柠檬苦素对大鼠肝纤维化的影响及机制初探
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篇名: | 柠檬苦素对大鼠肝纤维化的影响及机制初探 |
TITLE: | Effect of limonin on liver fibrosis in rats and preliminary exploration of its mechanism |
摘要: | 目的 探讨柠檬苦素对大鼠肝纤维化的影响及机制。方法将42只SD雄性大鼠随机分为正常组、模型组、阳性对照组(秋水仙碱0.1mg/kg)和柠檬苦素低、高剂量组(12.5、50mg/kg),正常组和模型组各9只,其余组各8只。除正常组外,采用四氯化碳构建肝纤维化模型。造模成功后,正常组及模型组灌胃水,其余各组灌胃相应药物,每日1次,持续4周。末次给药后,采用酶联免疫吸附测定法检测各组大鼠血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、透明质酸(HA)、层粘连蛋白(LN)含量;采用苏木精-伊红(HE)及Masson染色观察肝组织形态和肝纤维化变化;采用Westernblot法检测肝组织中蛋白激酶B(Akt)、磷酸化Akt(p-Akt)、E-钙黏蛋白(E-cadherin)、波形蛋白(vimentin)蛋白表达水平,并计算p-Akt/Akt比值。结果与正常组比较,模型组大鼠血清ALT、AST、HA、LN含量均显著升高(P<0.01),HE及Masson染色均观察到肝组织有大量纤维组织增生,肝组织中Akt、p-Akt、vimentin蛋白表达水平及p-Akt/Akt比值均显著升高(P<0.01),E-cadherin蛋白表达水平显著降低(P<0.01)。与模型组比较,柠檬苦素低、高剂量组大鼠肝组织的纤维增生程度均有所减轻,除柠檬苦素低剂量组的LN、E-cadherin指标外其余指标均显著逆转(P<0.05或P<0.01)。结论柠檬苦素能改善大鼠肝纤维化,作用机制可能与抑制Akt磷酸化及逆转上皮间质转化进程有关。 |
ABSTRACT: | OBJECTIVE To explore the effect and mechanism of limonin on liver fibrosis in rats. METHODS Forty-two SD male rats were randomly divided into normal group, model group, positive control group (colchicine, 0.1 mg/kg), low-dose group of limonin (12.5 mg/kg) and high-dose group of limonin (50 mg/kg), with 9 rats in normal group and model group, and 8 rats in other groups. Except for normal group, the model of liver fibrosis was established by carbon tetrachloride. After modeling, normal group and model group were given distilled water intragastrically, and administration groups were given relevant medicine intragastrically,once a day,for consecutive 4 weeks. After the last medication, the serum contents of alanine aminotransferase (ALT), aspartate aminotransferase (AST), hyaluronic acid (HA) and laminin (LN) in rats were detected by enzyme-linked immunosorbent assay (ELISA) method. HE and Masson staining were used to observe the pathological changes and fibrotic change of liver tissue. Western blot assay was used to detect the protein expressions of protein kinase B (Akt), phosphorylated-protein kinase B (p-Akt), E-cadherin and vimentin in liver tissue; ratio of p-Akt/Akt was calculated. RESULTS Compared normal group, the serum contents of ALT, AST, HA and LN in rats were all increased significantly in model group (P<0.01); HE and Masson staining showed that a large number of fibrous tissue hyperplasia, protein expressions of Akt, p-Akt and vimentin as well as p-Akt/ Akt ratio in liver tissue were significantly increased (P<0.01), while E-cadherin protein expression was significantly decreased (P<0.01). Compared with model group, the degree of fibrosis hyperplasia in liver tissue was relieved in high-dose group and low- dose group of limonin, and other indexes were all reversed significantly except for LN and E-cadherin in the low-dose group of limonin (P<0.01 or P<0.05). CONCLUSIONS Limonin can improve liver fibrosis in rats, the mechanism of which may be related to the inhibition of Akt phosphorylation and reversal of epithelial-mesenchymal transition process. |
期刊: | 2022年第33卷第24期 |
作者: | 肖玉洪,安祯祥 |
AUTHORS: | XIAO Yuhong,AN Zhenxiang |
关键字: | 柠檬苦素;肝纤维化;上皮间质转化;大鼠 |
KEYWORDS: | limonin; liver fibrosis; epithelial-mesenchymal |
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