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芒果苷对四氯化碳致肝纤维化小鼠模型的干预作用
Interventional effect of mangiferin on carbon tetrachloride-induced hepatic fibrosis in mice
文章发布日期:2019年03月07日  来源:  作者:王虎,程玉佳,王莉,等  点击次数:139次  下载次数:29次

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【摘要】:目的探讨芒果苷对四氯化碳(CCl4)诱导的肝纤维化小鼠模型的影响及其潜在的机制。方法将45只C57BL/6雄性小鼠随机分为3组:正常对照组(NC组,n=15)、肝纤维化模型组(CCl4组,n=15)和芒果苷预处理组(CCl4+M组,n=15)。全自动生化分析仪检测血清ALT和AST水平;苏木素-伊红(HE)和Masson染色观察肝组织病理学改变;ELISA检测血清IL-1β、IL-6和TNFα水平;Western Blot检测肝脏α-平滑肌肌动蛋白(α-SMA)、核因子-κB(NF-κB)、IL-1β、p62和微管相关蛋白1 Ⅰ/Ⅱ轻链3(LC3-Ⅰ/Ⅱ)的表达;荧光定量PCR检测肝脏中Ⅰ型胶原蛋白(Col-Ⅰ)和α-SMA mRNA的表达。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果HE和Masson染色结果显示CCl4+M组肝细胞变性坏死少见,胶原纤维明显减少;与CCl4组相比,芒果苷能显著降低血清ALT和AST水平(P值均<0.01);ELISA结果显示CCl4+M组血清中IL-1β、IL-6和TNFα水平与CCl4组相比明显降低(P值均<0.01);Western Blot结果显示CCl4+M组肝脏α-SMA、NF-κB、IL-1β和LC3-Ⅱ/Ⅰ的蛋白表达与CCl4组相比明显降低,p62蛋白明显升高(P值分别<0.01、<0.05、<0.01、<0.05、<0.05);荧光定量PCR结果显示CCl4+M组Col-Ⅰ和α-SMA mRNA的表达与CCl4组相比均明显降低(P值分别<0.05、<0.01)。结论芒果苷具有抑制CCl4致小鼠肝纤维化的作用,其机制可能与减轻炎症反应保护肝功能,抑制自噬反应降低肝星状细胞的活化有关。
【Abstract】:ObjectiveTo investigate the interventional effect of mangiferin on carbon tetrachloride (CCl4)-induced hepatic fibrosis in mice and the potential mechanism. MethodsA total of 45 male C57BL/6 mice were randomly divided into three groups: normal control group (NC group), liver fibrosis model group (CCl4 group), and mangiferin pretreatment group (CCl4+M group), with 15 mice in each group. An automatic biochemical analyzer was used to measure the serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST); HE staining and Masson staining were performed to observe liver pathological changes; ELISA was used to measure the serum levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNFα); Western Blot was used to measure the protein expression of α-smooth muscle actin (α-SMA), nuclear factor-kappa B (NF-κB), IL-1β, p62, and microtubule-associated protein 1 light chain 3 (LC3) in the liver; quantitative real-time PCR was used to measure the mRNA expression of type I collagen (Col-I) and α-SMA in the liver. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsHE staining and Masson staining showed a low proportion of mice with hepatocyte degeneration and necrosis and a significant reduction in collagen fibers in the CCl4+M group. Compared with the CCl4 group, the CCl4+M group had significant reductions in the serum levels of ALT and AST (both P<001). ELISA showed that compared with the CCl4 group, the CCl4+M group had significant reductions in the serum levels of IL-1β, IL-6, and TNF-α (all P<0.01). Western Blot showed that compared with the CCl4 group, the CCl4+M group had significant reductions in the protein expression of α-SMA, NF-κB, IL-1β, and LC3-II/I in the liver (P<0.01, P<0.05, P<0.01, and P<0.05) and a significant increase in the protein expression of p62 (P<0.05). Quantitative real-time PCR showed that compared with the CCl4 group, the CCl4+M group had significant reductions in the mRNA expression of Col-I and α-SMA (P<0.05 and P<0.01). ConclusionMangiferin can alleviate CCl4-induced hepatic fibrosis in mice, possibly by reducing inflammation to protect liver function and inhibiting autophagy to reduce the activation of hepatic stellate cells.
【关键字】:肝硬化, 实验性; 芒果苷; 小鼠, 近交C57BL; 四氯化碳; 自噬
【Key words】:liver cirrhosis, experimental; mangiferin; mice, inbred C57BL; carbon tetrachloride; autophagy
【引证本文】:WANG H, CHENG YJ, WANG L, et al. Interventional effect of mangiferin on carbon tetrachloride-induced hepatic fibrosis in mice[J]. J Clin Hepatol, 2019, 35(4): 808-812. (in Chinese)
王虎, 程玉佳, 王莉, 等. 芒果苷对四氯化碳致肝纤维化小鼠模型的干预作用[J]. 临床肝胆病杂志, 2019, 35(4): 808-812.

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