1.重庆市人民医院中医科,重庆 400014
2.重庆医科大学附属康复医院中医康复科,重庆 400050
3.重庆医科大学中医药学院中医药防治代谢性疾病重庆市重点实验室,重庆 400016
Received:11 March 2022,
Revised:2022-09-16,
Accepted:17 October 2022,
Published:28 March 2023
移动端阅览
姚太万,陈艳华,卜凡,等.电针对衰老模型大鼠睾丸AMPK/SIRT1/NF-κB信号通路的影响[J].重庆医科大学学报,2023,48(3):255-260.
Yao Taiwan,Chen Yanhua,Bu Fan,et al.Effect of electroacupuncture on the AMPK/SIRT1/NF-κB signaling pathway in testis of aging model rats[J].Journal of Chongqing Medical University,2023,48(3):255-260.
姚太万,陈艳华,卜凡,等.电针对衰老模型大鼠睾丸AMPK/SIRT1/NF-κB信号通路的影响[J].重庆医科大学学报,2023,48(3):255-260. DOI: 10.13406/j.cnki.cyxb.003185.
Yao Taiwan,Chen Yanhua,Bu Fan,et al.Effect of electroacupuncture on the AMPK/SIRT1/NF-κB signaling pathway in testis of aging model rats[J].Journal of Chongqing Medical University,2023,48(3):255-260. DOI: 10.13406/j.cnki.cyxb.003185.
目的
2
探讨电针对衰老模型大鼠睾丸腺苷酸活化蛋白激酶(adenosine monophosphate activated protein kinase,AMPK)、沉默信息调节因子1(silent information regulator of transcription 1,SIRT1)、核因子-κB(nuclear factor-κB,NF-κB)蛋白表达的影响及其延缓生殖衰老的作用机制。
方法
2
将40只雄性SD大鼠随机分为对照组(
n
=10)和造模组(
n
=30),造模组经腹腔注射D-半乳糖复制衰老模型大鼠,从造模成功的大鼠中随机筛选24只平均分为3组:模型组、药物组和电针组,每组8只。电针组取“肾俞”“关元”予电针治疗,每日治疗1次,每次15 min,治疗5 d休息2 d,连续治疗8周。药物组予腹部皮下注射丙酸睾酮注射液(7 mg/kg,每3日1次),连续8周。对照组和模型组给予腹部皮下注射0.9%氯化钠溶液,注射剂量及疗程同药物组。治疗前后检测大鼠血清总睾酮(total testosterone,TT)和游离睾酮(free testosterone,FT)水平,免疫组化法、Western blot检测大鼠睾丸AMPK、p-AMPK、SIRT1、NF-κB p65蛋白表达。
结果
2
治疗前,与对照组比较,模型组、药物组及电针组血清TT、FT水平降低(
P
=0.000,
P
=0.001;
P
=0.000,
P
=0.001;
P
=0.000,
P
=0.000);治疗后,与对照组比较,模型组、药物组及电针组血清TT、FT水平降低(
P
=0.000,
P
=0.000;
P
=0.009,
P
=0.025;
P
=0.006,
P
=0.022),与模型组比较,药物组及电针组血清TT、FT明显升高(
P
=0.001,
P
=0.004;
P
=0.001,
P
=0.004)。与对照组比较,模型组、药物组及电针组p-AMPK、SIRT1蛋白表达均明显降低(
P
=0.000,
P
=0.000;
P
=0.000,
P
=0.000;
P
=0.000,
P
=0.000),NF-κB p65蛋白表达明显升高(
P
=0.000,
P
=0.000,
P
=0.005)。与模型组及药物组比较,
电针组p-AMPK、SIRT1蛋白表达明显升高(
P
=0.000,
P
=0.000;
P
=0.023,
P
=0.002),NF-κB p65蛋白表达明显降低(
P
=0.000,
P
=0.000)。
结论
2
电针可能通过激活睾丸间质细胞(Leydig细胞)AMPK/SIRT1/NF-κB通路,抑制Leydig细胞炎性反应,从而改善血清TT、FT水平,这可能是电针延缓生殖衰老的作用机制之一。
Objective
2
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
2
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
2
Before treatment,compared with the control group,the levels of se
rum 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
2
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.
Pechersky A . Features of diagnostics and treatment of partial androgen deficiency of aging men [J ] . Central Eur J Urol , 2014 , 67 ( 4 ): 397 - 404 .
Traish AM . Negative impact of testosterone deficiency and 5α-reductase inhibitors therapy on metabolic and sexual function in men [J ] . Adv Exp Med Biol , 2017 , 1043 : 473 - 526 .
