Yao H M, Jia Y P, Xue Z, Guo M, Lyu J Y
Department of Cardiology, First Affiliated Hospital of Shanxi, Medical University, Taiyuan 030001, China.
Zhonghua Xin Xue Guan Bing Za Zhi. 2017 Apr 24;45(4):323-328. doi: 10.3760/cma.j.issn.0253-3758.2017.04.013.
To investigate the effect of natural active compounds apigenin (API) on the proliferation of rat aortic vascular smooth muscle cells (VSMCs) induced by lipopolysaccharide (LPS) and related mechanisms. VSMCs of primary cultured SD rats were obtained and the cytotoxic effects of API (0, 10, 20, 40 and 80 μmol/L) was explored by CCK-8 method. Impact of LPS (0, 0.1, 1, 10 and 100 μg/ml) on VSMCs proliferation and the impact of API (0, 10, 20, 40 μmol/L) on LPS (10 μmol)-induced VSMCs proliferation by CCK-8 methods. Using EdU and FCM method, we observed the effect of API on proliferation of VSMCs induced by LPS. VSMCs proliferation and cell cycle were also assessed by EdU method and FACS in 10 μg/ml LPS, 10 μg/ml LPS+ 40 μmol/L API and equal volume DMSO treated VSMCs. (1) CCK-8 cell vitality test showed that cell vitality was not affected by 0-40 μmol/L API, while cell vitality was significantly reduced by 80 μmol/L API (57%), which was significantly lower than in blank group (<0.05). (2) VSMCs proliferation was significantly promoted by 0.1, 1 and 10 μg/ml LPS and peaked in 10 μg/ml LPS stimulated VSMCs group, while VSMCs proliferation was significantly reduced in 100 μg/ml LPS stimulated group (<0.05 vs. blank group). (3) LPS (10 μm/ml) induced VSMCs proliferation was not affected by 10 μmol/L API, which was significantly inhibited by 20 and 40 μmol/L API (both <0.05 vs. LPS). (4) VSMCs proliferation assessed by EdU was significantly higher in LPS group than in blank group (<0.01), which could be significantly reduced by cotreatment with API (<0.01). (5) FACS results showed that percent of VSMCs in G0/G1 stage was significantly lower in LPS group compared to blank group (<0.05), which could be significantly increased post API treatment (<0.05 vs. LPS), while percent of VSMCs in S stage was significantly lower post API treatment in comparison with LPS group. API can significantly inhibit LPS-induced proliferation of VSMCs, partly through inhibiting mitosis and inducing G0/G1 cell cycle arrest.
探讨天然活性化合物芹菜素（API）对脂多糖（LPS）诱导的大鼠主动脉血管平滑肌细胞（VSMCs）增殖的影响及其相关机制。获取原代培养的SD大鼠VSMCs，采用CCK-8法探究API（0、10、20、40和80μmol/L）的细胞毒性作用。采用CCK-8法检测LPS（0、0.1、1、10和100μg/ml）对VSMCs增殖的影响以及API（0、10、20、40μmol/L）对LPS（10μmol）诱导的VSMCs增殖的影响。利用EdU和流式细胞术（FCM）方法，观察API对LPS诱导的VSMCs增殖的影响。采用EdU法和流式细胞分选术（FACS）评估10μg/ml LPS、10μg/ml LPS + 40μmol/L API及等量二甲基亚砜（DMSO）处理的VSMCs的增殖和细胞周期。（1）CCK-8细胞活力检测显示，0 - 40μmol/L API对细胞活力无影响，而80μmol/L API可使细胞活力显著降低（57%），显著低于空白组（<0.05）。（2）0.1、1和10μg/ml LPS可显著促进VSMCs增殖，在10μg/ml LPS刺激的VSMCs组中达到峰值，而100μg/ml LPS刺激组的VSMCs增殖显著降低（与空白组相比<0.05）。（3）LPS（10μm/ml）诱导的VSMCs增殖不受10μmol/L API影响，而20和40μmol/L API可显著抑制其增殖（两者与LPS相比均<0.05）。（4）通过EdU评估，LPS组的VSMCs增殖显著高于空白组（<0.01），与API共同处理可使其显著降低（<0.01）。（5）FACS结果显示，LPS组VSMCs在G0/G1期的百分比显著低于空白组（<0.05），API处理后可显著升高（与LPS相比<0.05），而与LPS组相比，API处理后VSMCs在S期的百分比显著降低。API可显著抑制LPS诱导的VSMCs增殖，部分是通过抑制有丝分裂和诱导G0/G1期细胞周期阻滞实现的。
