抗结核药物性肝损伤小鼠模型细胞角蛋白18与基质金属蛋白酶7的表达水平

Expression levels of cytokeratin 18 and matrix metalloproteinase-7 in mouse model with anti-tuberculous drug-induced liver injury

目的 分析抗结核药物性肝损伤(ATB-DILI)模型小鼠组织及血清中细胞角蛋白18(CK18)与基质细胞金属蛋白酶7(MMP-7)表达,探讨CK18与MMP-7在肝损伤发生中的临床价值。方法 6周龄SPF级雄性KM小鼠20只,体重18~24 g,按照随机区组法分为4组,每组5只。选择3组进行卡介苗(BCG)滴鼻使小鼠感染结核分枝杆菌,其中两组分别灌胃异烟肼(INH)45 mg/(kg·d)、利福平(RIF)90 mg/(kg·d),持续3周,记为INH 3周组及RIF3周组,剩余一组为BCG感染对照组;未经任何处理的为空白对照组。通过PCR检测小鼠肺组织BCG含量,病理观察小鼠肝损伤证实动物模型建立成功。比较各组肝脏指数、病变范围;比较各组小鼠血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、碱性磷酸酶(ALP)、γ-谷氨酰转移酶(GGT)、总胆红素(TBil)、CK18、MMP-7表达情况;比较各组小鼠肝脏组织CK18、MMP-7蛋白表达情况。结果 BCG感染对照组与空白对照组小鼠ALT、AST、ALP、GGT、TBil表达比较,差异无统计学意义(P>0.05);INH 3周组和RIF 3周组小鼠ALT、AST、GGT、TBil表达高于BCG感染对照组(P<0.05)。空白对照组、BCG感染对照组小鼠肝脏组织CK18、MMP-7蛋白表达比较,差异无统计学意义(P>0.05);INH 3周组、RIF 3周组小鼠肝脏组织CK18、MMP-7蛋白表达高于BCG感染对照组(P<0.05)。空白对照组、BCG感染对照组小鼠血清CK18、MMP-7表达比较,差异无统计学意义(P>0.05);INH 3周组、RIF 3周组小鼠血清CK18、MMP-7表达高于BCG感染对照组(P<0.05)。空白对照组、BCG感染对照组小鼠肝脏指数、病变范围比较,差异无统计学意义(P>0.05);INH 3周组、RIF 3周组小鼠肝脏指数、病变范围高于BCG感染对照组(P<0.05)。结论 CK18与MMP-7可能影响ATB-DILI,明确CK18与MMP-7表达水平,提前干预,可以降低ATB-DILI发生风险。

Objective To analyze the expression of cytokeratin 18(CK18) and matrix metalloproteinase-7(MMP-7) in tissues and serum of model mouse with anti-tuberculosis drug-induced liver injury(ATB-DILI),and to investigate the clinical value of CK18 and MMP-7 in the occurrence of liver injury.Methods Twenty 6-week-old SPF grade male KM mice,weighing 18-24 g,were divided into four groups according to random block method,with five mice in each group.Three groups were selected for Bacillus Calmette-Guérin(BCG) nasal drip to infect mice with Mycobacterium tuberculosis,and two groups were administrated with Isoniazid(INH) 45 mg/(kg·d) and Rifampicin(RIF) 90 mg/(kg·d),respectively,for three weeks,which were recorded as INH 3-week group and RIF 3-week group,and the remaining group was BCG infection control group;the untreated group was the blank control group.The BCG content of mouse lung tissue was detected by PCR,and the liver injury of mouse was observed by pathology.Liver index and lesion range were compared among all groups;the expressions of alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),γ-glutamyltransferase(GGT),total bilirubin(TBil),CK18,and MMP-7 in serum of all groups were compared;and the protein expressions of CK18 and MMP-7 in liver tissues of all groups were compared.Results There were no significant differences in the expressions of ALT,AST,ALP,GGT,and TBil between BCG infection control group and blank control group(P>0.05);the expressions of ALT,AST,GGT,and TBil in INH 3-week group and RIF 3-week group were higher than those in BCG infection control group(P<0.05).There were no significant differences in liver tissue protein expressions of CK18 and MMP-7 between blank control group and BCG infection control group(P>0.05);the liver tissue protein expressions of CK18and MMP-7 in INH 3-week group and RIF 3-week group were higher than those in BCG infection control group(P<0.05).There were no significant differences in serum expressions of CK18 and MMP-7 between blank control group and BCG infection control group(P>0.05);the serum expressions of CK18 and MMP-7 protein in INH 3-week group and RIF 3-week group were higher than those in BCG infection control group(P<0.05).There were no significant differences in liver index and lesion range between blank control group and BCG infection control group(P>0.05);and the liver index and lesion range of INH 3-week group and RIF 3-week group were higher than those of BCG infection control group(P <0.05).Conclusion CK18 and MMP-7 may affect ATB-DILI.Clarifying the expression levels of CK18 and MMP-7 and intervening in advance can reduce the risk of ATB-DILI.