Long Non-Coding RNA-MALAT1 Mediates Retinal Ganglion Cell Apoptosis Through the PI3K/Akt Signaling Pathway in Rats with Glaucoma (Retracted article. See vol. 55, pg. 383, 2021)
Long Non-Coding RNA-MALAT1 Mediates Retinal Ganglion Cell Apoptosis Through the PI3K/Akt Signaling Pathway in Rats with Glaucoma (Retracted article. See vol. 55, pg. 383, 2021)
复制标题
长非编码 RNA-MALAT1 通过 PI3K/Akt 信号通路介导青光眼大鼠视网膜神经节细胞凋亡
DOI：
10.1159/000484231
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发表时间：
2017-01-01
影响因子：
--
通讯作者：
Ji, Dan
中科院分区：
文献类型：
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作者：
Li, Hai-Bo;You, Qi-Sheng;Ji, Dan
背景/目标：本研究旨在探讨长链非编码RNA-MALAT 1（LncRNA-MALAT 1）对PI 3 K/Akt信号通路介导的青光眼大鼠视网膜神经节细胞（RGC）凋亡的影响。方法：分离培养RGCs，免疫细胞化学法检测抗大鼠Thy-1、Brn 3a和RBPMS单克隆抗体的表达。构建过表达载体MALAT 1-RNA激活（RNAa）、基因敲除载体MALAT 1-RNA干扰（RNAi）和对照载体MALAT 1-阴性对照（NC）。采用巩膜外静脉烧灼法建立慢性高眼压大鼠青光眼模型。RGC分为RGC对照组、RGC压力组、RGC压力+MALAT 1-NC组、RGC压力+MALAT 1-RNAi组和RGC压力+MALAT 1-RNAa组。将60只SD大鼠随机分为正常组、高眼压组、高眼压+MALAT 1-NC组、高眼压+MALAT 1-RNAa组和高眼压+MALAT 1-RNAi组。采用qRT-PCR和western blotting检测LncRNA-MALAT 1和PI 3 K/Akt的表达水平。末端脱氧核苷酸转移酶dUTP缺口末端标记法（TUNEL）和流式细胞术检测RGC凋亡。结果：免疫细胞化学显示培养的RGCs纯度达90%。与RGC压力+MALAT 1-NC组相比，RGC压力+MALAT 1-RNAi组MALAT 1表达水平升高，PI 3 K和Akt总蛋白水平降低，RGC凋亡减少，而RGC压力+MALAT 1-RNAi组这些表达水平逆转。高眼压模型组大鼠视网膜神经节细胞数及PI 3 K/Akt表达水平均低于正常组。高眼压+MALAT 1-RNAa组PI 3 K/Akt的mRNA和蛋白表达水平较低，但高于其他3个高眼压模型组。此外，高眼压+MALAT 1-RNAa组RGC数低于正常组，但高于其他3个高眼压模型组。结论：我们的研究为LncRNA-MALAT 1通过激活PI 3 K/Akt信号通路抑制青光眼RGC凋亡提供了证据。
Background/Aims: The aim of the present study is to investigate the effect of long non-coding RNA-MALAT1 (LncRNA-MALAT1) on retinal ganglion cell (RGC) apoptosis mediated by the PI3K/Akt signaling pathway in rats with glaucoma. Methods: RGCs were isolated and cultured, and monoclonal antibodies (anti-rat Thy-1, Brn3a and RBPMS) were examined by immunocytochemistry. An overexpression vector MALAT1-RNA activation (RNAa), gene knockout vector MALAT1-RNA interference (RNAi), and control vector MALAT1-negative control (NC) were constructed. A chronic high intraocular pressure (IOP) rat model of glaucoma was established by episcleral vein cauterization. The RGCs were divided into the RGC control, RGC pressure, RGC pressure + MALAT1-NC, RGC pressure + MALAT1-RNAi and RGC pressure + MALAT1-RNAa groups. Sixty Sprague-Dawley (SD) rats were randomly divided into the normal, high IOP, high IOP + MALAT1-NC, high IOP + MALAT1-RNAa and high IOP + MALAT1-RNAi groups. qRT-PCR and western blotting were used to detect the expression levels of LncRNA-MALAT1 and PI3K/Akt. Terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) and flow cytometry were used to detect RGC apoptosis. Results: Immunocytochemistry revealed that the cultured RGCs reached 90% purity. Compared with the RGC pressure + MALAT1-NC group, the RGC pressure + MALAT1-RNAa group exhibited elevated expression levels of MALAT1, lower total protein levels of PI3K and Akt and decreased RGC apoptosis, while these expression levels were reversed in the RGC pressure + MALAT1-RNAi group. RGC numbers and PI3K/Akt expression levels in the high IOP model groups were lower than those in the normal group. In the high IOP + MALAT1-RNAa group, the mRNA and protein expression levels of PI3K/Akt were reduced but higher than those in the other three high IOP model groups. Additionally, RGC numbers in the high IOP + MALAT1-RNAa group were lower than those in the normal group but higher than those in the other three high IOP model groups. Conclusion: Our study provides evidence that LncRNA-MALAT1 could inhibit RGC apoptosis in glaucoma through activation of the PI3K/Akt signaling pathway.