The role of recombinant human annexin A6 in the repair of early plasma membrane damage in mouse skeletal muscle after eccentric exercise
Objective To explore the role of exogenous recombinant human annexin A6(rhANX A6) intervention in early plasma membrane repair of skeletal muscles in mice with eccentric motion injury.Methods Fifty-six 8-week-old C57 male mice were randomly divided into a resting control group(C,n=8),an exercise+PBS injection group(EP,n=24) and an exercise+rhANX A6 injection group(ER,n=24).Two hours before the exercise,both EP and ER groups were injected 1 × PBS buffer and rhANX A6(0.8 mg/kg) intraperitoneally,respectively.Both EP and ER groups underwent a 60-min downhill centrifugal running on the running platform of 15° slope,with an initial speed of 5 m/min,increased by 1 m/min up to 17 m/min.At 2 h,6 h and 12 h after exercise,8 mice were executed to collect samples.The serum creatine kinase(CK) levels were detected using enzyme-linked immuno-sorbent assay,while the positive area of Xin actin-binding repeat-containing protein (XIRP),the co-localization of annexin A6(ANX A6),annexin A1 (ANX A1) and annexin A2(ANX A2),as well as the pore size of plasma membrane repair mediated by ANX A6 were observed using immunofluores-cence.Results Compared with group C,a significant increase was observed in the average serum CK level at all sampling points in EP group and 12 h after exercise in ER group(P<0.01),with that of EP group significantly higher than ER group at all sampling points(P<0.05).Moreover,the percentag-es of Xirp staining positive area in EP group and ER group were significantly higher group C at all time points(P<0.05),with that of EP group 2 h and 12 h after exercise significantly higher than ER group(P<0.01).Meanwhile,the co-localization coefficients of ANX A6 and A1 increased significantly at all sampling points in both EP and ER groups(P<0.01),with those of EP group 6 h and 12 h af-ter exercise significantly higher than ER group(P<0.01).What's more,compared with group C,the co-localization fluorescence points of ANX A6 with A1 or A2 of EP group at all times and of ER group 6 h and 12 h after exercise increased significantly,with those of EP group 6 h and 12 h after exercise significantly higher than ER group(P<0.01).At 6 h after eccentric exercise injury,ANX A1 and A2 were mainly involved in ANX A6-mediated plasma membrane repair of a'large wound' of about 10 μm.Conclusion Exogenous rhANX A6 injected 2 h before exercise significantly promotes the early-stage repair of plasma membrane damage in skeletal muscle after eccentric exercise and has a positive effect on the formation of micron-sized'patches'.
eccentric exerciserecombinant human annexin A6skeletal muscle injuryplasma membrane repair
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