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[电针干预通过激活睾丸间质细胞的ERK/Nrf2/HO-1信号通路提高老年大鼠睾酮水平]

[Electroacupuncture intervention increases testosterone level of aged rats by activating ERK/Nrf2/HO-1 signaling of Leydig cells].

作者信息

Zeng Ting-Ting, Yao Tai-Wan, Li Xue-Zhi, Tang Cheng-Lin

机构信息

Key Laboratory of Traditional Chinese Medicine for Prevention and Cure of Metabolic Diseases, College of Traditional Chinese Medicine of Chongqing Medical University, Chongqing 400016, China.

Department of Rehabilitation, Affiliated Rehabilitation Hospital of Chongqing Medical University, Chongqing 400050.

出版信息

Zhen Ci Yan Jiu. 2019 Aug 25;44(8):566-70. doi: 10.13702/j.1000-0607.180790.

Abstract

OBJECTIVE

To observe the effect of electroacupuncture (EA) on anti-oxidant function of Leydig cells in aged rats with low testosterone, so as to reveal its underlying mechanism of anti-aging of male reproduction.

METHODS

Eighteen 20 months old SD rats were divided into aged control, medication and EA groups(=6 in each group), and other 6 young male SD rats (2 months of age) were used as the youth control group. The rats of the youth and the aged control groups received subcutaneous injection of 0.9% normal saline (7 mg·kg·3 d) for 8 weeks, and those of the medication group received abdominal subcutaneous injection of Testosterone Propionate (7 mg·kg·3 d) for 8 weeks. EA (2 Hz/100 Hz, 1 mA) was applied to bilateral "Shenshu" (BL23) and "Guanyuan" (CV4) for 15 min, once daily for 8 weeks except the weekends. The levels of serum total testosterone (TT) and free testosterone (FT) were determined by enzyme linked immune sorbent assay (ELISA), the immunoactivity of phosphorylated extracellular signal-regulated protein kinase (p-ERK) was detected by immunohistochemistry, and the expression levels of ERK, p-ERK, nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) proteins in the testis tissues were determined by Western blot.

RESULTS

Before and after the treatment, the levels of serum TT and FT in the aged control group were significantly lower than those of the youth control group (<0.01). After the treatment, the serum TT and FT contents in the EA and medication groups were obviously higher than those in the aged control group (<0.01). Compared with the youth control group, the expression levels of ERK, p-ERK, Nrf2 and HO-1 proteins were significantly down-regulated in the aged control group (<0.01), while in comparison with the aged control group, the expression levels of ERK, p-ERK, Nrf2 and HO-1 proteins were significantly up-regulated in the medication and EA groups (<0.01). The therapeutic effect of EA was notably superior to that of medication in up-regulating the immunoactivity of p-ERK, and the expression levels of ERK, p-ERK, Nrf2 and HO-1 proteins (<0.01). No significant differences were found between the EA and medication groups in the increased levels of serum TT and FT after the intervention (>0.05).

CONCLUSION

EA intervention may increase testosterone level of the aged rats, which may be related to its effects in triggering ERK /Nrf2 /HO-1 signaling (anti-oxidative stress signal pathway) in the testis.

摘要

目的

观察电针(EA)对低睾酮老龄大鼠睾丸间质细胞抗氧化功能的影响,以揭示其延缓雄性生殖衰老的潜在机制。

方法

将18只20月龄的SD大鼠分为老龄对照组、药物组和电针组(每组6只),另取6只2月龄的雄性SD大鼠作为青年对照组。青年对照组和老龄对照组大鼠皮下注射0.9%生理盐水(7mg·kg·3d),连续8周;药物组大鼠腹腔皮下注射丙酸睾酮(7mg·kg·3d),连续8周。采用2Hz/100Hz、1mA的电针刺激双侧“肾俞”(BL23)和“关元”(CV4),每次15分钟,除周末外每日1次,连续8周。采用酶联免疫吸附测定(ELISA)法检测血清总睾酮(TT)和游离睾酮(FT)水平,采用免疫组织化学法检测磷酸化细胞外信号调节蛋白激酶(p-ERK)的免疫活性,采用蛋白质印迹法检测睾丸组织中ERK、p-ERK、核因子E2相关因子2(Nrf2)和血红素加氧酶-1(HO-1)蛋白的表达水平。

结果

治疗前后,老龄对照组血清TT和FT水平显著低于青年对照组(<0.01)。治疗后,电针组和药物组血清TT和FT含量明显高于老龄对照组(<0.01)。与青年对照组相比,老龄对照组ERK、p-ERK、Nrf2和HO-1蛋白表达水平显著下调(<0.01);与老龄对照组相比,药物组和电针组ERK、p-ERK、Nrf2和HO-1蛋白表达水平显著上调(<0.0主要观察指标:观察各组大鼠血清TT、FT水平,睾丸组织中ERK、p-ERK、Nrf2和HO-主要观察指标:观察各组大鼠血清TT、FT水平,睾丸组织中ERK、p-ERK、Nrf2和HO-1蛋白表达水平。

结论

电针干预可能提高老龄大鼠睾酮水平,其机制可能与激活睾丸组织中ERK/Nrf2/HO-1信号通路(抗氧化应激信号通路)有关。

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