尼古丁对大鼠主动脉平滑肌细胞舒缩功能的影响
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摘要
目的:观察尼古丁对大鼠主动脉血管平滑肌细胞(vascular smooth muscle cells,VSMCs)舒缩功能的影响。方法:体外培养大鼠原代主动脉VSMCs。用不同浓度尼古丁对大鼠主动脉VSMCs干预24 h后,用罗丹明-鬼笔环肽染细胞骨架,拍摄不同实验组的照片,通过测量不同细胞表面积的大小来反映细胞收缩水平;用胶原收缩法观察不同浓度的尼古丁对大鼠VSMCs收缩功能影响。结果:原代大鼠主动脉VSMCs被成功培养。用不同浓度(0.1μmol/L、1μmol/L、10μmol/L和100μmol/L)尼古丁处理VSMCs 24 h后,其骨架呈现显著收缩,细胞板状化明显增强,且呈浓度依赖性关系;10μmol/L为尼古丁对VSMCs的最适刺激浓度(P<0.01)。胶原收缩法也显示,10μmol/L尼古丁对大鼠主动脉VSMCs有收缩作用,随着作用时间的增加,其收缩效应逐渐增强,作用60 min收缩作用最显著(P<0.01)。结论:尼古丁对大鼠主动脉VSMCs具有较强收缩作用,其收缩作用具有浓度依赖性和时间依赖性。
AIM: To observe the effects of nicotine on systolic and diastolic function of rat aortic vascular smooth muscle cells(VSMCs). METHODS: The primary rat aortic VSMCs were cultured in vitro. After exposed to nicotine at different concentrations for 24 h, the cytoskeleton of the VSMCs was stained with rhodamine-phalloidin,the photographs of the VSMCs in different experimental groups were taken and the surface area was measured to reflect the cell contractility. Collagen contraction method was also used to determine the effect of nicotine on the contractility of rat aortic VSMCs. RESULTS: The primary rat aortic VSMCs were successfully cultured. After the VSMCs were treated with nicotine(0.1 μmol/L, 1 μmol/L, 10 μmol/L and 100 μmol/L) for 24 h, the skeleton showed a significant contraction, and the cell plate shape was obviously enhanced in a concentration-dependent manner. The results showed that 10 μmol/L was the optimal concentration of nicotine for VSMCs(P<0.01). The collagen contraction method also showed that 10 μmol/L nicotine contracted the rat aortic VSMCs. With the increase in the nicotine action time, the maximum contraction effect was observed at 60 min(P<0.01). CONCLUSION: Nicotine has a strong contractile effect on VSMCs of rat aorta, and its contractile effect is dependent on concentration and time.
AIM: To observe the effects of nicotine on systolic and diastolic function of rat aortic vascular smooth muscle cells(VSMCs). METHODS: The primary rat aortic VSMCs were cultured in vitro. After exposed to nicotine at different concentrations for 24 h, the cytoskeleton of the VSMCs was stained with rhodamine-phalloidin,the photographs of the VSMCs in different experimental groups were taken and the surface area was measured to reflect the cell contractility. Collagen contraction method was also used to determine the effect of nicotine on the contractility of rat aortic VSMCs. RESULTS: The primary rat aortic VSMCs were successfully cultured. After the VSMCs were treated with nicotine(0.1 μmol/L, 1 μmol/L, 10 μmol/L and 100 μmol/L) for 24 h, the skeleton showed a significant contraction, and the cell plate shape was obviously enhanced in a concentration-dependent manner. The results showed that 10 μmol/L was the optimal concentration of nicotine for VSMCs(P<0.01). The collagen contraction method also showed that 10 μmol/L nicotine contracted the rat aortic VSMCs. With the increase in the nicotine action time, the maximum contraction effect was observed at 60 min(P<0.01). CONCLUSION: Nicotine has a strong contractile effect on VSMCs of rat aorta, and its contractile effect is dependent on concentration and time.
引文
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[6] 黄捷,谢良地,许昌声,等.siRNA干扰热休克蛋白27基因对自发性高血压大鼠血管平滑肌细胞骨架及迁移的影响[J].中华高血压杂志,2010,18(10):936-939.
[7] 田年秀,王晓磊,王涛,等.芪卫颗粒含药血清通过caspase-12途径减轻高糖环境下肾足细胞损伤的机制研究[J].中华中医药杂志,2018,33(5):1858-1862.
[8] 曲明娟,徐伟艳,王晓安,等.尼古丁对大鼠血管平滑肌细胞增殖和凋亡的影响[J].解剖学报,2011,42(2):215-219.
[9] Boehme AK,Esenwa C,Elkind MS.Stroke risk factors,genetics,and prevention [J].Circ Res,2017,120(3):472-495.
[10] 张文通,李俊,吴玉婷,等.山柰酚-3-O-芸香糖苷对血管平滑肌细胞增殖、迁移及TGFBR1信号通路活化的影响[J].中国病理生理杂志,2018,34(5):832-838.
[2] Howard G,Wagenknecht LE,Burke GL,et al.Cigarette smoking and progression of atherosclerosis:the atherosclerosis risk in communities (ARIC) study[J].JAMA,1998,279(2):119-124.
[3] d’Adesky ND,de Rivero Vaccari JP,Bhattacharya P,et al.Nicotine alters estrogen receptor-beta-regulated inflammasome activity and exacerbates ischemic brain damage in female rats[J].Int J Mol Sci,2018,19(5):1330-1335.
[4] 张坤,秦小江,侯晓敏,等.杜鹃素通过降低Cx43的表达抑制AngⅡ诱导的VSMCs增殖[J].中国药理学通报,2018,34(3):401-407.
[5] 林京,张子力.改良植块贴壁法建立大鼠主动脉平滑肌细胞体外培养模型[J].中国病理生理杂志,2006,22(4):827-829.
[6] 黄捷,谢良地,许昌声,等.siRNA干扰热休克蛋白27基因对自发性高血压大鼠血管平滑肌细胞骨架及迁移的影响[J].中华高血压杂志,2010,18(10):936-939.
[7] 田年秀,王晓磊,王涛,等.芪卫颗粒含药血清通过caspase-12途径减轻高糖环境下肾足细胞损伤的机制研究[J].中华中医药杂志,2018,33(5):1858-1862.
[8] 曲明娟,徐伟艳,王晓安,等.尼古丁对大鼠血管平滑肌细胞增殖和凋亡的影响[J].解剖学报,2011,42(2):215-219.
[9] Boehme AK,Esenwa C,Elkind MS.Stroke risk factors,genetics,and prevention [J].Circ Res,2017,120(3):472-495.
[10] 张文通,李俊,吴玉婷,等.山柰酚-3-O-芸香糖苷对血管平滑肌细胞增殖、迁移及TGFBR1信号通路活化的影响[J].中国病理生理杂志,2018,34(5):832-838.