L-carnitine(左旋肉碱)在视网膜色素上皮细胞-间充质细胞转换过程中的作用及机制研究

Proliferative vitreoretinopathy（PVR） is the main cause of refractory blindness. The epithelial-mesenchymal transition (EMT) process of retinal pigment epithelium (RPE) cells is the key in the fibrosis process of PVR. Our recent metabolomic study demonstrated that L-carnitine (LC) was seriously down-regulated in vitreous of PVR patients. Further study indicated that LC could significantly depress the EMT process of RPE cells. But the mechanism is still unknown. It has been shown that LC inhibited fibrosis through PPARγ indirectly and PPARγ is in the downstream of MAPK pathway which is an important pathway in PVR process. So, we conjecture that LC may directly bind to the target of MAPK pathway which is one of the main signal pathways in PVR and regulate the EMT process of RPE cells in the end. In this study, we will adopt molecular biological tools combined with computer reverse docking technology and biacore technology, making deeply investigation on the effects and mechanisms of LC on EMT process of RPE cells. This study will supply a foundation for the small molecular intervention of PVR.

增生性玻璃体视网膜病变（PVR）是难治性致盲性眼病，视网膜色素上皮细胞（RPE）发生上皮-间充质细胞转换（EMT）是其纤维瘢痕化病理过程中的关键。我们前期代谢组学研究发现，L-carnitine（左旋肉碱，LC）在PVR患者玻璃体中含量明显减少，体外实验进一步证实LC可以抑制RPE细胞的EMT过程。但是LC调控RPE细胞EMT的机制尚不清楚。有研究表明，LC可间接调控PPARγ抑制纤维化，而PPARγ处于PVR过程中重要信号通路-MAPK信号通路的下游。据此，我们推测LC可能与MAPK信号通路中的靶蛋白结合，最终通过PPARγ抑制RPE细胞的EMT过程。本课题将采用分子生物学手段结合计算机模拟分子对接及biacore技术，从细胞水平和动物模型上深入调查LC对RPE细胞EMT的调控作用和作用机制。本研究将为PVR的小分子药物干预研究奠定基础。

增生性玻璃体视网膜病变（PVR）是难治性致盲性眼病，视网膜色素上皮细胞（RPE）发生上皮-间充质细胞转换（EMT）是其纤维瘢痕化病理过程中的关键。本研究发现，L-carnitine（左旋肉碱，LC）在体外可以明显抑制RPE细胞的迁移及EMT过程中上皮及间充质细胞标志物的变化。本研究显示MAPK通路中的ERK通路及JNK通路参与CL对EMT的抑制作用，同时PPARγ在其中起关键作用，并验证了LC的作用通路JNK/ERK-PPARγ-NF-kB/Snail。同时发现长链非编码RNA- MALAT1在RPE细胞EMT过程中的促进作用；发现BMP7能在体内及体外抑制RPE细胞的增殖及纤维化过程。本研究扩充了PVR研究的代谢组学及表观遗传学领域，并为PVR的小分子药物干预及预防研究奠定了基础。