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ObjectiveTo preliminarily predict the active components， targets， and signaling pathways of modified Shengjiangsan in the treatment of immunoglobulin A nephropathy （IgAN） based on network pharmacology， and to explore its underlying mechanism through molecular docking and experimental verification on animals. MethodThe active ingredients and related targets of modified Shengjiangsan were obtained from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP）， UniProt， SwissTargetPrediction， and literature review. IgAN-related targets were obtained from GeneCards and Online Mendelian Inheritance in Man （OMIM）. Cytoscape 3.9.0 was used to construct the regulation network of the related targets of Shengjiangsan and IgAN， and the protein-protein interaction （PPI） network was plotted by STRING. The common genes were analyzed for gene ontology （GO） functional annotation and Kyoto encyclopedia of genes and genomes（KEGG） pathway enrichment by Metascape. Key targets and main active ingredients were selected for molecular docking by AutoDockTools 1.5.6. The experimental model of IgAN was induced by bovine serum albumin（BSA， ig） combined with lipopolysaccharide （LPS， iv） and the complex of CCl4 and castor oil （sc） in rats. The model rats were treated with modified Shengjiangsan and benazepril hydrochloride for four weeks. The rats were sacrificed after drug administration. The levels of transforming growth factor-β1 （TGF-β1） and interleukin-6 （IL-6） in the serum and kidney tissues were detected by enzyme-linked immunosorbent assay（ELISA）， immunohistochemistry， Real-time quantitative polymerase chain reaction （Real-time PCR）， and Western blot. ResultA total of 105 active ingredients were obtained according to oral bioavailability（OB）， drug-likeness（DL）， and literature screening. There were 124 common genes and 59 core targets. Neurotrophic tyrosine receptor kinase 1 （NTRK1）， cullin-3 （CUL3）， tumor protein 53 （TP53）， epidermal growth factor receptor （EGFR）， exportin 1 （XPO1）， and other targets might be closely related to IgAN. As predicted by KEGG enrichment analysis， the treatment of IgAN with modified Shengjiangsan mainly involved the phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway， nuclear transcription factor-kappa B （NF-κB） signaling pathway， and cytokine-cytokine receptor interaction signaling pathway. As revealed by molecular docking， the main active ingredients in modified Shengjiangsan showed stable binding activities with NTRK1， CUL3， TP53， EGFR， and XPO1 in the core targets， indicating that it presumedly regulated inflammatory responses by affecting NTRK1， CUL3， TP53， EGFR， and XPO1 target proteins. The results of experimental verification on animals showed that the expression levels of cytokines TGF-β1 and IL-6 in the serum and kidney tissues of IgAN rats were significantly decreased by modified Shengjiangsan， suggesting that Shengjiangsan might inhibit excessive fibrosis， and inflammatory and immune responses by regulating signaling pathways such as cytokine-cytokine receptor interaction， PI3K/Akt， and NF-κB. ConclusionModified Shengjiangsan may treat IgAN through multiple targets and pathways. Its mechanism may be related to the inhibition of excessive fibrosis， and inflammatory and immune responses by affecting the expression of NTRK1， CUL3， TP53， EGFR， and XPO1 and the regulation of the cytokine-cytokine receptor interaction， PI3K/Akt， NF-κB， and other signaling pathways.
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IgA nephropathy (IgAN) is one of the common primary glomerulonephritis, which is also an important risk factor for end-stage renal disease. Kidney transplantation is the optimal treatment for end-stage renal disease induced by IgAN, whereas there is still a risk of recurrence of IgAN after kidney transplantation. At present, research progress upon IgAN recurrence after kidney transplantation is relatively lacking. The pathogenesis of IgAN recurrence remains elusive, and its pathological manifestations are not specific. The diagnosis of IgAN recurrence still depends on renal biopsy. Besides, no effective prevention and treatment are available for recurrent IgAN. In this article, research progress on IgAN recurrence after kidney transplantation was illustrated from the perspectives of pathogenesis, diagnosis, risk factors and treatment, aiming to provide reference for clinical prevention and treatment of IgAN recurrence after kidney transplantation and improve clinical prognosis of kidney transplant recipients.
