Shu Xiangbing, Cao Ying, Wu Yan, Chen Milian, Zhao Wenxia, Ji Guang, Zhang Li
Institute of Digestive Diseases, Shanghai University of Traditional Chinese Medicine, Shanghai, 200032, China; Department of Geratology, Baoshan Branch of Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Institute of Digestive Diseases, Shanghai University of Traditional Chinese Medicine, Shanghai, 200032, China.
J Ethnopharmacol. 2025 May 12;347:119719. doi: 10.1016/j.jep.2025.119719. Epub 2025 Apr 3.
Metabolic dysfunction-associated steatohepatitis (MASH) is the progressive form of metabolic dysfunction-associated steatotic liver disease (MASLD), and is currently the most prevalent chronic liver disease worldwide. Gegen-Qinlian decoction (GQD), a classical Traditional Chinese Medicine (TCM) formula from Treatise on Febrile Diseases, has been historically used to treat heat-dampness syndromes. Recent studies revealed that GQD is effective in treating MASH, but the underlying mechanisms remain unknown.
This study aims to evaluate the therapeutic effect of GQD on MASH and explore the potential mechanisms targeting the gut microbiota-bile acid (BA) axis.
Phytochemical profiling of GQD was performed using UPLC-Q-TOF-MS. MASH was induced in mice via a fructose-, palmitate-, and cholesterol-enriched (FPC) diet, followed by treatment with low-, medium-, or high-dose GQD. H&E and oil red O staining were utilized to examine the histological change, and serum lipids and enzymes were biochemically analyzed. 16SrDNA sequencing was applied to analyze the alteration of the gut microbiota, and the gas chromatography-mass spectrometry technique was introduced to investigate the fecal bile acid (BA) profile. Serum lipopolysaccharide (LPS) concentrations were analyzed by enzyme-linked immunosorbent assay. Intestinal tight junction proteins (ZO1, Occludin) and BA receptors (FXR, TGR5, and VDR) were detected by Western blot and immunofluorescence staining.
The quality of GQD was confirmed, and GQD treatment improved hepatic steatosis, reduced the content of liver triglyceride (20-40 % reduction, p < 0.01) and cholesterol (20-25 % reduction, p < 0.01) in FPC-induced MASH mice. High-dose GQD further decreased serum TC (3.97 ± 1.00 vs 5.51 ± 1.11, p < 0.05), LDL-c (0.53 ± 0.18 vs 1.07 ± 0.28, p < 0.01), ALT (31.90 ± 6.20 vs 47.90 ± 12.78, p < 0.05) and ALP (90.83 ± 13.46 vs 132.90 ± 23.67, p < 0.05) levels, suggesting the effects of GQD in counteracting metabolic inflammation. GQD treatment restored gut microbiota diversity and reversed gut dysbiosis by decreasing the abundance of pathogenic bacteria, resulting in reduced serum LPS while enhancing intestinal tight junction proteins (ZO1, Occludin). Concurrently, GQD treatment reshaped fecal BA profiles, increased intestinal TGR5/VDR expression, with BA shifts strongly correlating to microbiota changes.
GQD alleviated hepatic and metabolic disorders in MASH mice, possibly through reversing gut dysbiosis and modulating BA profile. Targeting the microbiota-BA axis represents a promising pattern for TCM prescriptions in treating MASH.
代谢功能障碍相关脂肪性肝炎（MASH）是代谢功能障碍相关脂肪性肝病（MASLD）的进展形式，目前是全球最普遍的慢性肝病。葛根芩连汤（GQD）是《伤寒论》中的经典中药方剂，历来用于治疗湿热证。最近的研究表明，GQD对治疗MASH有效，但其潜在机制尚不清楚。
本研究旨在评估GQD对MASH的治疗效果，并探索靶向肠道微生物群-胆汁酸（BA）轴的潜在机制。
采用超高效液相色谱-四极杆飞行时间质谱联用技术对GQD进行植物化学分析。通过富含果糖、棕榈酸和胆固醇（FPC）的饮食诱导小鼠发生MASH，随后用低、中、高剂量的GQD进行治疗。利用苏木精-伊红（H&E）染色和油红O染色检查组织学变化，并对血脂和酶进行生化分析。应用16S核糖体DNA（16SrDNA）测序分析肠道微生物群的变化，并采用气相色谱-质谱联用技术研究粪便胆汁酸（BA）谱。通过酶联免疫吸附测定法分析血清脂多糖（LPS）浓度。采用蛋白质免疫印迹法和免疫荧光染色法检测肠道紧密连接蛋白（ZO1、闭合蛋白）和BA受体（法尼酯X受体（FXR）、G蛋白偶联胆汁酸受体5（TGR5）和维生素D受体（VDR））。
确认了GQD的质量，GQD治疗改善了FPC诱导的MASH小鼠的肝脂肪变性，降低了肝脏甘油三酯含量（降低20%-40%，p<0.01）和胆固醇含量（降低20%-25%，p<0.01）。高剂量GQD进一步降低了血清总胆固醇（TC）（3.97±1.00对5.51±1.11，p<0.05）、低密度脂蛋白胆固醇（LDL-c）（0.53±0.18对1.07±0.28，p<0.01）、谷丙转氨酶（ALT）（31.90±6.20对47.90±12.78，p<0.05）和碱性磷酸酶（ALP）（90.83±13.46对132.90±23.67，p<0.05）水平，表明GQD具有对抗代谢炎症的作用。GQD治疗恢复了肠道微生物群的多样性，通过降低病原菌的丰度逆转了肠道生态失调，从而降低了血清LPS水平，同时增强了肠道紧密连接蛋白（ZO1、闭合蛋白）。同时，GQD治疗重塑了粪便BA谱，增加了肠道TGR5/VDR表达，BA的变化与微生物群的变化密切相关。
GQD可能通过逆转肠道生态失调和调节BA谱减轻MASH小鼠的肝脏和代谢紊乱。靶向微生物群-BA轴是中药方剂治疗MASH的一种有前景的模式。
