四君子汤含药血清对IEC-6细胞增殖及多胺调控NPM及p53表达的影响  

Effects of Sijunzi decoction-containing serum on IEC-6 cell proliferation and expression of NPM and p53 regulated by polyamines

作　　者：涂小华 杨光勇 杨欣 徐萌萌 邓颖 何光志 王慧 TU Xiao-hua;YANG Guang-yong;YANG Xin;XU Meng-meng;DENG Ying;HE Guang-zhi;WANG Hui(Dept of Functional Experiment,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China;Dept of Microbiology,Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China)

机构地区：[1]贵州中医药大学机能实验室,贵州贵阳550025 [2]贵州中医药大学微生物教研室,贵州贵阳550025

出　　处：《中国药理学通报》2021年第12期1762-1769,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金地区科学基金项目(No 81760864);贵州省教育厅青年科技人才成长项目(No黔教合KY字[2017]173);贵州中医药大学博士启动基金(No 20160035)。

摘　　要：目的考察四君子汤含药血清(sijunzi decoction containing serum,SJZDS)对小肠上皮细胞增殖、多胺含量、核仁磷酸蛋白(nucleophosmin,NPM)及p53表达的影响,以探讨四君子汤修复胃肠黏膜损伤的作用机制。方法采用SD大鼠制备SJZDS,MTT法检测细胞增殖,HPLC法检测细胞内多胺含量,RT-PCR法检测NPM及p53 mRNA表达,Western blot法检测NPM及p53蛋白表达。结果与空白对照组比较,10%、20%SJZDS可促进给药24 h后细胞增殖(P<0.01),可增加精脒含量及降低p53 mRNA表达(P<0.05,P<0.01),5%、10%、20%SJZDS可促进给药48 h及72 h后细胞增殖和提高腐胺含量(P<0.01),抑制p53蛋白表达(P<0.01)和NPM mRNA及蛋白表达(P<0.05,P<0.01)。与α-二氟甲基鸟氨酸组比较,5%、10%、20%SJZDS可促进结论SJZDS可促进小肠上皮细胞增殖,其机制与影响多胺调控生长相关基因NPM及p53表达有关。Aim To observe the effects of Sijunzi decoction-containing serum(SJZDS)on cell proliferation of small intestinal epithelial cells,polyamines contents and expression of nucleophosmin(NPM)and p53,in order to explore the mechanism of Sijunzi decoction in promoting gastrointestinal mucosa injury repair.Methods SJZDS was prepared from SD rats.IEC-6 cell proliferation was detected by MTT assay,the contents of cell polyamines were determined by HPLC,mRNA expressions of NPM and p53 were tested by real-time quantitative PCR,and NPM and p53 protein levels were determined by Western blot.Results Compared with control group,10%and 20%SJZDS promoted cell proliferation after administration for 24 h(P<0.01),enhanced spermidine content and decreased p53 mRNA level(P<0.05,P<0.01);5%,10%and 20%SJZDS accelerated cell proliferation after administration for 48 h and 72 h(P<0.01),increased the content of putrescine(P<0.01),inhibited the protein expression of p53(P<0.01)and lowered NPM mRNA and protein levels(P<0.05,P<0.01).Compared withα-difluoromethylornithine group,5%,10%and 20%SJZDS inhibited cell proliferation after administration for 48 h(P<0.01),decreased mRNA and protein levels of NPM and p53(P<0.05,P<0.01),10%and 20%SJZDS could promote cell proliferation after administration for 72 h and increase contents of putrescine and spermidine(P<0.01).Conclusions SJZDS can promote small intestinal epithelial cell proliferation,and the mechanism is related to its effect on growth-related genes NPM and p53 expression regulated by polyamines.

关 键 词：四君子汤 胃肠黏膜损伤修复 细胞增殖 多胺 核仁磷酸蛋白 p53 

分 类 号：R-332[医药卫生] R289.5