知识与财富的链接
目的:基于超声剪切波弹性成像技术探讨加味茵陈蒿汤(MYCHD)改善大鼠胆汁淤积性肝纤维化的作用机制。方法:选用24只7~8周龄SPF级雄性SD大鼠,按随机数字表法均分为4组:假手术组、模型组、低剂量MYCHD组(MYCHD-L组)、高剂量MYCHD组(MYCHD-H组)。采用胆总管结扎制备大鼠胆汁淤积性肝纤维化模型,假手术组大鼠仅暴露胆总管后关腹。自造模后7 d起,MYCHD低、高剂量组分别以7.74 g/kg、15.48 g/kg剂量灌胃MYCHD,每天1次,连续7 d。假手术组和模型组以同样方式给予饮用水。行超声剪切波弹性成像测定肝硬度值(LSM)后处死大鼠,采用苏木精-伊红(HE)和天狼星红染色法观察肝组织病理学变化,检测血清碱性磷酸酶(ALP)、γ-谷氨酰转移酶(GGT)、透明质酸(HA)、层粘连蛋白(LN)、III型前胶原(PC-Ⅲ)、Ⅳ型胶原(Col-Ⅳ)水平,分析LSM与大鼠肝组织纤维化评分及血清各项指标之间的相关性。结果:与假手术组比较,模型组大鼠LSM、肝组织病理学评分及天狼星红染色阳性比例增加,血清ALP、GGT、HA、LN、PC-Ⅲ和Col-Ⅳ水平升高(P <0.05)。与模型组比较,MYCHD-L组和MYCHD-H组大鼠LSM、肝组织病理学评分及天狼星红染色阳性比例减少,血清ALP、GGT、HA、LN、PC-Ⅲ和Col-Ⅳ水平降低(P <0.05);与MYCHD-L组比较,MYCHD-H组大鼠各指标均有改善,差异有统计学意义(P <0.05)。LSM与ALP、GGT、肝组织病理学评分、天狼星红染色阳性比例、血清HA和Col-Ⅳ水平呈强正相关(r分别为0.802、0.765、0.77、0.83、0.751和0.771,P <0.01),与LN和PC-Ⅲ呈中等正相关(r分别为0.485和0.60,P分别为0.01和0.02)。结论:MYCHD能减轻胆总管结扎诱导的大鼠胆汁淤积性纤维化,改善肝功能。无创的超声剪切波弹性成像技术测定肝硬度值可用于对肝纤维化程度的评估和药物疗效监测。
Objective To evaluate the value of ultrasound shear wave elastography (SWE) in dynamically monitoring the improvement of biliary liver fibrosis stiffness by modified Yinchenhao decoction (MYCHD). Methods Twenty-four 7-8-week-old male Sprague-Dawley (SD) rats under specific pathogen-free (SPF) conditions were selected to establish a bile duct ligation (BDL)-induced biliary liver fibrosis model. Rats were randomly divided into four groups: sham operation group (Sham group), model group (BDL group), model + low-dose MYCHD group (MYCHD-L group), and model + high-dose MYCHD group (MYCHD-H group). The Sham group underwent laparotomy with bile duct exposure but no ligation, followed by abdominal closure. The other groups underwent common bile duct ligation using 4-0 surgical suture. Seven days post-modeling, the MYCHD-L and MYCHD-H groups received MYCHD (7.74 g/kg and 15.48 g/kg, respectively) via gavage once daily for 7 days. The Sham and BDL groups received an equivalent volume of pure water. After gavage, liver stiffness measurements (LSM) were obtained using SWE. All rats were euthanized post-ultrasound. Liver histopathological changes were observed using Hematoxylin and Eosin (HE) and Sirius red staining. Serum liver function markers—alkaline phosphatase (ALP) and gamma-glutamyl transferase (GGT)—were analyzed using an automated biochemical analyzer. Serum levels of liver fibrosis markers—hyaluronic acid (HA), laminin (LN), type III procollagen (PC-Ⅲ), and type IV collagen (Col-Ⅳ)—were detected via ELISA. Correlations between LSM and serum markers (ALP, GGT) and liver fibrosis markers were analyzed. Results Compared to the Sham group, rats in the Model group (BDL) exhibited significantly increased hepatocyte injury, inflammatory cell infiltration, and collagen deposition area in liver tissue (P <0.05), alongside significantly elevated serum levels of ALP, GGT, HA, LN, PC-Ⅲ, and Col-Ⅳ (P <0.05). Furthermore, liver stiffness values (LSM) measured by SWE were significantly higher in the Model group (P <0.05). Compared to the BDL group, rats in both MYCHD dose groups showed significantly reduced hepatocyte injury, inflammatory cell infiltration, and collagen deposition area (P <0.05), as well as significantly decreased serum levels of ALP, GGT, HA, LN, PC-Ⅲ, and Col-Ⅳ(P <0.05). SWE measurements also revealed significantly lower LSM values in the MYCHD groups (P <0.05). Further analysis revealed that LSM exhibited a strong positive correlation with ALP, GGT, liver histopathological scores, Sirius red staining-positive area, and serum levels of HA and Col-Ⅳ (r=0.802,0.765,0.77,0.83,0.751,0.771, respectively; all P < 0.01), and a moderate positive correlation with LN and PC-Ⅲ (r= 0.485,0.60, P =0.01,0.02, respectively). Conclusion MYCHD significantly improves BDL-induced liver fibrosis. LSM is valuable for assessing the anti-fibrotic efficacy and therapeutic outcomes of MYCHD. This study provides new evidence and a visualized evaluation method for traditional Chinese medicine in anti-liver fibrosis treatment.