Coward R , McBride J . Recovery of spermatogenesis following testosterone replacement therapy or anabolic-androgenic steroid use [J ] . Asian J Androl , 2016 , 18 ( 3 ): 373 .
姚太万 , 刘 嘉 , 李学智 , 等 . 电针对睾酮低下老年大鼠睾丸间质细胞慢性炎性反应的影响 [J ] . 针刺研究 , 2018 , 43 ( 3 ): 163 - 168 .
Yao TW , Liu J , Li XZ , et al . Electroacupuncture intervention improves physical power of aged rats with low testosterone by reducing chronic inflammatory response of Leydig cells [J ] . Acupunct Res , 2018 , 43 ( 3 ): 163 - 168 .
曾婷婷 , 姚太万 , 李学智 , 等 . 电针对睾酮低下老年大鼠睾丸间质细胞抗氧化功能的影响 [J ] . 针刺研究 , 2019 , 44 ( 8 ): 566 - 570,582 .
Zeng TT , Yao TW , Li XZ , et al . Electroacupuncture intervention increases testosterone level of aged rats by activating ERK/Nrf2/HO-1 signaling of Leydig cells [J ] . Acupunct Res , 2019 , 44 ( 8 ): 566 - 570,582 .
Michalik A , Jarzyna R . The key role of AMP-activated protein kinase(AMPK)in aging process [J ] . Postepy Biochem , 2016 , 62 ( 4 ): 459 - 471 .
Cantó C , Jiang LQ , Deshmukh AS , et al . Interdependence of AMPK and SIRT1 for metabolic adaptation to fasting and exercise in skeletal muscle [J ] . Cell Metab , 2010 , 11 ( 3 ): 213 - 219 .
Kume S , Koya D . Autophagy:a novel therapeutic target for diabetic nephropathy [J ] . Diabetes Metab J , 2015 , 39 ( 6 ): 451 - 460 .
Sciarretta S , Hariharan N , Monden Y , et al . Is autophagy in response to ischemia and reperfusion protective or detrimental for the heart? [J ] . Pediatr Cardiol , 2011 , 32 ( 3 ): 275 - 281 .
Li FT , Chen Y , Li YJ , et al . Geniposide alleviates diabetic nephropathy of mice through AMPK/SIRT1/NF-κB pathway [J ] . Eur J Pharmacol , 2020 , 886 : 173449 .
宋亚男 , 袁 丁 , 张长城 , 等 . 竹节参总皂苷通过调节AMPK/Sirt1/NF-κB通路抑制衰老大鼠心肌细胞凋亡的作用研究 [J ] . 中国中药杂志 , 2017 , 42 ( 23 ): 4656 - 4660 .
Song YN , Yuan D , Zhang CC , et al . Effect of saponins extracted from Panax japonicus on inhibiting cardiomyocyte apoptosis by AMPK/Sirt1/NF-κB signaling pathway in aging rats [J ] . China J Chin Mater Med , 2017 , 42 ( 23 ): 4656 - 4660 .
赵凡凡 , 周玉枝 , 高 丽 , 等 . D-半乳糖致衰老大鼠模型的研究进展 [J ] . 药学学报 , 2017 , 52 ( 3 ): 347 - 354 .
Zhao FF , Zhou YZ , Gao L , et al . Advances in the study of the rat model of aging induced by D-galactose [J ] . Acta Pharm Sin , 2017 , 52 ( 3 ): 347 - 354 .
Haider S , Liaquat L , Shahzad S , et al . A high dose of short term exogenous D-galactose administration in young male rats produces symptoms simulating the natural aging process [J ] . Life Sci , 2015 , 124 : 110 - 119 .
Samad N , Imran I , Zulfiqar I , et al . Ameliorative effect of lithium chloride against d-galactose induced behavioral and memory impairment,oxidative stress and alteration in serotonin function in rats [J ] . Pharmacol Rep , 2019 , 71 ( 5 ): 909 - 916 .
Saleh DO , Mansour DF , Hashad IM , et al . Effects of sulforaphane on D-galactose-induced liver aging in rats:role of Keap-1/Nrf-2 pathway [J ] . Eur J Pharmacol , 2019 , 855 : 40 - 49 .
王德军 , 常小荣 , 严 洁 , 等 . 针刺足阳明胃经特定穴与非特定穴治疗功能性消化不良疗效比较 [J ] . 中国针灸 , 2012 , 32 ( 8 ): 703 - 708 .
Wang DJ , Chang XR , Yan J , et al . Comparative study on therapeutic effect between acupuncture at special acupoints and non-specific acupoints in Foot Yangming Meridian for functional dyspepsia [J ] . Chin Acupunct Moxibustion , 2012 , 32 ( 8 ): 703 - 708 .
