The Effect Of Acupuncture Through The TRP Channel On The Activity Of ?7?1 Integrin After Eccentric Exercise-induced Skeletal Muscle Injury

1.ObjectiveTo observe the protein changes of?7 integrin,?1 integrin,total FAK and phosphorylated p-FAK³⁹⁷ and the collagen area changes of extracellular matrix and endomysium recovery distribution changes in skeletal muscle of rats after an acute eccentric exercise,and to analyze the signal pathway of?7?1 integrin on the basement membrane remodeling of extracellular matrix of skeletal muscle,and to reveal the therapeutic mechanism of acupuncture effect on skeletal muscle injury.2.Methods72 healthy male Wistar rats are randomly divided into the control group?C?,exercise group?E?,exercise acupuncture group?EA?and exercise acupuncture impedance group?EAI?.All groups except group C are trained for 90 min of eccentric exercise:treadmill slope-16°,speed 16m·min^-1,5 min per group,18 groups in total,rest between groups 2 min.Acupuncture treatment:?1?after the exercise of group E,no recovery measures are applied.?2?In the EA group,the middle femoral muscle of rats is punctured once 40 min after exercise,and the effect of once acupuncture is observed.?3?Immediately after exercise,the EAI group is injected intraperitoneally with a physiological saline solution containing 2%GdCl₃?calcium channel blocker?(40?M·kg^-1),and acupuncture is performed 40 min later.The acupuncture treatment is the same as that of the EA group,and the blocking effect is observed once.The vastus internus muscle is collected from each group 1 h,48 h and 120 h after eccentric exercise training.The expressions of?7 integrin,?1integrin,total FAK and phosphorylated p-FAK³⁹⁷ are tested by Western blot,the changes of collagen area in extracellular matrix of skeletal muscle are observed by transmission electron microscopy,and the changes of endomysium recovery distribution in extracellular matrix are observed by scanning electron microscopy.3.Results3.1 Changes in the expression of?7 integrin and?1 integrin in skeletal muscle:?1??7 integrin:full-length?7 integrin?120 kDa?and cleaved?7 integrin?70 kDa?expression levels in each group andphase are consistent with those in the group C,without significant change.?2??1 integrin:Compared with group C,the peak value of?1 integrin in group E is significantly different at 48 h?P<0.05?,and EA group shows extremely significant increased at 1 h?P<0.001?and significantly increased at 48 h?P<0.05?.The EA group reaches its peak at 1 h,which shows a very significant increase compared with the E group?P<0.001?.Acupuncture advances the expression of?1integrin earlier.3.2 Changes in the expression of phosphorylated p-FAK³⁹⁷ and total FAK in skeletal muscle:?1?P-FAK³⁹⁷:Compared with group C,the expression of p-FAK³9797 in group E,EA and EAI are lower than that in group C,and show significant differences?P<0.05?.The expression of p-FAK³⁹⁷ in group E shows a trend of rising first and then decreasing.The expression of p-FAK³⁹⁷ reaches a peak at 1 h in the EA group,and increases significantly compared with the E group?P<0.05?.Compared with the EA group,the expression of p-FAK³⁹⁷ in the EAI group decreases significantly at 1 h and 48 h?P<0.05,P<0.05?.Acupuncture activates FAK activity in advance.?2?Total FAK:The expression level of total FAK in each group and phase remains the same as that in the group C,without significant changes.3.3 Changes in the area of the extracellular matrix collagen in skeletal muscle:Compared with group C,the number of collagen fibers and the area of collagen increase in group E,reaching a peak within 48 h,and the EA group inhibits collagen proliferation,showing a trend of rising first and then decreasing.Compared with the E group,the collagen area in the EA group is significantly decreased at 1 h and 48 h?P<0.001?.Compared with the EAI group,the EA group is significantly increased at 48 h?P<0.01?,but significantly decreased at 120 h?P<0.01?.3.4 Changes in recovery distribution of skeletal muscle endomysium:in group C,the collagen fiber network of skeletal muscle is uniformly and completely cross-linked,with random distribution.In the 120 h E group,skeletal muscle endomysium injury is not completely recovered,and there is a large collagen fiber cross-linking vacancy in part of the endomysium network.In the120 h EA group,the endomysium injury is reduced,and there is less collagen fiber cross-linking vacancy.In the 120 h EAI group,skeletal muscle endomysium injury is obvious,and the vacancy of collagen fiber cross-linking is more serious.4.Conclusion4.1 Acute eccentric exercise inhibits?7?1 integrin activity of skeletal muscle.4.2 Acupuncture can regulate the expression of skeletal muscle?7?1 integrin through TRP channels,promote skeletal muscle injury repair and remodeling of extracellular matrix.