Effect of renal fibrosis after macrophage depletion in C3-deficient unilateral ureteral obstruction mice / 中华肾脏病杂志

Objective@#To investigate the effect and mechanism of renal fibrosis after macrophage depletion in C3-deficient unilateral ureteral obstruction mice.@*Methods@#Renal interstitial fibrosis model was established by unilateral ureteral obstruction (UUO) in male C3-deficient mice and age-matched C57BL/6 WT mice (8-12 weeks of age). Mice were randomly divided into 4 groups, including sham operation in wild type group (WT/sham) (n=18), UUO operation in wild type group (WT/UUO) (n=18), sham operation in C3-deficient group (C3KO/sham) (n=18), and UUO operation in C3-deficient group (C3KO/UUO) (n=18). The expression of complement C3 was detected by immunohistochemical staining and renal interstitial macrophages were assessed by immunofluorescence staining. Tubulointerstitial fibrosis was observed by both HE staining and Masson staining after 14 days of UUO. Collagen accumulation and score of tubulointerstitial injury were obtained. Wild type and C3-deficient UUO mice were treated by liposome clodronate in early or late stage respectively and then interstitially infiltrated macrophages and renal fibrosis were analysed. Mice were sacrificed randomly at 3,7,14 days after UUO and obstructed kidneys were collected. Macrophage phenotype was detected by double-labeling immunofluorescence with F4/80 and iNOS for the M1, F4/80 and CD206 for the M2 macrophage subpopulation. iNOS, Arg-1 and CD206 were also detected by western blot.@*Results@#C3 deficient mice exhibited attenuated renal fibrosis, reduced collagen accumulation and tubulointerstitial injury score compared with WT mice (P<0.01). Meanwhile, macrophage depletion in early or late stage of UUO reduced renal fibrosis in WT mice, but had no effect on C3-deficient UUO mice. Decreased accumulation of M1 macrophages and expression of iNOS, increased accumulation of M2 macrophages and expression of Arg-1, CD206 were found in C3 deficient mice compared with WT mice in early stage of UUO (P<0.01).@*Conclusion@#Renal fibrosis is not reduced after depletion of macrophages in C3 deficient UUO mice due to the altered macrophage polarization.

Effect of hyperuricemia on the clinicopathological features and progression of IgA nephropathy / 中国实用内科杂志

OBJECTIVE: To investigate the clinical and pathological features in hyperuricemic IgA nephropathy,and the effect of hyperuricemia on progression of IgAN.METHODS: Patients with IgAN confirmed by renal biopsy were enrolled.Those patients were admitted to the First Affiliated Hospital of Fujian Medical University from January 2006 to December 2016.The relationship between uric acid and clinical and pathological changes was analyzed.Patients were followed up and the level of serum creatinine was measured.Primary endpoint was double of creatinine or end-stage renal disease(ESRD)or renal replacement therapy.Risk factors of progression in IgAN were assessed using Kaplan-Meier and Cox proportional hazards analyses.RESULTS: A total of 231 patients with IgAN either reached the endpoint or followed up for more than 2 years were enrolled.Among these patients,the prevalence of hyperuricaemia was 39.8%.There were significant differences in gender,systolic blood pressure,diastolic blood pressure,serum creatinine,blood urea nitrogen,24-hour urine protein,eGFR,degree of tubule atrophy/interstitial fibrosis between high(H-UA)group and normal(N-UA)uric acid group(P<0.05).A total of29 patients reached the endpoint.Univariate COX regression analysis showed that there were significant differences between the progressive group and the non-progressive group in glomerulosclerosis,renal tubular atrophy/interstitial fibrosis,24-hour urine protein quantification,hyperuricemia,anemia,hypertension,blood creatinine,and blood urea nitrogen(P<0.05).Kaplan-Meier survival curve suggested that renal survival was lower in H-UA IgAN group.Anemia,24-hour urine protein,glomerulosclerosis,and serum creatinine are independent risk factors for progression of Ig AN assessed by multivariate analysis.CONCLUSION: Hyperuricemia may be positively associated with severe clinical and pathological damage,more renal tubular atrophy/interstitial fibrosis lesions,and lower renal survival.

