组蛋白乳酸化通过HDAC2影响抗结核药物性肝损伤炎症的机制研究

Histone lactylation affects anti-tuberculosis drug-induced liver injury inflammation through HDAC2

目的探讨组蛋白乳酸化促进剂和抑制剂草氨酸钠对抗结核药物性肝损伤(ADLI)炎症的影响。方法体外培养AML12细胞,采用CCK8法确定适宜的促进剂和抑制剂培养浓度后,设置对照组、三联药组、乳酸组、草氨酸钠组、三联药+乳酸组和三联药+草氨酸钠组。采用RT-PCR、Western Blot法检测细胞中HDAC2 mRNA和蛋白表达水平,微板法检测上清液丙氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST)活性。结果乳酸的培养浓度取8mmoL/L;草氨酸钠的培养浓度取18mmoL/L。与对照组相比,三联药组细胞HDAC2蛋白和mRNA表达水平明显降低,差异有统计学意义(P<0.05);与三联药组相比,HDAC2表达水平在三联药+乳酸组显著升高,而在三联药+草氨酸钠组表达降低,差异有统计学意义(P<0.05);NF-κB与HDAC2表达相反。与对照组相比,三联药组AST、ALT活性显著升高,差异有统计学意义(P<0.05);与三联药组相比,三联药+乳酸组的上清AST、ALT酶活性显著降低(P<0.05)。结论组蛋白乳酸化的促进剂乳酸和抑制剂草氨酸钠能影响ADLI炎症有关蛋白的表达和酶活性,乳酸可以缓解ADLI炎症,提示组蛋白乳酸化可能是治疗ADLI的新途径。

Objective To explore the effects of histone lactylation promoters and inhibitor sodium oxamate on anti-tuberculosis drug-induced liver injury(ADLI)inflammation.Methods AML12 cells were cultured in vitro,and the appropriate concentrations of promoter and inhibitor were determined by CCK8 method.Then,the cells were divided into control group,triple drug group,lactate group,sodium oxamate group,triple drug+lactate group,and triple drug+sodium oxamate group.HDAC2 mRNA and protein expression levels were detected by RT-PCR and Western Blot.Activities of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in the supernatant were detected by microplate method.Results The culture concentration of lactate was 8mmol/L,and the culture concentration of sodium oxamate was 18mmol/L.Compared with the control group,the protein and mRNA expression levels of HDAC2 in the triple drug group were significantly lower(P<0.05);compared with the triple drug group,the expression levels of HDAC2 in the triple drug+lactate group were significantly higher(P<0.05),while the expression levels of HDAC2 in the triple drug+sodium oxamate group were lower(P<0.05).NF-κB was expressed opposite to HDAC2.Compared with the control group,the activities of AST and ALT in the triple drug group were significantly higher(P<0.05);compared with the triple drug group,the activities of AST and ALT in the triple drug+lactate group were significantly lower(P<0.05).Conclusion Histone lactylation promoters lactate and inhibitor sodium oxamate can affect the expression and enzyme activity of ADLI inflammation related proteins.Lactate can alleviate ADLI inflammation,indicating that histone lactylation may be a new pathway for treating ADLI inflammation.