| 研究実績の概要 |
In vitro studies were conducted to determine the protective effects of (P)RR monoclonal antibodies (mAb) in podocyte injury. In first step, we tried to focus on the development of podocyte injury model in mouse immortalized podocytes. After differentiation, podocytes were cultured for 72 hours in medium containing 5 mM D-glucose plus 20 mM D-mannitol (osmotic control); 25 mM D-glucose (high glucose); and 25 mM D-glucose (high glucose) with 200 microgram/mL (P)RR neutralizing mAb (clone 48_8). Quantitative real time PCR was performed to check the mRNA expression of podocyte injury markers: nephrin, podocin and synaptopodin. The gene expression data revealed that high glucose intervention reduced the mRNA expression of nephrin compared to the osmotic control group, although the level of gene expression was very small. In contrary, (P)RR mAb treatment caused the increase expression of nephrin mRNA, suggestive of the reduced podocyte injury. In case of podocin and synaptopodin the expression level is unsatisfactory. Therefore, we tried to establish the podocyte injury model in the human immortalized podocyte, AB 8/13 cells. Gene expression of podocin, nephrin and synaptopodin in this human podocyte cell line were also not satisfactory. Therefore, it is hard to conduct other experiments in the above mentioned podocyte injury model to test the effects of our neutralizing (P)RR mAb in detail. Therefore, the podocin data are in line with our hypothesis that (P)RR mAb has the potential to protect the podocytes against injurious effects.
|
