The Role Of Cdc42 Signal Protein In The Mechanism That ACTH Protected The Injuried Podocytes

Objective: Through observation and analysis of the expressed changes of nephrin, podocin, actin and cdc42 after adrenocorticotropic hormone(ACTH) effected on the podocytes those adriamycin(ADR) injuried, we studied the role of cdc42 signal protein in the mechanism that ACTH protected the injuried podocytes.Methods: 1. Podocytes recovery and culture: Firstly, rapidly melt the resurgent conditional immortalized mouse podocyte clone cells(E11) preserved in liquid nitrogen.Then the podocytes were cultured in 33℃, 3~4 days in one passage. Remove about90% podocytes and digest the podocytes by trypsin in T75 bottles. The differentiation of the podocytes was proceeding in 37℃ without gamma interferon conditions after 10 days.At this time observing the normal podocyte morphology, the cells in good condition could be used for our experiment.2.Experimental groups: All podocytes were divided into the following groups: normal control, ADR-induced group, ZCL278 intervention group, ACTH intervention group(low,middle and high concentration), ACTH and ADR intervention group(low, middle and high concentration), ACTH and ADR and ZCL278 intervention group(low, middle and high concentration).3.Detection index: RT-PCR and Western-blotting technology were used to examine the expression of nephrin, podocin, cdc42 and actin.Results: Compared with the control group, the m RNA and protein expression of nephrin and podocin in ADR-induced group decreased significantly, and the difference was statistically significant(P<0.05), meanwhile the m RNA and protein expression of cdc42 and actin did not significantly reduce. Compared with the control group, the m RNA and protein expression of cdc42 and actin in ZCL278 intervention group decreased significantly, the difference was statistically significant(P< 0.05), meanwhile the m RNA and protein expression of nephrin and podocin did not significantlyreduce. Compared with the control group, the m RNA and protein expression of nephrin and podocin in the middle ACTH intervention group treated group increased, and the difference was statistically significant(P< 0.05), and the m RNA and protein expression of cdc42 and actin did not significantly increased. Compared with low and high ACTH intervention group, the m RNA and protein expression of nephrin and podocin in the middle ACTH intervention group treated group increased, and the difference was statistically significant(P< 0.05). Compared the ADR-induced group, the m RNA and protein expression of nephrin and podocin in the middle ACTH and ADR intervention group increased obviously, and the difference was statistically significant(P<0.05), meanwhile the m RNA and protein expression of cdc42 and actin did not significantly increased. Compared with the control group, the m RNA and protein expression of cdc42 and actin in ACTH and ADR and ZCL278 intervention group decreased significantly, and the difference was statistically significant(P<0.05).Conclusions: The nephrin and podocin protein of podocytes is the basis that ACTH protected podocytes, but this protective effect is not affected by cdc42 signal protein.And we believe that our study can provide some ideas for further research.