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

doi:10.7506/spkx1002-6630-20220507-089

食品科学 ›› 2023, Vol. 44 ›› Issue (9): 82-87.doi: 10.7506/spkx1002-6630-20220507-089

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，谢琳琳，王鹤霖，迟晓星，张东杰

（1.黑龙江八一农垦大学食品学院，黑龙江大庆 163319；2.黑龙江八一农垦大学杂粮工程技术研究中心，黑龙江大庆 163319）

出版日期:2023-05-15 发布日期:2023-05-24
基金资助:
黑龙江省自然科学基金项目（LH2021H055）；黑龙江省自然科学基金研究团队项目（TD2020C003）； “十三五”国家重点研发计划重点专项（2018YFE0206300）

Genistein Alleviates Fatigue in Immunosuppressed Rats through the AMPK/SIRT1/PGC-1α Signaling Pathway

DONG Jiaping, XIE Linlin, WANG Helin, CHI Xiaoxing, ZHANG Dongjie

(1. College of Food, Heilongjiang Bayi Agricultural University, Daqing 163319, China; 2. Miscellaneous Grains Engineering Technology Research Centre, Heilongjiang Bayi Agricultural University, Daqing 163319, China)

Online:2023-05-15 Published:2023-05-24

摘要/Abstract

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、免疫抑制模型组与金雀异黄素低、中、高剂量组以及阳性对照组。除空白对照组外，其余组腹腔注射环磷酰胺40 mg/kg mb，连续3 d，建立免疫抑制大鼠模型。金雀异黄素低、中、高剂量组分别灌胃10、20、40 mg/kg mb金雀异黄素，阳性对照组灌胃贞芪扶正颗粒3.125 g/kg mb，空白对照组灌胃等量花生油。实验结束后，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）活力；酶联免疫吸附测定法检测大鼠血清中免疫球蛋白G（immunoglobulin G，IgG）、肿瘤坏死因子α（tumor necrosis factor α，TNF-α）质量浓度；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（adenosine monophosphate-activated protein kinase，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γ辅激活因子1α（peroxisome proliferator-activated receptor γ coactivator 1α，PGC-1α）和过氧化物酶体增殖物激活受体γ（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p-AMPK、SIRT1、PGC-1α和PPARγ的蛋白表达水平。结果：与免疫抑制模型组相比，补充GEN后极显著延长了大鼠力竭游泳时间（P＜0.01）；与免疫抑制模型相比，高剂量GEN能够显著降低血清中CK活力（P＜0.05）和LDH活力（P＜0.01），极显著提高大鼠血清中IgG、TNF-α质量浓度（P＜0.01），同时显著提高大鼠骨骼肌中p-AMPK、SIRT1、PGC-1α和PPARγ基因及蛋白表达水平（P＜0.05、P＜0.01）。结论：GEN具有缓解免疫低下大鼠疲劳的作用，其机制可能与激活骨骼肌AMPK/SIRT1/PGC-1α信号通路及改善大鼠运动耐力、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信号通路；过氧化物酶体增殖物激活受体γ

Abstract: Objective: To explore the effect of genistein (GEN) on relieving fatigue in immunosuppressed rats and the underlying mechanism. Methods: A total of 96 male SD rats were randomly divided into six groups with 16 rats in each group, including blank control (CG), immunosuppression model (MG), low-dose GEN (LG), medium-dose GEN (MG), high-dose GEN (HG) and positive control (PG). All rats except those in the blank control group were injected intraperitoneally with cyclophosphamide at a dose of 40 mg/kg mb for three days to establish an immunosuppressed rat model. The rats in the LG, MG and HG groups were gavaged with GEN at doses of 10, 20, and 40 mg/kg mb, respectively. The positive control group was gavaged with 3.125 g/kg mb of Zhenqi Fuzheng granule, and the blank control group was gavaged with an equal amount of peanut oil. After the experiment, the exhaustive swimming time of rats was recorded. The activities of creatine kinase (CK) and lactate dehydrogenase (LDH) in rat serum were detected by colorimetry. The concentrations of immunoglobulin G (IgG) and tumor necrosis factor-α (TNF-α) in rat serum were detected by enzyme-linked immunosorbent assay (ELISA). Real-time fluorescence quantitative PCR was used to detect adenylate-activated protein kinase (AMPK), silent information regulator 1 (SIRT1), peroxisome proliferator-activated receptorγ coactivator-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 the skeletal muscle of rats were detected by Western blot. Results: Compared with the MG group, GEN supplementation significantly prolonged the exhaustive swimming time of rats (P < 0.01), and high-dose GEN significantly decreased CK and LDH activities and increased the concentrations of IgG and TNF-α in the serum (P < 0.01), and significantly increased the gene and protein expression levels of AMPK, SIRT1, PGC-1α and PPARγ in the skeletal muscle of rats (P < 0.05 and P < 0.01). Conclusion: GEN relieves fatigue in immunosuppressed rats, and the underlying mechanism may be related to activating the AMPK/SIRT1/PGC-1α signaling pathway in the skeletal muscle and improving the exercise endurance, energy production and immunomodulatory capacity of rats.

Key words: genistein; fatigue; immunosuppression; AMPK/SIRT1/PGC-1α signaling pathway; peroxisome proliferators-activated receptor γ

中图分类号:

R151.1

董佳萍，谢琳琳，王鹤霖，迟晓星，张东杰. 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[J]. 食品科学, 2023, 44(9): 82-87.

DONG Jiaping, XIE Linlin, WANG Helin, CHI Xiaoxing, ZHANG Dongjie. Genistein Alleviates Fatigue in Immunosuppressed Rats through the AMPK/SIRT1/PGC-1α Signaling Pathway[J]. FOOD SCIENCE, 2023, 44(9): 82-87.

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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

Genistein Alleviates Fatigue in Immunosuppressed Rats through the AMPK/SIRT1/PGC-1α Signaling Pathway

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