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Objective To analyze the expression of serum anti M phospholipase A2 receptor (PLA2R)antibody in idiopathic membranous nephropathy (IMN),and to investigate its value in the diagnosis and evaluation of idiopathic membranous ne-phropathy.Methods One hundred and eighteen patients with biopsy-proved glomerular diseases were involved in this study, including 97 cases with IMN,21 cases with IgA nephropathy (IgAN)and 19 healthy people.ELISA was used to detect ser-um anti-PLA2R antibodies.Correlations of anti-PLA2R antibody level with laboratory parameters,including serum albumin, 24-hour urine protein of IMN patients were evaluated.Results The median of anti PLA2R antibody in IMN group,IgAN group and healthy group was 45.2(3.6~705.9)RU/ml,5.9(2.3~10.6)RU/ml and 1.2(0.6~9.3)RU/ml.The levels of serum anti PLA2R antibody in IMN group were higher than those in IgA nephropathy group and healthy control group.The difference was statistically significant (t=-5.027,-3.077;P=0.05).Among 97 cases with IMN,76 cases showed posi-tive anti-PLA2R antibodies (positive rate 78.35%).There was none patient showed positive anti-PLA2R antibody respec-tively in IgAN and healthy people.Furthermore,anti-PLA2R antibody level was negatively correlated with serum albumin (r=-0.453,P=0.000)and positively correlated with CREA,TC,ESR,24 hour urine protein (r=0.233,0.234,0.363, 0.586;P=0.004,0.217,0.021,0.000)in IMN patients.Conclusion Serum anti PLA2R antibody may be used as a IMN specific marker for the diagnosis of IMN,and it has important reference value for evaluating the severity of IMN.
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Objective To evaluate the therapeutic effects of Thesium chinensis Turcz extracts on IgA nephropathy (IgAN) rats .Methods The hot water extracted solution of Thesium chinensis Turcz flow through the AB‐8 macroreticular res‐in column repeatedly ,then being eluted with different contents of ethanol to get four part of extract .The IgAN model of SD rats were established by oral intake of bovine serum albumin(BSA ,400 mg/kg) together with injection of lipopolysaccharide(LPS) and CCl4 .Beginning with 8th week ,the positive group(LGT ,10 mg/kg) ,water extract group(TT ,1 .0 g/kg) ,water soluble group(TTW ,0 .4 g/kg) ,high polarity group(TT20 ,0 .4 g/kg) and medium polarity group(TT50 ,0 .4 g/kg) rats were intragas‐tric administrated drug daily ,the normal group control and model control group rats were given with saline(n=8) .During the next 5 weeks treatment process ,the 24 h urine volume ,24 h urine protein concentration ,and quantity of erythrocyte in urine were observed .The levels of TP ,ALP ,ALT ,AST ,Scr and BUN in serum were determined .The expression of IgA deposition was measured by immunofluorescence staining ,and the pathological changes of kidney tissue were assayed after the therapy process .Results After 5 weeks therapy ,the 24 h urine protein concentration ,the quantity of erythrocyte in urine ,and the level of Scr and BUN of TT and TT50 groups were significantly lower than model control .The fluorescence intensities in the me‐sangial area were weaker than model control .Also ,the degrees of renal capsule expansion and mesangial matrix proliferations , and the kidney tubules damage of TT and TT50 groups were weaker than those of model control .Conclusion The water extract and its medium polarity fraction of Thesium chinensis Turcz show good therapeutic effects on IgAN rats .
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Objec0tive To investigate the molecular mechanism of the mal-production of IgA and IgA1 by tonsillar mononuclear cells (TMCs) in IgA nephropathy (IgAN) patients by measuring the mRNA expression of Iα-Cα germline transcript and the mRNA and protein expression of activated induced cytidine deaminase (AID) in cultured TMCs stimulated with lipopolysaccharide (LPS) or hemolytic streptococcus (HS) in IgAN patients as well as the chronic tonsilitis patients. Methods Twent-seven IgAN patients admitted into our hospital from Jan.2009 to Feb.2010 were enrolled.Twent-seven patients with chronic tonsillitis without renal disease were selected as control.Tonsillar mononuclear cells were isolated by density gradient centrifugation in lymphocyte separation medium.The amount of IgA or IgA1 secreted in the culture supernatants was determined by specific enzyme-linked immunosorbent assay (ELISA).Expressions of Iα-Cα germline transcript and AID mRNA were examined by real-time PCR.The AID protein was determined by Western blot. Results The production of IgA and IgA1 protein,especially the ratio of IgA1/IgA and the expression of AID protein in TMCs were significantly increased in IgAN group compared with chronic tonsillitis group (all P＜0.05).The IgA and IgA1 level of stimulated TMCs were obviously increased in patients with IgAN compared with control group (P＜0.05).And the expressions of Iα-Cα mRNA,AID mRNA and AID protein were up-regulated significantly in stimulated TMCs (all P＜0.05). Conclusions Both LPS and HS can induce the production of IgA and IgA1 and up-regulate the expressions of AID and Iα-Cα in TMCs of IgAN patients.Our results indicate that the TMCs are capable of producing high level of IgA and IgA1 stimulated by LPS or HS,whuch may be due to the incression of AID and Iα-Cα.
