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血管平滑肌细胞表型转化在高盐诱导的大鼠颈动脉重构中的作用
  • ISSN号:1007-3949
  • 期刊名称:《中国动脉硬化杂志》
  • 时间:0
  • 分类:R363[医药卫生—病理学;医药卫生—基础医学]
  • 作者机构:[1]遵义医学院临床医学研究所心血管病研究所高血压研究室, [2]遵义医学院附属医院心内科,贵州省遵义市563003, [3]重庆市高血压研究所,重庆市400042
  • 相关基金:国家自然科学基金资助项目(81160041;81460077); 贵州省优秀科技教育人才省长专项基金项目[黔省专合字(2012)15号]; 贵州省国际科技合作计划项目[G字(2012)7038号]
中文摘要:

目的研究辣椒素对高盐诱导的大鼠血管平滑肌细胞(VSMC)增殖的抑制作用。方法组织贴壁法培养原代大鼠血管平滑肌细胞,根据MTT结果绘制大鼠血管平滑肌细胞的生长曲线,大鼠血管平滑肌细胞传至第4代去血清同步化24 h,不同浓度辣椒素(0.01μmol/L、0.1μmol/L、1μmol/L、10μmol/L、100μmol/L)进行高盐培养72 h,MTT比色法检测大鼠血管平滑肌细胞增殖情况,选取抑制血管平滑肌细胞增殖的辣椒素浓度进行后续实验。CCK-8检测渗透压对大鼠血管平滑肌细胞增殖的影响,流式细胞仪分析细胞增殖周期,免疫荧光染色法和Western blot检测增殖细胞核抗原(PCNA)的表达,实时荧光定量PCR检测瞬时受体电位家族香草醛1型受体(TRPV1)mRNA的表达,Western blot检测瞬时受体电位家族香草醛1型受体的蛋白表达。结果 MTT结果显示,辣椒素浓度为10μmol/L时大鼠血管平滑肌细胞增殖开始受抑制,100μmol/L时抑制作用增强(P〈0.05),选取10μmol/L辣椒素进行后续实验。CCK-8结果显示,高盐组细胞明显增殖,而甘露醇组、正常组和高盐+辣椒素组无差异。流式细胞仪测得高盐组G0/G1、G2/M期细胞比例减少(P〈0.05),S期细胞比例增多(P〈0.05);与高盐组比较,高盐+辣椒素组G0/G1、G2/M期细胞比例增加(P〈0.05),S期细胞比例下降(P〈0.05)。免疫荧光及Western blot结果显示,高盐组增殖细胞核抗原阳性细胞核增多,蛋白表达增加(P〈0.05),而高盐+辣椒素组增殖细胞核抗原阳性细胞核减少,蛋白表达减少(P〈0.05)。实时荧光定量PCR和Western blot结果显示高盐组瞬时受体电位家族香草醛1型受体mRNA及蛋白表达减少(P〈0.05),辣椒素组则增加(P〈0.05)。结论辣椒素可抑制高盐所致的大鼠血管平滑肌细胞增殖,其作用机制可能与激活瞬时受体电位家族香草醛1型受体表达有关。

英文摘要:

Aim To investigate the effect of capsaicin on the inhibition of rat vascular smooth muscle cells (VSMC) induced by high sodium.Methods Vascular smooth muscle cells were isolated from thoracic aorta of male Sprague-Dawley rats by tissue adherent method,and the growth curve of vascular smooth muscle cells was made according to MTT assay.The experimental procedures were performed with the fourth generation of vascular smooth muscle cells.Vascular smooth muscle cells were synchronized by serum starvation for 24 h before capsaicin and high sodium treatment.Vascular smooth muscle cells were treated with different concentration of capsaicin (0.01,0.1,1,10 and 100 μmol/L) for 72 h in high sodium.The effects of different concentration capsaicin and osmotic pressure on the proliferation induced by high sodium medium in vascular smooth muscle cells were evaluated by MTT assay and CCK-8,respectively.The cell cycle of vascular smooth muscle cells was analyzed by flow cytometry technology.The localization and protein expression of PCNA were employed by immunofluorecence staining and Western blot.The mRNA and protein expressions of TRPV1 were detemined by real-time PCR and Western blot.Results MTT results showed that when the concentration of capsaicin was 10μmol/L,the proliferation of rat vascular smooth muscle cells began to be inhibited,and the inhibitory effect was increased at 100 μmol/L (P〈0.05),so 10 μmol/L capsaicin was selected for subsequent experiments.CCK-8 results showed that the cells in high sodium group proliferated obviously,but there was no difference between mannitol group,normal group and capsaicin group.Capsaicin prevented notably the decrease of cell number in G0/G1 and G2/M stage and increase of cell number of S stage in vascular smooth muscle cells treated with Na+(159 mmol/L).PCNA protein level was also decreased by capsaicin.The decrease of mRNA and protein level of TRPV1 stimulated Na+(159 mmol/L) was alleviated by capsaicin.Conclusion Capsaicin can inhibit the proliferat

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期刊信息
  • 《中国动脉硬化杂志》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学技术协会
  • 主办单位:中国病理生理学会
  • 主编:姜志胜
  • 地址:湖南省衡阳市南华大学内
  • 邮编:421001
  • 邮箱:dmzzbjb@163.net
  • 电话:0734-8160765
  • 国际标准刊号:ISSN:1007-3949
  • 国内统一刊号:ISSN:43-1262/R
  • 邮发代号:42-165
  • 获奖情况:
  • 中国科学技术论文统计源期刊,中国科学引文数据库来源期刊,中国学术期刊综合评价数据库来源期刊
  • 国内外数据库收录:
  • 美国化学文摘(网络版),中国中国科技核心期刊,中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:17016