Abstract
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.
Highlights
Diabetic nephropathy (DN) is one of the common microvascular complications of diabetes mellitus, with glomerular mesangial cell (MC) proliferation, thickening of glomerular basement membrane (GBM), and accumulation of extracellular matrix as the main histological features
The A values of MC cells at 12, 24, and 48 h were higher in the high glucose group (HGG) than in the control group (CNG), and were lower in the WMG, Qijin granules 1/2/3 groups than in the HGG, with the lowest A value in Qijin granules 3 group (p < 0.05), indicating that high glucose could induce excessive proliferation of MC cells, while Qijin granules inhibited the proliferation ability of cells in a dosedependent manner (Figure 1)
Yang et al high glucose could aggravate the inflammatory response while Qijin granules could downregulate NF-κB and MCP-1 expression in a dosedependent manner (Figure 3)
Summary
Diabetic nephropathy (DN) is one of the common microvascular complications of diabetes mellitus, with glomerular mesangial cell (MC) proliferation, thickening of glomerular basement membrane (GBM), and accumulation of extracellular matrix as the main histological features. To explore whether Qijin granules can delay the progression of DN, this study created the high glucose environment in vitro and analyzed the effects of Qijin granules on the proliferation, apoptosis and expression of NF-κB and MCP-1 in MC cells induced by high glucose, aiming to provide theoretical and experimental basis for the clinical treatment of early DN. (China Center For Type Culture Collection (Wuhan University), Wuhan, China); D-glucose (Shanghai Baoman Biotechnology Co. Ltd, Shanghai Other materials used include China); Qijin granules Kit, radioimmunoprecipitation assay (RIPA) lysate (Shanghai Biolab, Shanghai, China); Mouse antirat Bax, Cyt-C, Caspase-3, TGF-β1, p-Smad, Smad monoclonal antibodies (CST Inc, Danvers, MA, USA). HBZY-1 was routinely cultured in DMEM containing 10 % fetal bovine serum, 100 mg/L streptomycin, and 100 U/mL penicillin in an incubator (37 °C, 5 % CO2), and the medium was changed every 2 - 3 days.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
