小半夏汤对化疗致胃肠黏膜炎模型大鼠胃肠组织NF-κB、IL-1β、TNF-α mRNA和蛋白表达水平的影响

Effect of Xiaobanxia Decoction on NF-κB,IL-1β,TNF-α mRNA and Protein Expression Levels in Sinuses Ventriculi and Ileum of Gastrointestinal Mucositis Model Rats Induced by Chemotherapy

目的:观察小半夏汤对顺铂化疗致胃肠黏膜炎模型大鼠胃窦和回肠组织中核因子κB(NF-κB)、白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α) mRNA和蛋白表达水平的影响,探讨小半夏汤防治化疗性胃肠黏膜炎的机制。方法:雄性Wistar大鼠112只随机分为7组,每组16只。空白对照组和顺铂模型组每日予20 mL/kg蒸馏水灌胃,小半夏汤正常对照组每日予1.6 g/kg小半夏汤灌胃,奥美拉唑组每日予2.4 mg/kg奥美拉唑灌胃,小半夏汤低、中、高剂量组每日分别予0.8 g/kg、1.6 g/kg、3.2 g/kg小半夏汤灌胃。各组上述剂量分早、晚两次给药,连续给药5 d。给药第3天,除空白对照组和小半夏汤正常对照组腹腔注射等容积生理盐水,其他各组均腹腔注射顺铂6 mg/kg造模。造模后24 h和72 h取各组大鼠胃窦和回肠,采用RT-PCR、Western blot法检测胃肠组织中NF-κB、IL-1β、TNF-αmRNA和蛋白表达水平。结果:造模后24 h和72 h,与空白对照组比较,顺铂模型组大鼠胃窦和回肠组织中NF-κB、IL-1β、TNF-α的mRNA表达水平均显著升高(P<0.01或P<0.001);同时间点小半夏汤高剂量组胃窦和回肠组织中NF-κB、IL-1β、TNF-α的mRNA表达水平显著低于顺铂模型组(P<0.01或P<0.001)。造模后24 h和72 h,顺铂模型组大鼠胃窦组织中NF-κB、IL-1β、TNF-α的蛋白表达水平均显著升高(P<0.01或P<0.001),回肠组织中除TNF-α蛋白表达水平无明显变化外,NF-κB和IL-1β的蛋白表达水平亦显著升高(P<0.001);同时间点小半夏汤各剂量组大鼠胃窦和回肠组织中NF-κB、IL-1β、TNF-α的蛋白表达均不同程度低于顺铂模型组。结论:小半夏汤防治化疗性胃肠黏膜炎的机制可能与抑制胃肠道炎性因子表达有关。

Objective:To observe the effect of Xiaobanxia Decoction(XBXD) on nuclear factor-κB(NF-κB),interleukin-1β(IL-1β),tumor necrosis factor-ɑ(TNF-α) mRNA and protein expression levels in sinuses ventriculi and ileum of gastrointestinal mucositis model rats induced by cisplatin,so as to explore the mechanism of XBXD in preventing and treating gastrointestinal mucositis induced by chemotherapy. Methods:A total of112 male Wistar rats were randomly divided into control group,XBXD normal control group,cisplatin model group,omeprazole group,XBXD low dose group,XBXD middle dose group and XBXD high dose group,with 16 rats in each group. The rats in the control group and cisplatin model group were given distilled water 20 mL/kg a day by oral gavage. The rats in the XBXD normal control group were given XBXD1.6 g/kg a day by oral gavage. The rats in the omeprazole group were given omeprazole 2.4 mg/kg a day by oral gavage. The rats in the XBXD low,middle and high dose groups were given XBXD 0.8 g/kg,1.6 g/kg,3.2 g/kg a day by oral gavage respectively. The above dose was given to the rats at two separate time(morning and evening) and the medicines were given to the rats for 5 consecutive days. On the third day,the rats in the control group and XBXD normal control group were given equal volume of physiological saline and the rats in the other groups were given cisplatin 6 mg/kg to establish the gastrointestinal mucositis model. Twenty-four hours and seventy-two hours after modeling,the mRNA and protein expression levels of NF-κB,IL-1β,TNF-α in rats’ sinuses ventriculi and ileum were measured by PCR and Western Blot. Results:Twenty-four hours and seventy-two hours after modeling,compared with the control group,the NF-κB,IL-1β,TNF-α mRNA expression levels in rats’ sinuses ventriculi and ileum of the cisplatin model group were increased significantly(P<0.01 or P<0.001). At the same time,the NF-κB,IL-1β,TNF-α mRNA expression levels in rats’ sinuses ventriculi and ileum of the XBXD high dose group were significantly lower than those of the cisplatin model group(P<0.001). Twenty-four hours and seventy-two hours after modeling,the NF-κB,IL-1β,TNF-α protein expression levels in rats’ sinuses ventriculi of the cisplatin model group were increased significantly(P<0.01 or P<0.001). The NF-κB and IL-1β protein expression levels in rats’ ileum of the cisplatin model group were also increased significantly(P<0.001),but the TNF-α protein expression level in rats’ ileum of the cisplatin model group did not change obviously. At the same time,the NF-κB,IL-1β,TNF-α protein expression levels in rats’ sinuses ventriculi and ileum of XBXD low,middle and high dose groups were all lower than those of the cisplatin model group to varying degrees. Conclusion:The mechanism of XBXD in preventing and treating gastrointestinal mucositis induced by chemotherapy may be related to the inhibition of inflammatory factor expression.