Effect Of Resveratrol On Soft Tissue Injury Of Hind Limb Induced By Agkistrodon Venom In Rats

Objective: To investigate the therapeutic effect of resveratrol on muscle injury induced by agkistrodon halys venom to explore the mechanism of its therapeutic effect.Methods: 60 healthy SD rats were randomly divided into 4 groups: control group,snake venom group,RES groupand inhibitor group,with 15 rats in each group.The toxic model was reproduced by injecting venom(10mg/kg)into the tibialis anterior muscle of the hind leg of rats.The tibialis anterior muscle of the control group was injected with equal amount of normal saline.The rats in the snake venom group,anti-venom serum was injected into the tail vein immediately after snake venom injection for 3 consecutive days(350U/kg).The rats in the RES group,antivenin was injected into the tail vein within 10 min after snake venom injection and RES was given by gavage.The antivenin was administered continuously for 3 days and RES was given by gavage for 7 days.The rats in the inhibitor group,EX527(5mg/kg)was injected into the tail vein for 30 min,then antivenin was injected into the tail vein and RES was gavaged immediately.Antivenin was administered continuously for 3 days,RES was gavaged for 7 days,EX527 was injected into the tail vein 30 min before administration of RES.The changes of bilateral leg circumference and muscle injury area were measured daily in each group.The serum levels of interleukin-6(IL-6)were detected by enzyme-linked immunosorbent assay(ELISA).The tibialis anterior muscle tissue was sacrificed to observe the pathological changes of the muscle,and the expression level of SIRT1 protein was detected by western blot(WB).Results:(1)After 1,3 and 7 days of intervention,compared with the control group,necrosis wounds were formed in snake venom group,resveratrol group and inhibitor group(P<0.05).Compared with snake venom group and inhibitor group,the necrosis wound surface formed in RES group was significantly smaller(P<0.05),and the leg circumference was also significantly decreased(P<0.05).(2)Compared with the control group,the activities of CK,LDH and AST in each group were significantly increased(P<0.05),and the values of CK,LDH and AST in each group were gradually increased with the prolongation of time in the first 3 days,and the expression level on the 7th day was lower than that on the 3rd day.Compared with snake venom group and inhibitor group,the expression levels of CK,LDH and AST in RES group were significantly decreased(P<0.05).(3)Compared with control group,serum IL-6 level in each group was significantly increased(P<0.05).On day 7,IL-6 expression level in RES group was significantly lower than that in snake venom group and inhibitor group(P<0.05).(4)Compared with the control group,the destruction of striated muscle structure,disorder of muscle cell arrangement and inflammatory cell infiltration were observed in all groups.Compared with snake venom group and inhibitor group,RES intervention significantly reduced the damage of striated muscle structure,disorder of muscle cell arrangement and inflammatory cell infiltration.(5)The expression of SIRT1 was significantly up-regulated after RES intervention(P<0.05),but was inhibited after EX527.Conclusion: RES can significantly reduce soft tissue edema and necrosis of hind limbs in rats induced by agkistrodon venom,and the mechanism may be related to the activation of SIRT1 signaling pathway by RES.