相关作用机制尚不清楚。目的:探讨鼠妇对于骨桥蛋白诱导的血管平滑肌细胞增殖影响的相关分子机制。方法:按常规方法培养SD大鼠胸腹主动脉血管平滑肌细胞,选取3-5代细胞用于实验。实验分为8组:对照组、骨桥蛋白组、鼠妇水提取物(0.5,1.0,2.0 g/L)+骨桥蛋白组、鼠妇乙酸乙酯提取物(0.25,0.5,1.0 g/L)+骨桥蛋白组。加入相应药物培养24 h后,收集细胞,采用Western bolt检测PCNA、ILK蛋白表达水平。结果与结论:(1)在鼠妇水提取物及乙酸乙酯提取物的作用下血管平滑肌细胞PCNA蛋白表达下降,其中0.5 g/L的鼠妇水提取物组和1.0 g/L的鼠妇乙酸乙酯提取物组抑制作用最明显,分别为骨桥蛋白组的77.8%和74.1%;(2)低质量浓度(0.5 g/L)的鼠妇水提取物抑制ILK表达作用明显,是骨桥蛋白组的81.4%;鼠妇乙酸乙酯提取物作用下血管平滑肌细胞ILK的表达水平没有明显改变;(3)结果表明,鼠妇水提取物及乙酸乙酯提取物均可抑制骨桥蛋白诱导的血管平滑肌细胞增殖,且鼠妇水提取物该作用可能与ILK蛋白通路有关。BACKGROUND： Pillbugs have been proved to exert an effect on the proliferation and migration of vascular smooth muscle cells （VSMCs） through MTT assay and scratch assay, but the mechanisms are poorly understood. OBJECTIVE： To investigate the molecular mechanisms underlying pillbugs influencing osteopontin-induced VSMC migration and proliferation. METHODS： VSMCs from Sprague-Dawley rat thoraco-abdominal aorta were cultured in a common medium, and 3-5 generations of cells were selected. There were eight groups： control, osteopontin, pillbug water extract （0.5, 1.0, and 2.0 g/L） with osteopontin, and pillbug ethyl acetate extract （0.25, 0.5, and 1.0 g/L） with osteopontin groups. After 24 hours of culture, the expression levels of proliferating cell nuclear antigen （PCNA） and integrin kinase （ILK） were detected by western blot assay. RESULTS AND CONCLUSION： Pillbug water and ethyl acetate extracts both inhibited the expression of PCNA, especially when cultured in 0.5 g/L pillbug water extract （77.8% of the osteopontin group） and 1.0 g/L ethyl acetate extract （74.1% of the osteopontin group）. 0.5 g/L pillbug water extract significantly downregulated ILK level induced by osteopontin, which was 81.4% of the osteopontin group, while pillbug ethyl acetate extract exposed no influence on ILK expression. These results reveal that pillbug water and ethyl acetate extracts both are able to inhibit osteopontin-induced VSMC proliferation, and the role of the pillbug water extract maybe associated with ILK protein pathway.

关 键 词：甲虫 骨桥蛋白质 增殖细胞核抗原 组织工程 组织构建 鼠妇 骨桥蛋白 PCNA ILK 

分 类 号：R318[医药卫生—生物医学工程]