茯苓酸调节Hippo信号通路对二乙基亚硝胺诱导的肝癌大鼠的治疗作用研究

Therapeutic effect of pachymic acid on diethylnitrosamine induced hepatoma in rats by regulating Hippo signaling pathway

关键词:  茯苓酸  Hippo  二乙基亚硝胺  肝癌  治疗作用

Key words:  pachymic acid  Hippo  diethylnitrosamine  liver cancer  therapeutic effects

摘要:

[目的]探讨茯苓酸调节Hippo信号通路对二乙基亚硝胺(DEN)诱导的肝癌大鼠的治疗作用。[方法]取SD大鼠随机分为对照组、模型组、茯苓酸低剂量(3.5 mg/kg)组、茯苓酸中剂量(7 mg/kg)组、茯苓酸高剂量(14 mg/kg)组、氟尿嘧啶(50 mg/kg)组,每组10只,模型组和给药组大鼠通过腹腔注射DEN诱导建立肝癌模型,对照组大鼠腹腔注射等剂量0.9%氯化钠溶液,以茯苓酸和氟尿嘧啶分组处理后检测各组大鼠血清肝功能指标谷丙转氨酶(ALT)、谷草转氨酶(AST)水平;检测各组大鼠体质量变化、肝脏指数、肝表面癌结节数、最大癌结节体积;以苏木精-伊红(HE)染色检测各组大鼠肝组织病理形态并比较其炎症细胞浸润数;以试剂盒检测大鼠血清炎症因子白细胞介素(IL)-6、IL-17、肿瘤坏死因子-α(TNF-α)与氧化应激因子超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、丙二醛(MDA)水平;以蛋白免疫印迹法检测各组大鼠肝组织Hippo信号通路相关蛋白[Yes相关蛋白(YAP)、LATS1]表达。[结果]与对照组比较,模型组大鼠肝组织发生明显癌变,给药结束后体质量、血清SOD、CAT水平显著降低(P＜0.05),血清ALT、AST、IL-6、IL-17、TNF-α与MDA水平、肝脏指数、肝表面癌结节数、最大癌结节体积、炎症细胞浸润数、肝组织LATS1蛋白表达及p-YAP1/YAP1显著升高(P＜0.05);与模型组比较,茯苓酸低、中、高剂量组、氟尿嘧啶组大鼠肝组织癌变症状均减轻,给药结束后体质量、血清SOD、CAT水平升高(P＜0.05),血清ALT、AST、IL-6、IL-17、TNF-α与MDA水平、肝脏指数、肝表面癌结节数、最大癌结节体积、炎症细胞浸润数、肝组织LATS1蛋白表达及p-YAP1/YAP1降低(P＜0.05),且茯苓酸各组呈剂量依赖性(P＜0.05);茯苓酸高剂量组与氟尿嘧啶组大鼠比较,各指标比较差异无统计学意义(P＞0.05)。[结论]茯苓酸可降低Hippo信号通路活性,抑制DEN诱导的炎症和氧化应激,减轻大鼠肝组织损伤并缓解其癌变症状,改善大鼠肝功能。

Abstract:

[Objective] To investigate the therapeutic effect of pachymic acid on diethylnitrosamine(DEN) induced hepatoma in rats by regulating Hippo signal pathway. [Methods] SD rats were randomly grouped into a control group,a model group,low-dose(3.5 mg/kg) pachymic acid group, middle dose (7 mg/kg) pachymic acid group, high-dose( 14 mg/kg) pachymic acid group, and fluorouracil(50 mg/kg) group,with 10 rats in each group. The liver cancer model was induced by intraperitoneal injection of DEN in the model group and the administration group rats,while the control group rats were intraperitoneally injected with an equal dose of 0.9% sodium chloride solution. After treatment with pachymic acid and fluorouracil,the serum levels of alanine transaminase(ALT) and aspartate transaminase(AST) were detected;the weight changes,liver index,number of liver surface cancer nodules,and maximum cancer nodule volume of rats in each group were measured;hematoxylin-eosin(HE) staining was applied to detect the pathological morphology of liver tissue of rats in each group and compare the number of inflammatory cell infiltration;the serum levels of inflammatory factors such as interleukin(IL)-6,IL-17,tumor necrosis factor-α(TNF-α) and oxidative stress factors such as superoxide dismutase(SOD), catalase(CAT) and malondialdehyde(MDA) were measured with the kits;Western Blot was applied to detect the expression of Hippo signaling pathway related proteins [Yes-related protein(YAP),LATS1] in the liver tissues of rats in each group. [Results] Compared with the control group,the liver tissue of the model group rats showed obvious carcinogenesis,weight after administration,the levels of serum SOD and CAT were obviously reduced (P＜0.05), and the levels of serum ALT,AST,IL-6,IL-17,TNF-α and MDA,liver index,number of liver surface cancer nodules,maximum cancer nodule volume,number of inflammatory cell infiltratio,the protein expression of LATS1 in liver tissue,and p-YAP1/YAP1 were obviously increased(P＜0.05);compared with the model group,the canceration symptoms of liver tissue of rats in the low,medium and high dose pachymic acid groups and the fluorouracil group were alleviated,weight after administration,and the levels of serum SOD and CAT were increased(P＜0.05),and the levels of serum ALT,AST,IL-6,IL-17,TNF-α and MDA,liver index,number of liver surface cancer nodules,maximum cancer nodule volume,number of inflammatory cell infiltratio,the protein expression of LATS1 in liver tissue,and p-YAP1/YAP1 were decreased(P＜0.05),pachymic acid groups were dose-dependent(P＜0.05);there was no obvious difference in each index between the high dose pachymic acid group and the fluorouracil group(P＞0.05). [Conclusion]Pachymic acid can reduce the activity of Hippo signaling pathway,inhibit DEN induced inflammation and oxidative stress,alleviate liver tissue damage and canceration symptoms in rats,and improve liver function in rats.