两种不同状态系膜细胞的培养及其功能研究

Culture of mesangial cells in two different states and a study on their functions]

OBJECTIVE: To make a comparision of cell function between the mesangial cells from rats with diabetic mellitus (DMC) and the mesangial cells stimulated by high glucose (HMC) and to explore the feasibility and advantage of culturing mesangial cells (MC) from diabetic rats in studies on the pathogenesis of diabetic nephropathy. METHODS: DMC were obtained from rats with streptozotocin-induced diabetes and were cultivated in normal medium. The MC from normal rats were divided into HMC high glucose and NMC cultured in normal medium as control. The cellular proliferation was assessed using 3H-thymidine incorporation. Production of fibronectin (FN) was analysed by ELISA. And with the use of laser scanning confocal microscopy, the calcium concentration (Ca2+]i) and the response of Ca2+]i to Angiotensin II (Ang II) were measured. RESULTS: The 3H-TdR incorporation efficiency of DMC was significantly higher than that of CMC; the 3H-TdR incorporation efficiency of HMC was lower than that of CMC (P < 0.05). The secretion of FN in DMC and HMC was higher than that in CMC (P < 0.01 and P < 0.05 respectively). The response of Ca2+]i to Ang II in DMC and HMC was decreased (P < 0.05), and the decrease was more significant in DMC than in HMC. CONCLUSIONS: The function and biologic character of mesangial cells cultured from diabetic rats are more similar to those of mesangial cells from diabetic in vivo. The result suggest that establishing a diabetes model first and starting the culture of MC next is probably a good method for investigating the mesangial cells in diabetic nephropathy.