Study On The Mechanism Of Glucocorticoid Promoting Apoptosis Of Mice Hepatocytes Through Mitochondrial Pathway

Objective:GC has been widely used in the treatment of connective tissue diseases such as SLE due to its powerful anti-inflammatory effect to control disease activity.However,the treatment process is often accompanied by recurrence of disease activity and organ damage,and the cause has not been elucidated.Numerous studies have confirmed that GC can induce apoptosis,but the mechanism is unknown.The aim of this study was to explore the possible pathogenesis of GC mitochondrial pathway regulation promoting apoptosis,and to explore the therapeutic effect of mitochondrial targeted antioxidant on GC pro-apoptotic effects.Method : The effect of GC on apoptosis of AML12 cells was investigated by direct intervention with DEX;Gc-treated AML12 hepatocytes were treated with HPOB to inhibit GR translocation into mitochondria,and the effect of GR mitochondrial translocation on apoptosis of AML12 cells was studied.The effect of GC on the apoptosis of AML12 cells was verified by silencing BAX by si RNA.Mito Q was used to intervene GC-treated AML12 hepatocytes to investigate the effect of antioxidant intervention on the level of apoptosis in AML12 cells.Results:1.Gc-induced GR translocation into mitochondria in AML12 cells increased,the expression of mitochondrial genes MT-CO1 and MT-CO3 decreased,the activity of complex IV decreased,the MMP decreased,the level of ROS increased,the expression of pro-apoptotic gene BAX increased,and the expression of anti-apoptotic gene Bcl-2decreased,Annexin v-pe positive cells were increased,cytochrome C in the cytoplasm was increased,and cytochrome C in the mitochondria was decreased.2.HPOB inhibited mitochondrial translocation of GR,increased the expression of mitochondrial genes MT-CO1 and MT-CO3,increased the activity of complex IV,increased MMP,decreased ROS level,decreased the expression of pro-apoptotic gene BAX,increased the expression of anti-apoptotic gene Bcl-2,decreased cytochrome C in the cytoplasm,and increased cytochrome C in the mitochondria,Annexin v-pe positive cells were reduced.3.After si RNA gene silencing BAX,mitochondrial gene MT-CO1 and MT-CO3 expression increased,complex IV activity increased,MMP increased,proapoptotic gene BAX expression decreased,anti-apoptotic gene Bcl-2 expression increased,cytochrome C decreased in cytoplasm,cytochrome C increased in mitochondria.Annexin v-pe positive cells decreased.4.The antioxidant Mito Q targeted inhibition of ROS,increased mitochondrial genes MT-CO1 and MT-CO3 expression,increased complex IV activity,increased MMP,decreased ROS level,decreased pro-apoptotic gene BAX expression,increased anti-apoptotic gene Bcl-2 expression,decreased cytochrome C in the cytoplasm,and increased cytochrome C in the mitochondria.Annexin v-pe positive cells were reduced.Conclusions:GC induces apoptosis in the mitochondrial pathway,which may be a new pathogenesis of organ damage.The combination of Mito Q and GC can reduce the pro-apoptotic effect of GC in the mitochondrial pathway,suggesting that antioxidant therapy may be an effective method for the treatment of connective tissue diseases.