ABSTRACT
@#AIM:To establish the hypoxia induced endothelial-mesenchymal transition(EndoMT)model of endothelial cells, and to investigate the effect and mechanism of Pirfenidone(PFD)on inhibiting the subretinal fibrosis progression.<p>METHODS: Primary cultured human umbilical vein endothelial cells(HUVEC), 4-7 passages were used for experiments after cell identification. CoCl<sub>2</sub> induced hypoxia to establish the transformation model of endothelial cells into fibroblasts. CCK-8 was performed to detect cell proliferation rate and chose the optimal drug concentration. All cells were divided into 4 groups: control group(FBS-free), CoCl<sub>2</sub>(200μmol/L)group, CoCl<sub>2</sub>+0.3mg/mL PFD group, CoCl<sub>2</sub>+0.6mg/mL PFD group. The protein expression of CD31, VE-cadherin, α-SMA, FSP1, p-p38 and p38 were detected by Western blot. Double immunofluorescence labeling method was used to observe the CD31/α-SMA expression. Wound healing assay detected the cell migration. The q-PCR was applied to detect the mRNA levels of TGF-β1 and SNAI1.<p>RESULTS: Compared with CoCl<sub>2</sub> group, PFD increased cell proliferation rate and inhibited cell migration significantly under hypoxia(<i>P</i><0.05). PFD decreased the protein expression of the mesenchymal markers α-SMA and FSP1, and increased the protein level of the endothelial markers CD31 and VE-cadherin(<i>P</i><0.05). Double immunofluorescence results showed that PFD could reduce the expression of α-SMA and increase the level of CD31(<i>P</i><0.05). In the process of EndoMT, the p38 protein expression level was stable(<i>P</i>>0.05). PFD down-regulated significantly the high protein expression of p-p38, and high mRNA expression of TGF-β1 and SNAI1 compared with control group(<i>P</i><0.05). There was no significant difference between the 0.3 and 0.6mg/mL PFD groups in all results above.<p>CONCLUSION: PFD can inhibit the formation of fibrosis in endothelial cells. TGF-β/p38MAPK signaling pathway might be one of the mechanisms that PFD regulates EndoMT progression. PFD will be expected to become a potential new sight on the treatment of subretinal fibrosis.
ABSTRACT
AIM:To analyze the characteristics of optical coherence tomography ( OCT ) in diabetic optic neuropathy ( DON ) retrospectively. ●METHODS:Retrospective study. A total of 175 cases of type ll diabetes with fundus lesions from Dec. 2013 to Dec. 2015 were selected and the clinical information was collected. These cases were diagnosed by consultation between Departments of Ophthalmology and Endocrinology in the Second Affiliated Hospital of Xi′an Jiao Tong University. The results of body examination were recorded and cases were examined by color fundus photography, fluorescein fundus angiography ( FFA) and OCT. All these data were analyzed. ●RESULTS: A total of 49 cases ( 90 eyes, 25. 7%) were diagnosed DON through FFA which manifested abnormal fluorescence in optic papilla. Results of OCT showed:among 90 eyes of DON patients, 15 eyes ( 16. 7%) had normal optic nerve form; 20 eyes(22. 2%) of excavation of optic disc became smaller or disappeared, with prelaminar tissue and peripapillary retinal nerve fiber layer (RNFL) swelling;26 eyes (28. 9%) manifested optic cup deep and cup/disc ratio increasing;18 eyes (20. 0%) had tissue hyperplasia in the hollow or on the surface of optic disc; 11 eyes(12. 3%) had symptoms including vitreous traction optic papilla and optic disc rim rising. DON eyes which had similar fluorescence features could manifest different tissue morph by OCT. ●CONCLUSION: FFA defines DON by change of blood circulation in optic nerve. However, OCT can show differences of tissue morph of optic nerve that FFA fails to do. So OCT can manifest the causes and sites of optic neuropathy more clearly and also provide basis for treatment. The advantages of OCT are conducive to reviews and curative effect tracking among DON patients and these advantages including noninvasive, convenient, inexpensive and repeatable.