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Objective To explore the expression of BID protein and the relation ship of cell proliferation with apoptosis at different stages of IgAN.Methods The expression of BID protein、PCNA and FN in renal tissue was detected by immunohistochemistry,TUNEL and light microscopy were used to detect apoptotic cells.Results With the progress of IgAN,the expression of BID protein、PCNA、the number of apoptotic cells and FN increased gradually(P
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BACKGROUND: Vascular endothelial growth factor (VEGF) is a potent enhancer of microvascular permeability and a selective endothelial cell growth factor. In human kidney, VEGF is expressed mainly in glomerular visceral epithelial cells. We investigated the relationship between serum levels of VEGF and factors reflecting the severity of disease including histological patterns in order to elucidate the relevance of VEGF in the pathogenesis of IgA nephropathy. METHODS: Serum VEGF was studied using a sandwich ELISA from 21 patients with IgA nephropathy. Histological patterns are classified to 5 grades by WHO classification and frequencies of crescent and glomerular sclerosis, degree of interstitial fibrosis were recorded. Serum concentrations of creatinine, albumin, IgA, amounts of 24 hour urine protein excretion, and creatinine clearances are also evaluated. RESULTS: Serum VEGF levels were significantly correlated with histological grade(r=0.471, p < 0.05), frequency of cellular crescent(r=0.485, p < 0.05), degree of interstitial fibrosis(r=0.562, p < 0.01), and 24 hour urine protein excretion(r=0.439, p < 0.05), and inversely with serum albumin concentration(r=-0.594, p < 0.01). Studies in 17 patients without crescent formation revealed that only serum albumin concentration showed significant correlation with serum VEGF level. CONCLUSION: Serum VEGF concentration is mainly correlated with cellular crescent formation reflecting activity of the disease rather than chronic structural changes such as glomerular sclerosis or interstitial fibrosis. Elevated serum VEGF concentration seems to be due to the release of relatively large amounts of stored VEGF from damaged visceral epithelial cells. Serum VEGF concentration may be a useful marker to evaluate the degree of acute renal injury, especially cellular crescent formation.
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Humans , Acute Kidney Injury , Capillary Permeability , Classification , Creatinine , Endothelial Cells , Enzyme-Linked Immunosorbent Assay , Epithelial Cells , Fibrosis , Glomerulonephritis, IGA , Immunoglobulin A , Kidney , Podocytes , Sclerosis , Serum Albumin , Vascular Endothelial Growth Factor AABSTRACT
Objective The expression of IL-13 in plasma and renal tissue was studied to eluciate the molecular pathogenesis of Mesangial proliferative glomerulonephritis (MsPGN). Methods Plasma level of IL-13 in patients with MsPGN and in normal control groups were examined by ELISA. The expression of IL-13 in patients with MsPGN and in control group were examined by immune histochemistry ABC technique in renal tissue.Results Plasma levels of IL-13 in patients with MsPGN were significantly higher than that in control group,and the highest was found in IgAN;IL-13 expressions in glomerulus and tubulointerstitial zone in patients with MsPGN were higher than that in control group, and IL-13 expression in tubulointerstitial zone was more strong,but it was not distinctive difference between non-IgA MsPGN and IgAN. Conclusion IL-13 may play an important role in the molecular pathogenesis of MsPGN.
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PURPOSE: We studied to find out apo-E genotype polymorphism in minimal change nephrotic syndrome(MCNS) and IgA nephropathy(IgAN) and to determine the relationship between apo-E genotype and clinical course of MCNS. MATERIALS AND METHOD: 43 MCNS patients and 15 IgAN patients were examined for apo-E polymorphism. 50 healthy blood donors were examined for apo-E genotype as control. Genomic DNA was prepared from peripheral blood leukocytes according to standard procedures. RESULTS: As compared with control group, e4 allele frequency was significantly increased in MCNS (P<0.01). However, in IgAN e2 allele frequency, however, was 2.6 times higher than normal control (P<0.01). The frequency of e4 allele of frequent relapser group was 4.6 times higher than normal control and was 2 times higher than infrequent relapser group. CONCLUSION: We think that apo-E typing might be one of the parameters, which should be considered to predict the course of MCNS in children. MCNS with risky HLA profile and E4/4 genotype could indicate the need for a longer steroid dministration. And apo-E genotype needs to be considered for the evaluation of therapeutic responses to other drugs.