松果菊苷对大鼠Leydig细胞衰老模型AXL和wnt2b基因的影响
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摘要
目的探讨松果菊苷对大鼠Leydig细胞衰老模型AXL和wnt2b基因的影响。方法选用原代大鼠睾丸间质(Leydig)细胞为研究对象,主要分为正常组,衰老组和松果菊苷用药组,采用实时荧光定量聚合酶链式反应(PCR)和甲基化特异性PCR检测AXL和wnt2b基因的表达水平和甲基化水平。结果松果菊苷用药组AXL mRNA表达水平明显低于衰老组(P<0.05),wnt2b甲基化水平明显高于衰老组(P<0.05)。结论松果菊苷可以通过降低AXL mRNA表达水平,提高wnt2b基因的甲基化水平,抑制Leydig细胞的衰老状态。
Objective To investigate the effects of echinacoside on AXL and wnt2 b genes in aging Leydig cell model of rats.Methods The primary rat Leydig cells were selected as the study subjects. They were divided into normal, aging and echinacoside groups. Real-time fluorescence quantitative PCR and methylation-specific PCR were used to detect the expression and the methylation levels of AXL and wnt2 b genes. Results The expression level of AXL mRNA in the echinacoside treatment group was lower than that of the aging group(P<0.05), however the methylation level of wnt2 b was significantly increased(P<0.05).Conclusions Echinacoside can inhibit the senescence of Leydig cells by decreasing the mRNA expression level of AXL and increasing the methylation level of wnt2 b gene. However, the specific mechanism of action remains to be further studied.
Objective To investigate the effects of echinacoside on AXL and wnt2 b genes in aging Leydig cell model of rats.Methods The primary rat Leydig cells were selected as the study subjects. They were divided into normal, aging and echinacoside groups. Real-time fluorescence quantitative PCR and methylation-specific PCR were used to detect the expression and the methylation levels of AXL and wnt2 b genes. Results The expression level of AXL mRNA in the echinacoside treatment group was lower than that of the aging group(P<0.05), however the methylation level of wnt2 b was significantly increased(P<0.05).Conclusions Echinacoside can inhibit the senescence of Leydig cells by decreasing the mRNA expression level of AXL and increasing the methylation level of wnt2 b gene. However, the specific mechanism of action remains to be further studied.
引文
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9 Bormann F,Rodríguez-Paredes M,Hagemann S,et al.Reduced DNA methylation patterning and transcriptional connectivity define human skin aging[J].Aging Cell,2016;15:563-71.
10 Zampieri M,Ciccarone F,Calabrese R,et al.Reconfiguration of DNA methylation in aging[J].Mech Ageing Dev,2015;151:60-70.
11 Heidelbaugh JJ.Management of erectile dysfunction[J].Am Fam Physician,2010;81(3):305-12.
12 Chen FF,Song FQ,Chen YQ,et al.Exogenous testosterone alleviates cardiac fibrosis and apoptosis via Gas6/Axl pathway in the senescent mice[J].Exp Gerontol,2019;119:128-37.
13 Shen LX,Zhou S,Glowacki J.Effects of age and gender on WNT gene expression in human bone marrow stromal cells[J].J Cell Biochem,2009;106(2):337-43.
14 Lombardi APG,Royer C,Pisolato R.Physiopathological aspects of the Wnt/β-catenin signaling pathway in the male reproductive system[J].Spermatogenesis,2013;3(1):1-8.
15 Takase HM,Nusse R.Paracrine Wnt/β-catenin signaling mediates proliferation of undifferentiated spermatogonia in the adult mouse testis[J].Proc Natl Acad Sci U S A,2016;113(11):E1489-E1497.
16 Koch S,Acebron SP,Herbst J,Hatiboglu G,Niehrs C.Post-transcriptional Wnt signaling governs epididymal sperm maturation[J].Cell,2015;163:1225-36.
17 Theka I,Sottile F,Cammisa M,et al.Wnt/β-catenin signaling pathway safeguards epigenetic stability and homeostasis of mouse embryonic stem cells[J].Sci Rep,2019;9:948.
