Regulation effect of leech on farnesol X receptor pathway in rat model of intrahepatic cholestasis
LIU Yuqing1 CAI Xin1 QIN Renwu2 WANG Yao3 LUO Lei4 YANG Fan5
1.School of First Clinical Medicine, Hubei University of Chinese Medicine, Hubei Province, Wuhan 430061, China;
2.Department of Infectious Disease, Yichang Traditional Chinese Medicine Hospital, Hubei Province, Yichang 443003, China;
3.Department of Infectious Disease, Renmin Hospital of Wuhan University, Hubei Province, Wuhan 430060, China;
4.Department of Digestive, the Second People’s Hospital of Three Gorges University, Hubei Province, Yichang 443000, China;
5.Department of Liver Disease, Hubei Provincial Hospital of Traditional Chinese Medicine, Hubei Province, Wuhan 430061, China
Abstract：Objective To study the regulatory effect of leech on farnesol X receptor (FXR) pathway in rat model of intrahepatic cholestasis. Methods Forty SPF male SD rats aged eight weeks were selected and divided into normal group, model group, ursodeoxycholic acid (UDCA) group, leech high-dose group and leech low-dose group according to random number table method, with eight rats in each group. The UDCA group was intragastrically administered with UDCA solution at a dose of 60 mg/kg, and the leech high-dose and low-dose groups were intragastrically administered with Leech Granules at a dose of 50 mg/kg and 20 mg/kg, once a day, for consecutive seven days，the normal group and the model group were given the same amount of normal saline. Except normal group, rats in other groups were given 100 mg/kg α-isothiocyanate (ANIT) on the fifth day of intragastric administration for model formation. Gastric administration for three days, once a day, the normal group was given the same amount of normal saline. Blood samples were collected from the abdominal aorta 24 h after the last administration, and serum levels of alanine aminotransferase (ALT), ascorbic aminotransferase (AST), total bilirubin (TBil) and total bile acid (TBA) were detected. Hepatic tissue was collected for microscopic examination by HE staining to observe the pathological conditions of the liver. The mRNA and protein expressions of FXR and its downstream small heterodimer chaperone receptor (SHP), uridine diphosphate glucuronide transfer 2B4 (UGT2B4) and bile salt output pump (BSEP) were detected by real-time fluorescence quantitative PCR and Western blot. Results The levels of ALT, AST and TBil in serum of leech high-dose group were lower than those in model group, with statistically significant differences (all P ＜ 0.05). There was no statistical significance in TBA level between leech high-dose group and model group (P ＞ 0.05). The levels of serum AST, TBA and TBil in leech low-dose group were lower than those in model group, with statistically significant differences (all P ＜ 0.05). There was no significant difference in ALT level between leech low-dose group and model group (P ＞ 0.05). The serum AST level of UDCA group was lower than that of model group, and the difference was statistically significant (all P ＜ 0.05). There were no statistically significant differences in ALT, TBA and TBil levels between UDCA group and model group (P ＞ 0.05). Compared with model group, liver tissue damage of rats in leech high-dose and low-dose groups was alleviated. The mRNA expressions of FXR, BSEP, UGT2B4 and SHP in leech high-dose and low-dose groups and UDCA group were higher than those in model group, with statistically significant differences (all P ＜ 0.05). The protein expressions of FXR, BSEP, UGT2B4 and SHP in leech high-dose and low-dose groups were higher than those in model group, and the differences were statistically significant (all P ＜ 0.05). There was no statistically significant difference in protein levels between UDCA group and model group (P ＞ 0.05). Conclusion Leech can relieve intrahepatic cholestasis induced by ANIT and alleviate liver tissue damage. Leech can protect liver cells by upregulating FXR gene expression, promoting the upregulation of SHP, UGT2B4 and BSEP gene expression and inhibiting cholestasis pathway, thus providing effective drugs for clinical treatment of intrahepatic cholestasis.
刘玉青1 蔡昕1 覃仁武2 王瑶3 罗磊4 杨帆5. 水蛭对肝内胆汁淤积大鼠模型法尼醇X受体通路的调节作用[J]. 中国医药导报, 2021, 18(5): 4-9.
LIU Yuqing1 CAI Xin1 QIN Renwu2 WANG Yao3 LUO Lei4 YANG Fan5. Regulation effect of leech on farnesol X receptor pathway in rat model of intrahepatic cholestasis. 中国医药导报, 2021, 18(5): 4-9.
[1] 中华医学会肝病学分会.胆汁淤积性肝病诊断和治疗共识（2015）[J].中华消化杂志，2015，35（12）：39.
[2] Mato JM，Lu SC. Luole of S-Adenosyl-L-Methionine in Liver Health and Injury [J]. Hepatology，2007，45（5）：1306-1312.
