ABSTRACT
Objective To investigate the effect of all-trans retinoic acid(ATRA) on the expressions of Prohibitin1 (PHB1) and Prohi-bitin2 (PHB2) in hypoxic damage of renal tubular epithelial cells (RTEC).Methods Rat proximal tubular epithelial cell line NRK-52E culture was performed in the 37 ℃ 50 mL/L carbon dioxide incubator with mixture medium of fetal bovine serum and double-antibody(100 mL medium plus 5 mL fetal calf serum and 1 mL double-antibody).After 3 times cell propagation,the cells were divided into 3 groups:normal group,model group and ATRA intervention group.The normal group wasn't disposed,and the model group was put into vacuum tank filled with hypoxic gas(950 mL/L nitrogen and 50 mL/L carbon dioxide) to induce a hypoxic damage of RTEC.The ATRA intervention group was added 0.1 μmol/L ATRA and was treated with hypoxic gas as model group.After 24 h and 36 h,the mRNA expressions of PHB1,PHB2 and transforming growth factor-β1 (TGF-β1) were measured by using real-time PCR method,and the protein expressions of PHB1,PHB2 and TGF-β1 were detected by using Western blot method.Results 1.Compared with normal group,NRK-52E cells PHB1,PHB2 protein expressions and mRNA expressions at 2time points(24 h,36 h) were significantly decreased in model group and ATRA intervention group (all P ＜ 0.05),and the longer hypoxia time,the lower expression value.Compared with model group,NRK-52E cells PHB1 and PHB2 protein expressions and mRNA expressions in ATRA intervention group were increased significantly at 2 time points (all P ＜ O.05).2.Compared with normal group,NRK-52E cells TGF-β1 expressions and mRNA expressions at 2 time points(24 h,36 h) were significantly increased in model group and ATRA intervention group(all P ＜ 0.05),and the longer hypoxia time,the higher expression value;Compared with model group,NRK-52E cells TGF-β1 protein expressions and mRNA expressions in ATRA intervention group were decreased significantly at 2 time points(all P ＜ 0.05).Conclusions ATRA can significantly up-regulate the mRNA expressions and protein expressions of PHB1 and PHB2 in hypoxic damage of RTEC,and ATRA may have a protective effect against hypoxic damage.
ABSTRACT
Objective To investigate expression and significance of matrix metalloproteinase-2(MMP-2),MMP-9 and tissue inhibitor of metalloproteinase-1(TIMP-1) in rats with glomerular sclerosis made by doxorubicin.Methods Forty Wistar male rats(8-week-old) were randomly assigned into 2 groups:sham operated and model groups.Rats in model group were nephrectomized after anesthesia and injected with adriamycin(5 mg/kg) after 1 week.Rats in sham operated group was subjected to sham operation and injected with normal saline after 1 week through the tail vein.All rats were killed in the 12th week.Immuno-histochemistry was performed on renal tissue to detect Collagen Ⅳ(Col-Ⅳ),fibronectin(FN),MMP-2,-9 and TIMP-1.Results Immunohistochemistry staining indicated that expressions of MMP-2,-9 in model group decreased significantly compared to sham operated group(Pa
ABSTRACT
Objective To study the expression of ?-smooth muscle actin(?-SMA)in glomerulosclerosis rats and its relationship with renal function.Methods Forty healthy Wistar rats were equally divided into 2 groups including sham operated group and model control group.Rats in model groups were uninephrectomized and injected with daunorubicin(5 mg/kg)after 1 week through the tail vein.Twenty-four hours of urinary protein excretion,serum creatinine(Scr),blood urea nitrogen(BUN)were measured at the 12th week.Renal pathology was evaluated.Immunohistochemistry(SupervisionTM)was performed on renal glomeruli tissue to detect the expression of ?-SMA.Reverse transcription polymerase chain reaction(RT-PCR)was used to examine the expression levels of ?-SMA mRNA in glomeruli.SPSS 13.0 software was used to analyze the two variables.Results In model control group,the urinary protein,Scr,BUN significantly increased(Pa