枳实含药血清对大鼠胃窦平滑肌细胞收缩效应及细胞内钙离子浓度、钙调蛋白表达的影响  

Effect of drug-containing serum of immature bitter orange on cell contraction, intracellular Ca^(2+) concentration and calmodulin expression in rat antral smooth muscle cells

作　　者：李东鑫 凌江红[1] 王煜姣[1] Akarayosapong Pichamon 宁海恩 张智[1] 刘培凤[1] 

机构地区：[1]广西医科大学第一附属医院中医科,广西壮族自治区南宁市530021

出　　处：《世界华人消化杂志》2015年第8期1224-1230,共7页World Chinese Journal of Digestology

基　　金：广西自然科学基金资助项目;No.2013GXNSFAA019139;广西中医药民族医药继承创新工程攻关课题基金资助项目;No.GZGG13-04;广西高校科学技术研究重点项目基金资助项目;No2013ZD013~~

摘　　要：目的:观察单味中药枳实含药血清对大鼠胃窦平滑肌细胞收缩效应及细胞内Ca2+、钙调蛋白表达的影响.方法:按照血清药理学通法制备大鼠枳实含药血清,将血清分为枳实组(10%、20%、50%枳实含药血清)、空白血清对照组,酶解法提取胃窦平滑肌细胞并进行培养,倒置显微镜下观察各组对胃窦平滑肌细胞收缩效应,激光共聚焦显微镜技术检测各组加药前后胃窦平滑肌细胞内Ca2+浓度变化,免疫组织化学方法观察各组对胃窦平滑肌细胞内钙调蛋白的表达.结果:10%、20%、50%枳实含药血清对胃窦平滑肌细胞收缩及细胞内Ca2+荧光强度的影响呈药物剂量依赖关系,细胞收缩、细胞内Ca2+荧光强度随药物浓度的增高而增强(P<0.05),20%、50%枳实含药血清与空白血清对照组对胃窦平滑肌细胞收缩、细胞内Ca2+荧光强度差异有统计学意义(P<0.05),50%枳实组与空白血清对照组对胃窦平滑肌细胞内钙调蛋白表达差异有统计学意义(P<0.05).结论:枳实含药血清可以促进胃窦平滑肌细胞收缩,其机制可能与增加细胞内Ca2+浓度、调节钙调蛋白的表达有关.AIM：To observe the effect of drug-containing serum of single herb immature bitter orange on cell contraction,intracellular Ca^2＋concentration and calmodulin expression in rat antral smooth muscle cells.METHODS：Rat drug-containing serum of immature bitter orange was prepared using the routine method.The drug-containing serum was divided into different concentration groups（10%,20%,and 50%drug-containing serum）,and normal serum was used as a blank control.Antral smooth muscle cells were extracted using enzymatic method and cultured.The effect of each group of serum on antral smooth muscle cell contraction was observed under an inverted microscope.The changes of intracellular Ca^2＋concentration in the antral smooth muscle cells were tested using confocal laser scanning microscopy.Calmodulin expression in antral smooth muscle cells was detected using immunohistochemical method.RESULTS：The effect of drug-containing serum of immature bitter orange on antral smooth muscle cell contraction and intracellular Ca^2＋fluorescence intensity was dose-dependent.Cell contraction and intracellular Ca^2＋ fluorescence intensity increased with the increase of drug concentration（P〈0.05）.Antral smooth muscle cell contraction and intracellular Ca^2＋fluorescence intensity differed significantly between the 20%and 50%drug-containing serum groups and the blank control group（P〈0.05）.Antral smooth muscle intracellular calmodulin expression differed significantly between the 50%drug-containing serum group and the blank control group（P〈0.05）.CONCLUSION：The drug-containing serum of immature bitter orange can promote antral smooth muscle cell contraction possibly via mechanism related to the increase of intracellular Ca^2＋ concentration and the modulation of calmodulin expression.

关 键 词：枳实 含药血清 平滑肌细胞 钙离子 钙调蛋白 

分 类 号：R285.5[医药卫生—中药学]