[3] Festi D，Montagnani M，Azzaroli F，et al. Clinical Efficacy and Effectiveness of Ursodeoxycholic Acid in Cholestatic Liver Diseases [J]. Curr Clin Pharmacol，2007，2（2），155-177.
[4] 刘为民，姚乃礼.络病理论与肝纤维化关系探讨[J].中医杂志，2003，44（2）：85-87.
[5] 司远，车念聪，杜宇琼，等.浅析“肝络病”概念与特点[J].环球中医药，2006（2）：214-216.
[6] Hirschfield GM，Mason A，Luketic V，et al. Efficacy of obeticholic acid in patients with primary biliary cirrhosis and inadequate response to ursodeoxycholic acid [J]. Gastroenterology，2015，148（4）：751-761.
[7] Beuers U，Trauner M，Jansen P，et al. New paradigms in the treatment of hepatic cholestasis：from UDCA to FXR，PXR and beyond [J]. J Hepatol，2015，62（1 Suppl）：S25-S37.
[8] Moscovitz JE, Kong B, Buckley K，et al. Restoration of Enterohepatic Bile Acid Pathways in Pregnant Mice Following Short Term Activation of Fxr by GW4064 [J]. Toxicol Appl Pharmacol，2016，310：60-67.
[9] 吴以岭.络病治疗原则与通络药物[J].疑难病杂志，2005， 4（4）：213-215.
[10] 刘宝山.水蛭治疗瘀血发黄的临床体会[J].北京中医药，2008，27（2）：141-142.
[11] 王凤岐.中华名医特技集成[M].北京：中国医药科技出版社，1993.
[12] 刘维伊，孙建国，曹伟灵，等.法尼醇X受体拮抗剂及药理活性研究进展[J].中国临床药理学与治疗学，2020， 25（7）：764-774.
[13] 刘笑，王琰.胆汁酸的合成调控及其在生理与病理中的功能机制[J].遗传，2019，41（5）：365-374.
[14] 丁艳，熊小丽，尹薇，等.小异源二聚体伴侣受体在淤胆型肝炎模型中的表达及大黄素的干预作用[J].中药新药与临床药理，2016，27（4）：474-479.
[15] Kim KH，Choi S，Zhou Y，et al. Hepatic FXR/SHP Axis Modulates Systemic Glucose and Fatty Acid Homeostasis in Aged Mice [J]. Hepatology，2017，66（2）：498-509.
[16] 张峻崎，祁飞，徐加敏，等.核受体FXR调控SHP的机制及FXR-SHP轴在肝脏中作用的研究进展[J].现代生物医学进展，2018，18（3）：592-596.
[17] 陈浩，赵威，俞浩，等.京尼平苷酸对ANIT诱导胆汁淤积大鼠的酮洛芬肝代谢和胆汁排泄的影响及机制探究[J].中药新药与临床药理，2019，30（2）：147-155.
[18] 李尚恒，杜宏波，孟培培，等.尿苷二磷酸葡萄糖醛酸转移酶1A1与高胆红素血症的关系研究[J].中国临床医生杂志，2017，45（3）：33-35.
[19] 申弘，胡萌，魏泽辉，等.胆汁的生成、分泌、排泄及胆汁淤积发生机制[J].临床肝胆病杂志，2019，35（2）：431-437.
[20] 高飞，谈欣乐，闵秋霞，等.胆汁酸盐输出泵BSEP/ABCB11研究概述[J].华中科技大学学报：医学版，2017，46（1）：104-109.
[21] Charach G，Argov O，Geiger K，et al. Diminished bile acids excretion is a risk factor for coronary artery disease： 20-year follow up and long-term outcome [J]. Therap Adv Gastroenterol，2018，11：1756283X17743420.
[22] Telbisz ?魣， Homolya L. Recent advances in the exploration of the bile salt export pump（BSEP /ABCB11）function [J]. Expert Opin Ther Targets，2016，20（4）：501-514.
[23] Wang L，Wu G，Wu F，et al. Geniposide attenuates ANIT-induced cholestasis through regulation of transporters and enzymes involved in bile acids homeostasis in rats [J]. J Ethnopharmacol，2017，196：178-185.
[24] Ding L，Zhang B，Li J，et al. Beneficial effect of resveratrol on α-naphthylisothiocyanate induced cholestasis viar-egulation of the FXR pathway [J]. Mol Med Rep，2018， 17（1）：1863-1872.
[25] Meng Q，Chen XL，Wang CY，et al. Alisol B 23-acetate protects against ANIT-induced hepatotoxity and cholestasis，due to FXR-mediated regulation of transporters and enzymes involved in bile acids homeostasis [J]. Toxicol Appl Pharmacol，2015，283（3）：178-186.
2021, Vol. 18 
京公网安备 11010502046598号