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Objective To investigate the role of the group IB secretory phospholipase A2 (sPLA2-IB) and M-type phospholipase A2 receptor (PLA2R) in human podocyte injury and its possible signal transduction pathway.Methods Differentiated human podocytes were exposed to PBS or different sPLA2-IB concentration conditions (10-9,10-7,10-5 mol/L) for 2 hours.The wound healing assay was used to measure cell migration rate;Apoptosis in cultured human podocytes was assessed by Hoechst 33342 staining and flow cytometry;Western blot was used to analyze the protein expression of p-cPLA2α,p-p38,and p53.Then control siRNA or PLA2R siRNA were transfected to podocytes.Podocytes were divided into normal control group,negative control siRNA group and PLA2R siRNA group.Twenty four hous later,the cells were stimulated by 105 mol/L sPLA2-IB for 2 hours.The protein expression of p-cPLA2α,p-p38,and p53 were detected by Western blot.Results Compared to PBS control group,the migration ability of podocytes decreased when stimulated with sPLA2-IB (10-7mol/L-10-5 mol/L),and the apoptosis of podocytes increased in a concentration-dependent manner,the protein level of p-cPLA2α,p-p38 and p53 protein increased too.After the knockdown of PLA2R by PLA2R siRNA transfection,stimulated the podocytes with the same dosage of sPLA2-IB,the protein expression of p-cPLA2α,p-p38 and p53 all decreased.Conclusion sPLA2-IB stimulation can increase human podocyte apoptosis and decrease its migration ability.The possible mechanism might be through p38-cPLA2α-p53 pathway.
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Objective To investigate the role of the group IB secretory phospholipase A2 (sPLA2-IB) and M-type phospholipase A2 receptor (PLA2R) in human podocyte injury and its possible signal transduction pathway.Methods Differentiated human podocytes were exposed to PBS or different sPLA2-IB concentration conditions (10-9,10-7,10-5 mol/L) for 2 hours.The wound healing assay was used to measure cell migration rate;Apoptosis in cultured human podocytes was assessed by Hoechst 33342 staining and flow cytometry;Western blot was used to analyze the protein expression of p-cPLA2α,p-p38,and p53.Then control siRNA or PLA2R siRNA were transfected to podocytes.Podocytes were divided into normal control group,negative control siRNA group and PLA2R siRNA group.Twenty four hous later,the cells were stimulated by 105 mol/L sPLA2-IB for 2 hours.The protein expression of p-cPLA2α,p-p38,and p53 were detected by Western blot.Results Compared to PBS control group,the migration ability of podocytes decreased when stimulated with sPLA2-IB (10-7mol/L-10-5 mol/L),and the apoptosis of podocytes increased in a concentration-dependent manner,the protein level of p-cPLA2α,p-p38 and p53 protein increased too.After the knockdown of PLA2R by PLA2R siRNA transfection,stimulated the podocytes with the same dosage of sPLA2-IB,the protein expression of p-cPLA2α,p-p38 and p53 all decreased.Conclusion sPLA2-IB stimulation can increase human podocyte apoptosis and decrease its migration ability.The possible mechanism might be through p38-cPLA2α-p53 pathway.
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Objective To investigate association between serum uric acid (SUA),albuminuria and glomerular filtration rates (eGFR) in type 2 diabetic patients.Methods A total of 220 patients were enrolled in this cross-sectional study.According to urinary albumin excretion rates,patients were divided into 3 groups:normoalbuminuria (NAU) group,microalbuminuria (MAU) group,and macroalbumnuria group (MAAU).The first two groups were subdivided at SUA ＞ 420 μmol/L (＞ 357 μmol/L,female) into normouricemia group and hyperuricemia group,at eGFR ＞ 90 ml/min into high and low renal function groups.General information,blood biochemical results were collected to analyze the association between serum uric acid,eGFR,UAER and urine albumin quantification among different groups.Results The difference of SBP,duration of diabetes (DD),Scr,SUA and eGFR between every two groups were significant (P ＜ 0.05).SBP,DD,Scr and SUA were highest in subjects with macroalbumnuria,second in microalbuminuria group,and lowest in normoalbuminuria group,while eGFR was lowest in macroalbumnuria group and highest in normoalbuminuria group.Prevalence of hyperuricemia in macroalbumnuria group (56.9％) and microalbuminuria group (51.2％) were also significantly higher than that in normoalbuminuria group (17.5％) (all P ＜ 0.01).The difference of UAER in the subgroups of normouricemia and hyperuricemia was more significant in microalbuminuria group than in normoalbuminuria group.eGFR was significantly lower in hyperuricemia subgroups (P ＜0.01).Age and SUA were significantlg higher in subjects with low renal function compared with high eGFR (P ＜ 0.05).Linear regression analysis indicated SUA was negatively correlated with eGFR after adjusted age,DD and UAER (β =-0.430,P ＜ 0.01).Binary logistic regression analysis found that increased age,DD and SUA were risk factors of microalbuminuria β =1.092,95％ CI(1.025,1.163),P ＜ 0.01;β =1.005,95％CI(1.001,1.009),P ＜ 0.05;β =1.407,95％ CI(1.052,1.881),P ＜ 0.05)] and SUA,age were risk factors of early renal function decline [β =1.015,95 ％ CI(1.00,1.023),P ＜ 0.01;β =1.098,95％ CI(1.006,1.199),P ＜ 0.05].Conclusion SUA is independently associated with albumnuria and renal function decline in type 2 DM patients.
