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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