电针对睾酮低下老年大鼠Keap1-Nrf2/ARE信号通路的影响

Effects of Electroacupuncture on Keap1-Nrf2/ARE Signaling Pathway in Aged Rats with Low Testosterone

目的观察电针对中老年男性部分雄激素缺乏综合征(PADAM)模型大鼠睾丸间质细胞(Leydig细胞)的作用,并探讨电针抗雄性生殖衰老的可能作用机制。方法18只SD PADAM大鼠随机分为老年组、药物组、电针组,每组6只,SD青年大鼠6只作为青年组。青年组和老年组给予0.9%氯化钠溶液[7 mg/(kg·3d)]腹部皮下注射。药物组予丙酸睾酮注射液[7 mg/(kg·3d)]腹部皮下注射。电针组取"肾俞""关元"穴进行干预。治疗时间均为8周。观察治疗前后各组大鼠强迫游泳力竭时间,采用ELISA检测各组大鼠血清总睾酮(TT)和游离睾酮(FT)水平,采用免疫组化法检测Leydig细胞Keleh样环氧氯丙烷相关蛋白-1(Keap1)蛋白表达,Western Blot检测大鼠睾丸Keap1、核因子E2相关因子2(Nrf2)、醌氧化还原酶1(NQO1)及超氧化物歧化酶(SOD)蛋白表达。结果与青年组比较,老年组治疗前强迫游泳力竭时间、血清TT、FT水平及睾丸Nrf2、NQO1、SOD1蛋白表达降低(P<0.01),睾丸Keap1蛋白表达增加(P<0.01)。与老年组比较,电针组与药物组治疗后强迫游泳力竭时间、血清TT、FT及睾丸Nrf2、NQO1、SOD1蛋白表达升高(P<0.01),睾丸Keap1蛋白表达降低(P<0.01)。结论电针可改善睾酮低下老年大鼠血清TT、FT含量,提高老年大鼠抗疲劳能力,其作用机制可能与调控Keap1-Nrf2/ARE信号通路,降低老年大鼠睾丸组织氧化应激损伤有关。

Objective To observe the antioxidantive effect of electroacupuncture(EA)on Leydig cells of rats with partial androgen deficiency in the aging male(PADAM)and explore the underlying mechanism of anti-aging effect of EA.Methods Eighteen SD PADAM rats were randomly divided into aged group,medication group and EA group(6 in each group),and young SD rats(n=6)were selected as the youth group.The youth group and the aged group were given 0.9%sodium chloride solution[7 mg/(kg·3 d)]by abdominal subcutaneous injection.In the medication group,testosterone propionate[7 mg/(kg·3 d)]was injected subcutaneously.The EA group was treated by needling acupoints of Shenshu(BL23)and Guanyuan(CV4).The therapeutic course for all was 8 weeks.Before and after treatment,the exhaustive swimming time of rats in the forced swimming test was observed,and the levels of serum total testosterone(TT)and free testosterone(FT)levels were measured by enzyme-linked immunosorbent assay(ELISA).The Kelch-like ECH-associated protein-1(Keap1)expression in Leydig cells was detected by immunohistochemistry.The expression of NF-E2-related factor 2(Nrf2),Keap1,NAD(P)H:quinone oxidoreductase 1(NQO1)and superoxide dismutase(SOD)in tesis were assayed by Western Blot.Results Compared with the youth group,the duration of exhaustion swimming,serum TT and FT levels,and the protein expressions of Nrf2,NQO1 and SOD1 in testis decreased in the aged group before treatment(P<0.01),while the protein expression of KEAP1 in testis increased(P<0.01).Compared with the aged group,duration of exhaustion swimming,TT,FT,the protein expressions serum of Nrf2,NQO1 and SOD1 in testis increased in EA group and drug group after treatment(P<0.01),and the protein expression of KEAP1 in testis decreased(P<0.01).Conclusions EA can improve the serum TT and FT contents of the aged rats with low testosterone,and improve the anti-fatigue ability.The mechanism of action may be related to the regulation of Keap1-Nref2/Arf signaling pathway and the reduction of oxidative stress injury in the testicular tissue of the aged rats.