Effect of renal fibrosis after macrophage depletion in C3 - deficient unilateral ureteral obstruction mice / 中华肾脏病杂志

Objective To investigate the effect and mechanism of renal fibrosis after macrophage depletion in C3 - deficient unilateral ureteral obstruction mice. Methods Renal interstitial fibrosis model was established by unilateral ureteral obstruction (UUO) in male C3-deficient mice and age-matched C57BL/6 WT mice (8-12 weeks of age). Mice were randomly divided into 4 groups, including sham operation in wild type group(WT/sham)(n=18), UUO operation in wild type group(WT/UUO)(n=18), sham operation in C3-deficient group(C3KO/sham)(n=18), and UUO operation in C3-deficient group(C3KO/UUO)(n=18). The expression of complement C3 was detected by immunohistochemical staining and renal interstitial macrophages were assessed by immunofluorescence staining. Tubulointerstitial fibrosis was observed by both HE staining and Masson staining after 14 days of UUO. Collagen accumulation and score of tubulointerstitial injury were obtained. Wild type and C3-deficient UUO mice were treated by liposome clodronate in early or late stage respectively and then interstitially infiltrated macrophages and renal fibrosis were analysed. Mice were sacrificed randomly at 3,7,14 days after UUO and obstructed kidneys were collected. Macrophage phenotype was detected by double-labeling immunofluorescence with F4/80 and iNOS for the M1, F4/80 and CD206 for the M2 macrophage subpopulation. iNOS, Arg-1 and CD206 were also detected by western blot. Results C3 deficient mice exhibited attenuated renal fibrosis, reduced collagen accumulation and tubulointerstitial injury score compared with WT mice (P<0.01). Meanwhile, macrophage depletion in early or late stage of UUO reduced renal fibrosis in WT mice, but had no effect on C3-deficient UUO mice. Decreased accumulation of M1 macrophages and expression of iNOS, increased accumulation of M2 macrophages and expression of Arg-1, CD206 were found in C3 deficient mice compared with WT mice in early stage of UUO (P<0.01). Conclusion Renal fibrosis is not reduced after depletion of macrophages in C3 deficient UUO mice due to the altered macrophage polarization.

Effect of complement fragment C3a on the phenotype of mice podocytes in vitro / 中华肾脏病杂志

Objective To investigate the effect of complement C3a on mouse podocytes phenotype transformation.Methods Purified C3a recombinant protein was used to stimulate mature mouse podocytes.The expression of the mature podocyte markers synaptopodin,podocin,nephrin,CD2-associated protein (CD2AP) and the mesenchymal cell markers fibroblast specific protein 1 (FSP-1),α-smooth muscle actin (α-SMA) were detected by RT-PCR,Western blotting,immunochemistry and immunofluorescence,respectively.Some podocytes were transfected with integrin-linked kinase (ILK) siRNA before the administration of C3a,the expression of nephrin and α-SMA were accessed by Western blotting,and the expression of Snail and α-actinin 4 were accessed by Western blotting and immunochemical method.The migration ability of podocytes was observed by scratch test.Results Immunocytochemistry and immunofluorescence analysis showed that synaptopodin,podocin,nephrin,CD2AP were highly expressed by mature mouse podocytes.The expression of these podocyte markers could be markedly inhibited after 24 h of C3a (0.1 μmol/L) treatment,and accompanied by the induction of mesenchymal markers FSP-1 and α-SMA.Compared with control group,the mRNA levels of synaptopodin,podocin,CD2AP and nephrin were significantly repressed by the administration of C3a in a dose-dependent manner,whereas the transcription of FSP-1 and α-SMA were remarkably up-regulated by C3a treatment (P ＜ 0.05,respectively).Western blotting analysis also confirmed the decrease of synaptopodin,podocin,nephrin and CD2AP protein and the increase of FSP-1 and α-SMA protein were closely depend on the C3a concentration (P ＜ 0.05,respectively).To further assess the downstream of C3a,some podocytes were transfected with ILK siRNA before the administration of C3a.Compared with C3a group,the protein levels of nephrin and α-SMA were significantly changed by the administration of ILK siRNA (P ＜ 0.05,respectively).The expression of α-actinin 4 and Snail induced by C3a were inhibited by ILK knockdown (P ＜ 0.05,respectively),accompanied by a decline of cell migration potency.Conclusion Complement fragment C3a can induce transformation of mouse podocytes to mesenehymal cells,and ILK signaling pathway is involved in this cell type transformation.

Effects of bone marrow-derived mesenchymal stem cells on glomerular podocyte injured by lipopolysaccharide / 中华肾脏病杂志

Objective To observe the effects of bone marrow?derived mesenchymal stem cells (BMSC) on glomerular podocyte injured by lipopolysaccharide (LPS) and the expression of related protein. Methods Podocytes are divided into control group, BMSC group, LPS group and LPS plus BMSC group. After 24 hours of intervention, observing each experimental group podocyte form under inverted phase contrast microscope;detecting the expressions of mRNA and protein of nephrin, CD2AP, synaptopodin, and TRPC6 by RT?PCR and Western?blot. Results Compared with control group, expressions of nephrin, CD2AP, and synaptopodin in LPS group decreased (P<0.05) while that of TRPC6 increased (P<0.05); compared with LPS group, expressions of nephrin, CD2AP, and synaptopodin in LPS+MSC group increased (P<0.05) while that of TRPC6 decreased (P<0.05). Conclusion BMSC may relieve LPS?induced podocyte injury.

