A Potential Research Target for Scleral Remodeling: Effect of MiR-29a on Scleral Fibroblasts

作者全名："Yang, Qianying; Lv, Sha; Zhu, Huirong; Zhang, Liming; Li, Hua; Song, Shengfang"

作者地址："[Yang, Qianying; Lv, Sha; Zhu, Huirong; Zhang, Liming; Li, Hua; Song, Shengfang] Chongqing Med Univ, Dept Ophthalmol, Yongchuan Hosp, Chongqing, Peoples R China"

通信作者："Li, H; Song, SF (通讯作者)，Chongqing Med Univ, Dept Ophthalmol, Yongchuan Hosp, Chongqing, Peoples R China."

来源：OPHTHALMIC RESEARCH

ESI学科分类：CLINICAL MEDICINE

WOS号：WOS:000875571800009

JCR分区：Q2

影响因子：2.1

年份：2022

卷号：65

期号：5

开始页：566

结束页：574

文献类型：Article

关键词：miR-29a; Collagen I; PTEN; Myopia; Scleral remodeling

摘要："Introduction: The purpose of this study was to determine whether miR-29a regulates cell survival and apoptosis and the expression of phosphatase and tensin homolog deleted on chromosome 10 (PTEN), MMP-2, and collagen I in scleral fibroblasts. Methods: We transfected scleral fibroblasts with the miR-29a mimic and inhibitor. The effects of miR-29a on cell proliferation and apoptosis were determined using the CCK-8 assay and flow cytometry, respectively. Quantitative polymerase chain reaction (qPCR) was used to determine whether miR-29a regulates the mRNA levels of PTEN, MMP-2, and collagen I. The protein expression of PTEN, MMP-2, and collagen I was also assessed by western blot analysis. Results: The results of CCK-8 showed that, at 0, 24, 48, and 72 h after transfection, the relative optical density values in the mimic group were 0.233 +/- 0.005, 0.380 +/- 0.008, 0.650 +/- 0.040, and 0.906 +/- 0.032, and in the inhibitor group were 0.272 +/- 0.011, 0.393 +/- 0.029, 0.597 +/- 0.059, and 0.950 +/- 0.101, respectively. The flow cytometry results showed that the apoptosis rates of each group were as follows: the mimic group (0.043 +/- 0.007), the NC group (0.040 +/- 0.006), the inhibitor group (0.032 +/- 0.003), the inhibitor NC group (0.027 +/- 0.010), the lipofectamine group (0.027 +/- 0.005), and the blank group (0.031 +/- 0.009). The qPCR results indicated that in the mimic group, PTEN (0.795 +/- 0.182, p = 0.2783), MMP-2 (0.621 +/- 0.105, p = 0.0033), and COL1A1 (0.271 +/- 0.100, p = 0.0002) expression decreased, whereas in the inhibitor group, PTEN (1.211 +/- 0.100, p = 0.2614), MMP-2 (1.161 +/- 0.053, p = 0.1190), and COL1A1 (1.7040 +/- 0.093, p = 0.0003) increased. Western blot analysis showed that in the mimic group, the expression of PTEN (0.392 +/- 0.039, p < 0.0001), MMP-2 (0.577 +/- 0.017, p < 0.0001), and COL1A1 (0.072 +/- 0.006, p < 0.0001) protein decreased, whereas in the inhibitor group, PTEN (1.043 +/- 0.042, p = 0.9413), MMP-2 (1.397 +/- 0.075, p = 0.0002), and COL1A1 (1.935 +/- 0.081, p < 0.0001) expression increased. Conclusion: MiR-29a inhibits the expression of PTEN, MMP-2, and collagen I on scleral fibroblasts, which may provide a basis studies in sclera."

基金机构："Chongqing Municipal Science and Health Commission Joint Medical Program [Ycstc,2018nb0233]; Science and Technology Program of Yongchuan, Chongqing [YJSCX202005]; Postgraduate Innovative Program of Yongchuan Hospital of Chongqing Medical University; [2018MSXM114]"

基金资助正文："Funding for experimental materials was provided by Chongqing Municipal Science and Health Commission Joint Medical Program (2018MSXM114), Science and Technology Program of Yongchuan, Chongqing (Ycstc,2018nb0233) and Postgraduate Innovative Program of Yongchuan Hospital of Chongqing Medical University (YJSCX202005)."