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【Objective】 To explore the role and mechanism of TRPC in promoting extracellular matrix (ECM) deposition in rat glomerular mesangial cells (HBZY-1). Methods Immunofluorescence staining was performed to observe the distribution and expression of TRPC1 and TRPC6 in HBZY-1 cells. After AngⅡ stimulation, qRT-PCR and Western blotting were used to detect the mRNA and protein expressions of Gαq/PLCβ4/TRPC signaling pathway main proteins and ECM deposition indicators (α-SMA, collagenⅢ and fibronectin). By silencing the expressions of TRPC1 and TRPC6 by RNA interference, the expressions of ECM deposition indicators were detected. Changes in [Ca2+]i influx were determined through Fluo-4AM Ca2+ imaging. 【Results】 Both TRPC1 and TRPC6 were expressed in HBZY-1, and were mainly located in cell membrane and cytoplasm. After AngⅡ stimulation, Gαq/PLCβ4/TRPC signaling pathway was activated, and the mRNA and protein expressions of Gαq, PLCβ4, TRPC1 and TRPC6 were all increased (P<0.05). [Ca2+]i influx also increased (P<0.01), and the mRNA and protein expressions of ECM deposition indicators (α-SMA, ColⅢ and Fn) were upregulated (P<0.05). Silencing the expressions of TRPC1 and TRPC6 by RNA interference led to decreased [Ca2+]i influx (P<0.05), and downregulated mRNA and protein expressions of ECM deposition indicators in HBZY-1 cells (P<0.05). The results suggested that inhibition of TRPC expressions could inhibit AngⅡ induced ECM deposition in HBZY-1 cells, which might be associated with decreased [Ca2+]i influx. 【Conclusion】 TRPC may be a novel therapeutic target of renal fibrosis.
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【Objective】 To explore the role and mechanism of TRPC6 in apoptosis of glomerular mesangial cells (HBZY-1) induced by endoplasmic reticulum stress (ERS). 【Methods】 The experiment groups were classified as follows: normal control (NC), thapsigargin (TG), TG+SKF96365, and TG+TRPC6 siRNA groups. Transcription and protein expressions of TRPC6 and ERS related proteins (GRP78 and Caspase12) were detected by qRT-PCR and Western blotting. Additionally, cell apoptosis was measured by flow cytometry and Hoechst33258. Finally, Fluo-4 AM Ca2+ imaging technique was used to determine changes of intracellular calcium ( [Ca2+] i) by laser scanning confocal microscope. 【Results】 Morphological changes of apoptotic cells were characterized by nuclear enrichment or nuclear fragmentation, and the apoptosis rate was increased after TG stimulation. The expressions of TRPC6 and ERS related proteins (GRP78 and Caspase12) were elevated in TG group compared with NC group (P<0.05). Pre-incubation of HBZY-1 cells with SKF96365 and TRPC6 siRNA decreased cell apoptosis (P<0.05). The entry of [Ca2+] i also increased after TG stimulation (P<0.05). The expressions of TRPC6, GRP78 and Caspase12 were downregulated compared with TG group after treatment with SKF96365 and TRPC6 siRNA accompanied by decreased [Ca2+] i (P<0.05). 【Conclusion】 Taken together, this study suggests that inhibition of TRPC6 can alleviate TG-induced HBZY-1 cell apoptosis.