18 Chassot AA,Le Rolle M,Jourden M,et al.Constitutive WNT/CTNNB1 activation triggers spermatogonial stem cell proliferation and germ cell depletion[J].Dev Biol,2017;426(1):17-27.
19 Takase HM,Nusse R.Paracrine Wnt/β-catenin signaling mediates proliferation of undifferentiated spermatogonia in the adult mouse testis[J].Proc Natl Acad Sci U S A,2016;113(11):E1489-97.
2 韩飞,周孟良.过氧化氢诱导 HepG2 细胞产生氧化应激细胞模型的建立[J].食品科学,2011;32(5):55-7.
3 李朝晖,王芬,刘水平,等.过氧化氢诱导PCI2细胞损伤的模型的建立[J].法医学杂志,2007;23(3):191-2.
4 da Silva RA,Mariano FS,Planello AC.HaCaT anchorage blockade leads to oxidative stress,DNA damage and DNA methylation changes[J].Biochem Biophys Reports,2015;(2):94-102.
5 Wang Y,Wu W,Yao C.Elevated tissue cr levels,increased plasma oxidative markers,and global hypomethylation of blood DNA in male sprague-dawley rats exposed to potassium dichromate in drinking water[M].Wiley Periodicals,Inc.2015:1-11.
6 Sasaki M,Kajiya H,Ozeki S.Reactive oxygen species promotes cellular senescence in normal human epidermal keratinocytes through epigenetic regulation of p16INK4a[J].Biochem Biophys Res Commun,2014;452:622-8.
7 Ferioli M,Zauli G, Maiorano P,et al.Role of physical exercise in the regulation of epigenetic mechanisms in inflammation,cancer,neurodegenerative diseases,and aging process[J].J Cell Physiol,2019:epub.
8 孙静.何首乌饮及其单味药对大鼠Leydig细胞衰老的表观遗传基因调控作用[D].保定:河北大学,2018.
9 Bormann F,Rodríguez-Paredes M,Hagemann S,et al.Reduced DNA methylation patterning and transcriptional connectivity define human skin aging[J].Aging Cell,2016;15:563-71.
10 Zampieri M,Ciccarone F,Calabrese R,et al.Reconfiguration of DNA methylation in aging[J].Mech Ageing Dev,2015;151:60-70.
11 Heidelbaugh JJ.Management of erectile dysfunction[J].Am Fam Physician,2010;81(3):305-12.
12 Chen FF,Song FQ,Chen YQ,et al.Exogenous testosterone alleviates cardiac fibrosis and apoptosis via Gas6/Axl pathway in the senescent mice[J].Exp Gerontol,2019;119:128-37.
13 Shen LX,Zhou S,Glowacki J.Effects of age and gender on WNT gene expression in human bone marrow stromal cells[J].J Cell Biochem,2009;106(2):337-43.
14 Lombardi APG,Royer C,Pisolato R.Physiopathological aspects of the Wnt/β-catenin signaling pathway in the male reproductive system[J].Spermatogenesis,2013;3(1):1-8.
15 Takase HM,Nusse R.Paracrine Wnt/β-catenin signaling mediates proliferation of undifferentiated spermatogonia in the adult mouse testis[J].Proc Natl Acad Sci U S A,2016;113(11):E1489-E1497.
16 Koch S,Acebron SP,Herbst J,Hatiboglu G,Niehrs C.Post-transcriptional Wnt signaling governs epididymal sperm maturation[J].Cell,2015;163:1225-36.
17 Theka I,Sottile F,Cammisa M,et al.Wnt/β-catenin signaling pathway safeguards epigenetic stability and homeostasis of mouse embryonic stem cells[J].Sci Rep,2019;9:948.
18 Chassot AA,Le Rolle M,Jourden M,et al.Constitutive WNT/CTNNB1 activation triggers spermatogonial stem cell proliferation and germ cell depletion[J].Dev Biol,2017;426(1):17-27.
19 Takase HM,Nusse R.Paracrine Wnt/β-catenin signaling mediates proliferation of undifferentiated spermatogonia in the adult mouse testis[J].Proc Natl Acad Sci U S A,2016;113(11):E1489-97.