ABSTRACT
Objective To compared two classical rat models of nephrotic syndrome and to provide some reference data to researchers.Methods Thirty male SD rats were randomly divided into control group,puromycin aminonucleoside-induced nephrotic syndrome (PAN) group and adriamycininduced nephrotic syndrome (ADR) group.The body weight,twenty four hour proteinuria level,serum albumin concentration,cholesterol concentration,creatinine and urea concentration were measured.The renal pathology change was evaluated.The drug toxic effects,administration methods and the costs were also compared.Results There was no significant difference in body weight and hair color between control group and PAN group.Compared to control group,the body weight of the rats significantly decreased at day 15 and day 21 in ADR group (P ＜ 0.01),accompanied by epilation and diarrhea.Compared to control group,the 24-hour urinary protein levels increased significantly at day 10 (P ＜ 0.01),day 15 (P ＜ 0.01),and reached the peak level at day 15 (P ＜ 0.01),day 21 (P ＜ 0.01) in PAN group and ADR group respectively.Compared to control group,the serum albumin concentration decreased significantly at day 10 (P＜0.01),and return to normal level at day 15.The serum cholesterol concentration was increased significantly at day 10 (P ＜ 0.01) and return to normal at day 15 in PAN group.Compared to control group,the serum albumin concentration was decreased significantly at day 15 (P＜0.05) and return to normal at day 21 in ADR group.No significant difference of serum creatinine and serum urea nitrogen levels were found among three groups.Compared to control group,the width of foot process increased significantly at day10 (P ＜ 0.01) and day 15 (P ＜ 0.05) in PAN group and ADR group respectively.To successfully induce a nephrotic rat model (per 100 g),the cost of PAN group was 3.1 times of ADR group (578.10 yuan vs 186.94 yuan).Conclusions Nephrotic syndrome can be induced by both PAN and ADR.The administration of PAN via intraperitoneal injection is more convenient as compared to ADR via tail intravenous injection.Compared to ADR,PAN can induce nephrotic syndrome model more rapidly,with more consistent detection index,and less toxic effects,but its cost is more expensive.
ABSTRACT
Objective To explore the effect of uric acid (UA) on the rat glomerular mesangial cells (GMCs) and its possible mechanism in vitro. Methods Cultured rat GMCs were treated with various concentrations of UA (50 μmol/L, 100 μmol/L, 300 μmol/L) in the presence or absence of U0126, apocynin, rotenone. The incorporation of 3H-thymidine (3H-TdR) and cell counting were used to measure GMCs proliferation. GMCs cell-cycle was analyzed by flow cytometry. CyclinD1 and cyclin A2 expression were determined by real-time PCR and Western blotting analysis. ERK1/2 phosphorylation was detected by Western blotting. Reactive oxygen species (ROS) was measured by 2, 7-dichlorofluorescein diacetate (DCFDA) fluorescence. Results (1) Uric acid increased GMCs proliferation in dose-dependent manner compared with the control groups, as assessed by 3H-TdR incorporation and cell counting. GMCs proliferation induced by 300 μmol/L uric acid was increased by more than 1.5-fold assessed by both of the methods. (2) Uric acid decreased cell number in G1/G0 phase and increased cell number in S phase in dosedependent manner, as assessed by flow cytometry. (3) Uric acid iuduced cyclin D1 and cyclin A2 expression in dose-dependent manner. (4)Uric acid increased pospho-ERK1/2 in dose-dependent manner and ERK1/2 specific inhibitor U0126 could suppress uric acid-induced cell proliferation.The inhibition percentage of U0126 was 22% and 31% assassed by cell counting and 3H-TdR incorporation, respectively. (5) Uric acid increased ROS production in dose-dependent manner.NADPH oxidase inhibitor apocynin could also significantly inhibit uric acid-induced ROS production, ERK phosphorylation and cell proliferation. In contrast, rotenone had no effect on them.Conclusion Uric acid can stimulate rat GMCs proliferation, partially by the activation of ERK pathway via NADPH oxidase-derived ROS generation.