Wang Yanyan, Zhao Derrick, Su Lianyong, Tai Yun-Ling, Way Grayson W, Zeng Jing, Yan Qianhua, Xu Ying, Wang Xuan, Gurley Emily C, Zhou Xi-Qiao, Liu Jinze, Liu Jinpeng, Chen Weidong, Hylemon Phillip B, Zhou Huiping
Department of Microbiology and Immunology, Virginia Commonwealth University and Richmond Veterans Affairs Medical Center, 1220 East Broad Street, MMRB-5044, Richmond, VA, 23298-0678, USA.
School of Pharmaceutical Science, Anhui University of Chinese Medicine, Hefei, Anhui, China.
Cell Biosci. 2024 Jan 25;14(1):14. doi: 10.1186/s13578-024-01195-8.
Primary sclerosing cholangitis (PSC) is a chronic liver disease characterized by progressive biliary inflammation and bile duct injury. Berberine (BBR) is a bioactive isoquinoline alkaloid found in various herbs and has multiple beneficial effects on metabolic and inflammatory diseases, including liver diseases. This study aimed to examine the therapeutic effect of BBR on cholestatic liver injury in a PSC mouse model (Mdr2 mice) and elucidate the underlying mechanisms.
Mdr2mice (12-14 weeks old, both sexes) received either BBR (50 mg/kg) or control solution daily for eight weeks via oral gavage. Histological and serum biochemical analyses were used to assess fibrotic liver injury severity. Total RNAseq and pathway analyses were used to identify the potential signaling pathways modulated by BBR in the liver. The expression levels of key genes involved in regulating hepatic fibrosis, bile duct proliferation, inflammation, and bile acid metabolism were validated by qRT-PCR or Western blot analysis. The bile acid composition and levels in the serum, liver, small intestine, and feces and tissue distribution of BBR were measured by LC-MS/MS. Intestinal inflammation and injury were assessed by gene expression profiling and histological analysis. The impact on the gut microbiome was assessed using 16S rRNA gene sequencing.
BBR treatment significantly ameliorated cholestatic liver injury, evidenced by decreased serum levels of AST, ALT, and ALP, and reduced bile duct proliferation and hepatic fibrosis, as shown by H&E, Picro-Sirius Red, and CK19 IHC staining. RNAseq and qRT-PCR analyses indicated a substantial inhibition of fibrotic and inflammatory gene expression. BBR also mitigated ER stress by downregulating Chop, Atf4 and Xbp-1 expression. In addition, BBR modulated bile acid metabolism by altering key gene expressions in the liver and small intestine, resulting in restored bile acid homeostasis characterized by reduced total bile acids in serum, liver, and small intestine and increased fecal excretion. Furthermore, BBR significantly improved intestinal barrier function and reduced bacterial translocation by modulating the gut microbiota.
BBR effectively attenuates cholestatic liver injury, suggesting its potential as a therapeutic agent for PSC and other cholestatic liver diseases.
原发性硬化性胆管炎（PSC）是一种慢性肝病，其特征为进行性胆管炎症和胆管损伤。黄连素（BBR）是一种存在于多种草药中的生物活性异喹啉生物碱，对包括肝脏疾病在内的代谢性和炎症性疾病具有多种有益作用。本研究旨在探讨BBR对PSC小鼠模型（Mdr2小鼠）胆汁淤积性肝损伤的治疗效果，并阐明其潜在机制。
12至14周龄的Mdr2小鼠（雌雄均有）通过口服灌胃，每天接受BBR（50mg/kg）或对照溶液，持续8周。采用组织学和血清生化分析评估肝纤维化损伤的严重程度。通过全RNA测序和通路分析来鉴定BBR在肝脏中调节的潜在信号通路。通过qRT-PCR或蛋白质免疫印迹分析验证参与调节肝纤维化、胆管增殖、炎症和胆汁酸代谢的关键基因的表达水平。采用液相色谱-串联质谱法（LC-MS/MS）测定血清、肝脏、小肠和粪便中的胆汁酸组成和水平以及BBR的组织分布。通过基因表达谱分析和组织学分析评估肠道炎症和损伤。使用16S rRNA基因测序评估对肠道微生物群的影响。
BBR治疗显著改善了胆汁淤积性肝损伤，血清天冬氨酸转氨酶（AST）、丙氨酸转氨酶（ALT）和碱性磷酸酶（ALP）水平降低，且苏木精-伊红（H&E）染色、苦味酸天狼星红染色和细胞角蛋白19（CK19）免疫组化染色显示胆管增殖和肝纤维化减轻，证明了这一点。RNA测序和qRT-PCR分析表明纤维化和炎症基因表达受到显著抑制。BBR还通过下调 Chop、Atf4和Xbp-1的表达减轻了内质网应激。此外，BBR通过改变肝脏和小肠中的关键基因表达来调节胆汁酸代谢，从而恢复胆汁酸稳态，其特征为血清、肝脏和小肠中的总胆汁酸减少以及粪便排泄增加。此外，BBR通过调节肠道微生物群显著改善了肠道屏障功能并减少了细菌易位。
BBR有效减轻胆汁淤积性肝损伤，表明其作为PSC和其他胆汁淤积性肝病治疗药物的潜力。