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Objective To investigate the differentiation of rat bone marrow mesenchymal stem cells (MSCs) to renal tubular epithelial-like cells under different conditions. Methods MSCs were obtained from rat marrow. MSCs were isolated by gradient density centrifugation and plastic adherence and then purified. Surface markers were identified with flow cytometry after amplification in vitro. The purified MSCs of the third passage were cultured respectively as follows: (1) control group: DMEM medium with fetal bovine serum(FBS). (2) all-trans retinoic acid (ATRA) group: DMEM medium with FBS, ATRA and ischemic reperfusion-injured kidney tissue homogenate. (3)combination group: DMEM medium with FBS, ATRA, ischemic reperfusion-injured kidney tissue homogenate, epidermal growth factor (EGF) and bone morphogenetic protein 7 (BMP-7). After 7 days, the MSCs were collected for alkaline phosphatase (AKP) staining, cytokeratin-18 and E-cadherin immunocytochemical analysis. Results The positive rates of the third passage MSCs in CD44, CD90 and CD29 were 97.8%±0.9%, 96.8%±1.4% and 97.6%±2.4%,respectively, but in CD11b/c and CD34 were only 13.2%±0.6% and 1.2%±0.5%. The MSCs in control group were spindle. The MSCs in ATRA group were round and elliptic. The MSCs in combination group became cobblestone-like cells after 7 days. AKP staining showed that tubular epithelial-like cells from MSCs in control group were negative, some above cells in ATRA group were positive and number of above cells increased in combination group. Compared with negative control group, the ratios of cytokeratin-18 positive cells in ATRA group and combination group were respectively increassed by 29.47%±1.08% and 47.52%±2.13% (all P<0.05), the ratios of E-cadherin positive cells in ATRA group and combination group were respectively increased by 14.88%±2.46% and 36.15%±1.13% (all P<0.05). Conclusion MSCs may differentiate by renal tubular epithelial-like cells under the induction of ischemic reperfusion-injured kidney tissue homogenate and ATRA in vitro, which are further differentiated under the combined induction of EGF and BMP-7.
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Objective To assess the effects of early correction of anemia with recombinant human erythropoietin (rHuEPO) on the development and progression of left ventricular hypertrophy (LVH) in patients with mild-to-moderate chronic renal insufficiency (CRI) who are not on hemodialysis. Methods A total of 158 patients with serum creatinine from 147μmol/L to 400μmol/L were nrolled in this prospective, multicenter study. Eighty-six patients with hemoglobin (Hb)＜110g/L received rHuEPO treatment with a target Hb of ≥110g/L (Group A). Forty patients with comparable Hb concentration (＜110g/L) but did not receive rHuEPO (Group B) and 32 patients with Hb≥110g/L and without rHuEPO treatment (Group C) were served as controls. Left ventricular mass index (LVMI) was evaluated by echocardiography at baseline and every 3 months for 2 years. Results There was no difference in age, gender, etiology of renal failure, blood pressure and cardiovascular risk factors among the 3 groups. At baseline, the prevalence of LVH was 72.1% in group A,72.5% in group B and 59.4% in group C. LVMI was inversely correlated with Hb levels (r=0.70, P＜0.01). During the 2-year period, the mean LVMI decreased from 142.6±25.7g/m2 to 132.4±18.5 g/m2 in group A, while increased significantly in both group B and group C. The mean Hb concentration increased from 93.8±14.6g/L to 111.2±10.3g/L (P＜0.05) in group A, but tended to decrease in group B and group C. There was no significant change of the mean blood pressure, number of anti-hypertensive drugs and serum creatinine concentrations in all 3 groups. However, patients' serum creatinine doubled more often in group B and group C than in group A.Conclusions LVH was common in predialysis CRI patients and was associated with the severity of anemia. Early intervention with rHuEPO may reverse LVH in these patients.
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Objective To investigate the early impact of incremental peritoneal dialysis.Methods 109 new peritoneal dialysis patients treated with incremental peritoneal dialysis were included in the present study.Clinical data,dialysis adequacy and nutritional status were evaluated at the initiation of dialysis and at 3 months after PD.Results The peritoneal dialysis patients achieved good results in the dialysis adequacy and nutrition at the end of the 3rd month.The patients' comorbidities reduced.BP and blood work improved significantly during the first 3 months.Conclusion Our results suggest that incremental PD improve patients clinical status even at the early stage of dialysis.