壮肝逐瘀煎对肝纤维化大鼠miR-137、FasL及ACSL4表达的影响

Effect of Zhuangan Zhuyu Decoction on the Expression of miR-137, FasL, and ACSL4 in Hepatic Fibrosis Rats

摘要: 目的：观察壮肝逐瘀煎对肝纤维化大鼠miR-137、FasL及ACSL4表达的影响。方法：设立正常组，采用四氯化碳(CCl₄)注射法构建肝纤维化大鼠模型，造模成功后分成两组：模型组、中药组，每组10只大鼠，中药组给予壮肝逐瘀煎方灌胃，正常组和模型组给予等量的生理盐水灌胃，采用qPCR检测三组大鼠的miR-137的表达，miRNA测序分析miR-137是否是模型组和中药组差异miRNA，Western Blot检测三组大鼠的FasL、ACSL4的表达，HE染色观察三组大鼠肝脏组织的病理，明确壮肝逐瘀煎对肝纤维化大鼠miR-137、FasL及ACSL4表达的影响。结果：模型组的miR-137低于正常组(P < 0.05)，中药组的miR-137高于模型组(P < 0.05)；miRNA测序显示：与模型组相比，中药组的miR-137高于模型组(P < 0.05) (P < 0.05)；模型组的FasL、ACSL4高于正常组(P < 0.05)，中药组的FasL、ACSL4低于模型组(P < 0.05)；与正常组相比，模型组的HE染色显示肝小叶结构明显破坏，肝细胞排列紊乱，肝细胞广泛脂肪变性，胞浆内可见脂滴空泡，部分肝细胞呈气球样变。可见散在的点灶状坏死及桥接坏死，坏死区及汇管区可见大量淋巴细胞和成纤维细胞浸润。与模型组相比，中药组的肝小叶结构破坏程度明显减轻，肝细胞脂肪变性和气球样变减少，点灶状坏死少见。汇管区及小叶内淋巴细胞和成纤维细胞浸润明显减轻，纤维组织增生得到显著抑制。结论：壮肝逐瘀煎可上调miR-137表达，抑制FasL及ACSL4水平，能够显著减轻肝纤维化的组织损伤与炎症。

Abstract: Objective: To observe the effect of Zhuangan Zhuyu Decoction on the expression of miR-137, FasL, and ACSL4 in rats with hepatic fibrosis. Methods: A normal control group was established. A rat model of hepatic fibrosis was induced using carbon tetrachloride (CCl₄). After successful model establishment, the rats were divided into two groups: a model group and a Chinese herbal medicine group, with 10 rats in each group. The Chinese herbal medicine group was administered Zhuangan Zhuyu Decoction by gavage, while the normal control group and the model group received an equal volume of normal saline by gavage. The expression of miR-137 in the three groups was detected by qPCR. miRNA sequencing was performed to analyze whether miR-137 was a differentially expressed miRNA between the model group and the Chinese herbal medicine group. The expression levels of FasL and ACSL4 in the three groups were detected by Western blot. HE staining was used to observe the liver pathology of rats in the three groups. The aim was to clarify the effect of Zhuangan Zhuyu Decoction on the expression of miR-137, FasL and ACSL4 in rats with hepatic fibrosis. Results: The expression level of miR-137 in the model group was significantly lower than that in the normal control group (P < 0.05), while the miR-137 level in the Chinese herbal medicine group was significantly higher than that in the model group (P < 0.05). miRNA sequencing further revealed that, compared with the model group, the Chinese herbal medicine group exhibited a significantly higher expression of miR-137 (P < 0.05).The expression levels of FasL and ACSL4 in the model group were significantly higher than those in the normal control group (P < 0.05), whereas their levels in the Chinese herbal medicine group were significantly lower than those in the model group (P < 0.05).HE staining showed that, compared with the normal control group, the model group showed significantly disrupted hepatic lobule structure, disorganized arrangement of hepatocytes, extensive hepatocyte steatosis with lipid droplet vacuoles visible in the cytoplasm, and ballooning degeneration in some hepatocytes. Scattered spotty necrosis and bridging necrosis were observed, with abundant infiltration of lymphocytes and fibroblasts in the necrotic areas and portal areas. Compared with the model group, the Chinese herbal medicine group showed significantly alleviated disruption of hepatic lobule structure, reduced hepatocyte steatosis and ballooning degeneration, and rare spotty necrosis. Lymphocyte and fibroblast infiltration in the portal areas and lobules was markedly reduced, and fibrous tissue proliferation was significantly inhibited. Conclusions: Zhuanggan Zhuyu Decoction can upregulate the expression of miR-137, inhibit the levels of FasL and ACSL4, and significantly alleviate tissue damage and inflammatory fibrosis in liver fibrosis.