lncRNA NEAT1-let 7b-P21 axis mediates the proliferation of neural stem cells cultured in vitro promoted by radial extracorporeal shock wave

Kun Han; Nan Kang; Xiaotong Yu; Jie Lu; Yuewen Ma

doi:10.1016/j.reth.2022.06.006

lncRNA NEAT1-let 7b-P21 axis mediates the proliferation of neural stem cells cultured in vitro promoted by radial extracorporeal shock wave

Regen Ther. 2022 Jul 1:21:139-147. doi: 10.1016/j.reth.2022.06.006. eCollection 2022 Dec.

Authors

Kun Han¹, Nan Kang¹, Xiaotong Yu², Jie Lu³, Yuewen Ma¹

Affiliations

¹ Department of Rehabilitation Medicine, The First Hospital of China Medical University, Shenyang, Liaoning, PR China.
² Institute of Meta-Synthesis Medicine, Beijing, PR China.
³ Department of Human Anatomy, College of Basic Medical Sciences, China Medical University, Shenyang, Liaoning, PR China.

Abstract

In previous studies, we found radial extracorporeal shock wave (rESW), can promote the proliferation of neural stem cells (NSCs). Emerging evidence suggests that lncRNA NEAT1 can regulate NSCs proliferation. Whether lncRNA NEAT1 plays a role in the proliferation of NSC induced by shock waves is unclear. Cell Counting Kit-8（CCK 8) method was used to detect the proliferation of NSCs, and the relative protein and mRNA expression of related genes of Nestin, Cyclin D1 and P21 were detected by Western Blot and Quantitative real-time PCR (RT-qPCR) respectively. Immunofluorescence staining was used to observe the changes in the number of BrdU/nestin positive cells. Overexpression of NEAT1 and let 7 b in cells were used to explore whether rESW can rescue the decreased number of NSCs.We found that the optimal dose of R15 transmitter promoting NSCs proliferation is 1.5 bar, 500 pulse, 2 Hz. 1.2-1.5 bar showed a dose-dependent effect on the proliferation of NSCs, but it was negatively correlated with the proliferation effect of NSC when it was more than 1.5 bar. We revealed that let 7 b-P21 axis was involved in regulating the inhibition of NSC proliferation which was activated by NEAT1 in NSCs. In addition, we demonstrated that rESW treatment resulted in the decrease of NEAT1 expression, which was accompanied by the improved biological function including proliferation. Our results confirm that low-intensity rESW (1.5 bar，500 pulse，2 Hz) can promote the proliferation of NSCs through NEAT1-let 7 b-P21 axis.

Keywords: ANOVA, Analysis of variance; CCK 8, Cell Counting Kit-8; LncRNA, Long non-coding RNA; NEAT1; NSCs, Neural stem cells; Neat 1, nuclear-enriched abundant transcript 1; Neural stem cells; P21; TBI, traumatic brain injury; let 7b; q-PCR, Quantitative real-time PCR; rESW; rESW, radial extracorporeal shock wave.