消癌平注射液联合奥曲肽对H22荷瘤小鼠细胞凋亡的影响

摘要

Objective To observe the inhibitory effects of Xiaoaiping injection (XI) combined with octreotide on H22 tumor-bearing mice on H22 hepatic transplantable tumor and its optimal drug concentration, and to explore its primary mechanism. Methods The subcutaneous H22 hepatic transplantable tumor models were established and randomized into 8 groups after tumorigenesis: model group (0.4 ml/d normal saline), low vs moderate vs high dose of XI group (at the dose of 10, 20, and 40 g/kg), octreotide group (100 u,g/kg), and low vs moderate vs high dose of XI + octreotide group. All mice were intraperitoneally injected for 14 days. Thereafter, the tumor volume was calculated, the tumor growth curve drawn, the inhibitory rate calculated, the apoptosis detected by TUNEL assay, the expressions of apoptosis correlated protein B-cell lymphoma/leukemia-2 (Bcl-2) and Bel-associated x protein (Bax) determined by im-munohistochemical assay. Meanwhile, the activity of natural killer cell (NK) was determined. Results Compared with the model group, the drug use groups showed certain inhibition effects on the growth of H22 hepatocarcinoma. Notably, the inhibitory effects of the combined drug use groups were better than those of the single drug use group (P< 0.05), with the high dose of XI + octreotide showing the best effects. The high dose of XI and high dose of XI + octreotide group could up-regulate the apoptotic index (AI), down-regulate the Bcl-2 expression, up-regulate the Bax expression and increase the activity of NK cells. Notably, the effects of high dose of XI + octreotide group were significantly better than those of octreotide group and high dose of XI group. Conclusion High dose of XI+octreotide was the best drug concentration and its tumor inhibitory effects might be correlated with regulating apoptosis-related protein Bcl-2, Bax expression that induced tumor apoptosis and enhance the activity of NK cells in the spleen.%目的 观察消癌平注射液联合奥曲肽对H22肝癌移植瘤的抑瘤作用及其抑瘤的最佳药效浓度,并初步探讨其作用机制.方法 建立小鼠皮下H22肝癌移植癌模型,成瘤后将实验动物随机分为8组:分别为模型组(生理盐水0.4 ml/d),消癌平低、中、高剂量组(10、20、40 g/kg),奥曲肽组(100 μg/kg)及消癌平低、中、高剂量分别联合奥曲肽组.腹腔注射给药14 d.计算各瘤体体积,绘制肿瘤生长曲线,计算抑瘤率,采用脱氧核糖核酸末端转移酶介导的缺口末端标记法(TUNEL)检测肿瘤组织中的细胞凋亡情况,免疫组织化学方法测定凋亡相关蛋白B细胞淋巴瘤/白血病基因-2(Bcl-2),Bcl相关X连锁蛋白(Bax)的表达情况；同时测定脾脏自然杀伤细胞(NK)活性.结果 与模型组比较,各用药组对H22肝癌的生长均有一定的抑制作用,联合用药组的抑瘤作用优于单药组(P＜0.05),以高剂量消癌平加奥曲肽的作用最好.高剂量消癌平及高剂量消癌平加奥曲肽组可使凋亡指数(AI)升高,下调Bcl-2的表达,上调Bax的表达,升高脾脏NK细胞活性,其中高剂量消癌平加奥曲肽组作用明显优于奥曲肽组和高剂量消癌平组.结论 高剂量消癌平加奥曲肽为抑瘤的最佳药效浓度,其抑制肿瘤生长的作用可能与通过调控凋亡相关蛋白Bcl-2、Bax的表达诱导肿瘤细胞凋亡及增强脾脏NK细胞活性有关.