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Objective To observe the effect of rat bone marrow mesenchymal stem cells(BMSCs)on the apoptosis of rat peritoneal mesothelium cells(PMCs)induced by high glucose peritoneal dialysis fluid(PDF),and to explore its possible molecular mechanism.Methods The primary BMSCs and PMCs were extracted and identified.Apoptosis of PMCs was induced by high glucose PDF.Cell supernatant from BMSCs after 24 h of culture was collected as the conditioned medium(BMSCs-CM).PMCs were co-cultured with BMSCs by conditioned media or Transwell chambers.PMCs were randomly divided into the control group,the PDF group and the PDF+BMSCs-CM group.The viability of PMCs was measured by CCK-8 in each group.The depolarization of mitochondrial membrane potential was measured by JC-1 method.TUNEL staining was used to detect cell apoptosis.Western blot assay was used to detect the expression levels of apoptosis related proteins B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax),Cleaved cysteine aspartase-3(Cleaved Caspase-3)and pathway related protein serine/threonine protein kinase(Raf),mitogen-activated extracellular signal-regulated kinase(MEK),extracellular-signal regulated protein kinase(ERK)and their phosphorylated proteins in each group.Results Compared with the control group,the proliferative activity and mitochondrial membrane potential of PMCs were decreased in the PDF group,while the apoptosis rate and the ratio of Bax/Bcl-2,Cleaved Caspase-3/Caspase-3,p-Raf/Raf,p-MEK/MEK and p-ERK/ERK were increased(P＜0.05).Compared with the PDF group,the proliferative activity and mitochondrial membrane potential of PMCs were increased in the PDF+BMSCs-CM group,while the apoptosis rate and the ratio of Bax/Bcl-2,Cleaved Caspase-3/Caspase-3,p-Raf/Raf,p-MEK/MEK and p-ERK/ERK were decreased(P＜0.05).Conclusion BMSCs can reduce the apoptosis of PMCs induced by high glucose PDF,and its mechanism maybe related to inhibiting the activation of Raf/MEK/ERK signaling pathway.
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[Objective]To summarize the advances in traditional Chinese medicine research on protein-energy wasting(PEW) of chronic kidney disease(CKD).[Methods]Read the domestic and foreign literature about PEW in recent years.Collate,study and summarize etiology and pathogenesis,therapeutic principle and symptomatology of PEW,as well as the application of classical prescriptions,compound prescriptions,single herbs and traditional Chinese external treatments about PEW.[Results]The external cause of PEW is feelings of six vessel disease,and the internal cause is weak and damage of Zang and Fu viscera.The pathogenesis consists of deficiency of Ben and repletion of Biao.Deficiency of Ben means dysfunction of Zang and Fu viscera,lack of Qi and blood,Yin and Yang.The disease mainly locates in the spleen and kidney;the retention of damp and hot,phlegm turbidity,blood stasis and water poison,damages the renal collaterals,which constitute Biao of PEW.The chief principle of treatment is to support the right and expel the evil.Emphasis on dialectical therapy,and center on "nourishing kidney and strengthening spleen,benefiting qi and nourishing blood,activating blood and removing stasis,cleaning bowel and dispelling turbidity,detoxificating and reconciling collaterals".Deficiency and excess exist simultaneously in the symptomatology of PEW.Qi deficiency of spleen and kidney is the most common deficiency syndrome,damp and turbidity,damp and hot and blood stasis are seen most of the time in accompanied symptom.Classical prescriptions,single traditional Chinese herb,and external therapy of TCM will be uesd to create good application prospect in the TCM treatments of PEW,which effectively improve clinical symptoms,increase nutrition related indicators,reduce micro-inflammatory response,and finally raise living quality of patients.[Conclusion]Traditional Chinese medicine treatments of the PEW provide advantages and have made certain achievements,which are worth further studying.
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Objective To investigate the effects of AstragalosideⅣ (AST) on the expression of high glucose peritoneal dialysis solution (PDS)-induced transforming growth factor-β1 (TGF-β1), connective tissue growth factor (CTGF), vascular endothelial growth factor (VEGF) in cultured human peritoneal mesothelial cells. To discuss the regulating effect of AST on induced fibrosis cytokines.Methods The HMrSV5 cell line was cultivated to the fifth generation and divided into normal group (10%FBS cultivation solution), model group (4.25% PDS and 10% FBS cultivation solution) and the low, medium and high doses AST groups (4.25% PDS with a respective 10, 20, 40 μg/mL AST). MTT colorimetric assay was employed to detect cell activity and ELISA was applied to detect expression of TGF-β1, CTGF and VEGF in cultured supernatants.Results Except for 5 μg/mL group, HPMCs activity of high glucose plus different concentration AST groups were enhanced in different degrees, especially with 40, 45, and 50 μg/mL (P<0.05). The expression of TGF-β1, CTGF, and VEGF in model group increased. Compared with the control group, expression of the three AST groups significantly decreased and showed dose-effect relationship (P<0.05). Conclusion AST could reduce the expression of TGF-β1, CTGF and VEGF in high glucose-induced HPMCs and was useful in slowing down the progress of peritoneal fibrosis.
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The main characteristic in pathogenesis of chronic nephritis proteinuria is asthenia in origin and asthenia in superficiality,which is Qi deficiency of spleen and kidney seaing the deficient foundation,and pathogenic wind,dampness-heat,and toxic-stasis injuring the body for the whole process of disease.The main therapeutic method is treating from spleen,kidney,liver,and lung,accompanied by the methods of clearing heat,removing dampness,expelling wind,and tonifying kidney to control nocturnal emission.In addition.patients with severe chronic illness should add worm medicines and medicines for promoting blood circulation to remove stasis.
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Objective To observe the effect of Astragalus Injection(AI) on aquaporin-1(AQP-1) expression of peritoneal mesothelial cells in rats with peritoneal dialysis.Methods Twenty-five male SD rats were divided into the normal control group(N =7),model group(N =9) and AI group(N =9).Except the normal control group,the model group was given intraperitoneal injection of peritoneal dialysate(containing glucose 42.5 g/L),and AI group was given intraperitoneal injection of peritoneal dialysate(containing glucose 42.5 g/L) plus AI(20 mg/mL),25mL/d for 10 continuous days.On the 11th day of experiment,the amount of dialysis ultrafiltration(UF) and net UF,and the concentration ratio of dialysate and plasma sodium(D/PNa) were measured,peritoneal pathological changes were observed with light microscope,and the expression of AQP-1 in peritoneal mesothelial cells was detected with immunohistochemical method.Results When the peritoneal dialysate was kept in the abdominal cavity for 60 min,the amount of UF and Net UF was increased in AI group(P