高压氧联合阿霉素处理对裸鼠肝癌移植瘤HepG2细胞凋亡及免疫调节因子的影响

Effects of hyperbaric oxygen combined with Adriamycin on HepG2 cell apoptosis and immunomodulatory factors in transplanted hepatocellular carcinoma of nude mice

目的:探索高压氧(HBO)联合阿霉素(adriamycin,Adc)处理对裸鼠肝癌移植瘤HepG2细胞凋亡及免疫调节因子的影响。方法:构建裸鼠肝癌移植瘤模型成功后,按照实验设计分为4组:对照组、HBO组、Adc组及HBO+Adc组,每组5只。对照组正常饲养,HBO组在0.20 MPa压力下吸氧1 h;Adc组予以10 mg/kg Adc灌胃处理;HBO+Adc组首先在0.20 MPa压力下吸氧1 h,再予以10 mg/kg Adc灌胃处理。隔天操作处理1次,共处理21 d。停药后处死裸鼠,剥离肿瘤,测量瘤体体积和重量;TUNEL法检测瘤体HepG2细胞凋亡情况;Western blotting实验检测肝癌HepG2细胞凋亡相关蛋白的变化;ELISA法检测裸鼠肝癌移植瘤组织中免疫调节因子含量的变化。结果:处理21 d后,HBO+Adc组裸鼠肿瘤体积和重量均小于对照组、HBO组和Adc组( P<0.05);与其他3组比较,HBO+Adc组裸鼠移植瘤HepG2细胞凋亡比例最高( P<0.05);Bax、caspase-3、Cytochrome C蛋白表达水平显著升高,Bcl-2蛋白表达水平明显降低( P<0.01);HBO+Adc组IL-6明显降低,IL-18及IFN-γ明显升高( P<0.05)。 结论:HBO联用Adc处理可促进裸鼠肝癌移植瘤HepG2细胞凋亡,能够有效调节机体免疫调节因子,促进机体免疫功能的恢复,具有显著的增敏减毒作用。

Objective To explore the effects of adriamycin(Adc)combined with hyperbaric oxygen(HBO)on the apoptosis of HepG2 cells and immunomodulatory factors in transplanted hepatocellular carcinoma(HCC)tissues in nude mice.Methods The model of transplanted HCC was established in 20 nude mice,then the mice were divided into four groups:control group,HBO group,Adc group,and HBO+Adc group,with five mice in each group.The control group was reared normally.The HBO group was treated with HBO at 0.20 MPa for 1 h;the Adc group was treated with 10 mg/kg of Adc by gavage;the HBO+Adc group was treated with HBO at 0.20 MPa for 1 h and then given 10 mg/kg of Adc by gavage.All groups were treated once every other day for 21 days.After drug withdrawal,the tumors were dissected from the sacrificed mice,of which the tumor volume and weight were measured.The apoptosis of HepG2 cells was detected by TUNEL.The changes of apoptosis-related proteins in HepG2 cells were detected by western blotting.The changes of the contents of immunomodulatory factors in transplanted tumor tissues of nude mice were detected by ELISA.Results After 21 days,the tumor volume and weight in the HBO+Adc group were lower than those in the other three groups(P<0.05).The proportion of apoptotic cells in the transplanted tumor tissues of nude mice of the HBO+Adc group was the highest.In the HBO+Adc group,the protein expression levels of Bax,caspase-3,and Cytochrome C were significantly increased;the protein expression of Bcl-2 was significantly reduced(P<0.01);and the IL-6 was significantly reduced;while the IL-18 and IFN-γwere significantly increased(P<0.05).Conclusion The Adc combined with HBO can promote apoptosis of HepG2 cells in transplanted HCC tissues in nude mice,and effectively regulate the immunomodulatory factors to promote the recovery of the body’s immune function,suggesting obvious sensitizing and detoxifying effects.