ACTH4-10对阿霉素诱导体外足细胞损伤的保护作用

Protective effect of ACTH4-10 on adriamycin-induced podocyte injury

目的 通过观察促肾上腺皮质激素（ACTH4-10）对阿霉素（ADR）诱导体外培养的足细胞损伤后细胞增殖、细胞凋亡、nephrin和podocin表达的影响,初步探讨ACTH410对ADR诱导足细胞损伤的保护作用.方法 将复苏及培养好的足细胞分为5组：正常对照组,ADR处理组,低、中、高浓度ACTH4-10＋ADR处理组.正常对照组不予任何处理;ADR处理组予以1μmol/L ADR处理24 h;低、中、高浓度ACTH4-10＋ADR处理组分别先加入1、10、100 μg/L ACTH4-10干预lh后再加入1 μmol/L ADR处理24 h.然后采用细胞计数试剂盒（CCK-8）测定足细胞增殖变化;用TUNEL法来检测足细胞的凋亡;用实时定量PCR及Western印迹法检测nephrin和podocin的表达.结果 与正常对照组相比,ADR处理组的足细胞增殖受到抑制;nephrin和podocin的mRNA及蛋白表达降低,差异均有统计学意义（P＜0.05）;足细胞的凋亡率明显上升（38.14％比5.12％）.与ADR处理组相比,低、中、高浓度ACTH4-10＋ADR处理组随着ACTH4-10浓度的增加,足细胞增殖增加,nephrin和podocin的mRNA及蛋白表达水平相应升高;低、中、高浓度ACTH4-10＋ADR处理组的足细胞凋亡率（20.45％、17.39％、11.02％）较正常对照组上升,但与ADR处理组相比则足细胞凋亡率下降.结论 ACTH4-10能够维持足细胞裂孔膜蛋白分子nephrin和podocin的表达,对ADR诱导足细胞损伤具有保护作用,其保护作用具有ACTH4-10浓度依赖性.

Objective To observe the influence of adrenocorticotropic hormone （ACTH4-10） in the changes of podocyte proliferation,apoptosis and expression of nephrin and podocin on adriamycin （ADR）-induced podocyte injury and investigate the protective effect of ACTH4-10.Methods All podocytes were randomly divided into following groups：normal control,ADR-induced group and ACTH4-10 intervention group （low,middle and high concentration）.Normal control group was not treated,ADR-induced group was induced to set the model of podocyte injury by ADR （1 μmol/L） for 24 hours and ACTH4-10 intervention groups were intervened by 1 μg/L,10 μg/L and 100 μg/L ACTH4-10 for 1 hours respectively,prior to setting the model of podocyte injury.Cell counting kit （CCK-8） was used to detect the multiplication of podocytes and TUNEL apoptosis detection kit was used to detect podocyte apoptosis.Real-time PCR and Western blotting were used to examine the expression of nephrin and podocin.Results Compared with control group,podocyte proliferation and expression of nephrin and podocin was decreased significantly in ADR-induced group （P ＜ 0.05）,meanwhile podocyte apoptosis was increased obviously （38.14％ vs 5.12％）.Compared with ADR-induced group,podocyte proliferation and expression of nephrin and podocin was increased generally with concentration of ACTH4-10.Although podocyte apoptosis rates （20.45％,17.39％,11.02％） were increased in ACTH4-10 intervention group （low,middle and high concentration） while comparing with normal control group,podocyte apoptosis decreased obviously while comparing with ADR-induced group.Conclusions ACTH4-10 can stabilize the expression of nephrin and podocin on slid diaphragm,and has the protective effect on podocyte injury induced by ADR,while the effect depends on the concentration of ACTH4-10.