淫羊藿次苷I通过Akt信号通路上调Bmi1促进小鼠精原干细胞增殖

廖天龙; 楚雪桐; 杨小平

PDF(3277 KB)

湖南师范大学学报医学版 ›› 2023, Vol. 20 ›› Issue (1) : 13-17.

基础医学

淫羊藿次苷I通过Akt信号通路上调Bmi1促进小鼠精原干细胞增殖

廖天龙, 楚雪桐, 杨小平

作者信息 +

Mechanism of Icariin I promoting proliferation of mouse spermatogenic stem cells

LIAO Tianlong, CHU Xuetong, YANG Xiaoping

Author information +

文章历史 +

摘要

目的: 探究淫羊藿次苷I对小鼠精原干细胞增殖的促进作用及相关机制。方法: 使用MTT实验、EdU细胞增殖实验检测淫羊藿次苷I对小鼠精原干细胞增殖能力的影响;Western blot法检测淫羊藿次苷I对Bmi1、p-Akt和Akt蛋白的表达情况;RT-PCR实验检测相关基因的表达结果,最后通过抑制剂和siRNA证明p-Akt和Bmi1蛋白之间存在上下游的调控关系。结果: 与对照组相比,0.5-1µM淫羊藿次苷I促进小鼠精原干细胞增殖（P<0.05）;相比于对照组,0.5-1µM淫羊藿次苷I处理组p-Akt和Bmi1蛋白表达水平上升（P<0.05）;Akt抑制剂下调p-Akt和Bmi蛋白水平（P<0.05）;siBmi1对Akt mRNA水平无显著影响（P>0.05）。结论: 淫羊藿次苷I激活Akt信号通路上调Bmi1表达促进小鼠精原干细胞增殖。

Abstract

Objective The medicine icariin I is applied to mouse spermatogonial stem cells to explore the effect of the medicine on the proliferation function of the cells and the related mechanism for the effect. Methods The changes in the value-added ability of mouse spermatogenic stem cells in the presence of icariin I were detected by using the MTT and EdU cell proliferation methods. The changes of Bmi1, p-Akt, and Akt proteins in mouse spermatogenic stem cells in the presence of icariin I were detected by Western Blot experiment. The expression of related genes was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Finally, inhibitors and siRNA were used to explore regulatory relationship between p-Akt and Bmi1 protein. Results Icariin I at the concentration of 0.5-1µM could significantly enhance the proliferation of mouse spermatogenic stem cells as compared with that in the control group (P<0.05). Meanwhile, within the same concentration range, the expression levels of p-Akt and Bmi1 proteins in mouse spermatogenic stem cells were significantly increased under the action of drugs (P<0.05). P-Akt and Bmi1 protein levels were significantly down-regulated in the AKT inhibitor group, and the difference was statistically significant with the control group (P<0.05). There were no significant changes in Akt mRNA levels in the siBmi1 group compared with the control group (P>0.05). Conclusion Icariin I can enhance the proliferation of mouse spermatogonial stem cells, specifically by activating Akt signaling pathway to up-regulate the expression of Bmi1.

导出引用

廖天龙, 楚雪桐, 杨小平. 淫羊藿次苷I通过Akt信号通路上调Bmi1促进小鼠精原干细胞增殖[J]. 湖南师范大学学报医学版. 2023, 20(1): 13-17

LIAO Tianlong, CHU Xuetong, YANG Xiaoping. Mechanism of Icariin I promoting proliferation of mouse spermatogenic stem cells[J]. Journal of Hunan Normal University(Medical Science). 2023, 20(1): 13-17

