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Objective:To investigate the expression of microRNA-155 (miR-155) in serum and kidney of C57BLKS/db (db/db) mice and its role in the pathogenesis of diabetic kidney disease (DKD).Methods:The db/db mice ( n=24) were divided into 6, 8, and 10 weeks old groups ( n=8) with age increasing according to the random number table, and C57BL/6 mice of the same age were used as control group. The expression of miR-155 in mouse serum and kidney tissue was determined using real-time quantitative PCR. The mRNA and protein expression of Ets-1, eNOS, AGTR1 in renal tissues was verified by real-time quantitative PCR, Western blotting and immunohistochemistry. Results:Compared with the control group, the expression of miR-155 in serum of db/db mice at 6, 8 and 10 weeks of age were significantly increased (all P<0.01), and the increase of miR-155 was most obvious at 10 weeks of age ( P<0.01). Meanwhile the expression of miR-155 in kidney tissues of 6, 8 and 10 weeks old db/db mice was significantly up-regulated (all P<0.01), and the highest expression of miR-155 was at 10 weeks of age ( P<0.01). Immunohistochemistry showed that Ets-1, eNOS and AGTR1 were localized in glomerular endothelial cells. The results of real-time quantitative PCR showed that the mRNA expression of Ets-1, eNOS and AGTR1 were down-regulated in the kidney tissues of db/db mice at 6, 8 and 10 weeks of age compared to the control(all P<0.05), and the level of down-regulation was the most obvious at 10 week. Western blotting results showed that there was no significant change in Ets-1, eNOS and AGTR1 in 6-week-old db/db mice compared to the control group; the eNOS protein expression was down-regulated at 8 weeks of age ( P<0.05); the expression of AGTR1 protein was down-regulated ( P<0.05), and the protein expression of Ets-1 and eNOS were significantly down-regulated at 10-week age (both P<0.01). Conclusions:The expression of miR-155 in serum and kidney tissues of db/db mice increases during the progression of DKD, while the expression of miR-155 target genes Ets-1, eNOS and AGTR1 decreases with the progression of DKD. MiR-155 may participate in the development and progression of DKD by inhibiting its target genes Ets-1, eNOS and AGTR1, affecting endothelial cell function.
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Objective To investigate the relationship between carotid artery intima-media thickness and renal function in patients with diabetes mellitus.Methods 424 patients of type 2 diabetes without dialysis were enrolled in a cross-sectional study.According to their artery intima-media thickness (IMT),the patients were divided into normal group and higher IMT group.All patients according to UAER or 24h urinary protein were divided into normal proteinuria group,micro-proteinuria group and clinical proteinuria group.The biochemical examination,eGFR,and atherosclerotic plaque of different groups were compared.Pearson or spearman correlation was used to analyze the relationship between eGFR,IMT and other parameters.Risk factors for eGFR decline were analyzed by binary logistic regression.Results Compared with normal group,patients in the higher IMT group were older [(63.3±10.2) year vs (52.5 ± 10.6) year,P ＜ 0.05],and underwent longer duration of diabetes [(8.9±6.7) year vs (6.2±5.7) year,P ＜ 0.05].Their level of eGFR was decreased [(75.92±28.00) ml/min vs (91.64±24.05) ml/ min,P ＜ 0.05],while plaque incidence (71.3％ vs 18.3％,x2=112.42,P ＜ 0.01) and prevalence of hypertension (56.4％ vs 29.6％,x2=27.22,P ＜ 0.01) increased.Correlation analysis showed that IMT was positively correlated with age (r=0.503,P ＜ 0.01),duration of diabetes (r=0.204,P ＜ 0.01),24 h urine protein (rs=0.175,P ＜ 0.05),plaque (rs=0.562,P ＜ 0.01),and hypertension (rs=0.193,P ＜ 0.01),but negatively correlated with eGFR (r=-0.307,P ＜ 0.01).Logistic regression analysis showed that age,serum uric acid,24 h urine protein and carotid artery intima-media thickness were independent risk factors for eGFR decline [OR=1.115,95％CI(1.053,1.165),P ＜ 0.001;OR=1.008,95％CI (1.002,1.014),P=0.006;OR=1.492,95％ CI(1.170,1.903),P=0.001;OR=1.619,95％ CI(1.121,2.339),P=0.010].Conclusion Carotid artery intima-media thickness is an independent risk factor for kidney function decline in patients of diabetes.
