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目的:观察中药大柴胡汤联合中等强度有氧运动对大鼠阻塞性黄疸致重症肝损伤肝脏炎症细胞因子的影响。方法:40只SD大鼠随机分为4组:正常组(CG组),模型组(OJ组),中药组,中药+运动组。采用改良的胆总管结扎法构建大鼠阻塞性黄疸致重症肝损伤模型,两中药组于胆总管结扎术后3天进行大柴胡汤灌胃,每天(下午6时)灌胃1次,剂量2 ml/次,共12周。运动组进行坡度5%、速度16.8 m/min、每天45 min、每周5天、共12周的中等强度跑台训练。12周后大鼠禁食过夜,随后分离出肝组织并取材,对各组大鼠肝脏组织损伤情况进行显微结构形态学观察;免疫组化法检测肝组织Toll样受体4(TLR4)及髓样分化因子88(MyD88)蛋白表达水平;RT-PCR法检测肝组织肿瘤坏死因子-α(TNF-α)、白介素6(IL-6)、白介素10(IL-10)mRNA表达水平;生物化学分析法检测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)和总胆红素(TBIL)含量。结果:①HE染色显示:正常组肝细胞索排列整齐;模型组大鼠肝细胞大量纤维化,可见大片状肝细胞变性、坏死,乃至肝细胞索结构紊乱;中药组可见肝细胞少量点、片状坏死,肝小叶结构改变,肝索排列欠整齐;中药+运动组无明显组织坏死,肝小叶结构基本正常,肝索排列较整齐。②血清学结果:中药+运动组ALT、AST、TBIL水平较模型组显著降低(P<0.01),中药组ALT、AST水平也显著下降(P<0.05)。③免疫组织化学染色结果显示:中药+运动组大鼠肝组织TLR4、MyD88表达均显著低于模型组(P<0.01)。④RT-PCR结果显示:与模型组比较,中药+运动组大鼠血清IL-6、TNF-α表达降低(P<0.01),而IL-10表达升高(P<0.01)。结论:中等强度有氧运动联合中药大柴胡汤能调节肝脏炎症细胞因子的表达,增强阻塞性黄疸致重症肝损伤大鼠的免疫功能,从而明显减轻阻黄致大鼠肝损伤的程度。
Objective: To observe the effect of Dachaihu Decoction combined with moderate-intensity aerobic exercise on inflammatory cytokines in the liver of rats with obstructive jaundice-induced severe liver injury. Methods: Forty SD rats were randomly divided into 4 groups: normal group (CG group), model group (OJ group), TCM group and TCM + exercise group. The model of obstructive jaundice-induced severe liver injury was established by modified common bile duct ligation. Two Chinese herbal medicine groups were administered with dachaihu decoction 3 days after common bile duct ligation, and gavage was given daily (6 pm) ml / time for a total of 12 weeks. The exercise group performed moderate intensity treadmill training with 5% slope, 16.8 m / min speed, 45 min daily and 5 days a week for a total of 12 weeks. Rats were fasted overnight for 12 weeks. Liver tissues were isolated and harvested for microscopic morphological observation of liver tissue injury in each group. Toll-like receptor 4 (TLR4) and The expression of MyD88 protein was detected by RT-PCR. The mRNA expression of TNF-α, IL-6 and IL-10 in liver tissues was detected by RT-PCR. The levels of serum ALT, AST and TBIL were determined by chemical analysis. Results: (1) HE staining showed that the normal liver cells arranged in neat rows; the model group of rats with a large number of liver fibrosis, showing large degeneration of liver cells, necrosis, and even liver cell cord structure disorder; Chinese medicine group can be seen a small amount of hepatocytes Like necrosis, changes in the structure of the hepatic lobule, hepatic cord arranged in regular neat; Chinese medicine + exercise group no significant tissue necrosis, hepatic lobule structure is normal, the liver arranged more neatly. ② Serological results: The levels of ALT, AST and TBIL in TCM + exercise group were significantly lower than those in model group (P <0.01), and ALT and AST levels in TCM group were also significantly decreased (P <0.05). ③ The results of immunohistochemical staining showed that the expressions of TLR4 and MyD88 in liver of TCM plus exercise group were significantly lower than those in model group (P <0.01). The results of RT-PCR showed that the levels of IL-6 and TNF-α were decreased (P <0.01) and the expression of IL-10 was increased (P <0.01) compared with model group. Conclusion: Moderate-intensity aerobic exercise combined with Dachaihu decoction can regulate the expression of inflammatory cytokines in the liver and enhance the immune function of rats with obstructive jaundice-induced severe liver injury, and thus significantly reduce the extent of liver damage induced by the yellow-induced obstruction in rats.