中图分类号： R711.6

参考文献

[1] Dutta S, Biswas A, Sengupta P.Obesity, endocrine disruption and male infertility[J]. Asian Pac J Reprod, 2019, 8(5): 195.
[2] Zhang F, Liu M, Gao J.Alterations in synaptonemal complex coding genes and human infertility[J]. Int J Biol Sci, 2022, 18(5): 1933-1943.
[3] Serrano R, Garcia-Marin L J, Bragado MJ. Sperm Phosphoproteome: Unraveling Male Infertility[J]. Biology (Basel), 2022, 11(5): 659.
[4] Yang D, Liu HQ, Yang Z, et al.BMI1 in the heart: Novel functions beyond tumorigenesis[J]. EBioMedicine, 2021, 63: 103193.
[5] Abeyrathna P, Su Y.The critical role of Akt in cardiovascular function[J]. Vascul Pharmacol, 2015, 74: 38-48.
[6] Peltier J, O'Neill A, Schaffer DV. PI3K/Akt and CREB regulate adult neural hippocampal progenitor proliferation and differentiation[J]. Dev Neurobiol, 2007, 67(10): 1348-1361.
[7] Rafalski VA, Brunet A.Energy metabolism in adult neural stem cell fate[J]. Prog Neurobiol, 2011, 93: 182-203.
[8] 刘子琛, 徐英, 陈世忠. 淫羊藿次苷Ⅰ在大鼠体内的药动学和组织分布研究[J]. 中国药学杂志, 2008(11): 848-851.
[9] 訾慧, 李可强, 尚德阳, 等. 单体及复方中的淫羊藿苷在骨质疏松模型大鼠体内代谢及组织分布的比较[J]. 中国实验方剂学杂志, 2015, 21(6): 135-139.
[10] 訾慧, 范颖, 蒋宁, 等. 淫羊藿次苷Ⅰ及淫羊藿次苷Ⅱ通过BMP/Runx2/Osx信号通路促进大鼠骨髓间充质干细胞成骨分化的实验研究[J]. 中国骨质疏松杂志, 2019, 25(05): 690-695.
[11] Zhou S, Dong J, Xiong M, et al.UHRF1 interacts with snRNAs and regulates alternative splicing in mouse spermatogonial stem cells[J]. Stem Cell Reports, 2022, 17(8): 1859-1873.
[12] Yu J, Shen C, Lin M, et al.BMI1 promotes spermatogonial stem cell maintenance by epigenetically repressing Wnt10b/β-catenin signaling[J]. Int J Biol Sci, 2022, 18(7): 2807-2820 .
[13] Chen Z, Li L, Wu W, et al.Exercise protects proliferative muscle satellite cells against exhaustion via the Igfbp7-Akt-mTOR axis[J]. Theranostics, 2020, 10(14): 6448-6466.
[14] Ma Y, Sender S, Sekora A, et al.The Inhibitory Response to PI3K/AKT Pathway Inhibitors MK-2206 and Buparlisib Is Related to Genetic Differences in Pancreatic Ductal Adenocarcinoma Cell Lines[J]. Int J Mol Sci, 2022, 23(8): 4295.
[15] Carson SA, Kallen AN.Diagnosis and Management of Infertility: A Review[J]. JAMA, 2021, 326(1): 65-76.
[16] Bosch E, De Vos M, Humaidan P.The Future of Cryopreservation in Assisted Reproductive Technologies[J]. Front Endocrinol (Lausanne), 2020, 11: 67.
[17] Wang M, Wang Q, Du Y, et al.Vitamins combined with traditional Chinese medicine for male infertility: A systematic review and meta-analysis[J]. Andrology, 2020, 8(5): 1038-1050.
[18] Zhang Z, Yang J, Kong T, et al.Traditional Chinese Medicine syndrome elements of male infertility revealed by latent tree model analysis[J]. J Tradit Chin Med, 2018, 38(6): 926-935.
[19] 纪昕. 淫羊藿苷免疫调节作用的研究进展[J]. 河北医药, 2016, 38(05): 753-756.
[20] 李康梅, 黎明星, 陈泯锜, 等. 淫羊藿苷对生殖系统影响的研究进展[J]. 西部中医药, 2022, 35(04): 143-146.
[21] 张攀, 史娟, 刘力, 等. 淫羊藿苷及其衍生物的抗氧化活性研究[J]. 中国食品添加剂, 2016,(02): 85-88.
[22] 张浩, 郑玉玲, 丁辉. 淫羊藿苷治疗骨质疏松的相关通路研究进展[J]. 天津中医药大学学报, 2022, 41(04): 531-538.
[23] Dai X, Zhang Q, Yu Z, et al.Bmi1 Deficient Mice Exhibit Male Infertility[J]. Int J Biol Sci, 2018, 14(3): 358-368.
[24] Yang W, Wu Z, Yang K, et al.BMI1 promotes cardiac fibrosis in ischemia-induced heart failure via the PTEN-PI3K/Akt-mTOR signaling pathway[J]. Am J Physiol Heart Circ Physiol, 2019, 316(1): H61-H69.
[25] Su Z, Yang Z, Xu Y, et al.Apoptosis, autophagy, necroptosis, and cancer metastasis[J]. Mol Cancer, 2015, 14: 48.