In�vitro and in�vivo study of the expression of the Syk/Ras/c‑Fos pathway in chronic glomerulonephritis

Abstract

Chronic glomerulonephritis (CGN) is the most common form of glomerular disease; however, its associated molecular mechanisms remain unclear. Spleen tyrosine kinase (Syk) is a key mediator of B-receptor signaling on the surface of inflammatory cells. The primary target for R406 is Syk. The aim of the present study was to investigate the molecular mechanisms involved in a rat model of CGN induced by adriamycin (ADR) and in the rat glomerular mesangial cell line, HBZY-1, stimulated by lipopolysaccharide (LPS). CGN was induced in the rat models by two intravenous injections of ADR into the tail: 3.5 mg/kg ADR was given on the first day and 3.0 mg/kg on the fourteenth day. HBZY-1 cells were incubated with 0.5 µg/ml LPS for 48 h. The pathological alterations in the kidney tissues were observed by hematoxylin and eosin staining. The 24 h urinary protein, blood urea nitrogen (BUN) and creatinine levels were measured using an automatic biochemistry analyzer. The mRNA expression levels of Syk, Ras, mitogen activated protein kinase kinase (MEK), extracellular signal regulated kinase (ERK)1/2 and c-Fos was measured by reverse transcription-quantitative polymerase chain reaction. Subsequently, the protein levels of phosphorylated (p)-Syk, Ras, p-MEK1/2, p-ERK1/2 and c-Fos were measured by western blot analysis. In the model group, 24 h urinary protein, BUN and creatinine levels were increased when compared with the normal group (P<0.05). In addition, compared with the normal group, the mRNA and protein levels of the Syk/Ras/c-Fos pathway components in vitro and in vivo were markedly increased, inhibiting the abnormal cell viability of mesangial cells. In conclusion, the results of the present study suggested a potential role for the Syk/Ras/c-Fos signaling pathway in CGN, which indicated the necessity for further investigation at the clinical level.

18F-FDG PET/MRI Imaging in a Preclinical Rat Model of Cardiorenal Syndrome—An Exploratory Study

“…Caspase-3 is involved in the pathogenesis of doxorubicin-induced cardiomyopathy [ 59 , 60 ] and nephropathy [ 61 ]. As a more proximal step in the cell response to doxorubicin toxicity, p53 overexpression plays a major role in developing dilatative cardiomyopathy and chronic kidney disease [ 11 ] mainly through its’ genomic integrity monitoring activity [ 62 ].…”

MicroRNA-339-5p inhibits lipopolysaccharide-induced rat mesangial cells by regulating the Syk/Ras/c-Fos pathway

“…In order to clarify the relationship between miR-339-5p and CGN, HBZY-1 cells induced by LPS were used as CGN experimental model in this study [20]. qRT-PCR was used to detect the expression levels of miR339-5p in the Control group and the Model group.…”

“…In addition, Syk is an important part of the middle Syk/Ras/c-Fos signal pathway. Through bioinformatics prediction and analysis in the early stage, our research group found that miR-339 had the highest binding degree with Syk, and studied the in uence of Syk/Ras/c-Fos signaling pathway on HBZY-1 cells [20], but did not discuss the pathogenesis of CGN from the genetic level. Therefore, through the establishment of LPS-induced HBZY-1 cell model, this paper expounds the pathogenesis of CGN from the point of view of miR-339-5p and Syk/Ras/c-Fos signal pathway.…”

Exploring the mechanism of Jianpi Qushi Huayu Formula in the treatment of chronic glomerulonephritis based on network pharmacology

“…The rat glomerular mesangial cells line (HBZY-1) was obtained from the Cell Bank of FuHeng Biology (Shanghai, China) and incubated with Dulbecco's modified Eagle's/F-12 medium [10% (v/v) fetal calf serum and 1% (v/v) antibiotics mixture] in 95% air and 5% CO2 at 37 °C. The lipopolysaccharide (LPS)-stimulated HBZY-1 cell model, which is often used for in vitro study of chronic glomerulonephritis, has been mentioned in our previous study (Gao et al 2018). Our experiment was separated into three groups: the control group (HBZY-1 cells were treated with normal rat serum for 24 h), the model group (HBZY-1 cells were treated with 0.5 μg/mL LPS for 48 h and then added to normal rat serum for 24 h), and the JQHF treatment group (HBZY-1 cells were treated with 0.5 μg/mL LPS for 48 h and then added to JQHF-medicated serum for 24 h).…”