微小 RNA-199a-5p 通过靶向 SP1 基因调节糖尿病性白内障中的上皮间质转化。
microRNA-199a-5p regulates epithelial-to-mesenchymal transition in diabetic cataract by targeting SP1 gene.
机构信息
Eye Center, The Second Hospital of Jilin University, #218 Ziqiang Street, Changchun, Jilin, China.
Department of Ophthalmology, Shanghai General Hospital (Shanghai First People's Hospital), Shanghai Jiaotong University Medical School, #100 Haining Road, Shanghai, China.
出版信息
Mol Med. 2020 Dec 4;26(1):122. doi: 10.1186/s10020-020-00250-7.
BACKGROUND
As a common ocular complication of diabetes mellitus, diabetic cataract is becoming a leading cause of visual impairment. The progression of diabetic cataract progression involves epithelial-to-mesenchymal transition (EMT), the precise role of which remains to be investigated. As microRNAs (miRNAs) are suggested to be involved in the pathogenesis of many diseases, identification of aberrantly expressed miRNAs in diabetic lens epithelial cells (LECs) and their targets may provide insights into our understanding of diabetic cataract and potential therapeutic targets.
METHODS
Diabetic cataract capsules and LECs exposed to high glucose (25 mmol/L, 1-5 days) were used to mimic the model. Quantitative RT-PCR was performed to evaluate the differential expression of miRNA. Dual luciferase reporter assay was used to identify the binding target of miR-199a-5p. The expression of EMT-associated proteins was determined by immunofluorescence and Western blot analysis.
RESULTS
Our results showed the differential expression of miR-9, -16, -22, -199a and -204. MiR-199a was downregulated in diabetic cataract capsule and hyperglycemia-conditioned human LECs. Specific protein 1 could be directly targeted and regulated by miR-199a in LECs and inhibit EMT in diabetic LECs.
CONCLUSION
Our findings implied miR-199a could be a therapeutic target by regulating SP1 directly to affect EMT in diabetic cataract and provided novel insights into the pathogenesis of diabetic cataract.
背景
作为糖尿病的一种常见眼部并发症，糖尿病性白内障正在成为导致视力损害的主要原因。糖尿病性白内障的进展涉及上皮间质转化（EMT），其确切作用仍有待研究。由于 microRNAs（miRNAs）被认为参与了许多疾病的发病机制，鉴定糖尿病晶状体上皮细胞（LEC）中异常表达的 miRNAs 及其靶标可能有助于我们了解糖尿病性白内障和潜在的治疗靶点。
方法
使用高糖（25mmol/L，1-5 天）诱导的糖尿病白内障囊和 LEC 来模拟模型。采用定量 RT-PCR 评估 miRNA 的差异表达。双荧光素酶报告基因检测鉴定 miR-199a-5p 的结合靶标。通过免疫荧光和 Western blot 分析测定 EMT 相关蛋白的表达。
结果
我们的结果显示 miR-9、-16、-22、-199a 和 -204 的差异表达。miR-199a 在糖尿病性白内障囊和高糖条件下人 LEC 中下调。特异性蛋白 1 可在 LEC 中被 miR-199a 直接靶向和调节，并抑制糖尿病 LEC 中的 EMT。
结论
我们的研究结果表明，miR-199a 可以通过直接调节 SP1 成为一种治疗靶点，从而影响糖尿病性白内障中的 EMT，并为糖尿病性白内障的发病机制提供了新的见解。
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