缝隙连接在同型半胱氨酸介导的自发性高血压大鼠血管平滑肌细胞增殖中的作用  

Roles of gap junctions in proliferation mediated by Hcy in the spontaneous hypertensive rat vascular smooth muscle cells

作　　者：钟华[1,2] 胡清华[1,5] 王恩帮[1,2] 司军强[1,3] 孙志萍[4] 李增春[4] 何芳[1,2] 

机构地区：[1]新疆地方病与民族高发病教育部重点实验室,石河子832002 [2]新疆地方病与民族高发病教育部石河子大学医学院病理生理教研室,石河子832002 [3]新疆地方病与民族高发病教育部生理教研室,石河子832002 [4]新疆地方病与民族高发病教育部医学机能实验中心,石河子832002 [5]华中科技大学同济医学院病理生理学系,卫生部呼吸系统疾病重点实验室,湖北武汉430030

出　　处：《中国应用生理学杂志》2012年第4期289-293,共5页Chinese Journal of Applied Physiology

基　　金：石河子大学科学技术研究发展计划“自然科学与技术创新”重点项目(ZRKX2009ZD03-1);国家自然科学基金资助项目(30860099)

摘　　要：目的:探讨缝隙连接(GJ)是否参与同型半胱氨酸(Hcy)介导的自发性高血压(SHR)大鼠血管平滑肌细胞(VSMCs)增殖及可能的分子机制。方法:原代培养SHR VSMCs,细胞分四组:①对照组,②Hcy组,③缝隙连接阻断剂(18α-GA)组和④Hcy+18α-GA组。MTT法及流式细胞仪检测细胞的增殖活性,免疫荧光技术观察细胞中Cx43、Cx40蛋白表达及定位,Western blot法检测细胞中Cx43、Cx40蛋白表达,染料示踪分子传递法(划痕标记染料传输法)检测细胞的缝隙连接功能。结果:①与对照组相比,Hcy组MTT法测得A值及细胞周期测得S值增高(P<0.05),18α-GA组降低(P<0.05);与Hcy组比,TGFβ-1+18α-GA组A值及S值均降低(P<0.05)。②免疫荧光技术检测细胞中Cx43、Cx40蛋白表达呈阳性,两者共定位于胞浆。③与对照组相比,Hcy组Cx43、Cx40蛋白表达增强(P<0.05),18α-GA组Cx43、Cx40表达均减弱(P<0.05);与Hcy组比,Hcy+18α-GA组Cx43、Cx40表达均减弱(P<0.05)。④与对照组相比,Hcy组缝隙连接功能明显增强(P<0.05),18α-GA组缝隙连接功能明显减弱(P<0.05),与Hcy组比,Hcy+18α-GA组缝隙连接功能显著降低(P<0.05)。结论:Hcy通过上调SHR VSMCs Cx43和Cx40的蛋白表达,引起缝隙连接通讯功能的增强,促进了SHR VSMCs增殖。Objective： To investigate whether homocysteine（Hcy）participates the proliferation of the spontaneously hypertensive rat（SHR） vascular smooth musde cell（VSMCs） and the molecular mechanism. Methods： The rat＇s arota were removed. The primary SHR VSMCs were isolated and cultured in vitro, then the SHR VSMCs were divided into four groups： （1）control group, （2）Hcy group, （3）18α-glycyrrhetinic acid （GA） group, （4）Hcy ＋ 18α-GA group. We detected proliferation of the SHR VSMCs by MTT and flow cytometry. The expression and co-localization of the connexin（Cx）43 and Cx40 proteins in the SHR VSMCs were deteced by immunofluorescence. The expression of the Cx43 and Cx40 proteins in SHR VSMCs were detected by Western blot. The molecular dye transfer method （scrape dye transfer method） was applied to detect the gap junction function in the SHR VSMCs. Results： （1）The Cx43 and Cx40 proteins expression in the SHR VSMCs were positive, confocal microscopy supported the co-loesliization of Cx43 and Cx40 in the cytoplasm. （2）The S value deteced by cell cycle and A value detected by MTT in the Hcy group were increased obviously compared with those in the control group （ P 〈 0.05）, decreased in 18α-GA group（ P 〈 0.05）. Compared with the Hcy group, the S and A value in the Hcy ＋ 18α-GA group were significantly decreased, respectively （ P 〈0.05）. （3）The expression of Cx43 and Cx40 proteins in Hcy group were increased compared with the control group （ P 〈 0.05）, decreased in 18α-GA group （ P 〈0.05）. Compared with the Hcy group, the expression of Cx43 and Cx40 proteins in the Hcy ＋ 18α-GA group were significantly decreased, respectively （ P 〈 0.05）. （4）The function of gap junction detected by scrape dye transfer method in the Hcy group were enhanced compared with the control group （ P 〈 0.05）, weakened in the 18α-GA group （ P 〈 0.05）. Compared with the Hcy greup,the function of gap junction in the Hcy ＋ 18α-GA grou

关 键 词：同型半胱氨酸 缝隙连接 血管平滑肌细胞 增殖 

分 类 号：R543.5[医药卫生—心血管疾病]