Effect of electroacupuncture on the AMPK/SIRT1/NF-κB signaling pathway in testis of aging model rats

Abstract:

Objective To investigate the effect of electroacupuncture（EA） on the protein expression of adenosine monophosphate activated protein kinase（AMPK）， silent information regulator of transcription 1（SIRT1） and nuclear factor-κB（NF-κB） in testis of aging model rats and its mechanism of delaying reproductive aging.Methods Forty male Sprague-Dawley rats were randomly divided into the control group（n=10） and the model group（n=30）. The aging model rats were replicated by intraperitoneal injection of D-galactose in the model group. Twenty-four rats were randomly selected from the successful model rats and divided into three groups： the model group， the drug group and the EA group， with 8 rats in each group. In the EA group，“Shen yu” and “Guan yuan” were used for EA treatment， once a day，15 minutes each time，5 days of treatment and 2 days of rest，continuous treatment for 8 weeks. The drug group received abdominal subcutaneous injection of testosterone propionate（7 mg/kg once，once every 3 days） for continuous 8 weeks. The control group and model group were given abdominal subcutaneous injection of 0.9% sodium chloride solution. The injection dose and course of treatment were the same as those in the drug group. The levels of serum total testosterone（TT） and free testosterone（FT）were detected before and after treatment. The protein expression of AMPK，p-AMPK，SIRT1 and NF-κB p65 in rat testis were detected by immunohistochemistry and Western blot.Results Before treatment，compared with the control group，the levels of serum TT and FT in the model group，drug group and EA group were decreased（P=0.000，P=0.001；P=0.000，P=0.001；P=0.000，P=0.000）. After treatment，compared with the control group，the levels of serum TT and FT in the model group，drug group and EA group were decreased（P=0.000，P=0.000；P=0.009，P=0.025；P=0.006，P=0.022）. Compared with the model group，the levels of serum TT and FT in the drug group and EA group were significantly increased（P=0.001，P=0.004；P=0.001，P=0.004）. Compared with the control group， the protein expression of p-AMPK and SIRT1 in the model group，drug group and EA group was significantly decreased（P=0.000，P=0.000；P=0.000，P=0.000；P=0.000，P=0.000），and the expression of NF-κB p65 was significantly increased（P=0.000，P=0.000，P=0.005）. Compared with the model group and drug group，the protein expression of p-AMPK and SIRT1 in the EA group were significantly increased（P=0.000，P=0.000；P=0.023，P=0.002），and the protein expression of NF-κB p65 was significantly decreased（P=0.000，P=0.000）.Conclusion EA may inhibit the inflammatory response of Leydig cells by activating the AMPK/SIRT1/NF-κB pathway of Leydig cells， thereby improving the levels of serum TT and FT， which may be one of the mechanisms of EA in delaying reproductive aging.