血根碱对小鼠肠系膜动脉血管平滑肌收缩的抑制作用及分子机制  

The Inhibition Effect and Mechanism of Sanguinarine on the Contraction of Mouse Mesenteric Vascular Smooth Muscle

作　　者：肖敏 张军利[1,2] 王鹏 杨光明 潘扬[1,2] XIAO Min;ZHANG Junli;WANG Peng;YANG Guangming;PAN Yang(School of Pharmacy, Nanjing University of Chinese Medicine,Nanjing 210023 Jiangsu,China;Laboratory of Medical Fungi and Phyto-Biotech,Nanjing University of Chinese Medicine,Nanjing 210023 Jiangsu,China;Jiangsu Key Laboratory of Chinese Medicine Processing & Key Laboratory of State Administration of Traditional Chinese Medicine for Standardization of Chinese Medicine Processing,Nanjing 210023 Jiangsu,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：《中药新药与临床药理》2019年第1期7-13,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(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：目的研究血根碱(San)对小鼠肠系膜血管及血管平滑肌细胞(VSMCs)收缩的抑制作用;并观察其对RhoA相关蛋白激酶(ROCK1)表达的影响,探讨引起该抑制作用的作用机制。方法利用微血管张力测定仪(DMT),测定San对血管收缩工具药氯化钾(KCl)预收缩后微血管的张力影响,并计算血管平滑肌松弛作用的IC50值;进一步利用成像分析技术观测San对KCl预收缩VSMCs后的影响,最后通过蛋白免疫印迹以及细胞免疫荧光法测定San对VSMCs中ROCK1蛋白的表达水平的影响。结果与ROCK1抑制剂法舒地尔(Fas)比较(IC_(50)为14.34μmol·L^(-1)),San对KCl预收缩的肠系膜动脉的松弛作用较强,其IC50为1.098μmol·L^(-1),且呈现不(弱)可逆的趋势。进一步在细胞水平上验证,与模型组(KCl)VSMCs的长度(40.03±4.91)μm比较,药物组(KCl+San)VSMCs的长度(46.12±6.37)μm显著延长(P<0.01)。蛋白免疫印迹与免疫荧光结果发现San可显著抑制ROCK1蛋白表达(P<0.01)。结论 San对KCl诱导的肠系膜血管平滑肌收缩具有明显的抑制作用,其作用机制可能与调控ROCK1蛋白表达相关。Objective To study the inhibitory effect of sanguinarine(San) on the contraction of mouse mesentericvessel and vascular smooth muscle cells(VSMCs);and to observe its effect on the expression of RhoA-associatedprotein kinase(ROCK1) and to explore the mechanism of the inhibition. Methods The microvasculature tension ofVSMCs was determined using a microvessel tension meter(DMT) after being precontracted by vasocontractorpotassium chloride(KCl). The IC50 value of relaxing vascular smooth muscle was calculated and KCl-precontractedVSMCs was further observed using imaging analysis technique. Finally,the expression of ROCK1 protein in VSMCswas detected by Western Blot and immunofluorescence. Results Compared with ROCK1 inhibitor fasudil(Fas)(IC50was 14.34 μmol·L^-1),San had a stronger relaxation effect on mesenteric artery preconstricted by KCl with IC50 of 1.098 μmol·L^-1and showed no(weak) reversibility. Compared with the length of VSMCs(40.03±4.91 μm) in model group(KCl),the length of VSMCs(46.12±6.37) μm of drug group(KCl+San) was significantly prolonged(P < 0.01). Western Blot and immunofluorescence detection showed that San could significantly inhibit theexpression of ROCK1 protein(P < 0.01). Conclusion San has a significant inhibitory effect on KCl-inducedmesenteric vascular smooth muscle contraction,and its mechanism may be related to the down-regulation of ROCK1 protein.

关 键 词：微血管张力测定 血根碱 血管平滑肌 ROCK1 抑制作用 

分 类 号：R285.5[医药卫生—中药学]