Tacrolimus And Ciliary Neurotrophic Factor Promote The Regeneration Of Peripheral Nerve Injury

Objective:Related Research had shown that single tacrolimus(FK506)or single ciliary neurotrophic factor(CNTF)could promote the regeneration of peripheral nerve injury,but the two combination whether to promote the regeneration of peripheral nerve injury as well as the two had cooperativity was unclear.This study is to research the FK506 and CNTF on regeneration of peripheral nerve injury.Methods :80 healthy male SD rats(SPF)were selected for the first time.The rats were aged for about 8 weeks.The body weight was 200-220 g,and were kept in the 12 h day and night rhythm.The laboratory temperature was maintained at(23±2)?.The rats were randomly divided into the control group,the FK506 group,the CNTF group and the FK506+CNTF group,20 rats in each group.All rats were operated on the left thigh and the right thigh was normal control.Preoperative routine skin preparation,anesthesia with 10% chloral hydrate intraperitoneal injection of 35mg/kg,stable anesthesia after male SD rats were fixed on the operating table,iodophor disinfection,in rats after the left femoral lateral line longitudinal incision,since entered the muscle gap,blunt separation unit two biceps,expose the sciatic nerve was about 2.5cm in Pear myocutaneous holes under surgical microscope at approximately 1cm below the complete transection of sciatic nerve,with 11-0 suture epineurial suture line continuous suture needle 4,then sutured the muscles and skin wound closure.Postoperative prophylactic application of Cefamandole Nafate for Injection 100mg/kg intramuscular injection once,prevention of infection.four groups of SD rats were reared in the same environment.Two days after operation,all rats were treated with intramuscular injection of muscle below the femoral head in first day.The NS group were given 0.9% physiological saline 1mg/kg/d,The FK506 group were given FK506 0.5mg/kg/d;The CNTF group were given 100ng/kg/d;The FK506+CNTF group were given FK506 0.5mg/kg/d and 100ng/kg/d,consecutive 14 days.Postoperative using general observation,footprint analysis,wet weight of gastrocnemius muscle and nerve morphology,tissue electrical physiological examination(HE staining)and immunohistochemistry staining(S100a and NF200)evaluated the situation of nerve regeneration and functional recovery.Results:All the incisions of 80 rats healed well without infection.The FK506 group and the FK506+CNTF group maybe due to the side effects of immunosuppressive agents were killed.78 models got into the result analysis.In the FK506 group and the FK506+CNTF group,there was a small amount of diarrhea in rats,which gradually meliorated after second weeks.Within fourth week after modeling,the left foot of each group was found to be red and swollen.After sixth week,the left foot ulcers in the combined group were significantly lower than those in other groups.After eighth week,the left foot ulcer scar healing was observed in the combined treatment group,and the left foot ulcer surface of the NS group were reduced.At twelfth week,the left foot ulcers were healed in the four groups.Before drawing materials,observing the anastomosis around,the NS group and the CNTF group had a small amount of rats anastomosis which had the formation of neuroma,the formation was not found in the FK506 group and the FK506+CNTF group.All rats did not suture off.Early postoperative all rats left limb gangled,left knee disorderly flexed.At the twelfth week after operation,before drawing materials,the flexion disorder of the left knee joint in the combined treatment group significantly meliorated,and the pain responses were more sensitive.The NS group did not significantly meliorated and the reactions were slow.After modeling,at fourth,eighth,Twelfth week after sciatic nerve index were measured and compared in each group:normal gait foot was right foot after the claw toe at full stretch,and the operative side left foot footprints were that the left foot after the claw toe gangled and adducted.At fourth week,At eighth week and twelfth week,the sciatic nerve indexes of the FK506+CNTF group were significantly more decreased than the other three groups,the difference was statistically significant(P<0.05).Compared with the normal gastrocnemius muscle in the experimental group,the gastrocnemius of the experimental group was significantly reduced,and the degree of atrophy in the NS group was the most obvious.There was no significant difference between the FK506 group and the CNTF group.The combination group had the least degree of atrophy.In each group,the degree of gastrocnemius atrophy in experimental rats was gradually improved as time went on.And electrophysiology,HE staining and S100 a NF200 staining showed that the nerve regeneration and function recovery of the combination group is better than the other three groups,there was no significant difference between the FK506 group and the CNTF group,but was better than that of the NS group.Conclusions:Results suggest the combination of CNTF and FK506 can promote the regeneration of peripheral nerve injury,and they have cooperativity.