祛脂方调控ITGB6/IL23A/STAT3信号通路改善MASH相关肝纤维化

摘要/Abstract

摘要： 目的探讨祛脂方(QZF)是否具有改善代谢功能障碍相关脂肪性肝炎(MASH)相关纤维化的作用以及潜在机制。方法人肝星状细胞(LX-2)在不含胎牛血清的DMEM培养基饥饿培养24小时后,用10 ng/mL浓度的转化生长因子-β1(TGF-β1)处理24小时。随后,细胞用含200 μg/mL祛脂方的无血清DMEM培养基处理。采用Cell Counting Kit-8(CCK-8)法检测祛脂方对细胞的毒性,通过蛋白免疫印迹、实时荧光定量PCR反应和免疫荧光技术检测祛脂方是否逆转TGFβ-1诱导的LX-2细胞活化。体内实验中,选用6周龄C57BL/6 SPF小鼠,随机分为三组:对照组、CDAHFD组和祛脂方组(QZF组)。CDAHFD组和祛脂方组饲喂含62%千卡脂肪和0.1%甲硫氨酸且胆碱缺乏的饲料3周,对照组和CDAHFD组小鼠灌胃生理盐水,祛脂方组小鼠灌胃祛脂方(10 g/kg/d)。9周后小鼠安乐死,收集血浆和肝组织,进一步分析祛脂方的疗效。结果动物实验表明祛脂方可减轻MASH小鼠肝脏脂质积累和纤维化。细胞实验结果显示,祛脂方通过调节ITGB6/IL23A/STAT3信号通路,逆转了TGFβ-1诱导的LX-2细胞活化。此外,STAT3抑制剂也可能逆转TGFβ-1诱导的LX-2活化,进一步验证了祛脂方通过该信号通路改善MASH相关纤维化的作用。结论祛脂方不仅能改善MASH脂肪变性,还能减轻MASH相关纤维化。这表明,祛脂方有潜力作为一种新的治疗MASH的药物,其作用机制可能与调控ITGB6/IL23A/STAT3信号通路有关。

Abstract: Objective The purpose of this study was to explore whether Quzhi formula has the effect of improving metabolic dysfunction associated steatohepatitis (MASH) related fibrosis and the internal mechanism of its efficacy. Methods LX2 cells were starved for 24 hours with fetal bovine serum-free DMEM medium overnight and then treated with TGF-β1 at 10 ng/mL (Abcam, ab50036). Finally, the cells were treated with serum-free DMEM medium containing 200μg/mL Quzhi formula. The cytotoxicity of Quzhi formula was detected by cell counting kit-8 assay, then western blotting, quantitative reverse transcription-polymerase chain reaction and immunofluorescence were performed to measure whether Quzhi formula reversed the activation of LX-2 induced by TGF-β1. For in vivo experiment, C57BL/6 SPF mice aged 6 weeks were randomly divided into 3 groups: control group, MASH group and Quzhi formula group. The MASH group and Quzhi formula group were fed diets containing 62% kcal fat and 0.1% methionine and choline deficiency for 3 weeks. Mice in the control group and MASH group were administrated with normal saline, and mice in the Quzhi formula group were administrated with Quzhi formula (10g/kg/d). After 9 weeks, the mice were euthanized, plasma and liver tissues were collected, and the effect of Quzhi formula was analyzed. Results The animal experiment proved that the lipid accumulation and liver fibrosis in MASH mice could be alleviated by Quzhi formula. Cell experiments demonstrated that Quzhi formula reversed TGF-β1 induced LX-2 activation by regulating ITGB6/IL23A/STAT3 signaling pathway. In addition, STAT3 inhibitors also reversed LX-2 activation induced by TGF-β1, which further demonstrated that Quzhi formula improves MASH related fibrosis through the above pathway. Conclusion Quzhi formula can not only improve MASH steatosis, but also alleviate MASH related fibrosis. It is suggested that Quzhi formula can be used as a new drug to treat MASH.

Key words: Quzhi formula, MASLD, MASH, Liver fibrosis

陈慧洁, 彭舟, 张琴. 祛脂方调控ITGB6/IL23A/STAT3信号通路改善MASH相关肝纤维化[J]. 肝脏, 2025, 30(5): 675-682.

CHEN Hui-jie, PENG Zhou, ZHANG Qin. Quzhi formula improves liver fibrosis related to metabolic dysfunction associated steatohepatitis by regulating the ITGB6/IL23A/STAT3 pathway[J]. Chinese Hepatolgy, 2025, 30(5): 675-682.

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