何 任 . 养生与长寿 [J ] . 安徽中医学院学报 , 1982 , 1 ( 3 ): 43 - 44 .
He R . Health preservation and longevity [J ] . J Anhui Tradit Chin Med Coll , 1982 , 1 ( 3 ): 43 - 44 .
罗 磊 , 许晓瑾 . 电针对力竭游泳大鼠肾脏线粒体自由基代谢和线粒体功能的影响 [J ] . 中国针灸 , 2001 , 21 ( 6 ): 366 - 368 .
Luo L , Xu XJ . Effect of electroacupuncture on M etabolism of mitochondrial free radicals and mitochondrial function in kidney in rats of full swimming [J ] . Chin Acuponcture & Moxibustion , 2001 , 21 ( 6 ): 366 - 368 .
李 珉 , 张 卫 , 王 刚 , 等 . 电针涌泉穴对D-半乳糖致衰老模型大鼠的p53和bcl-2基因表达的影响 [J ] . 华西医科大学学报 , 2002 , 33 ( 4 ): 589 - 591 .
Li M , Zhang W , Wang G , et al . Influence of electro-acupuncture of YONGQUAN-POINT on the expression of p53 and bcl-2 in the aging model rat induced by D-dalactose [J ] . J West China Univ Med Sci , 2002 , 33 ( 4 ): 589 - 591 .
李丽红 , 李 丽 , 赵志恩 , 等 . 艾灸、中药对衰老模型大鼠线粒体DNA、血清IL-2、IL-6含量的影响 [J ] . 中国针灸 , 2008 , 28 ( 9 ): 681 - 684 .
Li LH , Li L , Zhao ZE , et al . Effects of moxibustion and Chinese herbs on contents of mitochondrial DNA,serum IL-2 and IL-6 in the aging model rat [J ] . Chin Acupunct & Moxibustion , 2008 , 28 ( 9 ): 681 - 684 .
杜 艳 , 赵利华 , 吴海标 , 等 . 艾炷灸对D-半乳糖衰老小鼠大脑皮层细胞周期蛋白依赖性激酶抑制因子、视网膜母细胞瘤抑制蛋白及c-fos的影响 [J ] . 针刺研究 , 2010 , 35 ( 4 ): 250 - 254,276 .
Du Y , Zhao LH , Wu HB , et al . Effects of moxibustion on the expression of cell cycle protein and retinoblastoma and c-fos in the cerebral cortex of senile mice [J ] . Acupunct Res , 2010 , 35 ( 4 ): 250 - 254,276 .
Burkewitz K , Weir HJM , Mair WB . AMPK as a pro-longevity target [J ] . Exp Suppl , 2016 , 107 : 227 - 256 .
Salminen A , Kaarniranta K . AMP-activated protein kinase(AMPK) controls the aging process via an integrated signaling network [J ] . Ageing Res Rev , 2012 , 11 ( 2 ): 230 - 241 .
Han XJ , Tai HR , Wang XB , et al . AMPK activation protects cells from oxidative stress-induced senescence via autophagic flux restoration and intracellular NAD + elevation [J ] . Aging Cell , 2016 , 15 ( 3 ): 416 - 427 .
Xie J , Zhang XM , Zhang L . Negative regulation of inflammation by SIRT1 [J ] . Pharmacol Res , 2013 , 67 ( 1 ): 60 - 67 .
Hubbard BP , Sinclair DA . Small molecule SIRT1 activators for the treatment of aging and age-related diseases [J ] . Trends Pharmacol Sci , 2014 , 35 ( 3 ): 146 - 154 .
Xu GQ , Zhao J , Liu HY , et al . Melatonin inhibits apoptosis and oxidative stress of mouse leydig cells via a SIRT1-dependent mechanism [J ] . Molecules , 2019 , 24 ( 17 ): 3084 .
El Assar M , Angulo J , Rodríguez-Mañas L . Oxidative stress and vascular inflammation in aging [J ] . Free Radic Biol Med , 2013 , 65 : 380 - 401 .
Stein S , Matter CM . Protective roles of SIRT1 in atherosclerosis [J ] . Cell Cycle , 2011 , 10 ( 4 ): 640 - 647 .
Wu L , Zhang AJ , Sun YJ , et al . Sirt1 exerts anti-inflammatory effects and promotes steroidogenesis in Leydig cells [J ] . Fertil Steril , 2012 , 98 ( 1 ): 194 - 199 .
0
Views
6
下载量
2
CSCD
Publicity Resources
Related Articles
Related Author
Related Institution
京公网安备11010802024621
