何首乌饮延缓大鼠睾丸间质细胞衰老与调控胰岛素/胰岛素样生长因子-1信号通路的关系

李少华; 姜丽萍; 刘靓云; 刘雯青; 朱娟; 牛嗣云; 齐峰

doi:10.16098/j.issn.052-1356.2020.05.019

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解剖学报 ›› 2020, Vol. 51 ›› Issue (5) : 751-757. DOI: 10.16098/j.issn.052-1356.2020.05.019

细胞和分子生物学

何首乌饮延缓大鼠睾丸间质细胞衰老与调控胰岛素/胰岛素样生长因子-1信号通路的关系

李少华²姜丽萍¹ 刘靓云¹ 刘雯青¹ 朱娟¹牛嗣云^1* 齐峰^3*

作者信息 +

Relationship between the senescence of Leydig cells and the regulation of insulin/insuli-like growth factor-1 signaling pathway in rats delayed by Heshouwuyin

LI Shao-hua² JIANG Li-ping¹ LIU Jing-yun¹ LIU Wen-qing¹ ZHU Juan¹ NIU Si-yun^1* QI Feng3*

Author information +

文章历史 +

摘要

目的探讨何首乌饮延缓大鼠睾丸间质（Leydig）细胞衰老与调控胰岛素/胰岛素样生长因子-1（IGF-1）信号通路的关系。方法采用氧化损伤的方法建立大鼠Leydig细胞衰老模型。用流式细胞术检测各组细胞周期的变化。应用Real-time PCR、免疫荧光和Western blotting技术检测Leydig细胞胰岛素/IGF-1信号传导通路关键基因的表达水平，并运用胰岛素受体/IGF受体特异性抑制剂BMS-754807处理细胞，用流式细胞术检测细胞凋亡水平。结果与正常组相比，衰老组β-半乳糖苷酶阳性细胞率可达60%（P＜0.05），G1 期所占比重明显增加， S期所占比重明显减少（P＜0.05）。胰岛素/IGF-1信号传导通路关键基因表达变化：衰老组胰岛素受体（INSR），胰岛素受体底物（IRS1）、IRS2、IGF1表达水平明显低于正常组（P＜0.05），胰岛素样生长因子结合蛋白3（IGFBP3）的 mRNA表达明显高于正常组（P＜0.05），何首乌饮可逆转上述现象。用 BMS-754807处理衰老的Leydig细胞后，Bcl-2 mRNA表达水平低于衰老组（P＜0.05），用何首乌饮和BMS-754807同时处理衰老的Leydig细胞，细胞凋亡率高于何首乌饮组（P＜0.05）。结论何首乌饮通过调控胰岛素/IGF-1信号传导通路延缓Leydig细胞衰老。

Abstract

Objective To investigate the relationship between the senescence of testicular Leydig cells and the regulation of insulin/insulin-like growth factor 1(IGF-1) signaling pathway in rats delayed by Heshouwuyin. Methods The oxidative damage technique was used to establish Leydig cell aging model. Flow cytometry was used to detect the changes of cell cycle in each group. Detection of mRNA and protein expression levels of key genes of insulin/IGF-1 signaling pathway in Leydig cells by Real-time PCR, immunofluorescence and Western blotting. Cells were treated with insulin receptor/IGF receptor specific inhibitor BMS-754807, flow cytometry was used to detect the apoptotic level of Leydig cells. Results Compared with the normal group, the proportion of β-galactosidase positive cells in the aging group was more than 60% (P<0.05), and the proportion of G1 phase increased significantly, while that of S phase decreased significantly (P<0.05); Changes in the expression of key genes of the insulin/IGF-1 signaling pathway: the expression levels of insulin receptor(INSR), insulin receptor substrate(IRS1), (IRS2), IGF-1 mRNA and protein in the aging group were significantly lower than those in the normal group（P＜0.05）, and the mRNA expression of insulin-like growth factor binding protein 3(IGFBP3) was significantly higher than in the normal group（P＜0.05）, Heshouwuyin could reverse the above phenomenon. After the treatment of senescent testicular Leydig cells with BMS-754807, the mRNA expression level of Bcl-2 was lower than that of the senescent group (P<0.05), and the cell apoptosis rate was higher than that of the senescent testicular Leydig cells treated with Heshouwuyin and BMS-754807at the same time (P<0.05). Conclusion Heshouwuyin delayed the senescence of testicular Leydig cells by regulating the insulin/IGF-1 signaling pathway.

