Expression of miR-195 and its target gene Bcl-2 in human intervertebral disc degeneration and their effects on nucleus pulposus cell apoptosis

被引：13
作者：
Lin, Xue-Lin [1 ]
Zheng, Zhao-Yun [1 ]
Zhang, Qing-Shan [1 ]
Zhang, Zhen [1 ]
An, You-Zhi [1 ]
机构：
[1] Shandong First Med Univ, Hosp Liaocheng 2, Dept Spinal Surg 2, Linqing 252600, Shandong, Peoples R China
来源：
JOURNAL OF ORTHOPAEDIC SURGERY AND RESEARCH | 2021年 / 16卷 / 01期
关键词：
miR-195; Bcl-2; Intervertebral disc degeneration; Nucleus pulposus cells; Apoptosis; TNF-ALPHA; DOWN-REGULATION; MICRORNA-195; MITOCHONDRIAL; PATHWAY; MIRNAS; GROWTH; MODEL;
D O I：
10.1186/s13018-021-02538-8
中图分类号：
R826.8 [整形外科学]; R782.2 [口腔颌面部整形外科学]; R726.2 [小儿整形外科学]; R62 [整形外科学（修复外科学）];
学科分类号：
摘要：
Objective To investigate the expression of miR-195 and its target gene Bcl-2 in intervertebral disc degeneration (IVDD) and its effect on nucleus pulposus (NP) cell apoptosis. Methods The expressions of miR-195 and Bcl-2 in NP tissues of IVDD patients were quantified by qRT-PCR and western blotting, respectively. NP cells were divided into blank group, TNF-alpha group, TNF-alpha + miR-NC group, TNF-alpha + siBcl-2 group, and TNF-alpha + miR-195 inhibitors + siBcl-2 group. Cell proliferation was detected by MTT assay, cell apoptosis evaluated by flow cytometry, and mitochondrial membrane potential (MMP) tested by JC-1 staining. Moreover, the function of miR-195 on IVDD in vivo was investigated using a puncture-induced IVDD rat model. Results IVDD patients had significantly increased miR-195 expression and decreased Bcl-2 protein expression in NP tissues. The expression of miR-195 was negatively correlated with the expression of Bcl-2 in IVDD patients. Dual-luciferase reporter gene assay indicated that Bcl-2 was a target gene of miR-195. In comparison with blank group, TNF-alpha group showed decreased cell proliferation and MMP, increased cell apoptosis, upregulated expression of miR-195, Bax, and cleaved caspase 3, and downregulated Bcl-2 protein, while these changes were attenuated by miR-195 inhibitors. Additionally, siBcl-2 can reverse the protective effect of miR-195 inhibitors on TNF-alpha-induced NP cells. Besides, inhibition of miR-195 alleviated IVDD degeneration and NP cell apoptosis in the rat model. Conclusion MiR-195 was significantly upregulated in NP tissues of IVDD patients, and inhibition of miR-195 could protect human NP cells from TNF-alpha-induced apoptosis via upregulation of Bcl-2.
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页数：11
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