白头翁皂苷B4对大鼠溃疡性结肠炎的作用机制实验研究  

Experimental Study on Pharmacological Mechanism of Anemoside B4 in Ulcerative Colitis Induced by Acetic Acid in Rats

作　　者：张勇 沈文华 查正霞 何鸾 刘艳丽[1] 许琼明[1] ZHANG Yong;SHENWen-hua;ZHA Zheng-xia;HE Luan;LIU Yan-li;XU Qiong-ming(School of Pharmaceutical Science of Soochow University,Suzhou Jiangsu 215123,China)

机构地区：[1]苏州大学药学院,江苏苏州215123

出　　处：《抗感染药学》2020年第10期1405-1410,共6页Anti-infection Pharmacy

基　　金：国家自然科学基金项目(编号:81273402);苏州市科技计划项目(编号:SYS2019032)。

摘　　要：目的:通过乙酸致大鼠溃疡性结肠炎造模的实验,探究白头翁指标性成分白头翁皂苷B4对大鼠溃疡性结肠炎的作用及其对组织病理学变化的影响。方法:取大鼠分为5组(即:正常组、模型组、白头翁皂苷B45 mg/kg组、白头翁皂苷B410 mg/kg组和美沙拉嗪组),每组10只;将大鼠麻醉后,采用直肠推注乙酸致其溃疡性结肠炎模型,在造模后第2天腹腔注射白头翁皂苷B4,连续给药7 d,每天对疾病活动指数(DAI)进行评分,待第7天处死大鼠,取血,进行生化检测;取结肠,称其质量,并进行病理学切片检查;采用Western blotting法实验考察各组大鼠结肠凋亡的相关蛋白表达及其对组织病理学变化的影响。结果:采用8%乙酸直肠推注致大鼠溃疡性结肠炎造模后,其体质量明显下降,DAI评分值明显升高,肠质量系数上升,结肠结构被破坏而出现明显的炎症浸润和细胞凋亡;不同剂量白头翁皂苷B4(5,10 mg/kg)能明显抑制乙酸引起的大鼠体质量下降,降低大鼠的DAI评分值;与模型组相比,不同剂量白头翁皂苷B4(5,10 mg/kg)能抑制大鼠的肠质量系数;并经病理切片模型组大鼠结肠结构得到明显改变,炎症浸润明显,凋亡细胞数增多;白头翁皂苷B4能明显减轻炎症、修复结肠结构,抑制了细胞凋亡和炎症浸润;经Western blotting法实验结果表明,乙酸模型组大鼠凋亡相关蛋白P53、cleaved caspase 3和Bax明显升高,抗凋亡蛋白Bcl-2明显下降;白头翁皂苷B4可抑制P53、cleaved caspase 3和Bax的升高,抑制Bcl-2的下降;并经ELISA实验结果表明,与模型组相比,不同剂量白头翁皂苷B4能抑制炎症因子IL-6、TNF-α的释放。结论:白头翁皂苷B4对实验性大鼠溃疡性结肠炎有显著地保护及修复溃疡的作用,其机制可能与通过抗炎和抑制细胞凋亡有关。Objective:To explorer the effect of anemoside B4,the index component of Pulsatilla chinensis(Bge.),on ulcerative colitis in rats and its influence on histopathological changes,modeling of ulcerative colitis in rats induced by acetic acid.Methods:All animals were divided into five groups(n=10 per group):normal group,model group,anemoside B4 group(5 mg/kg),anemoside B4 group(10 mg/kg),and mesalazine group(200 mg/kg).Experimental ulcerative colitis was induced through colitis perfusion of 8%acetic acid solution after the rats were anesthetized.After 24 hours,anemosied B4 was injected twice per day continuously for 7 days,disease activity index(DAI)and weight of rats were measured every day.On the 7 th day,the blood sample was obtained for biochemical detection,and weight of colon was measured and obtained for pathological examination.Western blotting was used to measure the level of the apoptosis proteins in colon and their effects on histopathological changes in each group of rats.Results:Compared with normal control group,acetic acid decreased the weight of rats and increased DAI scores and,index of weight-length of colon.Anemoside B4 could increase weight of rats induced by acetic acid and decreased index of weight-length of colon and DAI scores induced by acetic acid.Observing the histological feature of colon sections using H&E staining,the renal cortex of rats revealed the structures of intestinal mucosa were damaged with great inflammatory cell infiltration and increased apoptosis.While anemoside B4 significantly reduced inflammation and repaired the colon structure;inhibited cell apoptosis and inflammatory infiltration.Western blotting experiment results showed that expression of p53,cleaved caspase 3 and Bax were increased and Bcl-2 decreased.However,anemoside B4 decreased the level of p53,cleaved caspase 3 and Bax,while it increased Bcl-2 levels.ELISA experiment showed that compared with the model group,anemoside B4 could inhibit the release of inflammatory factors such as IL-6 and TNF-α.Conclusion:The ef

关 键 词：白头翁皂苷B4 溃疡性结肠炎 凋亡 炎症因子 

分 类 号：R961[医药卫生—药理学]