辣椒素对高盐致大鼠血管平滑肌细胞增殖的抑制作用

发布时间：2018-02-14 14:32

本文关键词： 高盐血管平滑肌细胞细胞增殖辣椒素　出处：《中国动脉硬化杂志》2017年06期 　论文类型：期刊论文

【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(P0.05),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(P0.05),S期细胞比例增多(P0.05);与高盐组比较,高盐+辣椒素组G0/G1、G2/M期细胞比例增加(P0.05),S期细胞比例下降(P0.05)。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(P0.05),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(P0.05)。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(P0.05),辣椒素组则增加(P0.05)。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。
[Abstract]:Objective to study the inhibitory effect of capsaicin on the proliferation of rat vascular smooth muscle cells (VSMC) induced by high salt. Methods Primary rat vascular smooth muscle cells (VSMC) were cultured by tissue adherent method and the growth curve of VSMC was plotted according to MTT results. Rat vascular smooth muscle cells (VSMCs) were transferred to the 4th generation and synchronized for 24 h. The proliferation of rat vascular smooth muscle cells (VSMCs) was detected by MTT colorimetric assay at 72 h after high salt culture. The concentration of capsaicin was 0.01 渭 mol / L ~ (0.1 渭 mol / L) and 10 渭 mol / L ~ (10) 渭 mol / L ~ (10 渭 mol / L) respectively. The effect of osmotic pressure on proliferation of vascular smooth muscle cells in rats was detected by CCK-8. Flow cytometry was used to analyze cell proliferation cycle. Immunofluorescence staining and Western blot were used to detect the expression of proliferating cell nuclear antigen (PCNA). Real time fluorescence quantitative PCR was used to detect the mRNA expression of vanillin type 1 receptor type 1 receptor in transient receptor potential family. Western blot was used to detect the protein expression of vanillin type 1 receptor family in transient receptor potential family. When the concentration of capsaicin was 10 渭 mol/L, the proliferation of vascular smooth muscle cells in rats was inhibited at 100 渭 mol/L. The results of 10 渭 mol/L capsaicin test. CCK-8 showed that the proliferation of vascular smooth muscle cells in high salt group was obvious, while that in mannitol group was higher than that in mannitol group. There was no difference between normal group and high salt capsaicin group. In high salt capsaicin group, the proportion of G _ 0 / G _ 1 G _ 2 / M phase increased and the proportion of P _ (0.05) / S phase decreased. The results of immunofluorescence and Western blot showed that the number of proliferating cell nuclear antigen positive cells increased in high salt group. Protein expression increased in P0.05, but decreased in high salt capsaicin group. The results of real-time fluorescence quantitative PCR and Western blot showed that the expression of vanillin type 1 receptor mRNA and protein decreased in high salt group, but increased in capsaicin group. Conclusion capsaicin can inhibit the high salt level. Proliferation of rat vascular smooth muscle cells, The mechanism may be related to the activation of the transient receptor potential family vanillin type 1 receptor expression.
【作者单位】：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;遵义医学院附属医院心内科;重庆市高血压研究所;
【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]
【分类号】：R544.1

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