山芝麻酸甲酯对肝纤维化小鼠的影响

Effects of methyl sesamate on hepatic fibrosis in mice

目的观察山芝麻酸甲酯对肝纤维化小鼠的影响及其机制研究。方法小鼠腹腔注射四氯化碳(CCl4)-橄榄油混合液体8周构建肝纤维化模型。按照体重将小鼠随机分为5组:正常组、模型组和低、中、高3个剂量实验组,每组8只。模型组和正常组给予0.9%NaCl,低、中、高3个剂量实验组分别给予山芝麻酸甲酯10,20和40 mg·kg^-1·d^-1。于造模后第5周开始灌胃给药,直至第14周结束。用酶联免疫吸附法检测血清谷丙转氨酶(GPT)、谷草转氨酶(GOT)、透明质酸(HA)和肝组织的羟脯氨酸(Hyp)含量,用蛋白质印迹检测肝组织中肉瘤蛋白(Ras)、细胞外调节蛋白激酶(ERK)和磷酸化细胞外调节蛋白激酶(p-ERK)的蛋白相对表达量(灰度值)。结果正常组、模型组和低、中、高3个剂量实验组血清GPT分别为(27.68±5.55),(158.54±11.74),(96.90±5.02),(62.97±7.66)和(43.52±2.65)U·L^-1;这5组的血清GOT分别为(27.42±3.05),(108.27±12.62),(75.42±3.77),(58.98±5.82),和(37.49±3.63)U·L^-1;这5组的血清HA分别为(115.17±10.73),(248.39±22.47),(194.12±22.32),(178.42±11.45)和(147.42±13.76)μg·L^-1;这5组的肝组织中Hyp水平分别为(292.09±19.61),(404.53±16.39),(384.43±18.27),(375.04±13.59),和(346.42±14.55)μg·g-1;这5组的Ras蛋白相对表达量分别为0.99±0.08,2.38±0.19,1.73±0.17,1.64±0.11和1.47±0.12;这5组的ERK蛋白相对表达量分别为1.05±0.02,1.02±0.11,1.03±0.12,1.08±0.08和1.00±0.11;这5组的p-ERK蛋白相对表达量分别为1.03±0.12,1.80±0.16,1.59±0.17,1.36±0.08和1.23±0.09。上述指标:模型组与正常组比较,差异均有统计学意义(均P<0.01);低、中、高3个剂量实验组与模型组比较,差异均有统计学意义(均P<0.01)。结论山芝麻酸甲酯对小鼠肝纤维化具有一定的抑制作用,且与Ras/ERK信号通路有关。

Objective Objective To observe the effect of methyl sesamolate on liver fibrosis in mice and its mechanism.Methods Liver fibrosis model was established by intraperitoneal injection of CCl4-olive oil mixed liquid for 8 weeks.Mice were randomly divided into 5 groups:normal group,model group and low,medium and high dose experimental groups with 8 mice in each group.The model group and the normal group were given 0.85%normal saline,and the low,medium and high dose experimental groups were given 10,20,40 mg·kg^-1·d^-1 methyl sesamolate,respectively;and the gastric administmiceion was started at the 5 th week after the model was established until the end of the 14 th week.Serum glutamic-pyruvic transaminase(GPT)and glutamic-oxaloacetic transaminase(GOT),hyaluronic acid(HA)and hydroxyproline(Hyp)in liver tissue were detected by enzyme-linked immunosorbent assay.The relative expression(gray value)of sarcoma protein(Ras),extracellular regulated protein kinases(ERK)and phosphorylated extracellular regulated protein kinases(p-ERK)in liver tissue were detected by Western blot.Results The serum GPT of normal group,model group,control group and low,medium and high dose experimental groups were(27.68±5.55),(158.54±11.74),(96.90±5.02),(62.97±7.66)and(43.52±2.65)U·L^-1,respectively;serum GOT in the five groups were(27.42±3.05),(108.27±12.62),(75.42±3.77),(58.98±5.82)and(37.49±3.63)U·L^-1,respectively;serum HA in the five groups were(115.17±10.73),(248.39±22.47),(194.12±22.32),(178.42±11.45)and(147.42±13.76)μg·L^-1,respectively;Hyp levels in liver tissues in the five groups were(292.09±19.61),(404.53±16.39),(384.43±18.27),(375.04±13.59)and(346.42±14.55)μg·g-1,respectively;the relative expression of Ras protein in the five groups were 0.99±0.08,2.38±0.19,1.73±0.17,1.64±0.11 and 1.47±0.12,respectively;the relative expression of ERK protein in the five groups were 1.05±0.02,1.02±0.11,1.03±0.12,1.08±0.08 and 1.00±0.11,respectively;the relative expressions of p-ERK protein in the five groups were 1.03±0.12,1.80±0.16,1.59±0.17,1.36±0.08 and1.23±0.09,respectively.Comparison between model group and normal group,the differences of the factors were statistically significant(all P<0.01);comparison between three dose experimental groups and model group,the differences of the factors were statistically significant(all P<0.01).Conclusion Methyl sesamolate had a certain inhibitory effect on liver fibrosis in mice and was related to Ras/ERK signal pathway.