Bu-Shen-Zhu-Yun decoction induces PRLR deubiquitination and JAK2/STAT5 activation via CSN5 in vitro.
Bu-Shen-Zhu-Yun decoction induces PRLR deubiquitination and JAK2/STAT5 activation via CSN5 in vitro.
复制标题
补肾逐云汤体外通过 CSN5 诱导 PRLR 去泛素化和 JAK2/STAT5 激活
DOI：
10.18632/aging.203426
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发表时间：
2021-08-23
期刊：
影响因子：
--
通讯作者：
Shang Y
中科院分区：
文献类型：
--
作者：
Feng H;Zhou H;Lu J;Zhang Q;Tang X;Shang Y
目的：探讨补肾助孕汤对高催乳素血症（HPRL）不孕症患者Kisspeptin表达的影响。方法：用BSZY-D处理SD大鼠，抽取脑脊液。对GT 1 -7细胞进行不同的处理。采用Western blot、RT-qPCR和免疫荧光法检测各组GT 1 -7细胞JAK 2和STAT 5的磷酸化水平以及PRLR和kisspeptin的表达。通过Western blot、RT-PCR、免疫共沉淀和海肾荧光素酶活性检测CSN 5和GATA 1的表达及其他分子特征。结果如下：HPRL组JAK 2、STAT 5磷酸化水平及PRLR、kisspeptin表达均明显降低，BSZY-D可逆转上述变化。此外，在HPRL组中检测到PRLR去泛素化的存在，这可以被shRNA-CSN 5逆转，表明BSZY-D通过靶向CSN 5发挥作用。HPRL组GATA 1和CSN 5启动子结合水平明显降低，而HPRL（BSZY-D/CSF）组GATA 1和CSN 5启动子结合水平明显升高（P < 0.05）。结论：补肾滋阴汤能提高GATA 1的转录活性，增加GATA 1与CSN 5的结合。BSZY-D参与PRLR的去泛素化，这有助于减轻HPRL不育的症状。
Purpose: To determine the effect of Bu-Shen-Zhu-Yun Decoction (BSZY-D) on the kisspeptin through JAK2/STAT5 signaling pathway in hyperprolactinemia (HPRL) infertility. Method: SD rats were treated with BSZY-D for cerebrospinal fluid (CSF) extraction. GT1-7 cells were subjected to different treatments. The phosphorylation levels of JAK2 and STAT5, and the expressions of PRLR and kisspeptin of GT1-7 cells in different groups were detected by western blot, RT-qPCR and immunofluorescence. The expressions of CSN5 and GATA1 and other molecular features were checked by western blot, RT-PCR, co-immunoprecipitation and renilla luciferase activity. Results: The phosphorylation levels of JAK2 and STAT5, and the expressions of PRLR and kisspeptin in the HPRL group were significantly decreased, and these changes could be reversed after BSZY-D treatment. In addition, the presence of PRLR deubiquitination was detected in the HPRL group, which could be reversed by shRNA-CSN5, suggesting that BSZY-D played a role through targeting CSN5. The binding level of GATA1 and CSN5 promoter in the HPRL group was significantly decreased, but elevated in the HPRL (BSZY-D/CSF) group (P < 0.05). Conclusion: BSZY-D improved the transcription activity of GATA1 and increased the binding of GATA1 and CSN5. BSZY-D was involved in the deubiquitination of PRLR, which contributes to alleviating the symptoms of HPRL infertility.