Danhongqing formula alleviates cholestatic liver fibrosis by downregulating long non-coding RNA H19 derived from cholangiocytes and inhibiting hepatic stellate cell activation

IF 4.2 2区 医学 Q1 INTEGRATIVE & COMPLEMENTARY MEDICINE
Meng Li , Yang Zhou , Hui Zhu , Lie-ming Xu , Jian Ping
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引用次数: 0

Abstract

Objective

This study explores the mechanism of action of Danhongqing formula (DHQ), a compound-based Chinese medicine formula, in the treatment of cholestatic liver fibrosis.

Methods

In vivo experiments were conducted using 8-week-old multidrug resistance protein 2 knockout (Mdr2-/-) mice as an animal model of cholestatic liver fibrosis. DHQ was administered orally for 8 weeks, and its impact on cholestatic liver fibrosis was evaluated by assessing liver function, liver histopathology, and the expression of liver fibrosis-related proteins. Real-time polymerase chain reaction, Western blot, immunohistochemistry and other methods were used to observe the effects of DHQ on long non-coding RNA H19 (H19) and signal transducer and activator of transcription 3 (STAT3) phosphorylation in the liver tissue of Mdr2-/- mice. In addition, cholangiocytes and hepatic stellate cells (HSCs) were cultured in vitro to measure the effects of bile acids on cholangiocyte injury and H19 expression. Cholangiocytes overexpressing H19 were constructed, and a conditioned medium containing H19 was collected to measure its effects on STAT3 protein expression and cell activation. The intervention effect of DHQ on these processes was also investigated. HSCs overexpressing H19 were constructed to measure the impact of H19 on cell activation and assess the intervention effect of DHQ.

Results

DHQ alleviated liver injury, ductular reaction, and fibrosis in Mdr2-/- mice, and inhibited H19 expression, STAT3 expression and STAT3 phosphorylation. This formula also reduced hydrophobic bile acid-induced cholangiocyte injury and the upregulation of H19, inhibited the activation of HSCs induced by cholangiocyte-derived conditioned medium, and decreased the expression of activation markers in HSCs. The overexpression of H19 in a human HSC line confirmed that H19 promoted STAT3 phosphorylation and HSC activation, and DHQ was able to successfully inhibit these effects.

Conclusion

DHQ effectively alleviated spontaneous cholestatic liver fibrosis in Mdr2-/- mice by inhibiting H19 upregulation in cholangiocytes and preventing the inhibition of STAT3 phosphorylation in HSC, thereby suppressing cell activation.

Please cite this article as: Li M, Zhou Y, Zhu H, Xu LM, Ping J. Danhongqing formula alleviates cholestatic liver fibrosis by downregulating long non-coding RNA H19 derived from cholangiocytes and inhibiting hepatic stellate cell activation. J Integr Med. 2024; 22(2): 188–198.

丹红清方通过下调来自胆管细胞的长非编码 RNA H19 和抑制肝星状细胞活化,缓解胆汁淤积性肝纤维化。
目的:探讨复方中药丹红清方治疗胆汁淤积性肝纤维化的作用机制:本研究探讨复方中药丹红清方(DHQ)治疗胆汁淤积性肝纤维化的作用机制:以8周龄多耐药蛋白2基因敲除(Mdr2-/-)小鼠为胆汁淤积性肝纤维化动物模型进行体内实验。口服DHQ 8周,通过评估肝功能、肝组织病理学和肝纤维化相关蛋白的表达来评价DHQ对胆汁淤积性肝纤维化的影响。采用实时聚合酶链反应、Western印迹、免疫组化等方法观察了DHQ对Mdr2-/-小鼠肝组织中长非编码RNA H19(H19)和信号转导和激活剂3(STAT3)磷酸化的影响。此外,还对胆管细胞和肝星状细胞(HSCs)进行了体外培养,以测定胆汁酸对胆管细胞损伤和 H19 表达的影响。构建了过表达 H19 的胆管细胞,并收集了含有 H19 的条件培养基,以测量其对 STAT3 蛋白表达和细胞活化的影响。同时还研究了 DHQ 对这些过程的干预作用。构建了过表达H19的造血干细胞,以测量H19对细胞活化的影响并评估DHQ的干预效果:结果:DHQ减轻了Mdr2-/-小鼠的肝损伤、导管反应和纤维化,抑制了H19表达、STAT3表达和STAT3磷酸化。该配方还能减轻疏水胆酸诱导的胆管细胞损伤和H19的上调,抑制胆管细胞源性条件培养基诱导的造血干细胞活化,降低造血干细胞活化标志物的表达。在人造血干细胞系中过表达H19证实,H19能促进STAT3磷酸化和造血干细胞活化,而DHQ能成功抑制这些效应:结论:DHQ通过抑制胆管细胞中H19的上调和防止抑制造血干细胞中STAT3磷酸化,从而抑制细胞活化,有效缓解了Mdr2-/-小鼠自发性胆汁淤积性肝纤维化。本文引用如前Li M, Zhou Y, Zhu H, Xu LM, Ping J. Danhongqing formula alleviates cholestatic liver fibrosis by downregulating long non-coding RNA H19 derived from cholangiocytes and inhibiting hepatic stellate cell activation.J Integr Med.2024; Epub ahead of print.
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来源期刊
Journal of Integrative Medicine-Jim
Journal of Integrative Medicine-Jim Medicine-Complementary and Alternative Medicine
CiteScore
9.20
自引率
4.20%
发文量
3319
期刊介绍: The predecessor of JIM is the Journal of Chinese Integrative Medicine (Zhong Xi Yi Jie He Xue Bao). With this new, English-language publication, we are committed to make JIM an international platform for publishing high-quality papers on complementary and alternative medicine (CAM) and an open forum in which the different professions and international scholarly communities can exchange views, share research and their clinical experience, discuss CAM education, and confer about issues and problems in our various disciplines and in CAM as a whole in order to promote integrative medicine. JIM is indexed/abstracted in: MEDLINE/PubMed, ScienceDirect, Emerging Sources Citation Index (ESCI), Scopus, Embase, Chemical Abstracts (CA), CAB Abstracts, EBSCO, WPRIM, JST China, Chinese Science Citation Database (CSCD), and China National Knowledge Infrastructure (CNKI). JIM Editorial Office uses ThomsonReuters ScholarOne Manuscripts as submitting and review system (submission link: http://mc03.manuscriptcentral.com/jcim-en). JIM is published bimonthly. Manuscripts submitted to JIM should be written in English. Article types include but are not limited to randomized controlled and pragmatic trials, translational and patient-centered effectiveness outcome studies, case series and reports, clinical trial protocols, preclinical and basic science studies, systematic reviews and meta-analyses, papers on methodology and CAM history or education, conference proceedings, editorials, commentaries, short communications, book reviews, and letters to the editor. Our purpose is to publish a prestigious international journal for studies in integrative medicine. To achieve this aim, we seek to publish high-quality papers on any aspects of integrative medicine, such as acupuncture and traditional Chinese medicine, Ayurveda medicine, herbal medicine, homeopathy, nutrition, chiropractic, mind-body medicine, taichi, qigong, meditation, and any other modalities of CAM; our commitment to international scope ensures that research and progress from all regions of the world are widely covered. These ensure that articles published in JIM have the maximum exposure to the international scholarly community. JIM can help its authors let their papers reach the widest possible range of readers, and let all those who share an interest in their research field be concerned with their study.
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