摘要
目的:研究雷帕霉素对DEN所致肝损伤的保护作用及其对肝癌细胞HepG2的抑制作用。方法:(1)动物肝损伤模型制备及分组:将60只小鼠随机分为对照组(每日用蒸馏水灌胃)、DEN组(隔日腹腔注射DEN 30 mg/kg)、雷帕霉素组(隔日腹腔注射DEN 30 mg/kg的同时给予每日腹腔注射雷帕霉素10 mL/kg)。实验共进行8周,实验结束后采用脱颈椎法处死小鼠,对小鼠肝组织进行石蜡包埋、切片,采用SABC法检测肝组织中HER-1及HER-2的含量。(2)肝癌细胞凋亡实验:将HepG2细胞培养至对数生长期,利用25 nmol/L及50 nmol/L雷帕霉素进行不同药物浓度的培养,培养48小时后,在荧光显微镜下观察吖啶橙(AO)染色后HepG2细胞凋亡的形态学特征。结果:与DEN组比较,对照组及雷帕霉素组肝脏组织中HER-1的表达水平较低(P<0.01);对照组与雷帕霉素组HER-1的表达无差异。与DEN组比较,对照组及雷帕霉素组肝脏组织中HER-2的表达水平较低(P<0.01);对照组与雷帕霉素组HER-2的表达无差异。雷帕霉素可以促进肝癌细胞HepG2的凋亡,随着药物浓度的升高,HepG2的凋亡程度也随之加重。结论:雷帕霉素可以显著降低肝脏损伤过程中HER-1、HER-2的表达,同时可以促进肝癌细胞HepG2的凋亡,进而起到预防肝肿瘤、促进肿瘤细胞凋亡的作用。
Objective:To study the protective effect of rapamycin on DEN induced liver injury and its inhibitory effect on hepatoma cells HepG2.Methods:(1)Animal liver injury model preparation and grouping:60 mice were randomly divided into control group(daily distilled water intragastric injection of DEN 30 mg/kg),DEN group(every other day intraperitoneal injection of DEN 30 mg/kg),and rapamycin group(every other day intraperitoneal injection of DEN 30 mg/kg and daily intraperitoneal injection of rapamycin 10 mL/kg).The experiment was carried out for a total of 8 weeks.After the experiment,the mice were sacrificed by cervical vertebra removal method.The liver tissues of mice were paraffin embedded and sliced,and the contents of HER-1 and HER-2 in the liver tissues were detected by SABC method.(2)hepatoma cell apoptosis experiment:HepG2 cells were cultured to logarithmic growth stage,and rapamycin was cultured at 25 nmol/L and 50 nmol/L at different drug concentrations.After 48 h of culture,morphological characteristics of apoptosis of HepG2 cells were observed under fluorescence microscope after acridine orange(AO)staining.Results:Compared with DEN group,the expression level of HER-1 in liver tissues of control group and rapamycin group was lower(P<0.01).There was no difference in HER-1 expression between the control group and rapamycin group.Compared with DEN group,the expression level of HER-2 in liver tissues of control group and rapamycin group was lower(P<0.01).There was no difference in HER-2 expression between the control group and the rapamycin group.Rapamycin can promote the HepG2 apoptosis of hepatoma cells,and with the increase of drug concentration,the HepG2 apoptosis degree is also aggravated.Conclusions:Rapamycin can significantly reduce the expression of HER-1 and HER-2 in the process of liver injury,and promote the apoptosis of HepG2 in hepatoma cells,so as to prevent liver tu mors and promote the apoptosis of tumor cells.
作者
冯伟
李兆廷
FENG Wei;LI ZhaoTing(Linyi Mental Health Center,Linyi Shandong 276005)
