健脾化湿颗粒对大鼠结肠上皮细胞OCLN、AQPs及平滑肌细胞cAMP、AC的影响  

Effects of Jianpi Huashi Granules on OCLN and AQPs of colonic epithelial cells and cAMP and AC of colonic smooth muscle cells

作　　者：王迎寒[1] 刘玉玲[1] 陈光晖[1] 胡楠[1] 张晓峰[1] WANG Ying⁃han;LIU Yu⁃ling;CHEN Guang⁃hui;HU Nan;ZHANG Xiao⁃feng(Hebei Provincial Key Laboratory for Research and Development of Traditional Chinese Medicine,Institute of Chinese Materia Medica,Chengde Medical University,Chengde 067000,China)

机构地区：[1]承德医学院中药研究所河北省中药研究与开发重点实验室

出　　处：《中成药》2020年第2期323-328,共6页Chinese Traditional Patent Medicine

基　　金：河北省高等学校科学技术研究项目(YQ2014002);河北省高校重点学科建设项目(冀教高[2013]4);承德医学院高层次人才科研启动基金项目(201602)

摘　　要：目的探究健脾化湿颗粒对大鼠结肠上皮细胞OCLN、AQPs及平滑肌细胞cAMP、AC的影响。方法采用CCK⁃8法检测不同浓度健脾化湿颗粒对上皮细胞的存活率影响;利用Western blot检测健脾化湿颗粒对上皮细胞OCLN、AQP3、AQP4蛋白表达的影响;分别用健脾化湿颗粒及Ach、阿托品干预平滑肌细胞后,利用显微标尺测量平滑肌细胞长度,ELISA法检测平滑肌细胞cAMP、AC的水平。结果健脾化湿颗粒(≤1 mg/mL)处理上皮细胞24 h后,细胞存活率≥90%。与正常组比较,健脾化湿颗粒上调上皮细胞OCLN、AQP3,AQP4蛋白表达,促进结肠平滑肌的舒张,下调结肠平滑肌AC、cAMP的水平(P<0.05,P<0.01)。健脾化湿颗粒可抑制Ach引起的平滑肌细胞收缩,及cAMP、AC水平的上调;与阿托品协同作用促进平滑肌细胞舒张,及cAMP、AC水平的下调(P<0.05,P<0.01)。结论健脾化湿颗粒可能通过调节离体大鼠结肠OCLN、AQP3、AQP4蛋白表达及平滑肌细胞舒缩功能来改善D⁃IBS腹泻症状。AIM To investigate the effects of Jianpi Huashi Granules(JHG)on OCLN and AQPs of colonic epithelial cells and cAMP and AC of colonic smooth muscle cells.METHODS CCK⁃8 method was used to detect the effect of different concentrations of JHG on the survival rate of epithelial cells.Western blot was applied to iden⁃tifying the effect of JHG on the expression of OCLN,AQP3 and AQP4 in epithelial cells.The colonic smooth mus⁃cle cells treated respectively with JHG+Ach,and JHG+atropine were measured the length by microscopic scale,and their levels of cAMP and AC detected by ELISA.RESULTS The epithelial cells underwent 24 h JHG treat⁃ment(≤1 mg/mL)were observed with≥90%cellular viability.Compared with the normal group,JHG groups displayed up⁃regulation in the expression of OCLN,AQP3 and AQP4 in epithelial cells,promotion in the relaxation of colonic smooth muscle,and down⁃regulation in the levels of AC and cAMP in colon smooth muscle(P<0.05,P<0.01).JHG inhibited Ach⁃induced smooth muscle cell contraction and up⁃regulation of cAMP and AC levels.And the synergistic effect of JHG and atropine was found given the significant promotion of the relaxation of smooth muscle cells and the down⁃regulation of cAMP and AC(P<0.05,P<0.01).CONCLUSION JHG may improve diarrhea symptoms in D⁃IBS by regulating the expression of colonic tight junction protein,AQP3,AQP4 protein of colonic epithelial cells and the diastolic function of smooth muscle cells of rats.

关 键 词：健脾化湿颗粒 大鼠结肠上皮细胞 AQPS 大鼠结肠平滑肌细胞 CAMP AC 

分 类 号：R285􀆰[医药卫生—中药学]