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Pathological analysis of kidney tissues from adriamycin-treated and normal rats: (A) Normal and (B) model groups. In the model group, (a, arrows) casts (protein) in the lumen, (b, arrows) infiltration of inflammatory cells and (c, arrows) incrassation of the basal lamina were observed. Compared with the normal group, degeneration of renal tubule epithelial cells, infiltration of inflammatory cells and casts (protein) in the lumen were observed in the model group (magnification, x200).
Source publication
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 mec...
Context in source publication
Context 1
... HE staining is presented in Fig. 1. Rats from the control group invariably exhibited normal glomerular structure and glomerular basement membrane thickness, clear Bowman's capsule structure and convoluted tubule structure, and opened capillary loops. However, in the model group, there were incrassations of the capillary loops and Bowman's capsule. In addition, degeneration of renal tubule epithelial cells, infiltration of inflammatory cells and casts (protein) in the lumen were also observed, which was in agreement with the authors previous research and indicated that the CGN model was successfully established ...
Citations
... However, little is known about the m6A modifications in LncRNAs of CGN. To investigate whether RNA methylation plays a role in the CGN development, we established a model of lipopolysaccharide-induced mesangial cells proliferation and inflammation, a widely used CGN model in vitro [31,32]. Thus, we established a MMCs proliferation and inflammation model to preliminarily investigate whether LncRNA methylation plays a role in CGN development. ...
Background
Increasing evidence indicates that N6-methyladenosine (m6A) modification of mRNAs has been shown to play a critical role in the occurrence and development of many diseases, while little is known about m6A modification in long non-coding RNAs (LncRNAs). Our study aims to investigate the potential functions of LncRNA m6A modifications in lipopolysaccharide (LPS)-induced mouse mesangial cells (MMCs), providing us with a new perspective on the molecular mechanisms of chronic glomerulonephritis (CGN) pathogenesis.Methods
Differentially methylated LncRNAs were identified by Methylated RNA immunoprecipitation sequencing (MeRIP-seq). LncRNA–mRNA and LncRNA-associated LncRNA–miRNA–mRNA (CeRNA) networks were constructed by bioinformatics analysis. Furthermore, we utilized gene ontology (GO) and pathway enrichment analyses (KEGG) to explore target genes from co-expression networks. In addition, the total level of m6A RNA methylation and expression of methyltransferase and pro-inflammatory cytokines were detected by the colorimetric quantification method and western blot, respectively. Cell viability and cell cycle stage were detected by cell counting kit-8 (CCK-8) and flow cytometry.ResultsIn total, 1141 differentially m6A-methylated LncRNAs, including 529 hypermethylated LncRNAs and 612 hypomethylated LncRNAs, were determined by MeRIP-seq. The results of GO and KEGG analysis revealed that the target mRNAs were mainly enriched in signal pathways, such as the NF-kappa B signaling pathway, MAPK signaling pathway, Toll-like receptor signaling pathway, and apoptosis signaling pathway. In addition, higher METTL3 expression was found in CGN kidney tissues using the GEO database. METTL3 knockdown in MMC cells drastically reduced the levels of m6A RNA methylation, pro-inflammatory cytokines IL6 and TNF-α, and inhibited cell proliferation and cycle progression.Conclusions
Our findings provide a basis and novel insight for further investigations of m6A modifications in LncRNAs for the pathogenesis of CGN.
... Common pathologies of proliferative and inflammatory glomerular diseases in humans and experimental animals include mesangial cell proliferation (Cao et al., 2015;Singh et al., 2016;Kurihara and Sakai 2017), which may lead to excessive deposition of extracellular matrix, glomerulosclerosis, and loss of renal function. LPS is considered to be one of the strong stimulators of RMCs, and it can be used as an inducer of glomerular cell viability (Gao et al., 2018). Therefore, LPS was employed to mimic CGN in cells. ...
From Astragalus membranaceus (Fisch.) Bge.var. mongholicus (Bge.) Hsiao, astragaloside IV (AS-IV), a saponin can be purified and is considered traditional Chinese medicine. The purpose of this study was to evaluate the AS-IV-mediated mechanism on chronic glomerulonephritis (CGN). A cationic bovine serum albumin-induced CGN rat model was established and 10, 15, or 20 mg/kg of AS-IV was administered to measure renal function and inflammatory infiltration. Influences of AS-IV on proliferation, cell cycle, and inflammation of LPS-induced rat mesangial cells (RMCs) were determined. The results demonstrated that AS-IV alleviated renal dysfunction, renal lesions, and inflammation in CGN rats. AS-IV prolonged the G0-G1 phase, shortened the S phase, and inhibited cell proliferation and inflammation in RMCs. AS-IV can promote miR-181d-5p expression to inhibit CSF1. miR-181d-5p promotion or CSF1 suppression could further enhance the therapeutic role of AS-IV in CGN rats, while miR-181d-5p silencing or CSF1 overexpression abolished the effect of AS-IV. In conclusion, AS-IV by mediating the miR-181d-5p/CSF1 axis protects against CGN.
