The immunological regulation effects of human umbilical cord mesenchymal stem cells on RF/6A cultured in high glucose

Abstract

Objective To observe the immunological regulation effects of human umbilical cord mesenchymal stem cells (hUCMSC) on glucose-damaged rhesus retinal vascular endothelial cells (RF/6A). Methods hUCMSC and RF/6A were co-culture according to 1: 1 ratio in the co-culture system (Transwell plates), hUCMSC cells were added to upper chamber, while the lower chamber containing 25mmol/L glucose and RF/6A. There were three groups including RF/6A blank control group, high glucose treated RF/6A group, and high glucose treated RF/6A with hUCMSC co-culture group. MTT was used to measure the RF/6A cell viability. Western blot was used to to detect protein level of Foxp3. Enzyme-linked immunosorbent assay (ELISA) was used to detect the concentration of interleukin (IL)-17. Results MTT assay revealed that at the first day, the survival rate of the three groups had no significant difference (F=0.030, P>0.05). On day 3 and day 7, the cell viability of the high glucose group was significantly lower than that of the control group(t=36.072, 27.890; P<0.05), the cell viability of the high glucose treated RF/6A with hUCMSC co-culture group was higher than that of high glucose group (t=36.072, 19.650; P<0.05). Western blot analysis showed that Foxp3 in high glucose RF/6A group was significantly lower than that in the control group at day 7 after culture (t=7.826, P<0.05) and high glucose RF/6A with hUCMSC group (t=19.936, P<0.05). ELISA showed that IL-17 in the high glucose group, high glucose with hUCMSC co-culture group was significantly higher than that of the control group (F=1 267.503, P<0.05), while IL-17 in the hUCMSC co-culture group was significantly lower than that in high glucose group (t=17.386, P<0.05). Conclusion hUCMSC can regulate the expression of Foxp3 and IL-17 to increase the proliferative ability of RF/6A, which was suppressed by high glucose. Key words: Mesenchymal stem cells; Endothelial cells/immunology; Immunomodulation; Animal experimentation