Herbal Fumigation Of Rat Denervated Experimental Study Of Skeletal Muscle Injury And Repair

Objective:To study the mechanism of Xun xi yi haofang on the recovery of denervated skeletal muscle.Methods:90 male SD rats,10%Chloral Hydrate abdominal anesthesia,adopt emoribus internus incision,in the right femoribus internus severed femoral nerve.These rats were randomly divided into the following four groups after modeling successed and each group have 30 rats:Xun xi yi haofang group,Salvia miltiorrhiza group and model group.30 rats in control group.Experimental animals in each group after 7 days,14 days and 21 days were killed,to draw the materials from rectus femoris venter musculi tissue, conventional HE staining,to observe ultrastructural of denervated skeletal muscle ultramicrostructure with transmission electron microscopy after changes.NGF and bFGF immunohistochemical staining,and skeletal muscle homogenates of Na ~+-K~+ -ATP changes.Results:1.HE staining Results:After one week,early denervated skeletal muscle in majority of myofilament inocomma were still rules, can be seen staining weaken.Xun xi yi haofang group morphological integrity.Salvia miltiorrhiza group myocyte membrane shrinkage.After two weeks,surround of muscle fibers without proliferation of collagen fibers.Salvia miltiorrhiza group myocyte invaginated.Model group and the control group muscle spindle atrophy.After three weeks,Xun xi yi haofang group,myocyte thiner,model group myofilament fragmentation and derangemenin,the volum of cellular radually shrinked,stain deepened.Control group myocyte shrinkage,irregular margins relatively larger than control group,surrounding of muscle fibers appeared more fibroblast and the proliferation of collagen fibers.2.Acetylcholine activity:After one week,acetylcholine activity in every group is weak,Salvia miltiorrhiza group and model group compared to control group(P＜0.05)Xun xi yi haofang group compared to control group(P＞0.05).After two week,acetylcholine activity in Xun xi yi haofang group increased,but compared with the control group was not significant(P＞0.05),model group and Salvia miltiorrhiza group staining weaken, compared to control group(P＜0.01).After the 3 week,Xun xiyi haofang group acetylcholinesterase activity strong,and closer to normal,stained uniform, myoceptor structure rules,Similar in elliptical,or oval-shaped structure,compared to control group(P＞0.05);Model group and Salvia miltiorrhiza group acetylcholinesterase activity weak,compared to control group(P＜0.01).3.Immunohistochemical staining:(1)The expression of NGF:After seven days,Xun xi yi haofang group,muscle spindle myocyte staining positive, fibroblasts,vascular endothelial cells were weak,Salvia miltiorrhiza group muscle spindle myocyte were weakly positive.Model group muscle spindle smaller,there was no obvious staining.After 14 days,Xun xi yi haofang group,myocyte cellular nucleus not smooth,staining positive,fibroblast proliferation and staining positive strongly,vascular endothelial cells positive staining,muscle spindle no obvious staining,Salvia miltiorrhiza group myocyte cellular nucleus invagination,positive staining and fibroblasts positive staining.Model group and control group muscle spindle atrophy,negative staining.After 21 days,Xun xiyi haofang group,staining thin,endocytoplasmic reticulum expansion and weak positive,Salvia miltiorrhiza group myocyte cytolemma shrinkage and no staining.(2)expression of bFGF: After seven days,Xun xi yi haofang group myocyte staining positive,Salvia miltiorrhiza group and model group fibroblast positive staining.After 14 days,Xun xi yi haofang group skeletal muscle basilemma staining strong positive,fibroblast cells staining positive,a small amount of myocyte staining positive in muscle spindle,Salvia miltiorrhiza group nucleolemma staining was positive,fibroblasts stained positive in muscle spindle,model group positive weak.After 21 days,Xun xi yi haofang group skeletal muscle basilemma staining positive,fibroblasts staining strongly positive in the nerve endings,Salvia miltiorrhiza group with mitochondrial vacuolization positive cells scattered,endotheliocyte positive staining in muscle spindle,model group and control group not.5.Skeletal muscle homogenates Na~+-K~+-ATP results:After seven days,Na~+-K~+-ATP in all group there were no statistically significant(P＞0.05)compared with each other.After 14 days Xun xi yi haofang group compared with the control group(P＜0.05).After 21 days,Xun xi yi haofang group compared with control group(P＜0.01),and the content of Na~+-K~+-ATP enzyme in Xun xi yi haofang group was the highest.Conclusion:1.The repair ofdenervated skeletal muscle injury related to the changes of NGF and bFGF,skeletal muscle will atrophy if loss the nutrition of neurotrophic factor(NGF),bFGF can promote regeneration of the motorius neurofibra,plays an important role to the formation of neuromuscular junction,can promote recovery of muscle function after nerve injury.2.Xun xi yi haofang can delay degeneration of neuromuscular motor endplates(Motor endplate,MEP)and the effector.3.Xun xi yi haofang can improve denervated skeletal muscle nutrition supply and delayed skeletal muscle atrophy.