Protective Effect Of ACTH_4-10 On Adriamycin-induced Podocyte Injury

Objective：To observe the influence of adrenocorticotropic hormone (ACTH4-10) in thechanges of podocyte activity, apoptosis and expression of nephrin and podocin onadriamycin-induced podocyte injury and investigate the protective effect ofACTH4-10.Methods：1. Podocytes recovery and culture：the resurgent conditional immortalizedmouse podocyte clone (MPC5) cells were cultured in33℃．More than80%coverage couldbe achieved in about3days．Then the condition was replaced to37℃，and the cells woulddifferentiate into mature podocytes in10-14days later. At this time observing thenormal podocyte morphology, the cells in good condition could be used for experiment.2. Experimental groups: All podocytes were randomly divided into following groups:normal control, ADR-induced group and ACTH4-10intervention group (low, middle andhigh concentration). Normal control group was not treated, ADR (1μmol/L) was used toinduce the model of podocyte injury and then the podocytes were intervened by1ng/ml,10ng/ml and100ng/ml ACTH4-10for1hours prior toADR-induced24hours.3. Detection index: Cell counting Kit (Cell Counting Kit-8, CCK-8) was used to detect theactivity of podocytes and TUNEL (TdT-mediated dUTP nick-end labeling) apoptosisdetection kit was used to detect podocyte apoptosis. RT-PCR and Western-blottingtechnology were used to examine the expression of nephrin and podocin.Results: Compared with control group，podocyte activity and expression of nephrin andpodocin decreased significantly in ADR-induced group(P＜0.05), meanwhile podocyteapoptosis also increased obviously. Compared with ADR-induced group, podocyte activityincreased obviously in ACTH4-10intervention group (low concentration)(P＜0.05), andexpression of nephrin and podocin increased slightly(P＞0.05). podocyte activity increasedsignificantly in ACTH4-10intervention group (middle concentration)(P＜0.05), mRNA and protein of nephrin increased greatly(P＜0.05), and mRNA of podocin increased slightly (P＞0.05), while the increasing of protein was more significant than mRNA (P＜0.05).podocyte activity increased obviously in ACTH4-10intervention group (high concentration)(P＜0.05), and expression of nephrin and podocin increased greatly (P＜0.05). Althoughapoptosis of podocyte increased while comparing with normal control group, podocyteapoptosis decreased obviously while comparing with ACTH4-10intervention group (low,middle and high concentration).Conclusions: ACTH4-10can stabilize expression of nephrin and podocin on sliddiaphragm, and has the protective effect on podocyte injury induced by ADR, while theeffect size depends on the concentration ofACTH4-10.