Mitochondrial Separation Protein Inhibitor Inhibits Cell Apoptosis In Rat Lungs During Intermittent Hypoxia

Objective:In the experiment,the mechanism of mitochondrial cytochrome C pathway involved in the apoptosis of intermittent hypoxic cells and the effect of mitochondrial division inhibitor(Mdivi-1)on the apoptosis of intermittent hypoxic lung tissue were investigated?It provides a new target for studying the pathogenesis of OSAHS and seeking new prevention and treatment?Methods:32 healthy male SD rats were randomly divided into 4 equal size groups:control group(n=8,group A)?IH group(n=8,group B)?IH+25 mg/kg Mdivi-1 group(n=8,group C)?IH+50 mg/kg Mdivi-1group(n=8,group D)?The rats in group B?C?D were given nitrogen and compressed air,and the control group were given normoxic?Four weeks later,the rat model of intermittent hypoxic SD was established?The pathological changes and scores of HE staining,the level of apoptosis,and the expression of Drp1?Cyt-C?Caspase-9?Caspase-3?Bcl-2 m RNA and its protein detected by the RT-q PCRand WB in the lung tissue of each group were compared?WTRL1 cells were randomly divided into five groups:control group?IH group?IH +10 ? M Mdivi-1 group?IH +50 ? M Mdivi-1 group?IH+ 100 ? M Mdivi-1 group?The control group was exposed to normal chamber(CO2 5%,O2 21%),IH group?IH +10 ? M Mdivi-1 group?IH +50 ? M Mdivi-1 group?IH+ 100 ? M Mdivi-1 group were cultured in the chamber(CO2 5%,O221%,30 min;CO2 5%,O2 8% 30 min)given with nitrogen?oxygen and carbon dioxide,and this process was repeated for 12 h?The treatment groups were incubated with Mdivi-1for 30 min prior to IH culture?The expression of m RNA and protein in Drp1?Cyt-C?Caspase-9?Caspase-3?Bcl-2 of WTRL1 cells at 12 h after modeling was detected by RT-q PCR and Western Blot?After logarithmically cultured,WTRL1 cells were randomly divided into five equal size groups:control group(group A)?IH for 6 h group(group B)?IH for 12 h group(group C)? IH for 24 h group(group D)?IH for 48 h group(group E)?The control group was exposed to normal chamber(CO2 5%,O2 21%)for 48 h,WTRL1cells in group B?C?D?E were cultured in the chamber(CO2 5%,O2 21%,30 min;CO25%,O2 8% 30 min)given with nitrogen?oxygen and carbon dioxide at 6 h,12 h,24 h and48 h,respectively?After modeling,the apoptosis rate of WTRL1 cells in each group was detected by flow cytometry?Results:Following IH exposure for 4 weeks,the lung tissues of SD rats exhibited interstitial lesions,while Mdivi-1 reduced these pulmonary interstitial lesions?Bcl-2 m RNA and protein expression levels were decreased,however Caspase-3?Caspase-9 and Drp1 m RNA and protein expression levels were increased?Following Mdivi-1 intervention,Bcl-2m RNA and protein expression levels were increased while Caspase-3?Caspase-9 and Drp1 m RNA and protein expression levels were decreased(P<0.05)?After exposure to IH for 12 h,the apoptosis rate of WTRL1 cells in rats increased gradually with the IH time(P<0.05)? Bcl-2 m RNA and protein expression levels were decreased,whereas Caspase-3?Caspase-9?Cyt-C and Drp1 m RNA levels were increased,and Caspase-3?Caspase-9 and Drp1 protein expression levels were increased?After Mdivi-1 intervention,Bcl-2 m RNA and protein expression levels were increased but Caspase-3?Caspase-9?Cyt-C and Drp1 m RNA levels were decreased along with Caspase-9?Cyt-C and Drp1 protein expression levels which were decreased(P<0.05)?Conclusion: Mdivi-1 exerts protective effects against IH-induced apoptosis in lung tissue of SD rats,And the underlying mechanism may be through inhibiting Cyt-C-dependent mitochondrial apoptosis pathway.