Effect Of Acupuncture And Moxibustion On Chronic Inflammatory Response And NLRP3 Inflammasome Of Leydig Cells In Aged Rats With Androgen Deficiency

ObjectiveTo investigate the effect of acupuncture and moxibustion on the serum total testosterone(TT)and free testosterone(FT)levels,the nuclear factor kappa B(NF-κB)chronic inflammation pathway and the NLRP3 inflammasome pathway of leydig cells in aged rats with androgen deficiency,and to explore the possible mechanism of delaying the aging of leydig cells.MethodsThe serum TT and FT were detected in 8 young(2 months old)male SD rats,and the 95% confidence interval was calculated.FT and TT were detected in 50 aged(20 months old)male SD rats.Forty aged rats were randomly divided into five groups according to their body weight: acupuncture and moxibustion group(AM),mild moxibustion group(MM),electroacupuncture group(EA),medicine group(MC),and old control group(OC).Eight young rats were randomly selected as the young control group(YC),with 8 rats in each group.The EA group took "Shenshu"(BL23)"Guanyuan"(CV4)acupoints for treatment once a day,15 minutes each time,total treatment for 8 weeks.The acupoints in the AM and MM groups were same with the EA group.The treatment time and course were the same as those of the EA group.The MC group received a subcutaneous injection of testosterone propionate(7 mg/kg every third day)for 8 weeks.OC group and YC group were given 0.9% sodium chloride solution subcutaneously in the abdomen.The injection dose and course were the same as those of the MC group.The time of exhaustive swimming in each group was observed before and after treatment.Serum TT and FT levels were measured by enzyme-linked immunosorbent assay(ELISA).The expression of Caspase-1 protein in Leydig cells was detected by immunohistochemistry.NF-κB p 65,NLRP3,Caspase-1,IL-1 β,TNF-α,and COX-2 protein and mNRA expression was detected by Western blot and Real-time quantitative polymerase chain reaction detecting system(RT-qPCR).Results1.Before treatment,the exhausted swimming time of OC group was lower than that of YC group(P<0.01);after treatment,the exhausted swimming time of AM group,MM group,EA group and MC group was higher than that of OC group and lower than that of YC group(P<0.01).Before treatment,serum TT and FT levels in OC group were lower than those in YC group(P<0.01);after treatment,serum TT and FT levels in AM group,MM group,EA group and MC group were higher than OC group and lower than YC group(P <0.01).2.Compared with the YC group,the protein and mNRA expression of NF-κB p 65,NLRP3,Caspase-1,COX-2,IL-1 β and TNF-α in the OC group was significantly up-regulated(P<0.01).Compared with OC group,the protein and mNRA expression of NF-κB p 65,NLRP3,Caspase-1,COX-2,IL-1β and TNF-α were significantly down-regulated in AM,MM and EA groups(P<0.01).There was no significant difference between MC group and OC group(P>0.05).ConclusionAM,MM and EA groups can improve serum TT,FT levels and exhausted swimming time in aged rats with androgen deficient,and can reduce the inflammatory response of testis in aged rats.This may be one of the mechanisms of acupuncture and moxibustion to improve the male reproductive senescence.