四逆汤上调p53蛋白表达诱导肝癌细胞凋亡研究

Study on Sini Decoction induced Hepatoma carcinoma cell apoptosis through upregulating the expression of p53 protein

目的:以中医温阳法代表方四逆汤为研究对象,研究其对p53蛋白表达诱导肝癌细胞凋亡的影响,为提高肝癌中西医结合治疗效果提供实验依据。方法:采用含药血清方法,整体小鼠移植瘤实验,建立肝癌动物模型后,随机分为空白组、四逆汤组、阳性组共3组,每组10只,每日按0.2mL/10g标准定时胃饲给药1次。四逆汤组:3.7g生药/kg;空白组:同法灌胃等量0.9%氯化钠溶液;阳性组:顺铂4mg/kg,腹腔注射,每周给药1次,连续给药3周。用Tunel法检测细胞凋亡,Western Blot测定p53蛋白表达,从分子水平分析细胞凋亡的机制。结果:①四逆汤组对肝癌细胞具有抑制作用(P<0.05)。②四逆汤组在抑制肝癌组织生长的同时,对胸腺及脾脏起到了一定的保护作用。③Tunnel法测凋亡细胞结果表明,四逆汤可诱导小鼠肝癌细胞凋亡。④四逆汤明显上调肝癌组织p53蛋白的表达(P<0.01)。结论:温阳法对肝癌有抑制作用,同时具有免疫保护作用,降低肿瘤恶性程度的作用;促进肝癌组织p53蛋白的表达,四逆汤诱导细胞凋亡中的作用可能与p53蛋白的表达存在密切关系。

Objective： The research represents Sini Decoction as the study object. This project is to study the effect of Warm Yang to cancer cell apoptosis, and supplying theoretical and experimental evidence for the combination of Chinese and western medicine.Methods： Pharmaceutical serum method and mice-transplanted tumor experiments was applied to evaluate Warm Yang treatment. Animal model of liver cancer was established and randomized into blank group, Sini group, positive control group, with 10 rats in each group. According to the standard of 0.2mL/10g daily, Animal model was intragastric infusion on schedule. Sini group： 3.7g/kg; blank group： equivalent with 0.9% sodium chloride; positive group： Cisplatin 4mg/kg, intraperitoneal injection, Once a week, was administered for 3 weeks. Tunel was used to examine cell apoptosis .Western-blot was used to examine p53 protein expression to analysis the associated mechanism of cell apoptosis. Results： The research showed that （~）Warm Yang treatment inhibited the proliferation of liver cancer cell in vivo. @Sini Decoction had a protective effect on the immune organs. @The results of Tunnel show that Sini Decoction can be induced apoptosis of hepatoma cells in vivo. @Sini Decoction was markedly upregulated expression of p53 protein in hepatocellular carcinoma tissues. Conclusion： The above results suggested that Warm Yang treatment showed inhibitory effect on the hepatoma ,the role of immune protection and of reducing the degree of malignancy. Warming Yang induced cell apoptotic could be closely related with upregulation expression of p53.