ABSTRACT
Objective:To explore the effects of LncRNA HCG18 on endoplasmic reticulum stress and autophagy via regulating miR-185-5p/AGER axis in diabetic nephropathy (DN) .Methods:The kidney tissues of patients with DN were collected and the podocyte injury model induced by high glucose (HG) was established. The expression of HCG18 in renal tissue in DN patients and cell model was detected. The localization and expression of HCG18 in cells were determined. The regulatory relationship between HCG18 and miR-185-5p, miR-185-5p and AGER was testified. QRT-PCR and western blot were used to detect the expression of endoplasmic reticulum stress related factors (CHOP, XBP1) and autophagy related factors (Beclin-1, p62) .Results:Compared with non-DN patients, HCG18 was overexpressed in renal tissue of DN patients ( P<0.05) . Compared with normal glucose (NG) group, mRNA and protein expression of endoplasmic reticulum stress related factors (CHOP, XBP1) were overexpressed but mRNA and protein expression of autophagy related factors Beclin-1 were inhibited, p62 mRNA and protein expression were increased (all P<0.05) . HCG18 regulated the miR-185-5p/AGER axis and played a biological role. Knocking down of HCG18 reduced endoplasmic reticulum stress, activated autophagy and reduced podocyte injury, but this effect can be partially reversed by miR-185-5p inhibitors. Conclusion:HCG18 regulates the miR-185-5p/AGER signal axis and promotes DN progression through regulating endoplasmic reticulum stress and autophagy.
ABSTRACT
Objective:To investigate the relationship between the DNA methylation of APC gene and the occurrence and development of diabetic nephropathy (DN) in peripheral blood, and to analyze its effect on gene transcription and protein expression levels.Methods:From Mar. 2018 to Mar. 2020, 96 patients with DN and 100 healthy examinees in Department of Nephrology, Hangzhou First people’s Hospital Affiliated to Zhejiang University were selected as the research objects, and the methylation level, mRNA level and protein expression level of CpG island of APC gene in the two groups were detected.Results:The methylation level of APC-1 island in DN patients was 3.05%±0.65%, which was significantly higher than that in healthy controls 2.34%±0.56% ( t=5.231, P<0.001) ; The methylation level of APC-2 island was 1.65%±0.37%, which was significantly higher than that of the healthy control group 1.17%±0.29% ( t=4.381, P<0.001) ; Pearson correlation analysis showed that the methylation level of APC-1 island was significantly positively correlated with the ration of serum creatinine and uric acid (all P<0.05) , and the methylation level of APC-2 island was significantly positively correlated with uric acid ( P<0.05 ) ; The methylation level of APC-1 island in DN patients was negatively correlated with the relative expression of mRNA ( r=-0.426, P=0.019) and the level of APC protein ( r=-0.569, P=0.004) . The expression of miRNA-135b in rats in the overexpression lentivirus group was significantly higher than that in the no-virus group ( t=5.432, P<0.001) and DN group ( t=5.812, P<0.001) ; The APC methylation level of rats in the miRNA-135b overexpression lentivirus group was also significantly higher than that in DN group ( t=6.217, P<0.001) and the rats in the no-virus group ( t=6.446, P<0.001) . Conclusion:miRNA-135b may participate in the occurrence and development of DN by up-regulating DNA methylation of APC gene and reducing mRNA and protein expression levels.
ABSTRACT
Objective To investigate the changes of microelement in curde and calcined pyritum, and approach the process mechanism of pyritum. Methods The crude and calcined pyritum were extracted in water decocting. Subsequently, the content of ten elements was determined by inductively coupled plasma emission spectrometry (ICP) and compared. Results After being calcined, the content of Pb in pyritum decreased, while the content of the other nine elements investigated all increased in different degrees. Conclision After being calcined, the content of ten elements in pyritum changed largely.