Effects Of Electroacupuncture On Oxidative Stress ERK/Nrf2-ARE Pathway Of Leydig Cells In Aged Rats With Low Testosterone

ObjectiveTo observe the effects of electro-acupuncture on serum testosterone content and ERK / Nrf2-ARE Antioxidant stress pathway of leydig cells in aged rats with low testosterone,in order to improve the antioxidant function of Leydig cells in aged testosterone-lower rats,so as to improve the low testosterone aged rats total testosterone(TT)and free testosterone(FT)content as a starting point,explore the related mechanism of electro-acupuncture in delaying the senescence of leydig cells.MethodsThe serum TT and FT of 6 male SD young rats(2 months old)were measured,and the 95% confident interval was calculated.The serum FT and TT of 24 male SD old rats(20 months old)were detected,and they were included in the treatment when they were below this interval.The 18 aged rats included in the experiment were randomly divided into agedcontrol group(AC),drug group(DR)and electro-acupuncture group(EA)according to their body weight,with 6 rats in each group,Eight young rats were used as the young control group(YC).In the EA group,bilateral "Shenshu"(BL23)and "Guanyuan"(CV4)acupoints were intervened,once a day,15 minutes each time,continuous treatment five days a week with two days off,total treat for 8 weeks.The DR group received a abdominal subcutaneous injection of testosterone propionate injection(7 mg·kg-1·3d-1)for 8 weeks.YC group and AC group were given subcutaneous injection of sodium chloride solution,dose and course of treatment were the same as DR group.The time of swimming exhaustion was observed before and after treatment.Serum TT and FT levels were tested with enzyme-linked immunosorbent assay.The activity of phosphorylated extracellular signal regulating protein kinase(p-ERK)and Kelch-like ECH-associated protein-1(Keap1)in Leydig cells was detected by immunohistochemistry(IHC).The protein expressions of rat testicular Nrf2,ERK,p-ERK,Keap1,HO-1,NQO1,SOD1 were detected with Western blotting.ResultsBefore treatment,the swimming exhaustion time of AC group was lower than that of YC group(P<0.01),and after treatment,the swimming exhaustion time of DR group and EA group was higher than that of AC group(P< 0.01).Before treatment,serum TT and FT levels of AC groupwere lower than that of YC group(P< 0.01),and serum TT and FT levels of DR group and EA group were higher than that of AC group(P<0.01)after treatment.Compared with the YC group,the protein expression of Nrf2,ERK,p-ERK,HO-1,NQO1 and SOD1 in the testes of the AC group was significantly down-regulated(P<0.01),and the protein expression of Keap1 was significantly up-regulated(P<0.01).Compared with the AC group,the protein expression of Nrf2,ERK,p-ERK,HO-1,NQO1 and SOD1 in the DR group and EA group was significantly up-regulated(P<0.01),and the protein expression of Keap1 was significantly down-regulated.ConclusionElectroacupuncture can improve TT and FT levels in serum of aged rats with low testosterone,and its mechanism may be related to the regulation of ERK/ Nrf2-ARE antioxidant stress pathway,the reduction of oxidative stress damage in testicular tissues of aged rats,and the improvement of male reproductive senescence.