Clinical efficacy and safety of sequential treatment with alprostadil and beraprost sodium for chronic renal failure induced by chronic glomerulonephritis / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the clinical efficacy and safety of sequential treatment with alprostadil and beraprost sodium for chronic renal failure caused by chronic glomerulonephritis.</p><p><b>METHODS</b>Sixty-three patients with chronic renal failure due to chronic glomerulonephritis, after receiving a 2-week-long conventional treatment, were randomly divided into alprostadil group (n=20, with alprostadil injection at 10 µg/d for 2 weeks), sequential treatment group (n=21, with alprostadil injection at 10 µg/d for 2 weeks and oral beraprost sodium at 20 µg three times a day for 12 weeks), and strengthened sequential treatment group (n=22, with alprostadil injection at 20 µg/d for 2 weeks and a double dose of oral beraprost sodium for 12 weeks). Urinary albumin excretion rate (UAER), cystatin C (Cys C), blood urea nitrogen, creatinine, fibrinogen, D-dimer, prothrombin time (PT), and platelets were tested before and after the treatment, and the changes in urinary albumin discharge rate, serum creatinine, and glomerular filtration rate were determined.</p><p><b>RESULTS</b>The patients in strengthened sequential treatment group showed a significantly decreased change rate of urinary albumin discharge rate (P<0.01) than those in the other two groups. In the two sequential treatment groups, especially the strengthened treatment group, the change rate of glomerular filtration rate increased significantly compared with that in alprostadil group (P<0.01). Strengthened sequential treatment resulted also in significantly decreased increment of serum creatinine compared that in the other 2 groups (P<0.01). After 14 weeks of treatment, fibrinogen and D-dimer were decreased in all the 3 groups (P<0.05) to a comparable level between the 3 groups (P>0.05), and prothrombin time (PT) or platelet showed no significant changes (P>0.05).</p><p><b>CONCLUSION</b>Sequential treatment with alprostadil and beraprost sodium can improve the glomerular filtration rate and decrease urine albumin excretion rate, serum creatinine increase rate, and lower blood fibrinogen and D-dimer levels, thus delaying the progression of chronic renal failure caused by chronic glomerulonephritis. This therapy shows a dose-related effect with good clinical safety.</p>

Erythropoietin inhibits complement 3a-induced renal tubular epithelial to mesenchymal transition / 中华肾脏病杂志

Objective To investigate the effects of erythropoietin (EPO) on complement 3a (C3a)-induced renal tubular epithelial to mesenchymal transition. Methods The HK-2 cells were divided into 6 groups namely control group,EPO group,TGF-β group,TGF-β+EPO group,C3a group and EPO+C3a group.The mRNA and protein expressions of α-SMA,E-cadherin and C3 were investigated by RT-PCR,Western blot and immunofluorescence respectively. Results Compared with control group and EPO group,the mRNA and protein expressions of α-SMA in HK-2 cells were up-regulated after the intervention of C3a or TGF-β (all P＜0.05).On the contrast,the mRNA and protein expressions of E-cadherin were down-regulated(P＜0.05),the mRNA and protein expressions of C3 were enhanced (all P＜0.05).However,all those above effects of C3a or TGF-β were inhibited after the intervention of EPO (all P＜0.05). Conclusion EPO is capable of suppressing the epithelial to mesenchymal transition induced by C3a.

Effects of atorvastin on function of glomerular endothelial cells in rats with chronic renal failure / 中国药学杂志

