栀子柏皮汤对胆酸饲料诱导胆汁淤积症小鼠肝保护机制研究  

Study on liver protection mechanism of Zhizi Bopi Decoction on cholestasis mice induced by cholic acid feeding

作　　者：李玥 宋国超 徐晓青 邹斌 赵婧 翁凤仪 成思 金静怡 严东明 谭波 裘福荣 LI Yue;SONG Guochao;XU Xiaoqing;ZOU Bin;ZHAO Jing;WENG Fengyi;CHENG Si;JIN Jingyi;YAN Dongming;TAN Bo;QIU Furong(Laboratory of Clinical Pharmacokinetics,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Clinical Laboratory,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)

机构地区：[1]上海中医药大学附属曙光医院临床药代动力学实验室,上海201203 [2]上海中医药大学附属曙光医院检验科,上海201203

出　　处：《上海中医药大学学报》2022年第S01期154-160,共7页Academic Journal of Shanghai University of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81874346);上海市中医临床重点实验室项目(20DZ2272200);上海中医药大学预算内项目(2021LK080)

摘　　要：目的:研究栀子柏皮汤对含1%胆酸(CA)饲料诱导胆汁淤积症小鼠的肝保护作用及其机制。方法:C57BL/6雄性小鼠随机分为3组,即正常对照组、1%CA模型组、栀子柏皮汤组,每组5只。栀子柏皮汤组灌胃给予栀子柏皮汤(6 g/kg,每10 g小鼠体质量0.1 ml),每日1次,连续10 d。正常对照组和1%CA模型组分别灌胃等体积0.9%NaCl溶液。所有组前3 d喂食正常饲料,于第4天灌胃后将1%CA模型组及栀子柏皮汤组换为含1%CA饲料饲养,直至实验结束,正常对照组仍继续用普通饲料饲养。干预结束后,检测小鼠血浆碱性磷酸酶(ALP)和丙氨酸转氨酶(ALT)活性;HE染色观察小鼠肝脏病理改变;LC-MS/MS检测小鼠肝脏胆汁酸含量;Western blot法测定小鼠肝脏Nod样受体家族含pyrin结构域蛋白3(NLRP3)炎性小体、衔接子(ASC)和消皮素D的N端剪切体(GSDMD-N)蛋白表达。结果:与1%CA模型组比较,栀子柏皮汤组显著降低血浆ALT和ALP活性,缓解肝脏坏死及汇管区炎性细胞浸润,显著减少肝内总胆汁酸、牛磺胆酸(TCA)和牛磺脱氧胆酸(TDCA)的含量,并能下调NLRP3、ASC和GSDMD-N的蛋白表达,差异均有统计学意义(P<0.05)。结论:栀子柏皮汤对1%CA饲料诱导肝内胆汁淤积症小鼠具有肝保护作用,其机制可能与缓解肝内胆汁酸蓄积以及抑制NLRP3炎性小体活化、减少细胞焦亡有关。Objective:To study the hepatoprotective effect of Zhizi Bopi Decoction on cholestasis mice induced by 1%cholic acid(CA)feeding,and explore the mechanism.Methods:C57BL/6 male mice were randomly divided into 3 groups(n=5):normal control group,1%CA model group,Zhizi Bopi Decoction group.Zhizi Bopi decoction group was given Zhizi Bopi Decoction by gavage(6 g/kg,body weight 0.1 ml per 10 g),once a day,for consecutive 10 days.Normal control group and 1%CA model group were given 0.9%NaCl solution by gavage.All groups were fed normal feed for the first 3 days,and 1%CA model group and Zhizi Bopi Decoction group were changed to 1%CA diet on the 4th day after intragastric administration until the end of the experiment.The normal control group continued to feed with ordinary feed.After intervention,plasma alkaline phosphatase(ALP)and alanine aminotransferase(ALT)activities were detected,and liver histopathology was performed by H&E staining.Hepatic bile acid contents were measured by LC-MS/MS,and expression of NOD-like receptors family pyrin domain containing 3(NLRP3)inflammasome,adaptor(ASC)and N-terminal clipping body of gasdermin D(GSDMD-N)in mouse liver was quantified by Western blot.Results:Compared with 1%CA model group,the activities of ALT and ALP in plasma in Zhizi Bopi Decoction group were significantly decreased,liver necrosis and inflammatory cell infiltration were alleviated,the contents of hepatic total bile acid,taurocholic acid(TCA)and taurodeoxycholic acid(TDCA)in liver were significantly decreased,and the protein expressions of NLRP3,ASC and GSDMD-N were down-regulated.The differences were statistically significant(P<0.05).Conclusion:Zhizi Bopi Decoction can protect the liver of mice against 1%CA feeding-induced cholestatic liver injury,which may be related to alleviating hepatic bile acid accumulation,inhibiting the activation of NLRP3 inflammasome and reducing pyroptosis.

关 键 词：栀子柏皮汤 胆汁淤积症 胆汁酸 NLRP3炎性小体 细胞焦亡 

分 类 号：R285.5[医药卫生—中药学]