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Hyperglycemic kidney injury (HKI) is a common complication of diabetic patients. We examined the relationship between HKI and the abnormal expression of 5-hydroxytryptamine (5-HT) system induced by hyperglycemia in type 2 diabetes mellitus (T2DM). In animal experiments, a T2DM model was established in mice by feeding a high-fat diet with intraperitoneal injection of streptozotocin. The mice were treated with the 5-HT2A receptor (5-HT2AR) antagonist sarpogrelate hydrochloride (SH) and 5-HT synthesis inhibitor carbidopa (CDP) (respectively or in combination). In cell culture experiments, human glomerular mesangial cells (HMC) were stimulated with D-glucose (D-Glu), and 5-HT2AR, 5-HT synthesis, and 5-HT degradation were inhibited by SH, CDP, or monoamine oxidase A (MAO-A) inhibitor clorgyline. Periodic acid-Schiff (PAS) staining and Masson staining, immunohistochemistry and Western blot, fluorescent probe, and enzyme linked immunosorbent assay (ELISA) and enzyme reagent were respectively used to detect histopathology, protein expression, intracellular reactive oxygen species (ROS), and biochemical indexes. The animal experiments were in accordance with the regulations of the Animal Ethics Committee of China Pharmaceutical University. The results showed that 5-HT2AR, 5-HT synthases, and MAO-A were expressed in glomerular basement membrane and kidney tubular epithelial cells of mouse kidney and HMC. The expression of these proteins was significantly up-regulated in T2DM mice or when HMC cells were exposed to high concentration of D-Glu. HKI, characterized by abnormal renal function, glomerular swelling, and glomerular basement membrane thickening and fibrosis, is closely associated with an increase in kidney 5-HT2AR, 5-HT synthesis, and 5-HT degradation. Among them, 5-HT2AR can mediate the expression of 5-HT synthases and MAO-A; MAO-A can catalyze the degradation of 5-HT to increase the production of mitochondrial ROS, leading to the phosphorylation of nuclear factor kappa B (NF-κB) with the production of inflammatory cytokines, and the up-regulation of matrix metalloproteinase-2 (MMP-2) and α-smooth muscle actin (α-SMA) with the production of collagens. SH and CDP can effectively treat HKI, and the combination of SH and CDP has a clear synergistic effect.
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Objective To investigate the effect of different concentration of uric acid on proliferation and phentyic change of HBZY-1 cells in vitro. Methods The morphological change of HBZY-1 cells exposed to uric acid was observed under the inverted microscope and transmission electronmicroscope. MTT bioassay was used to assess the effect of uric acid on the proliferation of HBZY-1 cell. ELISA method was used to assess the level of angiotensin Ⅱ in HBZY-1 cell incubated with different concentration of uric acid for different treatment duration.The protein and mRNA expression of α-smooth muscle actin (α-SMA) , transforming growth factor-β1 (TGF-β1) and fibronectin in HBZY-1 cells was measured by Western blot and RT-PCR assay, respectively. Results Uric acid (0.4 mmol/L, 24 h) could induce phentypic change of messangial cell from triangle to fibroblast-like morphology.Uric acid could significantly stimulate the proliferation of HBZY-1 cells for 24 h in a concentration-dependent manner (P < 0.05). Uric acid could stimulate the secration of angiotensin Ⅱ in a dose-and time-dependent manner.RT-PCR and Western blot results showed that the markers of myofibroblast transdifferentiation, including a-smooth muscle actin (α-SMA) , transforming growth factor-β1 (TGF-β1) and fibronectin, were significantly upregulated in HBZY-1 cell treated with over 0.1 mmol/L uric acid for 48 h compared with the control group (P < 0.05, respectively). Conclusion Uric acid can induce the proliferation and phentypic change of HBZY-1 in vitro.
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Objective To observe the influence of serum containing esculentoside A(EsA) on the proliferation of glomerular mesangial cell (GMC) and the activation of ERK1/2-AP-1 pathway of glomerular mesangial cell induced by IL-1β.Methods SD rats were gavaged by different doses of EsA(5,10,20,40 mg/kg) for getting medicated sera.The control group was set (gavage by 0.5sodium carboxymethylcellulose);the EsA medicated serum was used to treat rGMC.The control serum group was set.The influence of EsA medicated serum in each group on rGMC proliferation was detected by MTT;the rGMC was divided into the blank control group,IL-1β single action group,IL-1β+ EsA double action group,IL-1β+ U016 double action group and IL-1β+ U0126 + EsA combined action group,which were synchronized and then cultured for 48 h.Western blot was used to detectthe expression of p-ERK1/2 and AP-1 an the imaging analysis was performed.Results The EsA medicated serum(5-10 mg/kg gavage) inhibited the cellular proliferation(P＜0.05 or P＜0.01);the IL-1β group promoted the expression of p-ERK1/2 and AP-1 in rGMC(P＜ 0.05),after acting on rGMC in the IL-1β+EsA double action group,IL-1β+U0126 double action group and IL-1β+U0126+EsA combined action group,the expression of p-ERK1/2 and AP-1 was decreased(P＜0.05).Conclusion Serum containing EsA (5～10 mg/kg gavage) significantly inhibits the rGMC proliferation;EsA inhibits IL-1β induced rGMC proliferation,its action pathway on ERK1/2-AP-1 is one of mechanisms for inhibiting rGMC proliferation.
