莲子心新碱对肠系膜血管平滑肌收缩的抑制作用及分子机制  

The Inhibitory Effect and Mechanism of Neoliensinine on the Contraction of Mesenteric Vascular Smooth Muscle

作　　者：张军利 肖敏 王鹏[1,2] 张弦 潘扬[1,2] 杨光明[1,3] ZHANG Jun-li;XIAO Min;WANG Peng;ZHANG Xian;PAN Yang;YANG Guang-ming(School of Pharmacy,Nanjing University of Chinese Medicine,Nanjing,210023,China;Laboratory of Medical Fungi and Phyto Biotech,Nanjing University of Chinese Medicine,Nanjing,210023,China;Key Laboratory of State Administration of Traditional Chinese Medicine for Standardization of Chinese Medicine Processing,Jiangsu Key Laboratory of Chinese Medicine Processing,Nanjing,210023,China)

机构地区：[1]南京中医药大学药学院,江苏南京210023 [2]南京中医药大学药用菌与中药生物技术研究所,江苏南京210023 [3]江苏省中药炮制重点实验室,国家中医药管理局中药炮制标准重点研究室,江苏南京210023

出　　处：《南京中医药大学学报》2019年第3期313-318,共6页Journal of Nanjing University of Traditional Chinese Medicine

基　　金：国家自然科学基金(81373295);江苏高校优势学科建设工程资助项目(PAPD);江苏高校品牌专业建设工程资助项目(PPZY2015A070)

摘　　要：目的 研究莲子心新碱(Neo)对小鼠肠系膜血管及血管平滑肌细胞(VSMCs)收缩的抑制作用;并通过观察其对肌球蛋白轻链(MLC20)磷酸化和RhoA相关蛋白激酶(ROCK1)表达的影响,初步探讨引起该抑制的作用机制。方法 采用离体微血管张力测定仪观察高钾溶液(KCl)对经Neo预处理后肠系膜血管收缩的影响,并通过Urea/glycerolPAGE法检测其MLC20磷酸化水平的变化。同时,利用成像分析技术考察高钾溶液对经Neo预处理后VSMCs收缩的影响,并通过细胞免疫荧光法测定VSMCs中ROCK1蛋白的表达水平。结果 经Neo预处理后的肠系膜血管能明显抑制高钾溶液引起的收缩,其IC50值为6.019μmol/L;且可阻断高钾溶液产生MLC20磷酸化。另外,与模型组(KCl)VSMCs的长度(40.94±1.94)μm相比,药物组(Neo+KCl)VSMCs的长度(44.95±5.15)μm显著延长(P<0.01),其ROCK1蛋白的表达也明显下降。结论 Neo对肠系膜血管平滑肌收缩具有明显的抑制作用,其作用机制可能与阻断平滑肌MLC20磷酸化和下调其ROCK1蛋白表达有关。OBJECTIVE To observe the inhibition effect of neoliensinine (Neo) on the contraction of mesenteric vascular smooth muscle and vascular smooth muscle cells (VSMCs) of mice,and to explore the mechanism of the effect by observing phosphorylation of myosin light chain (MLC20) in vascular smooth muscle and expression of ROCK1 (RhoA associated protein kinase) in VSMCs. METHODS The microvascular tension tester was used to evaluate the inhibition effect of Neo pretreatment on mesenteric vessel contraction stimulated by high potassium solution,and Urea/glycerol PAGE was applied to detect its change of phosphorylation of MLC20 . Meanwhile,image analyzer was used to detect the effect of Neo on VSMCs contraction induced by high potassium solution,and cell immunofluorescence assay was adopted to detect its expression of ROCK1 protein in VSMCs. RESULTS Neo had a significant inhibitory effect on the contraction of mesenteric vessel induced by high potassium solution with IC50 of 6.019 μmol/L and 95% confidence intervals from 5.299~6.838 μmol/L,and could block the phosphorylation of MLC20 induced by high potassium. In addition,compared with the VSMCs length of model group (KCl)(40.94± 1.94 )μm,that of treatment group (Neo+KCl) were prolonged (44.95±5.15)μm significantly ( P < 0.01 ),and Neo reduced the expression of ROCK1 protein in VSMCs. CONCLUSION Neo could inhibit the contraction of mesenteric vascular smooth muscle,and its mechanism might be related to blocking the phosphorylation of smooth muscle MLC20 and down regulating the expression of ROCK1 protein in VSMCs.

关 键 词：莲子心新碱 平滑肌收缩 抑制作用 MLC20磷酸化 ROCK1 

分 类 号：R285.5[医药卫生—中药学]