摘要
目的 探究补脾益肠丸(Bupi Yichang Pills)对溃疡性结肠炎小鼠的抗炎作用及潜在作用机制.方法 采用葡聚糖硫酸钠(DSS)建立溃疡性结肠炎模型,同步给予低、中、高剂量补脾益肠丸(1.5、3.0、6.0 g·kg-1·d-1)及美沙拉嗪(300 mg·kg-1·d-1),观察小鼠一般表现并称质量,苏木素-伊红染色法观察小鼠结肠组织病理损伤情况,实时荧光定量PCR(qPCR)、酶联免疫吸附法(ELISA)法测定炎性细胞因子[肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β、IL-6、IL-10、IL-35、转化生长因子(TGF)-β1]水平,qPCR、蛋白免疫印迹(Western Blot)法检测葡萄糖转运体及糖酵解代谢激酶的mRNA及蛋白表达水平.结果 低、中、高剂量补脾益肠丸明显下调结肠炎小鼠的疾病活动指数(P＜0.05,P＜0.01);其体质量、结肠长度明显增大,而结肠质量、结肠质量指数、单位结肠质量指数均明显减小(P＜0.05,P＜0.01),且结肠组织溃疡形成及炎性细胞浸润明显改善;中、高剂量补脾益肠丸明显下调结肠炎小鼠结肠组织促炎性细胞因子TNF-α、IL-1β、IL-6 的含量及mRNA表达水平(P＜0.01),而抗炎性细胞因子 IL-10、IL-35、TGF-β1 的含量及其 mRNA 表达水平被明显上调(P＜0.01);进一步研究发现在mRNA和蛋白水平上,高剂量补脾益肠丸可明显下调结肠组织中葡萄糖转运体(Glut1、Glut2、Glut4)和糖酵解激酶(HK2、Aldolase A、PKM2)的表达(P＜0.01).结论 补脾益肠丸能有效缓解DSS诱导的小鼠溃疡性结肠炎,其作用机制与改善糖酵解代谢途径及调节炎性细胞因子表达有关.
Abstract
Objective To investigate the anti-inflammatory effects of Bupi Yichang Pills on mice with experimental colitis and its potential mechanism of action.Methods Dextran sulfate sodium(DSS)was used to model the experimental colitis,and low-,medium-and high-doses of Bupi Yichang Pills(1.5,3.0,6.0 g·kg-1·d-1)and Mesalazine(300 mg·kg-1·d-1)were fed at the same time.Mice were observed for general behavior and weighed.Hematoxylin-eosin staining was used to observe the pathological injury of colonic tissues.qPCR and ELISA were used to detect the levels of inflammatory cytokines(TNF-α,IL-1β,IL-6,IL-10,IL-35 and TGF-β1),qPCR and Western Blot were used to detect the mRNA and protein levels of glucose transporters and glycolytic kinases.Results Low-,medium-and high-doses of Bupi Yichang Pills significantly down-regulated disease activity index in colitis mice(P＜0.05,P＜0.01).The body mass and colon length were significantly increased,while colon mass,colon mass index and unit colon mass index were significantly reduced(P＜0.05,P＜0.01),and ulcer formation and inflammatory cell infiltration in colonic tissue were significantly improved.In addition,medium-and high-doses of Bupi Yichang Pills significantly down-regulated the mRNA levels and concentrations of pro-inflammatory cytokines including TNF-α,IL-1β and IL-6(P＜0.01),while significantly up-regulated the mRNA levels and concentrations of anti-inflammatory cytokines such as IL-10,IL-35 and TGF-β1(P＜0.01).We further found that high-dose of Bupi Yichang Pills significantly down-regulated the mRNA and protein expressions of glucose transporters(Glut1,Glut2,Glut4)and glycolytic kinases(HK2,Aldolase A,PKM2)in colonic tissue(P＜0.01).Conclusions Bupi Yichang Pills effectively alleviates DSS-induced experimental colitis,and its specific mechanism of action is related to the improvement of glycolytic metabolic pathways and the regulation of inflammatory cytokine expression.
基金项目
国家自然科学基金(82260863)
江西省自然科学基金(20232BAB206172)
江西省中医药局科技项目(2022A344)
江西省卫生计生委科技项目(202110121)
江西省教育厅科学技术研究项目(GJJ201239)
江西省教育厅科学技术研究项目(GJJ2200961)
江西中医药大学博士启动金项目(2019WBZR011)
NETL
NSTL
万方数据