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To investigate the effect of estrogen receptor (ER) agonist 17β-estradiol and G protein-coupled estrogen receptor 1 (GPER) agonist fulvestrant on masangial cell fibrogenesis under protein kinase C (PKC),we quantified type Ⅳ collagen (COL4A1),fibronectin (FN1),connective tissue growth factor (CTGF) and transforming growth factor-β1 (TGFβ1) gene transcription and semi-quantified phosphorylation of Akt signal upon Phorbol 12-myristate 13-acetate stimulation (which increased COL4A1,FN1,CTGF and TGFβ1 gene transcription to 2.5-0.5,1.4 ±0.2,26 ± 11 and 1.9 ±0.3 times compared with baseline,P ＜0.05) when incubated with the two drugs.It was found that 17β-estradiol and fulvestrant down-regulated COL4A1,FN1,CTGF and TGFβ1 genes transcription (P ＜0.05) and Akt signaling under PKC activation via ER and GPER.ER and GPER agonists are beneficial in protecting the mesangial cells from fibrogenic stimuli by inhibiting PKC signaling and excessive extracellular matrix production.
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Objective:To evaluate the effects of rapamycin on the proliferation,apoptosis and cell cycle of glomerular mesangial cells induced by high glucose,and to explore its significance in the prevention and treatment of diabetic nephropathy. Methods:The rat GMC HBZY-1 was divided into four groups:control group,high glucose group,the first group of high glucose plus rapamycin,the second group of high glucose plus rapamycin. CCK-8 assay was used to detect the proliferation of cells, flow cytometry was introduced to evaluate the apoptosis and cell cycle of HBZY-1,Real-time PCR was used to detect the mRNA of AngiotensinⅡ(ANGⅡ),transfor ming growth factor beta1 ( TGF-β1 ) and vascular endothelial growth factor ( VEGF ) . Results: The proliferation level of HBZY-1 induced by high glucose was significantly increased,and the level of apoptosis decreased,and the expression level of ANGⅡ,TGF-β1 and VEGF was increased. Rapamycin significantly inhibited,and there was a dose dependent,and down regulated the expression of ANGⅡ,TGF-β1,and VEGF. For the cell cycle,the S phase cells in the high glucose group were significantly higher than those in the normal group (P<0. 05),and the S phase cell proportion was decreased after rapamycin intervention (P<0. 05). Conclusion:Rapamycin can inhibit the proliferation of HBZY-1 in high glucose,promote its apoptosis and lead to G1/S arrest,and down regulate the expression of ANGⅡ,TGF-β1 and VEGF.
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MicroRNA could participate in a variety of physiological and pathological processes via iden-tifying the target genes by their seed sequences and regulating gene expression at post-transcriptional level. Glo-merular mesangial cell is one of the most chief cells of the glomerulus and plays an important role in the renal fi-brosis. The mesangial cellular proliferation and dysfunction may appear once attacked by pathogenic factors. Then,the extracellular matrix increases in the stimulation of inflammatory cytokines,eventually,resulting in glo-merular sclerosis and renal fibrosis. Reports have shown that series of microRNA could take part in the process of injury of mesangial cells and lead to renal fibrosis. MicroRNA could be a potential therapeutic target for early chronic kidney diseases.
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Objective To investigate the effects of Hui-hui Gan-song Yin(HGY) on the accumulation of extracellular matrix of rat glomerular mesangial cells(MCs) induced by high glucose.Methods The 40 SD rats were randomly divided into normal control group(distilled water), glurenorm group(10 mg/kg), HGY high-dose group(10 g/kg) and HGY low-dose group(5 g/kg), 10 rats in each group.The rats in each group were treated with corresponding drugs, twice a day.After 3 days, the serum containing each drug were prepared to culture rat MCs in vitro.The MCs were divided into the normal control group( 10% serum of rats in normal control group ) , high glucose group ( 30 mmol/L glucose +10% serum of rats in normal control group), glurenorm group(30 mmol/L glucose+10% serum of rats in glurenorm group), HGY high-dose group(30 mmol/L glucose+10% serum of rats in HGY high-dose group) and HGY low-dose group(30 mmol/L glucose+10% serum of rats in HGY low-dose group).The fibronectin(FN), ColⅠand ColⅣ levels were detected by Western blot.Results Compared with normal control group, the expression of FN, ColⅠand ColⅣ in high glucose group increased(P<0.01).The HCY suppressed the protein expression of FN, ColⅠand ColⅣ significantly(P<0.05).Conclusion The serum containing HGY could suppressed protein expression of FN , ColⅠand ColⅣ and inhibit the accumulation of extracellular matrix of MCs induced by high glucose, which could protect glomerulus and delay the development of diabetic nephropathy.