导出引用

李少华姜丽萍刘靓云刘雯青朱娟牛嗣云齐峰. 何首乌饮延缓大鼠睾丸间质细胞衰老与调控胰岛素/胰岛素样生长因子-1信号通路的关系[J]. 解剖学报. 2020, 51(5): 751-757 https://doi.org/10.16098/j.issn.052-1356.2020.05.019

LI Shao-hua JIANG Li-ping LIU Jing-yun LIU Wen-qing ZHU Juan NIU Si-yun QI Feng. Relationship between the senescence of Leydig cells and the regulation of insulin/insuli-like growth factor-1 signaling pathway in rats delayed by Heshouwuyin[J]. Acta Anatomica Sinica. 2020, 51(5): 751-757 https://doi.org/10.16098/j.issn.052-1356.2020.05.019

中图分类号： Q291

参考文献

［1］ Heemst DV, Insulin, IGF-1 and longevity［J］. Aging Dis, 2010, 1(2):147-157.
［2］ Cawthon PM, Schousboe JT, Harrison SL, et al. Sex hormones, sex hormone binding globulin, and vertebral fractures in older men［J］. Bone, 2016, 84:271-278.
［3］ Xu J, Xia LL, Song N, et al. Testosterone, estradiol, and sex hormone-binding globulin in Alzheimer’s disease: a meta-analysis［J］. Curr Alzheimer Res, 2016, 13(3): 215-222.
［4］ Zhu Z, Huang F, Wang F, et al. Morinda officinalis polysaccharides stimulate hypothalamic gnrh secretion in varicocele progression［J］. Evid Based Complementary Alternat Medi, 2017, 2017: 9057959.
［5］ Wen W, Chen J, Ding L, et al. Astragaloside exerts anti-photoaging effects in UVB-induced premature senescence of rat dermal fibroblasts through enhanced autophagy［J］. Arch Biochem Biophys, 2018, 657: 31-40.
［6］ Chen JB, Wang YJ, Guo ShN, et al. Heshouwuyin attenuates hippocampal neuron damage induced by beta-amyloid protein ［J］. Acta Anatomica Sinica, 2015, 46(2): 175-181. (in Chinese)
陈靖博，王玉娟，郭胜男，等. 何首乌饮减轻β-淀粉样蛋白诱导的海马神经元损伤的作用机制［J］. 解剖学报, 2015, 46(2): 175-181.
［7］ Wang XJ, Niu SY, Zhang YQ, et al. Protective effect of HSWY on the reproductive functions of male aging rats［J］. Lishizhen Medicine and Materia Medica Research, 2011, 22(3): 532-535. (in Chinese)
王小杰，牛嗣云，张艳青，等. 何首乌饮对雄性衰老大鼠生殖功能的保护作用［J］. 时珍国医国药, 2011, 22(3): 532-535.
［8］ Niu S, Chen J, Duan F, et al. Possible mechanism underlying the effect of Heshouwuyin, a tonifying kidney herb, on sperm quality in aging rats［J］. BMC Complement Altern Med, 2014, 14: 250.
［9］ Hui ChH, Liu HK, Du YF, et al. Screening of differentially expressed genes regulated by Heshouwuyin in testicular tissues of aged rats［J］. Lishizhen Medicine and Materia Medica Research, 2017, 28(7): 1583-1586. (in Chinese)
回陈红，刘昊坤，杜云帆，等. 衰老大鼠睾丸组织受何首乌饮调控差异表达基因的筛选［J］. 时珍国医国药, 2017, 28(7): 1583-1586.
［10］ Zhen XL, Cui XY, Liu XL, et al. Effect of Heshouwuyin on the mRNA expression of p53, Bcl-2 and BAX in testicular Leydig cells of rats in vitro［J］. Chinese Journal of Anatomy, 2019, 42(2): 121-124. (in Chinese)
甄晓兰，崔晓燕，刘雪莉，等. 何首乌饮对离体大鼠睾丸Leydig细胞p53、Bcl-2和BAX mRNA表达的影响［J］. 解剖学杂志, 2019, 42(2): 121-124.
