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Objective To investigate the effects of vitamin E (VE) on renal ischemia/reperfusion (I/R) injury of rats.Methods A total of 18 male Wistar rats were randomly divided into sham-operated group,I/R group,VE + I/R group,and each group of 6 rats.All the animals were killed at the end of 24 h of reperfusion.Nephridial tissue were examined by light microscopy,and the level of blood urea nitrogen(BUN) and serum creatinine (SCr) were measured.The protein expressions of tumor necrosis factor α (TNF-α) were detected by Western blotting.Results Compared with sham-operated group,tubulointerstitial pathological injury in I/R group was significantly aggravated,which was shown by HE and PAS stain.Compared with I/R group,the degree of morphological changes as well as renal dysfunction in VE + I/R group were obviously lessened.Meanwhile,the levels of BUN,SCr in I/R group,VE + I/R group were (10.13 ± 2.14) mmol/L and (7.67 ± 1.63) mmol/L,(80.33 ±7.15) μmol/L and (63.67 ±5.40) μ mol/L,significantly higher than those in shamed-operated group ((3.85 ± 0.21) mmol/L,(48.67 ± 3.61) μmol/L;P ＜ 0.05).And the level of BUN and SCr in VE + I/R group were significant lower than those in I/R group(P ＜0.05).Western Blotting showed that the protein expressions of TNF-α in VE + I/R group were obviously lower compared with those in group of I/R without VE treatment (P ＜ 0.05).Conclusion Vitamin E can attenuate over-expressions of TNF-α in kidney following I/R,thus protect against structural damages and renal dysfunction in I/R rat models.
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Objective To investigate the effect of intermedin (IMD) on renal ischemia/ reperfusion (I/R) injury after the up-regulation of IMD. Methods A total of 24 healthy Wistar male rats were randomly divided into four groups,sham-operated group,I/R group,IMD gene transfection +I/R group and empty plamid +I/R group.All the animals were killed at the end of 24 h of reperfusion.Histological changes and renal function were estimated.The expression and site of IMD were determined by Immunohistochemistry method,semi-quantitative RT-PCR and Western blotting.The protein expressions of endothelin 1 (ET-1),tumor necrosis factor αt (TNF-α) were detected by Western blotting. Results Compared with sham-operated group,tubulointerstitial pathological injury was significant aggravated in I/R group (7.6±2.3) and empty plamid +I/R group (7.0±1.8),and such injury was improved in IMD+I/R group (1.5±0.8) (P＜0.05).Compared with I/R group and empty plamid +I/R group,the renal dysfunction of IMD +I/R group was obviously lessened [BUN:(7.73±1.03) mmol/L vs (10.13±2.14) mmol/L,(9.77±1.92) mmol/L; Scr:(58.50±3.27) μmol/L vs (80.33±7.15) μmol/L, (75.67±7.58) μmol/L,all P＜0.05].IMD expression was weak in the plasma of tubulointerstitial cells in sham-operated group,and was up-regulated in I/R group. Compared with I/R group, immunohistochemical IMD expression increased obviously (262.03±67.89 vs 175.57±48.06,P＜0.01).The mRNA expression of IMD in IMD+I/R group was up-regulated significantly by 60.7％,66.1％ and the protein expression of IMD in IMD+I/R group increased significantly by 51.4％,55.9％ as compared to I/R and empty plasmid +I/R group.Meanwhile,the protein expressions of ET-1 and TNF-αt in IMD+I/R group were obviously lower compared with those in I/R group (ET-1:0.08±0.02 vs 0.17±0.02; TNF-α:0.21±0.04 vs 0.35± 0.02,all P＜0.05). Conclusion IMD gene transfected into kidneys of rats prior to I/R surgery can attenuate the over-expressions of both ET-1 and TNF-o in I/R injured rat kidneys as well as the damages to the structure and function of the kidneys.
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Objective To observe the effects of intermedin (IMD) on nitric oxide synthetase (NOS) in renal ischemia-reperfusion (IR) rat models and the action mechanism.Methods A total of 24 rats were divided into four groups (n =6 each).Group Ⅰ underwent right nephrectomy one week prior to the exposure of left renal pedicles,but did uot receive any I/R.Group Ⅱ underwent right nephrectomy one week prior to left renal I/R surgery.Group Ⅲ underwent right nephrectomy and left renal IMD-pCDNA3.1 ( + ) transfection by ultrasound-mircobubbles and renal I/R surgeries were performed one week after gene transfection.Group Ⅳ was treated with the same way as group Ⅲ except that empty control vector was transfected.All the animals were killed at the end of 24 h of reperfusion.The expression and site of IMD were determined by using immunohistochemistry.Serum levels of BUN and creatinine were determined.The kidney formaldehyde-fixed and paraffin-embedded sections were stained with HE and PAS by standard methods and then histological changes were analyzed semiquantitatively.The mRNA expression levels of endothelial NOS (eNOS),inducible NOS (iNOS) and neuronal NOS (nNOS) in the kidneys of the four groups were detected by using RT-PCR.The protein expression levels of the three NOS mentioned above in the kidneys were semiquantitatively analyzed by Western blotting.Results IMD was weakly expressed in the plasma of tubulointerstitial cells in sham-operated group; whereas IMD expression in the kidneys subject to I/R was increased.Moreover,as compared with I/R group,IMD expression levels were obviously increased (P＜0.01 ).The degree of morphological changes as well as renal dysfunction in group Ⅲ was obviously lessened as compared with group Ⅱ.The mRNA and protein expression levels of eNOS in group Ⅲ were notably increased as compared with group Ⅱ,while the mRNA and protein expression levels of iNOSin group Ⅲ were obviously reduced as compared with I/R group not transfeeted with IMD (P＜0.05).Meanwhile,there were no significant differences in the mRNA and protein expression levels of nNOS among groups Ⅱ,Ⅲ and Ⅳ.Conclusion IMD gene in the kidneys of rats can promote the expression of eNOS and attenuate over-expression of iNOS in the kidneys following I/R,thus protecting against tubulointerstitial damages and renal dysfunction in rat I/R models.
