摘要
目的:探究清肝益脾胶囊通过抑制肝细胞铁死亡进而缓解抗结核药物性肝损伤的相关机制。方法:利用异烟肼(INF)和利福平(RFP)构建ATB-DILI小鼠动物模型。C57BL/6小鼠随机分为PBS对照组(NC组)、ATB-DILI组和Protectant组,分别给小鼠灌胃PBS、INH+RFP和INH+RFP+清肝益脾胶囊。实验采用血清酶学检测ALT、AST、ALP含量,流式细胞术检测小鼠的TMRE、ROS、脂质过氧化水平和Fe^(2+)浓度,RNA-seq测序检测基因表达情况,qPCR检测谷胱甘肽过氧化酶4(GPX4)和酯酰辅酶A合成酶长链家族成员4(ACSL4)的表达,Western blot检测GPX4和ACSL4蛋白表达,考察并记录小鼠体质量及生存状况,最后取小鼠肝脏作H&E染色行病理分析。结果:与ATB-DILI组小鼠相比,Protectant组小鼠生存期延长,生存率增加(P=0.001 3);Protectant组小鼠血清中ALT、ALP、AST含量较ATB-DILI组显著降低,有统计学差异(P=0.020 5,P=0.001 6,P=0.034 5);Protectant组小鼠脂代谢相关通路中的基因下调,其中,Fasn和Scd1显著下调(P=0.039 1,P=0.041 7);Protectant组小鼠的TMRE、ROS、脂质过氧化水平和Fe^(2+)浓度增加得到改善;与ATB-DILI组小鼠相比,Protectant组小鼠ACSL4蛋白表达显著降低(P=0.000 03)。结论:清肝益脾胶囊可以通过下调脂代谢相关通路中Fasn、Scd1、GPX4和ACSL4的表达,减少ALT、AST、ALP表达,降低肝脏铁死亡,从而改善肝脏脂代谢紊乱,缓解抗结核药物性肝损伤,显著提高模型小鼠的生存率。
OBJECTIVE To investigate the mechanism of Qinggan Yipi Capsules in alleviating anti-tuberculous drug-induced liver injury(ATB-DILI) by inhibiting hepatocyte iron death.METHODS Mouse model of ATB-DILI was established by intragastric administration of isoniazid and rifampicin.C57BL/6 mice were randomly divided into PBS control group(NC group),ATB-DILI group and Protectant group,and then given PBS,INH+RFP and INH+RFP+Qinggan Yipi capsules,respectively.The contents of ALT,AST and ALP were detected by serum enzymology.The levels of TMRE,ROS,lipid peroxidation and Fe^(2+) concentration were detected by flow cytometry.RNA sequencing was used to detect gene expression.The expression of glutathione peroxidase 4(GPX4) and ester acyl-CoA synthetase long chain family member 4(ACSL4) were detected by qPCR.The protein expression of GPX4 and ACSL4 were detected by Western blot.Body weight and survival status of mice were investigated and recorded.Finally,livers of mice were taken for pathological analysis after HE staining.RESULTS Compared with ATB-DILI group,Protectant group prolonged the survival time and increased the survival rate(P=0.001 3).The contents of ALT,ALP and AST in serum of Protectant group were significantly lower than those of ATB-DILI group(P=0.020 5,P=0.001 6,P=0.034 5).Genes in lipid metabolism related pathways were down-regulated in the Protectant group,among which Fasn and Scd1 were significantly down-regulated(P=0.039 1,P=0.041 7).The variation of TMRE,ROS,lipid peroxidation and Fe^(2+) concentration in Protectant group were improved.Compared with ATB-DILI group,the expression of ACSL4 protein in Protectant group was significantly decreased(P=0.000 03).CONCLUSION Qinggan Yipi Capsules can down-regulate the expression of Fasn,Scd1,GPX4 and ACSL4 in lipid metabolism related pathways,reduce the expression of ALT,AST and ALP,and improve the ferroptosis and dyslipidemia in liver,and then alleviate ATB-DILI and significantly improve the survival rate of model mice.
作者
季阿芳
宋全
潘韵芝
杨薇
程军平
JI Afang;SONG Quan;PAN Yunzhi;YANG Wei;CHENG Junping(Nanjing University of Chinese Medicine,Jiangsu Nanjing 210023,China;Department of Pharmacy,Infectious Disease Hospital Affiliated to Soochow University,Jiangsu Suzhou 215131,China)
出处
《中国医院药学杂志》
CAS
北大核心
2023年第13期1458-1464,共7页
Chinese Journal of Hospital Pharmacy
基金
第五批姑苏卫生人才培养项目(编号:GSWS2019069,GSWS2020094)
苏州市科技计划项目:民生科技科技示范工程(编号:SS202011)
苏州市科技计划项目:医疗卫生科技创新应用基础研究(编号:SKJY2021139)
江苏省药学会-天晴医院药学基金项目(编号:Q202027)。