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Objective To explore the function of MDA on diabetic nephropathy .Methods Glomerular mesangial cells ( GMC) were pretreated with MDA at a final concentrations of 0μmol/L, 1μmol/L, 5μmol/L, 10μmol/L and 50 μmol/L.MTT assay was used to examine the viability of GMC ) .AnnexinV-FITC was used to evaluate effect of MDA on cell apoptosis .RT-PCR and western blot were used to analyze the expression of Nrf 2, HO-1 andγGCL.Results MDA treatment inhibited GMC viability in a dose-dependent manner .MDA at the concentration of more than 5 μmol/L induced mass production of ROS in GMC ( P<0.05 ) .In addition , antioxygen of tBHQ may relieve MDA-induced reduction of cell viability .MDA inhibited the expression of HO-1 , γGCLand Nrf2 ( P <0.05 ) .Conclusions MDA inhibites GMC viability and promotes the cell apoptosis by ROS production through in-hibiting Nrf2/HO-1-γGCL.
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Objective To observe the effects of one kind of angiotensin converting enzyme inhibitor (ACE1) drugs fosinopril (FOS) on transforming growth factor β1 (TGF-β1)and β1- integrin( Itg-31 ) expression in rat glomerular mesangial cells (GMC)induced by lipopolysacchatide (LPS).Methods We established the cultured glomerular mesangial cells of rat in vitro and passages 3 ～ 10 of cells were used in the experiment after identification.The experiment included the following groups:Control group,LPS induced group (LPS group) and FOS intervened group.According to the different concentrations of FOS,FOS intervened group was divided into high,middle and low dose FOS groups,which were FOS1 group,FOS2 group and FOS3 group respectively.The changes of TGF-β1 protein secretion was detected by the enzyme-linked immunosorbent-assay; The changes of TGF-β1 and Itg-β1 mRNA expression was detected by quantitative real-time RT-PCR.Results (1) TGF-β1protein secretion in rat GMC at 6h,12h,24h three time points:They were 958.55 ± 34.67 ( ng/L),1052.05 ±48.59( ng/L),1166.06 + 35.39 (ng/L) respectively in Control group.They were 1342.12 + 39.87 ( ng/L),1432.31 + 39.33 (ng/L) and 1 537.77 + 43.79 (ng/L) respectively in LPS group,which were higher significantly than those in Control group ( all P ＜ 0.01 ).They were 779.58 ± 48.64 ( ng/L),878.33 ± 29.50 (ng/L) and 962.57 ±31.94( ng/L) in FOS1 group,989.311±73.56(ng/L),1073.29±66.89(ng/L) and 1210.75 ±61.68(ng/L) in FOS2 group,1 253.78 ±45.32( ng/L),1 348.18 ±45.81 (ng/L) and 1450.06 ±46.24( ng/L) in FOS3 group respectively,which were lower significantly in all FOS intervened groups than that in LPS group (all P＜O.01).(2)TGF-β1 mRNA expressions in rat GMC at6h,12h,24h three time points were higher significantly than that in Control group.TGF-β1 mRNA expressions were lower significantly in all FOS intervened groups than that in LPS group.( 3 ) Itg-β1 mRNA expressiones in rat GMC at 6h,12h,24h three time points were higher significantly than that in Control group.Itg-β1 expressions were lower significantly in all FOS intervened groups than that in LPS group.Conclusions LPS can induce the increase of TGF-β1 secretion and mRNA expression.FOS can inhibit the TGF-β1 secrection and mRNA expession in GMC as dose-dependent manner,at the same time down regulated the Itg-β1 mRNA expression iuduced by LPS.All above supply the theoretical evidence for the renal protection of FOS by non-hemodynamics mechanism.