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Objective To investigate the expression and clinical significance of serum microRNA (miRNA) expression profiling in the occurrence and progression of diabetic nephropathy.Methods The miRNA expression profiling was detected by miRNA TaqMan Low Density Array chip from 10 patient with diabetic nephropathy,10 diabetes patients with normoalbuminuria and 10 health control.Real-time quantitative PCR was applied to verify the result of miRNA array in serum samples of 66 patients with diabetic nephropathy (36 patients with microalbuminuria,30 patients with macroalbuminuria),40 diabetes patients with normoalbuminuria and 40 health control.And the relationship of differetial expression with clinical features was analyzed.Results miR-150-5p,miR-155-5p,miR-30e-5p and miR-3196 being validated by real-time quantitative PCR differentially expressed in 3 groups of serum samples from the diabetes patients with microalbuminuria (n=36),with normoalbuminuria (n=40) and health control (n=40) (P ＜ 0.05).Serum miR-150-5p (P=0.005) and miR-155-5p (P=0.006) changed significantly between diabetes patients with microalbuminuria (n=36) and with macroalbuminuria (n=30).Compared with the diabetes patients with microalbuminuria,serum miR-150-5p and miR-155-5p increased by 2.3 and 1.5 times in macroalbuminuria group,respectively.Estimated glomerular filtration rate and urinary albumin excretion rate significantly correlated with serum miR-150-5p and miR-155-5p level.Conclusions miR-150-5p and miR-155-5p may be involved in the process of pathological mechanisms of diabetic nephropathy.Serum miR-150-5p and miR-155-5p may be regarded as potential biomarkers to diagnosis the occurrence and development of diabetic nephropathy.
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Objective:This paper aimed to investigate the effects of isocorydinone on cell proliferation in SiHa human cervical carcinoma cell lines. Methods:Different concentrations of isocorydione (100, 200, 400, 800, and 1200 μmol/L) were used to treat SiHa human cervical carcinoma cells in vitro for 24, 48, and 72 h. Methyl thiazolyl tetrazolium (MTT) assays were conducted to determine the inhibitory action of isocorydione. Flow cytometry was performed to detect the cell cycle in SiHa human cervical carcinoma cells af-ter treatment with 400 μmol/L isocorydione. Hoechst 33342 staining was used to observe the micro-morphological changes of SiHa cell nucleus after the treatment. The expression of Bcl-2, Bax, and caspase-3 proteins in cervical carcinoma SiHa cell lines was determined using western blot analysis. Results: MTT assays showed that isocorydione inhibits the proliferation of SiHa cells in a dose- and time-dependent manner (P<0.05). The flow cytometry results showed that SiHa cervical carcinoma cells treated with different concen-trations of isocorydione exhibited increased cell cycle. Compared with the control group, Hoechst 33342 staining showed that SiHa cells became narrow, with nuclear pyknosis and fragmentation, and formed an apoptotic body after treatment with 400 μmol/L isocoryd-ione for 48 h. Furthermore, western blot analysis proved that isocorydione significantly inhibited the proliferation of SiHa cell lines, and the expression of Bax protein was increased. By contrast, the expression of Bcl-2 protein decreased gradually. Consequently, the ra-tio of Bax/Bcl-2 increased, as well as the expression of caspase-3 protein. Conclusion:Isocorydione exhibited an overt inhibitory ac-tion on SiHa cells. Isocorydione promoted the occurrence of cell apoptosis, which may be associated with related proteins of mitochon-drial apoptotic pathway.