Exploring The Repair Of Plantamajoside Based On Transcriptone Sequencing Effect Of Exhaustive Skeletal Muscle Injure

Objective: The protective effect and mechanism of Plantamajoside(PMS)on exhaustive exercise-induced skeletal muscle injury were investigated.Methods : Six-week-old male SD rats were randomly divided into control group(Control group),exhaustive exercise group(Model group),exhaustive exercise +low-dose PMS group(ML group),exhaustive exercise + high-dose PMS group(MH group),6 rats in each group.The control group was fed normally,the ML group was given a dose of 7 mg / kg / d,and the MH group was given a dose of 28 mg / kg / d for two weeks.Exhaustive skeletal muscle injury model was established in the second week of the model group,ML group and MH group.The initial speed was 10 m / min,lasted for 10 min,then added to 15 m / min for 10 min,and then added to 20 m / min exercise to exhaustion,training for 7 days.The next day after the training,the blood was taken after intraperitoneal anesthesia,and the gastrocnemius muscle was quickly separated.HE staining was used to observe skeletal muscle injury;the oxidative stress of skeletal muscle was observed by ROS staining.Differential genes were screened by transcriptome sequencing in Control group and Model group.Serum creatine kinase,lactate dehydrogenase,superoxide dismutase,glutathione peroxidase activity and malondialdehyde content were determined by ELISA.The protein expression of PI3 K,AKT,Keap1,Nrf2,HO-1,Bcl-2,Bax,Caspase-3,Caspase-9,FBXO31,CDKN1 A and CCND1 in skeletal muscle was determined by Western Blot.Results:(1)Compared with the Control group,the staining results in the Model group showed that the arrangement of muscle fibers was significantly disordered,the red fluorescence intensity of the nucleus was enhanced,and the apoptosis was increased.The results of ELISA showed that the contents of CK,LDH and MDA in the serum of the Model group were significantly increased,and the activities of SOD and GSH-Px were significantly decreased.WB results showed that the expression of Keap1,Bax,Caspase-3 and Caspase-9 protein in gastrocnemius muscle of Model group increased significantly,while the expression of Nrf2,HO-1,Bcl-2,PI3 K and AKT protein decreased significantly.(2)By analyzing the transcriptome genes of the Model group and the Control group,205 up-regulated genes and 135 down-regulated genes were found.FBXO31,CDKN1 A and CCND1 were further screened by GO analysis.Western Blot was used to predict gene expression.It was found that the level of CCND1 protein in the Model group was significantly down-regulated,and the levels of FBXO31 and CDKN1 A protein were up-regulated,which was consistent with the sequencing results.(3)Compared with the Model group,the gastrocnemius deformity cells in the ML and MH groups were reduced,arranged more neatly,the red fluorescence staining of the nucleus was reduced,and the apoptotic cells were reduced;the expression of CK,LDH and MDA in serum decreased,and the activity of SOD and GSH-Px increased.The expression of Nrf2,HO-1,Bcl-2,PI3 K and AKT protein in gastrocnemius muscle increased,and the expression of Keap1,Bax,Caspase-3 and Caspase-9 protein decreased.In addition,the level of CCND1 protein in gastrocnemius muscle increased after PMS intervention,and the levels of FBXO31 and CDKN1A protein were down-regulated.(4)Compared with ML group and MH group,HO-1 protein expression was found after low dose PMS intervention.Conclusion:(1)Through gene sequencing,FBXO31,CDKN1 A and CCND1 were significantly enriched.(2)The expression of CDKN1 A and CCND1 protein may be regulated by up-regulating the expression of PI3 K,AKT,Nrf2,HO-1 and other proteins after PMS intervention in exhaustive exercise model,thus reducing the expression of apoptotic proteins in skeletal muscle of exhaustive exercise rats.(3)Low-dose PMS may have a better effect on the repair of skeletal muscle injury in exhaustive exercise rats.