OBJECTIVE: To investigate the effects of atorvastin on function of glomerular endothelia cells in rats with chronic renal failure (CRF). METHODS: Twenty-eight male SD rats were randomly divided into 4 groups: sham operation group (control group), CRF group (model group), 8 mg · kg-1 · d-1 atorvastin treatment group (low-dosage group) and 16 mg · kg-1 · d-1 atorvastin treatment group (high-dosage group). The model of chronic renal failure was established by a two stage 5/6 nephrectomy procedure. The atorvastin treatment groups were given atorvastin by intragastric administration, and the other 2 groups were given sodium chloride. Serum creatinine (Scr), blood urea nitrogen (BUN), urine protein, total cholesterol (TCHO), triglycerides (TG), low density liporotein (LDL), AST, ALT and creatine kinase (CK) were measured after 8 weeks. The renal morphologic changes were e-valuated on periodic acid-schiff (PAS) stained sections. The CD34 and CD31 expressions in glomerulus were detected by immunohisto-chemistry method. The mRNA of ET1, eNOS and VEGF were detected by RT-PCR. RESULTS: The Scr, BUN and urine protein levels in atorvastin treatment groups were significantly lower than those in CRF model group (P < 0.05). Reanl pathological injuries were improved by atorvastin treatment in dose-dependent manner. There were no significant differences in TCHO, TG, LDL, AST, ALT and CK levels among the 4 groups. The expressions of CD34 and CD31 protein in glomerulus, and the expressions of eNOS and VEGF mRNA in renal tissue were higher in atorvastin treatment groups than those in CRF model group. The expression of ET-1 mRNA in renal tissue were lower in atorvastin treatment groups than those in CRF model group. CONCLUSION: It might be through promoting renovation of glomerulus capillary endothelium and improving function of glomerular endothelial cells that atorvastin ameliorates renal pathological injury and renal function in rats with chronic renal failure. Copyright 2012 by the Chinese Pharmaceutical Association.

Effects of erythropoietin On glomerular endothelial cells function in rats with chronic renal failure / 中华肾脏病杂志

Objective To investigate the effects of erythropoietin(EPO)on the function of glomerular endothelial cells in rats with chronic renal failure(CRF). Methods The CRF model was established by a two stage 5/6 nephrectomy procedure in rats.Experimental rats were randomly divided into four groups:sham operation group (control group),CRF group,CRF rats treated with 30 U/kg EPO(low-dosage group)and with 50 U/kg EPO (high-dosage group).CRF rats received EPO by hypodermic injection for 6 weeks and then were sacrificed.Serum creatinine(Scr),blood urea nitrogen fBUN),urine protein,haematoglobin (Hb) and blood pressure were measured.The renal morphologie changes were evaluated on periodic acid-schiff (PAS) stained sections.The CD34 and CD31 expressions in glomerulus were detected by immunohistochemistry method.The mRNA of endothelin 1(ET-1),endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF) were detected by RT-PCR. Results The expressions of CD34 and CD31 protein in glomerulus,and the expressions of eNOS and VEGF roRNA in renal tissue were higher in EPO treatment group than those in CRF model group(all P<0,05).The expression of ET-1 mRNA in renal tissue was lower in EPO treatment group than that in CRF model group.In addition,the Scr,BUN,urine protein and blood pressure in EPO treatment group were significantly lower than those in CRF model group (all P<0.05).Haematoglobin in EPO treatment group was higher than that in CRF model group (P<0.05).Reanl pathological injury wss improved by EPO treatment in dose-dependent manner. Conclusion EPO can ameliorate renal pathological injury and renal function in rats with chronic renal failure,maybe through promoting the renovation of glomerular capillary endothelium and improving the function of glomerular endothelial cells.

Effects of erythropoietin on peripheral blood endothelial progenitor cells from rats with chronic kidney disease / 中华肾脏病杂志

Objective To investigate the effects of erythropoietin (EPO) on the number and function of peripheral blood endothelial progenitor cells (EPCs) from rats with chronic renal failure (CRF). Methods The model of chronic renal failure was established by a two-stage 5/6nephrectomy procedure in rats. Experimental rats were randomly divided into four groups (n =7,respectively): sham operation group, CRF group, CRF rats treated with 30 U/kg EPO (low-dosage group) and with 50 U/kg EPO (high-dosage group). CRF rats were given EPO by hypodermic injection for 6 weeks, then EPCs were isolated by density gradient centrifugation from peripheral blood mononuclear cells. The ability of cell proliferation, adhesion and vasculogenesis in vitro was further observed. Results Compared to sham operation group, the ability of cell proliferation,adhesion and vasculogenesis in vitro in CRF rats was remarkably decreased (P＜0.05, respectively).Such ability was promoted significantly in dose-dependent manner by EPO treatment (P＜0.05,respectively). Conclusion EPO can improve the number and ability of endothelial progenitor cells from rats with chronic renal failure.

Relationship between serum concentrations of interleukin-6 (IL-6), soluble IL-6 receptor and the activity of Graves′ ophthalmopathy / 中华内分泌代谢杂志

Serum concentrations of interleukin-6 (IL-6), soluble IL-6 receptor (sIL-6R) were determined in patients with Graves′ ophthalmopathy (GO), Graves′disease without GO (NGO) and controls. Patients with GO treated by corciosteroids were assessed by the clinical activity score (CAS). The results suggested that serum levels of both IL-6, sIL-6R and CAS could reflect the activity of GO and predict the outcome of corcicosteroid therapy.