1-磷酸鞘氨醇对体外培养的人视网膜色素上皮细胞α-SMA表达的影响

目的 研究1-磷酸鞘氨醇对人视网膜色素上皮细胞α-平滑肌肌动蛋白表达的影响.方法 对RPE细胞常规培养,取传3～4代生长良好的细胞用于实验,以106个/ml接种于24孔培养板中,加入含1-磷酸鞘氨醇无血清培养液,使1-磷酸鞘氨醇终浓度为1、1×10-1、1×10-2、1×10-3、1×10-4 μg/ml,加入不含1-磷酸鞘氨醇的无血清培养液为对照组,采用免疫荧光法分析1-磷酸鞘氨醇对α-平滑肌肌动蛋白表达的影响.结果 1、1×10-1、1×10-2 μg/ml 1-磷酸鞘氨醇组中表达α-平滑肌肌动蛋白阳性的细胞与对照组相比差异具有明显统计学意义（P=0.000、0.000、0.000） 而1×10-3、1×10-4 μg/ml组表达α-平滑肌肌动蛋白阳性细胞与对照组相比差异无明显的统计学意义（P=0.832、0.781）.结论 α-平滑肌肌动蛋白主要在RPE细胞的胞浆中表达.随着1-磷酸鞘氨醇浓度的增加,RPE细胞α-平滑肌肌动蛋白的表达增强.

Influence of S1 P on RPE cell a - SMA expression and RPE cell - induced collangen gel contraction in human

Objective To observe the influence of Sphingosine - 1 - phosphate （ S1 P） on expression of a - smooth muscle acin（ a- SMA） in human retinal pigment epithelium（ vitreoretinopathy RPE）cells and RPE cell- induced contration of type 1 collagen gel. Methods RPE cells on conventional culture, take 3 - 4 on behalf of the mass of ceils grow well for the experiments to 106/ml were inoculated into 24 - well plate by adding serum - free medium containing S1P to S1P final concentration of 1,1 × 10 - 1,1 × 10 - 2,1 × 10 - 3,1 × 10 - 4 ug/ml, adding without S1 P in serum - free culture medium for the control group, using immunofluorescence analysis of SIP on the expression of a - SMA. Results 1,1 × 10 -1, 1 × 10 -2 ug/ml S1P group to express a -SMA- positive cells compared with the control group a clear statistical significance （P = 0. 000,0. 0000,000）, and 1 × 10-3, 1 × 10-4 ug/ml group to express × -SMA positive ceils compared with the control group no statistically significant（ P = 0. 832,0. 781 ）. Conclusion a - SMA mainly expresses in the cytoplasm of RPE cells. With the increasing in the concentration of SIP to enhance RPE cell a -SMA expression.