［11］ Chen XN, Liu Y, Ji JF. Advances in the clearance of senescent cells in diseases related to aging and aging［J］. Progress in Biochemistry and Biophysics, 2019, 46(12): 1150-1161. (in Chinese)
陈祥宁，刘洋，纪俊峰. 清除衰老细胞在衰老与老龄化相关疾病中的研究进展［J］. 生物化学与生物物理进展, 2019, 46(12): 1150-1161.
［12］ Yuan LL. Research progress on the relationship between cell cycle and signaling pathway and cell senescence［J］. Practical Gerontology, 2018, 32(12): 1109-1111. (in Chinese)
苑莉莉. 细胞周期及信号通路与细胞衰老关系的研究进展［J］. 实用老年医学, 2018, 32(12): 1109-1111.
［13］ Taniguchi CM, Emanuelli B, Kahn CR. Critical nodes in signalling pathways: insights into insulin action［J］. Nat Rev Mol Cell Biol, 2006, 7(2): 85-96.
［14］ Blair T , Moore SF, Williams CM, et al. Phosphoinositide 3kinases p110α and p110β have differential roles in insulinlike growth factor-1-mediated Akt phosphorylation and platelet priming［J］. Arterioscler Thromb Vasc Biol, 2014, 34(8): 1681-1688.
［15］ Lavan BE, Fantin VR, Chang ET, et al. A novel 160-kDa phosphotyrosine protein in insulin-treated embryonic kidney cells is a new member of the insulin receptor substrate family［J］. J Biol Chem, 1997, 272(34): 21403-21407.
［16］ Anjali G, Kaur S, Lakra R, et al. FSH stimulates IRS-2 expression in human granulosa cells through cAMP/SP1, an inoperative FSH action in PCOS patients［J］. Cell Signal, 2015, 27(12): 2452-2466.
［17］ Lei L, Han F, Cui Q, et al. IRS2 depletion inhibits cell proliferation and decreases hormone secretion in mouse granulosa cells［J］. J Reprod Develop, 2018, 64(5): 409-416.
［18］ Roser JF. Regulation of testicular function in the stallion: an intricate network of endocrine, paracrine and autocrine systems［J］. Anim Reprod Sci, 2008, 107(3-4): 179-196.
［19］ Cao Y, Lindstrm S, Schumacher F, et al. Insulin-like growth factor pathway genetic polymorphisms, circulating IGF1 and IGFBP3, and prostate cancer survival［J］. J Natl Cancer Inst, 2014, 106(6):dju085.
［20］ Lue Y，Swerdloff R，Liu Q，et al. Opposing roles of insulin-like growth factor binding protein 3 and humanin in the regulation of testicular germ cell apoptosis［J］. Endocrinology, 2010, 151(1): 350-357.
［21］ Vassilieva I, Kosheverova V, Vitte M, et al. Paracrine senescence of human endometrial mesenchymal stem cells: a role for the insulin-like growth factor binding protein 3.［J］.Aging (Albany NY), 2020, 12(2): 1987-2004.
［22］ Wang K, Lin C, Wang C, et al. Silencing Kif2a induces apoptosis in squamous cell carcinoma of the oral tongue through inhibition of the PI3K/Akt signaling pathway［J］. Mol Med Rep, 2014, 9(1): 273-278.
［23］ Hou X, Huang F, Macedo LF, et al. Dual IGF-1R/InsR inhibitor BMS-754807 synergizes with hormonal agents in treatment of estrogen-dependent breast cancer［J］. Cancer Res, 2011, 71(24): 7597-7607.