N-乙酰半胱氨酸活性炭缓释微囊对大鼠肝纤维化中细胞内成纤信号分子表达的影响

Effect of N-Acetylcysteine Activated Carbon Sustained-release Microcapsules on the Expression of Intracellular Fibroblast Signaling Molecules in Rat Liver Fibrosis

关键词:  N-乙酰半胱氨酸活性炭缓释微囊  肝纤维化  成纤信号分子

Key words:  N-acetylcysteine ativated carbon sustained-release microcapsules (ACNAC)  hepatic fibrosis  fibroblast signaling molecules

摘要:

目的 研究口服乙酰半胱氨酸活性炭缓释微囊（N-acetylcysteine activated carbon microcapsules，ACNAC）在大鼠肝纤维化治疗中肝细胞内成纤信号分子表达的影响。方法 以明胶包裹吸附乙酰半胱氨酸（N-acetylcysteine，NAC）的载药活性炭制备得到ACNAC，以四氯化碳（CCl₄）制备大鼠肝纤维化模型，ACNAC高、中、低剂量组（80，40，20 mg·kg^-1·d^-1，按照乙酰半胱氨酸计算）进行灌胃给药，NAC、水飞蓟宾为阳性对照药物。治疗8周后处死大鼠，通过大鼠肝功能指标、病理切片、免疫组织化学法评价ACNAC对胞内信号分子表达中的影响。结果 CCl₄大鼠肝纤维化模型制备成功，其大鼠肝组织病理切片HE染色和网状纤维染色表明其肝纤维化程度为Ⅲ~Ⅳ期，免疫组化显示胞内TGF-β₁/Tβ-R₁和Smad2/3大量表达。ACNAC、NAC和SM对肝纤维化大鼠血清ALT、AST及Tbil有显著的治疗作用，ACNAC高剂量组效果明显优于NAC和SM （P<0.05）。ACNAC各治疗组大鼠肝组织病理切片表明肝纤维化好转，免疫组化显示胞内TGF-β₁/Tβ-R₁和Smad2/3表达减少，ACNAC高剂量组表达具显著性差异（P<0.05）。结论 ACNAC对大鼠肝纤维化胞内成纤信号分子TGF-β₁/Tβ-R₁和Smad2/3有明显抑制作用，对抗大鼠肝纤维化的疗效明显，且较等剂量的NAC及对照药物水飞蓟宾有更显著的疗效。

Abstract:

OBJECTIVE To study the effects of N-acetylcysteine activated carbon sustained-release microcapsule (ACNAC) by oral administration on hepatic fibroblast signaling molecule expression in rats with hepatic fibrosis. METHODS ACNAC was prepared by gelatin entrapping with acetylcysteine-loaded activated carbon. The rat model of liver fibrosis was prepared with carbon tetrachloride (CCl₄). The rats were given ACNAC high, middle and low dose of 80 mg·kg^-1·d^-1, 40 mg·kg^-1·d^-1, 20 mg·kg^-1·d^-1 (calculated according to acetylcysteine), NAC and silybin as the positive control drug by gavage administration. Rats were sacrificed after 8 weeks of treatment. The effects of ACNAC on the expression of intracellular signal molecules were evaluated by rat liver function, pathological sections and immunohistochemistry. RESULTS CCl₄-induced rat liver fibrosis model was successfully established. The pathological changes of liver tissue were observed by HE staining and reticular fiber staining. The degree of hepatic fibrosis was grade Ⅲ-Ⅳ, and immunohistochemistry showed that TGF-β₁/Tβ -R1 and Samd2/3 were abundantly expressed. ACNAC, NAC and SM had significant therapeutic effects on serum ALT, AST and Tbil in rats with hepatic fibrosis. The ACNAC high dose group was superior to NAC and SM (P<0.05). The pathological sections of liver tissue with treatment groups showed improvement of hepatic fibrosis. Immunohistochemistry showed that TGF-β1/Tβ-R1 and Samd2/3 expression were decreased in the NCSM group, and the NCSM high dose group (80 mg·kg^-1·d^-1) expression was significantly different (P<0.05). CONCLUSION ACNAC could significantly inhibit the expression of TGF-β₁/Tβ-R₁ and Smad2/3 in rat liver fibrosis. The effect of ACNAC on liver fibrosis was significantly improved, and has a better therapeutic effect than of NAC and the control drug silybin.