Cell Cycle-Dependent Interleukin 1 Gene Expression by Cultured Glomerular Mesangial Cells

Purpose: To investigate the effects of Qijin granules on high glucose-induced proliferation and apoptosis in rat glomerular mesangial cells (MC).Methods: MC cells from rats were passaged and cultured, and randomly divided into control group (CNG), high glucose group (HGG), Western medicine group (WMG, high glucose + Benazepril + Gliquidone), and Qijin granules 1/2/3 group (high glucose + different doses of Qijin granules). Mesangial cells proliferation was measured using MTT assay. The NF-κB, MCP-1 and inflammatory factors in supernatant were determined by ELISA. Apoptosis rate and cell cycle were assessed by flow cytometry. The apoptosis-related TGF-β1/Smad signaling pathway-related protein expressions were measured by Western blot.Results: The A-value and early apoptosis rate, apoptosis rate and S-phase percentage, and protein expressions of NF-κB, MCP-1, IL-6, IL-2, TNF-ɑ, Bax, Cyt-C, caspase-3, TGF-β1, and p-Smad3 of MC cells in the HGG at 12 h, 24 h and 48 h were higher than those in the CNG. The above indices were lower in the WMG, and Qijin granules 1/2/3 groups than in the HGG. The Bcl-2, Smad7 protein expression level and the percentage of G1 and G2/M phase were lower in the HGG than in the CNG, and the above indeices were higher in the WMG and Qijin granules 1/2/3 group than in HGG.Conclusion: Qijin granules can dose-dependently inhibit high glucose-induced proliferation and apoptosis in rat MC cells, block the cell cycle and reduce inflammatory responses. This may be related to the regulation of NF-κB, MCP-1 and TGF-β1/Smad signaling pathways. These findings provide theoretical and experimental basis for the clinical treatment of early diabetic nephropathy.

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Mitogenic signaling of thrombin in mesangial cells: role of tyrosine phosphorylation

AJP Renal Physiology ◽

10.1152/ajprenal.1994.267.4.f528 ◽

1994 ◽

Vol 267 (4) ◽

pp. F528-F536 ◽

Cited By ~ 1

Author(s):

G. Grandaliano ◽

G. G. Choudhury ◽

P. Biswas ◽

H. E. Abboud

Keyword(s):

Gene Expression ◽

Dna Synthesis ◽

Tyrosine Phosphorylation ◽

Mesangial Cells ◽

Cellular Proteins ◽

Chain Gene ◽

Protein Tyrosine Phosphorylation ◽

Glomerular Mesangial Cells ◽

Human Glomerular Mesangial Cells

Thrombin elicits multiple biological effects on a variety of cells. We have previously shown that thrombin is a potent mitogen for human glomerular mesangial cells. This mitogenic effect of thrombin is associated with activation of phospholipase C (PLC) and induction of platelet-derived growth factor (PDGF) gene expression. The thrombin receptor, which belongs to the guanine nucleotide binding protein (G protein)-coupled receptor family, has recently been shown to induce rapid tyrosine phosphorylation of cellular proteins. In the present study, we investigated the role of protein-tyrosine phosphorylation in mediating the cellular responses elicited by thrombin in human glomerular mesangial cells. Amino acid labeling followed by immunoprecipitation with phosphotyrosine antibodies demonstrate that thrombin stimulates tyrosine phosphorylation of a set of cellular proteins. Treatment of mesangial cells with thrombin followed by immunoblotting with phosphotyrosine antibodies showed three major bands of tyrosine-phosphorylated proteins approximately 130, 70, and 44-42 kDa. Phosphorylation of these proteins was inhibited by two tyrosine kinase inhibitors, herbimycin A and genistein. Both compounds inhibited DNA synthesis and PDGF B-chain gene expression but had no effect on inositol phosphates production or increases in cytosolic calcium in response to thrombin. These data demonstrate that protein-tyrosine phosphorylation is not required for thrombin-induced PLC activation with inositol phosphate formation and subsequent intracellular calcium release, but it is an absolute requirement for thrombin-induced DNA synthesis and PDGF B-chain gene expression.

