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• 营养卫生 •    下一篇

金雀异黄素通过AMPK/SIRT1/PGC-1α信号通路缓解免疫抑制大鼠的疲劳作用

董佳萍1,2,谢琳琳2,王鹤霖3,迟晓星3,张东杰3   

  1. 1.
    2. 黑龙江八一农垦大学
    3. 黑龙江八一农垦大学食品学院
  • 收稿日期:2022-05-07 修回日期:2023-03-24 出版日期:2023-05-15 发布日期:2023-05-24
  • 通讯作者: 迟晓星 E-mail:chixiaoxing@sina.com
  • 基金资助:
    黑龙江省自然科学基金项目;黑龙江省自然科学基金研究团队项目;国家重点研发计划项目

Genistein alleviates the fatigue of immunosuppressed rats through AMPK/SIRT1/PGC-1α signaling pathway.

  • Received:2022-05-07 Revised:2023-03-24 Online:2023-05-15 Published:2023-05-24
  • Contact: Xiao-Xing CHI E-mail:chixiaoxing@sina.com

摘要: 摘 要:目的:探讨金雀异黄素(genistein,GEN)缓解免疫抑制大鼠的疲劳作用及其作用机制。方法:96只雄性SD大鼠随机分为6组,每组16只,分别为空白对照组(CG)、免疫抑制模型组(MG)、金雀异黄素低剂量组(LG)、中剂量组(MG)、高剂量组(HG)以及阳性对照组(PG)。除空白对照组外,其他组腹腔注射环磷酰胺40 mg/kg,连续3 d,建立免疫抑制大鼠模型。金雀异黄素低、中、高剂量组分别以10 mg/kg、20 mg/kg、40 mg/kg的金雀异黄素灌胃,阳性对照组以贞芪扶正颗粒3.125 g/kg灌胃,空白对照组等量花生油灌胃。试验结束后,记录大鼠力竭游泳时间;采用比色法检测大鼠血清中肌酸激酶(creatine kinase,CK)、乳酸脱氢酶(lactate dehydrogenase,LDH)含量;酶联免疫吸附测定法检测大鼠血清中免疫球蛋白G(immunoglobulin G,Ig-G)、肿瘤坏死因子-α(tumor necrosisfactor-α,TNF-α)含量;采用实时荧光定量技术检测大鼠骨骼肌中腺苷酸活化蛋白激酶(AMPK)、沉默信息调节因子1(silent information regulator 1,SIRT1)、过氧化物酶增殖活化受体辅激活因子-1α(peroxisome proliferator-activated receptorγcoactivator-1α,PGC-1α)和过氧化物酶体增殖物激活受体(peroxisome proliferators-activated receptor-γ,PPARγ)mRNA表达水平;采用蛋白免疫印迹法检测大鼠骨骼肌中AMPK、SIRT1、PGC-1α和PPARγ的蛋白表达水平。结果:补充GEN后,与MG相比,GEN-HG显著延长大鼠力竭游泳时间(P<0.01),显著降低血清中CK和LDH含量(P<0.01),显著提高大鼠血清中Ig-G、TNF-α含量(P<0.01),同时显著提高大鼠骨骼肌中AMPK、SIRT1、PGC-1α和PPARγ基因及蛋白表达水平(P<0.01)。结论:GEN具有缓解免疫低下大鼠疲劳的作用,其机制可能是通过提高骨骼肌AMPK/SIRT1/PGC-1α通路作用、提高大鼠运动耐力、能量产生及免疫调节能力有关。

关键词: 金雀异黄素, 疲劳, 免疫抑制, AMPK, SIRT1, PGC-1α, PPARγ

Abstract: Abstract: Objective: To explore the effect of genistein (GEN) on relieving fatigue in immunosuppressed rats and its mechanism. Methods: 96 male SD rats were randomly divided into 6 groups with 16 rats in each group, which were blank control group (CG), immunosuppression model group (MG), GEN low-dose group (LG), GEN medium-dose group (MG), GEN high-dose group (HG) and positive control group (PG). Except the blank control group, the other groups were injected intraperitoneally with cyclophosphamide 40 mg/kg for 3 days to establish an immunosuppressed rat model. GEN-LG, GEN-MG and GEN-HG groups were gavaged at the doses of 10 mg/kg, 20 mg/kg, and 40 mg/kg, respectively.The positive control group was gavaged at 3.125 g/kg of Zhenqi Fuzheng granules, and the blank control group was gavaged with equal amount of peanut oil. After the experiment, the exhaustive swimming time of rats was recorded. The contents of creatine kinase (CK) and lactate dehydrogenase (LDH) in serum of rats were detected by colorimetry. The contents of immunoglobulin G (Ig-G) and tumor necrosis factor-α (TNF-α) in serum of rats were detected by enzyme-linked immunosorbent assay. Real-time fluorescence quantitative technology was used to detect adenylate-activated protein kinase (AMPK), silent information regulator 1 (SIRT1), peroxisome proliferator-activated receptorγcoactivator-1α (PGC-1α) and peroxisome proliferators-activated receptor-γ(PPARγ) mRNA expression levels. The protein expression levels of AMPK, SIRT1, PGC-1α and PPARγ in skeletal muscle of rats were detected by western blot. Results:After supplementation of genistein, compared with MG, GEN-HG significantly prolonged the exhaustive swimming time of rats (P<0.01), significantly decreased the contents of CK and LDH in serum (P<0.01), significantly increased the contents of Ig-G and TNF-α in serum (P<0.01), and also significantly increased the gene and protein expression levels of AMPK, SIRT1, PGC-1α and PPARγ in skeletal muscle of rats (P<0.01). Conclusions: GEN had the effect of relieving fatigue in immunosuppressed rats, and its mechanism may be related to the improvement of AMPK/SIRT1/PGC-1α pathway in skeletal muscle, exercise endurance, energy production and immunomodulatory ability of rats.

Key words: genistein, fatigue, immunosuppression, AMPK, SIRT1, PGC-1α, PPARγ

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