JAK2/STAT3通路在雌激素缺乏相关骨髓间充质干细胞衰老中的作用初探

doi:10.3969/j.issn.1005-4979.2020.05.005

摘要/Abstract

摘要：

目的： 探究非受体型蛋白酪氨酸激酶2/信号转导与转录激活子3（janus kinase 2/signal transducer and activator of transcription 3， JAK2/STAT3）信号通路在雌激素缺乏所致骨髓间充质干细胞（bone marrow mesenchymal stem cells, BMSCs）衰老中的作用。方法： 提取小鼠BMSCs，并用适宜浓度的过氧化氢（H₂O₂）诱导其衰老。进一步添加雌激素类似物17β-雌二醇（17β-estradiol，E2）或JAK通路抑制剂（鲁索替尼），用实时荧光定量聚合酶链反应（quantitative polymerase chain reaction, qPCR）、Western印迹法（Western blot）、β-半乳糖苷酶（SA-β-Gal）染色法检测其衰老指标及JAK2/STAT3信号通路的改变。结果： H₂O₂诱导的BMSCs衰老可被雌激素抑制，且伴随着JAK2/STAT3信号的下调，而进一步抑制JAK2/STAT3信号通路也可以挽救BMSCs的衰老。结论： JAK2/STAT3信号通路是雌激素调控BMSCs的衰老靶点，且抑制该通路可以挽救BMSCs的衰老。

关键词: 细胞衰老, 酪氨酸激酶2/信号转导与转录激活子3, 骨髓间充质干细胞, 雌激素

Abstract:

Objective: To investigate the role of janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signaling pathway in bone marrow mesenchymal stem cells(BMSCs) senescence caused by estrogen deficiency. Methods: BMSCs of mice were extracted and treated with H₂O₂ in appropriate concentration to be senescent. Estrogen analog 17β-estradiol(E2) or JAK pathway inhibitor （ruxolitinib） was further added, and senescence indicators and changes of JAK2/STAT3 signaling pathway were detected by quantitative polymerase chain reaction(qPCR), Western blot and SA-β-gal staining. Results: The senescence of BMSCs induced by H₂O₂ can be inhibited by estrogen and accompanied with the down-regulation of JAK2/STAT3 signaling. Further suppression of JAK2/STAT3 signaling pathway can also rescue BMSCs senescence. Conclusion: The JAK2/STAT3 signaling pathway is a target of estrogen-related BMSCs senescence, and inhibition of JAK2/STAT3 can rescue BMSCs senescence.

Key words: cellular senescence, janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3), bone marrow mesenchymal stem cells （BMSCs）, estrogen

中图分类号:

R782

武文婧, 傅稼耀, 魏玉, 马鹏飞, 吴珺华. JAK2/STAT3通路在雌激素缺乏相关骨髓间充质干细胞衰老中的作用初探[J]. 《口腔颌面外科杂志》, 2020, 30(5): 288-294.

WU Wenjing, FU Jiayao, WEI Yu, MA Pengfei, WU Junhua. A Preliminary Study on the Role of JAK2/STAT3 Signaling Pathway in Estrogen-related BMSCs Senescence[J]. 《Journal of Oral and Maxillofacial Surgery》, 2020, 30(5): 288-294.

https://journal06.magtech.org.cn/Jweb_joms/CN/Y2020/V30/I5/288

图/表 5

表1 qPCR引物序列表

Table 1 Primer sequences of qPCR

基因	引物序列（5′-3′）
p21	上游引物：CCTGGTGATGTCCGACCTG
	下游引物：CCATGAGCGCATCGCAATC
p53	上游引物：CTCTCCCCCGCAAAAGAAAAA
	下游引物：CGGAACATCTCGAAGCGTTTA
白细胞介素6（interleukin6，IL6）	上游引物：TAGTCCTTCCTACCCCAATTTCC 下游引物：TTGGTCCTTAGCCACTCCTTC
肿瘤坏死因子α（tumor necrosis factor α，TNFα）	上游引物：CCCTCACACTCAGATCATCTTCT 下游引物：GCTACGACGTGGGCTACAG
白细胞介素1α（interleukin1α，IL1α）	上游引物：GCACCTTACACCTACCAGA 下游引物：AAACTTCTGCCTGACGAGC
核因子κB（nuclear factor kappa B，NFκB）	上游引物：ATGGCAGACGATGATCCCTAC 下游引物：TGTTGACAGTGGTATTTCTGGTG
雌激素受体α（estrogen receptor α，Erα）	上游引物：CCTCCCGCCTTCTACAGGT 下游引物：CACATTTGGGCTTGCAGTCTG
雌激素受体β（estrogen receptor β，Erβ）	上游引物：CTGTGATGAACTACAGTGTTCCC 下游引物：CACATTTGGGCTTGCAGTCTG
磷酸甘油醛脱氢酶（glyceraldehyde phosphate dehydrogenase，GAPDH）	上游引物：AGGTCGGTGTGAACGGATTTG 下游引物：TGTAGACCATGTAGTTGAGGTCA

图1 H₂O₂诱导BMSCs衰老及JAK2/STAT3通路激活注：A. 对照组和H₂O₂组p53、p21基因的表达；B. 对照组和H₂O₂组p53、p21的蛋白表达；C. 对照组和H₂O₂组SA-β-gal染色结果（×20）；D. 对照组和H₂O₂组SASP因子的表达；E. 对照组和H₂O₂组JAK2、STAT3、PJAK2、PSTAT3的蛋白表达。*表示与对照组相比，P<0.05。

Figure 1 H₂O₂ induced senescence of BMSCs and activation of JAK2/STAT3 pathway

图2 雌激素可以挽救H₂O₂诱导的BMSCs衰老注：A. qPCR检测加入E2后衰老基因p53、p21的表达；B. 各组SA-β-gal染色结果（×20）；C. 各组SASP因子的表达情况；D. Western印迹法检测各组衰老蛋白及JAK2/STAT3的改变。*表示与对照组相比，P<0.05；#表示与H₂O₂组相比，P<0.05。

Figure 2 Estrogen rescued H₂O₂-induced senescence of BMSCs

图3 雌激素通过JAK2/STAT3影响BMSCs衰老注：A. 各组雌激素受体ERα、ERβ的表达；B. 各组JAK2/STAT3的蛋白表达。*表示与H₂O₂组相比， P<0.05；#表示与H₂O₂+E2组相比，P<0.05。

Figure 3 Estrogen affected BMSCs senescence through JAK2/STAT3 signaling

图4 抑制JAK2/STAT3可以缓解H₂O₂诱导的BMSCs衰老注：A. Western印迹法检测加入鲁索替尼后JAK2/STAT3的改变；B. 各组p53、p21的基因表达；C. 各组p53、p21的蛋白表达；D. 各组SA-β-gal染色结果（×20）；E. 各组SASP因子的表达。*表示与对照组相比，P<0.05；#表示与H₂O₂组相比，P<0.05。

Figure 4 Inhibition of JAK2/STAT3 alleviated H₂O₂-induced BMSCs senescence

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