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Objective To construct eukaryotic expression vector encoding rat IMD gene and deliver it into rat renal tissue via ultrasound-mircobubbles. Methods IMD gene was inserted into pCDNA3.1 ( + )between Hind Ⅲ and EcoRI enzyme sites. The recombinant plasmid designated as IMD-pCDNA 3.1 wasconfirmed by restrictive enzyme digestion and sequencing. Eighteen male Wistar rats were randomized into 3groups, which were treated with no transfection, empty vector transfection and IMD transfection, respectively, in renal tissue via ultrasound-microbubbles. RT-PCR and Western blotting were applied to detect the expression level of IMD. Results Enzyme- digestion and sequencing data showed that IMD-pCDNA 3.1 was correctly constructed. The differences in ALT, AST, BUN and SCr were not significant; No obvious damage in the glomerular, tubular and interstitial was observed in all the treated groups;Compared with non-transfection group and empty vector-transfection group, IMD mRNA and protein expression in IMD transgenic renal tissue were significantly increased. Conclusion IMD-pCDNA 3.1 expression vector was successfully constructed and well expressed in rat kidney.
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Objective To evaluate the clinical significance of detecting urinary natural killer(NK) cells in patients with general types of glomerulonephritis. Methods The contents of urinary NK cells from 54 patients with glomerulonephritis were measured by flowcytometry,while all patients were classified into two groups including acute proliferation group and none-acute proliferation group by renal biopsy results. The content of urinary NK cells was compared between the two groups. Results The content of urinary NK cells in acute proliferative glomerular disease group were( 14. 8 ±3. 3)% (30 cases) ,which was significantly higher than that of(21. 6 ±2. 9)% (24 cases) in the non-acute proliferative glomerular patients(P<0.05). Conclusions Decreasing of the contents of NK cells in urine may be an indirect indicator of the activity of glomerulonephritis.
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Objective To investigate the protective effect of intermedin(IMD)on renal ischemia reperfusion injury(IRI)and its mechanism. Methods A total of twenty-four male Wistar rats were randomly divided into four groups: control group, IRI group, empty plasmid group and IMD group. After remove of right kidney, plasmid was transfected into the kidney by ultrasonic microbubbles technology, and IRI model was made after 1 week. Renal pathology was observed by PAS staining. Renal tissue superoxide dismutase(SOD), myeloperoxidase(MPO), caspase-3 activity, and malondialdehyde(MDA)content were detected by colorimetric method. The intercellular cell adhesion molecule-1(ICAM-1), endothelin 1(ET-1)and P-selection expression of renal tissue were detected by immunohistochemical method. Apoptosis of renal tubular cell was detected by TUNEL.Results Compared with control group, tubulointerstitial pathological injury was significant aggravated in IRI group(P<0.01);compared with IRI group, IMD pretreatment significantly alleviated the degree of renal injury(P<0.01). Compared with control group, in IRI group, SOD activity was significantly decreased(P<0.05), MPO activity, caspase-3 activity, MDA production and the expression of ICAM-1, P-selection, ET-1 were increased significantly(all P< 0.01). Compared with IRI group, IMD pretreatment significantly increased SOD activity(P <0.05), decreased the MPO activity, caspase-3 activity, MDA production and the expression of ICAM-1, P-selection, ET-1 (all P<0.01). The apoptosis rate of renal tubular epithelial cells in IRI group was significantly higher than that in control group(34.83%±8.75% vs 3.33%±0.47%, P<0.01), while the apoptosis rate of IMD group(20.67%±7.71%)was significantly lower than that of IRI group. There was no difference of above indexes between empty plasmid group and IRI group. Conclusions IMD pretreatment protects against renal IRI. The mechanism may be at least partly related to the clearance of oxygen free radicals, the improvement of lipid peroxidation, inflammatory cell infiltration and cell apoptosis, leading to the decrease of the production of reactive oxygen species caused by oxidative stress.
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Objective To investigate the relationship among interstitial infiltrating inflammatory cells (including CD4+, CD8+, CD25+, MAC387+ and 27E10+), renal function and pathological changes in IgA nephropathy (IgAN). Methods This study included 36 IgAN patients. PAS and immunohistochemistry method were applied to stain paraffin sections. The surface of interstitial tubules, the amount of CD4+, CD8+, CD25+, MAC387+, 27E10 + cells per unit were counted(cells/mm2), together with pathological changes (including glomerulosclerosis, tubular atrophy and interstitial sclerosis) and renal function during and after biopsy were determined by using image analysis system and SPSS software pack. Results In the renal tubulointerstitium, the number of CD4+ cells correlated with interstitial fibrosis (r=0.38 ,P