... 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]. ...
Cardiorenal syndrome (CRS) denotes the bidirectional interaction of chronic kidney disease and heart failure with an adverse prognosis but with a limited understanding of its pathogenesis. This study correlates biochemical blood markers, histopathological and immunohistochemistry features, and 2-deoxy-2-fluoro-D-glucose positron emission tomography (18F-FDG PET) metabolic data in low-dose doxorubicin-induced heart failure, cardiorenal syndrome, and renocardiac syndrome induced on Wistar male rats. To our knowledge, this is the first study that investigates the underlying mechanisms for CRS progression in rats using 18F-FDG PET. Clinical, metabolic cage monitoring, biochemistry, histopathology, and immunohistochemistry combined with PET/MRI (magnetic resonance imaging) data acquisition at distinct points in the disease progression were employed for this study in order to elucidate the available evidence of organ crosstalk between the heart and kidneys. In our CRS model, we found that chronic treatment with low-dose doxorubicin followed by acute 5/6 nephrectomy incurred the highest mortality among the study groups, while the model for renocardiac syndrome resulted in moderate-to-high mortality. 18F-FDG PET imaging evidenced the doxorubicin cardiotoxicity with vascular alterations, normal kidney development damage, and impaired function. Given the fact that standard clinical markers were insensitive to early renal injury, we believe that the decreasing values of the 18F-FDG PET-derived renal marker across the groups and, compared with their age-matched controls, along with the uniform distribution seen in healthy developing rats, could have a potential diagnostic and prognostic yield in cardiorenal syndrome.
... Mesangial cells play a prominent role in the process of autoimmune disorders and inflammatory diseases of the kidney, and mesangial cell-mediated glomerulonephritis is a frequent cause of ESRD (Yu et al. 2019;Guo et al. 2017). To explore whether m6A modification plays a role in the occurrence and progression of CGN, we established an LPS-induced mesangial cells proliferation and inflammation model, a common CGN model that has been widely used in vitro (Shushakova et al. 2007;Gao et al. 2018). Subsequently, genome-wide m6A analysis of unstimulated and stimulated MMC cells was performed to measure the correlation between m6A and CGN. ...
N6-Methyladenosine (m6A) is the most prevalent internal modification of messenger RNA (mRNA) in eukaryotes. The underlying molecular mechanisms of m6A modification in chronic glomerulonephritis (CGN) remain unexplored. Here, we performed methylated RNA immunoprecipitation sequencing (MeRIP-seq) and RNA sequencing (RNA-seq) analyses to assess the alterations of epitranscriptome-wide m6A profile in lipopolysaccharide (LPS)-induced mouse mesangial cells (MMC). The results of our data showed 2153 significantly differential m6A peaks and 358 significantly differentially expressed genes. Furthermore, integrated analysis from MeRIP-seq and RNA-seq identified a total of 64 genes with differential m6A modification and expressed levels, of which 5 genes displayed hypermethylation and upregulation, 42 genes displayed hypermethylation and downregulation, 11 genes displayed hypomethylation and upregulation, and 8 genes displayed hypomethylation and downregulation. Many of them (including Fosl1, Sorbs1, Ambp, Fgfr3, Nedd9, Fgg, Trim13, Fgf22, Mylk, and Muc6) are implicated in the regulation of the immune and inflammatory response. Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analysis found that differential 64 genes were mainly enriched in fatty acid oxidation, apoptosis signaling pathway, complement and coagulation cascades, and PPAR signaling pathway. Together, our study provided a new perspective on the understanding of molecular features of m6A modification in CGN pathogenic pathogenesis.
... 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. ...
Purpose: Chronic glomerulonephritis (CGN) is a disease that occurs in the glomeruli. The mechanism of CGN is thought to be involved in a range of inflammatory responses. MicroRNA-339-5p (miR-339-5p) has been reported to be involved in inflammatory responses in many diseases. However, the role of miR -339-5p in CGN remains unclear. The purpose of this study was to investigate the role of miR-339-5p in lipopolysaccharide (LPS) -induced nephritis injury in vitro.
Methods: The RNA expression of miR-339-5p and Syk/Ras/c-Fos pathway was detected by qRT-PCR, the protein expression and localization of Syk/Ras/c-Fos pathway was detected by western blot and immunofluorescence (IF), and the targeted binding of miR-339-5p to Syk was detected by double luciferase. Cell viability and cell cycle were detected by cell counting kit-8 (CCK-8) and flow cytometry. The concentrations of inflammatory cytokines IL-1β, IL-10, IL-6 and TNF-α were detected by enzyme linked immunosorbent assay (ELISA).