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Objective To study whether Fcα/μ receptor(Fcα/μR)can mediate complement killing of human glomerular mesangial cells.Methods Fcα/μR cDNA contained plasmid,pcDNA3.1-Fcα/μR was transfected into a human glomerular mesangial cell(NHMC).Fcα/μR expression was detected by Western blot and laser scanning eonfocal microscopy.Binding of IgM-immune complexes(IgM-IC)to the Fcα/μR on cell membrane was detected by flowcytometry and laser scanning confocal microscopy.Killing of cells by complement was shown by Trypan blue exclusion assay.Results NHMC cells transfected with Fcα/μR could bind IgM and IgM-IC.After treatment with complement,added IgM-IC,the death rate of pcDNA3.1-Fcα/μR transfected cell was significant higher than the control groups of wild type cell,pcDNA3.1 transfected cell and the pcDNA3.1-Fcα/μR transfected cell without IgM-IC.Conclusion IgM-IC can bind to the Fcα/μR expressed NHMC cells and mediate complement killing of the cells.
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Objective To observe the effects of serum containing the medicine of Tujian Mixture on phospho-Akt and phospho-PTEN of glomerular mesangial cell (MC) cultured in high concentrations of glucose. Method The glomerular mesangial cells from SD rat were divided into six groups:the normal control group, the group of MC cultured in high concentrations of glucose, and four other groups cultured in high concentrations of glucose + different concentrations of rat’s serum containing Tujian Mixture. After 72 hours, the content level of phospho-Akt (Thr308) and phospho-PTEN (Ser380) in MC was detected by using sandwich ELISA. Results The level of phospho-Akt (Thr308) in the group of MC cultured in high concentrations of glucose was significant higher than that in the normal control group (P
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PURPOSE: Excess proliferation of mesenchymal cells such as vascular smooth muscle cells and glomerular mesangial cells, cause transplant vascular sclerosis and glomerulosclerosis, which are typical pathological lesions of chronic allograft dysfunction. Mycophenoic acid (MPA) and rapamycin (RPM) were recently reported to have strong anti-proliferative potentials toward vascular smooth muscle cells. However, the potential effects of these drugs, either alone or in combination, on glomerular mesangial cells, remain to be reported. METHODS: Primary cultured mesangial cells, from Sprague-Dawley rats, were isolated, and stimulated with 10ng/ml of PDGF. The test drugs MPA and RPM were administered at various concentrations, either alone or in combination, 15 minutes before the addition of the PDGF. The cell proliferation was assessed by [3H]-thymidine incorporation. RESULTS: The PDGF effectively stimulated the proliferation of the mesangial cells. The MPA inhibited the proliferation in a dose-dependent manner. In comparison to the stimulated control, the MPA (above 500 nM) showed a significant inhibitory effect. The IC50 of the MPA, against PDGF-stimulated mesangial cell proliferation, was between 500 nM and 1microM. The RPM, at 10 nM, showed a significant inhibitory effect. In a linear regression analysis, the RPM was supposed to suppress the mesangial proliferation in a dose-dependent manner (P<0.05). The pattern of inhibition for the MPA and RPM combination was very similar to that of either the MPA or the RPM alone. Both the MPA and RPM were shown to independently suppress the mesangial proliferation from a multiple regression analysis (R2=0.415, P<0.001). CONCLUSION: We demonstrated that MPA and RPM significantly inhibited the proliferation of glomerular mesangial cells, and that these effects were well maintained when used in combination. Our data indicate that both MPA and RPM have unique potentials in preventing the development of transplant mesangial proliferation in renal transplant recipients.
Subject(s)
Animals , Rats , Allografts , Cell Proliferation , Inhibitory Concentration 50 , Linear Models , Mesangial Cells , Muscle, Smooth, Vascular , Mycophenolic Acid , Rats, Sprague-Dawley , Sclerosis , Sirolimus , TransplantationABSTRACT
Objective To investigate the effect of insulin (INS) on nuclear factor-kappa B (NF-KB) activation in glomerular mesangial cells(GMC) of the Zucker rats,and the correlation between the activity of NF-kB induced by insulin to the ages and genotype of Zucker rats. Methods (1) Four groups of cultured GMCs(O3m,O10m,L3m and L10m) from the Zucker obese rats(3 months old and 10 months old) and Zucker lean rats(3 months old and 10 months old) were stimulated by insulin. (2) Electrophoretic mobility shift assay (EMSA) was used to detect the activity of NF-KB. Gel supershift assay was used to detect the subunit of NF-KB dimer. (3) The protein of NF-KB p65 in cytoplasm and cytoblast was analysed by Western Blot. Results (1) NF-KB activity in 4 groups GMCs was significantly higher than that in control group after induced of INS ( F=219. 65 P
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Objective To investigate the effect of recombinant rat augmenter of liver regeneration (rrALR) on the biological activities of rat glomerular mesangial cell(GMC) with or without IL-1?. Methods GMC proliferation and the level of TGF-? protein were determined by 3H-TdR incorporation and ELISA. Results The stimulating activity of IL-1? to rat GMC was enhanced by rrALR with the concentrations from 0.005 pg/ml to 0. 1ng/ml. But rrALR could not promote the proliferation of GMC without IL-1?. Conclusion rrALR may be a new mediator of inflammation, which promote proliferation and TGF-? secretion in rat GMC stimulated by IL-1?.