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Spatial transcriptome profiling by MERFISH reveals subcellular RNA compartmentalization and cell cycle-dependent gene expression

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1912459116 ◽

2019 ◽

Vol 116 (39) ◽

pp. 19490-19499 ◽

Cited By ~ 76

Author(s):

Chenglong Xia ◽

Jean Fan ◽

George Emanuel ◽

Junjie Hao ◽

Xiaowei Zhuang

Keyword(s):

Gene Expression ◽

Cell Cycle ◽

Spatial Information ◽

Detection Efficiency ◽

Expression Profiles ◽

Regulation Of Gene Expression ◽

Rna Profiling ◽

Spatially Resolved ◽

Cycle Dependent

The expression profiles and spatial distributions of RNAs regulate many cellular functions. Image-based transcriptomic approaches provide powerful means to measure both expression and spatial information of RNAs in individual cells within their native environment. Among these approaches, multiplexed error-robust fluorescence in situ hybridization (MERFISH) has achieved spatially resolved RNA quantification at transcriptome scale by massively multiplexing single-molecule FISH measurements. Here, we increased the gene throughput of MERFISH and demonstrated simultaneous measurements of RNA transcripts from ∼10,000 genes in individual cells with ∼80% detection efficiency and ∼4% misidentification rate. We combined MERFISH with cellular structure imaging to determine subcellular compartmentalization of RNAs. We validated this approach by showing enrichment of secretome transcripts at the endoplasmic reticulum, and further revealed enrichment of long noncoding RNAs, RNAs with retained introns, and a subgroup of protein-coding mRNAs in the cell nucleus. Leveraging spatially resolved RNA profiling, we developed an approach to determine RNA velocity in situ using the balance of nuclear versus cytoplasmic RNA counts. We applied this approach to infer pseudotime ordering of cells and identified cells at different cell-cycle states, revealing ∼1,600 genes with putative cell cycle-dependent expression and a gradual transcription profile change as cells progress through cell-cycle stages. Our analysis further revealed cell cycle-dependent and cell cycle-independent spatial heterogeneity of transcriptionally distinct cells. We envision that the ability to perform spatially resolved, genome-wide RNA profiling with high detection efficiency and accuracy by MERFISH could help address a wide array of questions ranging from the regulation of gene expression in cells to the development of cell fate and organization in tissues.

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Soluble Immune Complexes Stimulate Production of Interleukin-1 by Cultured Rat Glomerular Mesangial Cells

American Journal of Nephrology ◽

10.1159/000168290 ◽

1991 ◽

Vol 11 (2) ◽

pp. 138-143 ◽

Cited By ~ 5

Author(s):

Koichi Matsumoto ◽

Michinobu Hatano

Keyword(s):

Immune Complexes ◽

Mesangial Cells ◽

Interleukin 1 ◽

Glomerular Mesangial Cells ◽

Soluble Immune Complexes

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Gene expression profile of endothelin-1-induced growth in glomerular mesangial cells

AJP Cell Physiology ◽

10.1152/ajpcell.00105.2003 ◽

2003 ◽

Vol 285 (5) ◽

pp. C1109-C1115 ◽

Cited By ~ 14

Author(s):

Rangnath Mishra ◽

Patrick Leahy ◽

Michael S. Simonson

Keyword(s):

Gene Expression ◽

Growth Factor ◽

Cell Growth ◽

Gene Expression Profile ◽

Expression Profile ◽

Kinase Inhibitor ◽

Mesangial Cells ◽

Growth Control ◽

Glomerular Mesangial Cells ◽

Regulated Gene Expression

Endothelin (ET)-1 is a vasoconstrictor and mitogen involved in vascular remodeling. Changes in gene expression that underlie control of cell growth by ET-1 remain poorly characterized. To identify pathways of growth control we used microarrays to analyze ET-1-regulated gene expression in human mesangial cells, an important ET-1 vascular target cell in vivo. Statistical assessment of differential expression (significance analysis of microarrays) revealed upregulated transcripts for growth factors [heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF), fibroblast growth factor (FGF), interleukin (IL)-6] and downregulated transcripts for genes that inhibit growth (BAX, p27KIP1, DAD1). Consistent with the gene expression profile, quantitative RT-PCR and Western blotting confirmed induction of HB-EGF by ET-1. To test a functional role for HB-EGF in ET-1 signaling, we showed that exogenous HB-EGF stimulated phosphorylation of ErbB1 and growth of mesangial cells. ET-1-induced proliferation was blocked by an ErbB1 receptor-selective kinase inhibitor and by a specific ErbB1 receptor-neutralizing antibody. Proliferation in response to ET-1 was also inhibited by neutralizing antisera against human HB-EGF. Together, these results provide data for modeling ET-1 pathways for growth control and suggest a specific role for HB-EGF gene induction in mesangial cell growth in response to ET-1.

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