Results: LPS increased HBZY-1 cell viability, decreased G2 phase, promoted cell proliferation and inflammatory cytokine release. Overexpression of miR-339-5p can inhibit HBZY-1 cell viability, decreased the expression of Syk/Ras/c-Fos signaling pathway, down-regulate the expression level of inflammatory cytokines, increase G2 phase, and inhibit cell proliferation.
Conclusion: miR-339-5p inhibits the proliferation and inflammation of rat mesangial cell through Syk/Ras/c-Fos signal pathway.
... Lipopolysaccharide (LPS) stimulation can induce GMC proliferation, causing inflammation and oxidative stress that favor CGN development. 13 Thus, LPS was used to induce HBZY-1 cell proliferation. After the cells had been pretreated with serum-free DMEM medium containing 100 µg/mL of LPS for 24 hours, the medium was removed and 1.5 ml of YSHS Granule DMEM solution (100 µg/mL) was added and incubated for 1 hour. ...
Yi-Shen-Hua-Shi (YSHS) Granule is a well-known patented herbal drug for treating chronic glomerulonephritis (CGN) in China. Its chemical compositions and anti-CGN components are not fully understood. This study aimed to establish the chemical profile and to identify the anti-CGN components of YSHS Granule. For these purposes, a HPLC-Q-TOF-MS/MS method was developed. A total of 105 peaks were detected in the mass spectrum of the Granule. Of these, 99 compounds were tentatively identified as terpenoids, flavonoids, coumarins, alkaloids, phenols, and other types of compounds, and 15 were further identified with reference substances. To screen bioactive compounds, a cell membrane immobilized chromatography (CMIC) method was used. Lipopolysaccharide (LPS)-challenged rat glomerular mesangial cells (HBZY-1) were incubated with YSHS Granule (100 μg/mL), and the binding components to the cell membrane were extracted and analyzed using the established HPLC-Q-TOF-MS/MS method. Seven potential bioactive components that bound to HBZY-1 cell membranes were detected and identified as calycosin-7- O-β-D-glucoside, 6-gingerol, naringin, ginsenoside Re, poncirin, liquiritigenin, and isoliquiritigenin. The outcomes of the present study provide a chemical basis for clinical use of the Granule in managing CGN, and provide quality control markers for the Granule’s production and use.
Diabetic kidney disease (DKD) is a major public health issue because of its refractory nature. Ferroptosis is a newly coined programmed cell death characterized by the accumulation of lipid reactive oxygen species (ROS). However, the prognostic and diagnostic value of ferroptosis-related genes (FRGs) and their biological mechanisms in DKD remain elusive. The gene expression profiles GSE96804, GSE30566, GSE99339 and GSE30528 were obtained and analyzed. We constructed a reliable prognostic model for DKD consisting of eight FRGs (SKIL, RASA1, YTHDC2, SON, MRPL11, HSD17B14, DUSP1 and FOS). The receiver operating characteristic (ROC) curves showed that the ferroptosis-related model had predictive power with an area under the curve (AUC) of 0.818. Gene functional enrichment analysis showed significant differences between the DKD and normal groups, and ferroptosis played an important role in DKD. Consensus clustering analysis showed four different ferroptosis types, and the risk score of type four was significantly higher than that of other groups. Immune infiltration analysis indicated that the expression of macrophages M2 increased significantly, while that of neutrophils and mast cells activated decreased significantly in the high-risk group. Our study identified and validated the molecular mechanisms of ferroptosis in DKD. FRGs could serve as credible diagnostic biomarkers and therapeutic targets for DKD.
Diabetic kidney disease (DKD) is a major public health issue because of its refractory nature. Ferroptosis is a newly coined programmed cell death characterized by the accumulation of lipid reactive oxygen species (ROS). However, the prognostic and diagnostic value of ferroptosis-related genes (FRGs) and their biological mechanisms in DKD remain elusive. The gene expression profiles GSE96804, GSE30566, GSE99339 and GSE30528 were obtained and analyzed. We constructed a reliable prognostic model for DKD consisting of eight FRGs (SKIL, RASA1, YTHDC2, SON, MRPL11, HSD17B14, DUSP1 and FOS). The receiver operating characteristic (ROC) curves showed that the ferroptosis-related model had predictive power with an area under the curve (AUC) of 0.818. Gene functional enrichment analysis showed significant differences between the DKD and normal groups, and ferroptosis played an important role in DKD. Consensus clustering analysis showed four different ferroptosis types, and the risk score of type four was significantly higher than that of other groups. Immune infiltration analysis indicated that the expression of macrophages M2 increased significantly, while that of neutrophils decreased significantly in the high-risk group. Our study identified and validated the molecular mechanisms of ferroptosis in DKD. FRGs could serve as credible diagnostic biomarkers and therapeutic targets for DKD.