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Objective To study the mechanism of traditional Chinese medicine Compound Xueniaoting (CXNT) in treating glomerulus diseases with the main pathological changes of proliferation in mesangial cells. Methods CXNT and rat liver microsomes were incubated together, and then the incubated CXNT was added into cultured glomerular mesangial cells (GMC) in vitro. The proliferation of GMC was observed by MTT assay, the levels of interleukin-6 (IL-6) and endothelin-1 (ET-1) were determined by radioimmunoassay, and content of lactic dehydrogenase (LDH) was determined by velocity assay. ResultsCXNT (2, 4, 8 mg/mL) metobolized by rat liver microsomes could all inhibite the proliferation of GMC and production of IL-6 and ET-1 in a dose-dependent manner. Conclusion CXNT can inhibite the proliferation of GMC and restrain the secretion of IL-6 and ET-1, this function may be one of CXNT mechanisms in treating some mesangial proliferative glomerulonephritis.
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Objective To explore the molecular and cellular mechanism of Zaohuang Capsule No.3 (ZHC3) in treating chronic renal failure through observing the effect of ZHC3 on the human glomerular mesangial cells (GMC). Methods The proliferation of human GMC fostered in ZHC3 was detected by MTT, and the fibronectin (FN), interleukin 6 (IL-6) and transforming growth factor ?1 (TGF-?1) secretion was detected by ELISA. Results ZHC3 significantly inhibited GMC proliferation (P
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Objective To observe the changes of extracellular matrix(ECM) production and hypertrophy of glomerular mesangial cells(GMCs) induced by high glucose(HG),and the inhibitory role of myriocin(ISP-1).Methods GMC cultured with normal glucose(5.6 mmol/L D-Glucose,NG),HG(25.2 mmol/L D-Glucose) and HG plus ISP-1(100 mg/L) for different durations(0,24,48,(72 h)).The sizes of GMC were indicated by forward scatter intensity,measured by flow cytometery,and the levels of fibronection(FN),collagen Ⅳ(Col Ⅳ),laminin(LN),precollagen Ⅲ(Pcol Ⅲ) and hyaluronic acid(HA) in the supernatant of cultured GMC were detec-(ted) by ELISA.Results Compared with NG,HG could induce GMC hypertrophy(P
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AIM: To investigate the influence of Astragalus Injection on morphology,cell cycle and oxidative stress of rat's glomerular mesangial cells(GMC) cultured with advanced glycation end products(AGEs). METHODS: GMC were incubated in culture medium containing AGEs in the presence of Astragalus Injection and aminoguanidin for 48 h.At the same time,the normal and control groups were established.Then the cultured GMC was stained by mixed fluorescence liquid and observed under fluorescence microscope.Cell cycle of GMC was analyzed using flowcytometry.The activity of SOD、MDA and GSH-PX in GMC supernatant were measured by test kit.The level of ROS was detected by flowcytometry. RESULTS: Morphology analysis showed that the morphology and structure of normal GMC were normal.The structure of most cells in AGEs was unclear,cell counts increased markedly and they grew intensively.Cell cycle analysis showed that cell percentage of S phase increased and G_0/G_1 reduced.The level of ROS,MDA remarkably increased,and SOD,GSHPX activity reduced.Whereas Astragalus Injection was added,cell morphology tended to be basically normal and cell counts decreased,the percentage of S phase also decreased.The level of ROS,MDA and the SOD,GSH-PX activity restored in comparison with the control groups. CONCLUSION: Astragalus Injection can prevent GMC from lesion caused by AGEs.Astragalus Injection may protect GMC from retarding the progression of diabetic nepropathy by partially inhibiting the occurrence of oxidative stress.