| Objective: In recent years,podocyte injury has received extensive attention in the pathogenesis of IgA nephropathy(IgAN).In this study,we have examined the expression of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and Wilm’s tumor 1(WT1)in renal tissue of IgA nephropathy and the levels of TNF-α and IL-6 in urine,and collected Clinical and pathological data in order to explore the role of TNF-α and IL-6 in podocyte injury and the development of IgA nephropathy.Methods: Forty patients with Ig A nephropathy diagnosed by renal biopsy and clinical and pathological data in the Second Hospital of Hebei Medical University were enrolled from December 2015 to December 2016.Except for secondary IgAN including allergic purpura,autoimmune diseases,cirrhosis,psoriasis,ankylosing spondylitis and combined with other kidney disease,such as diabetic nephropathy.The patient’s pathological indicators weregraded by Ig A nephropathy Lee Grading,and with reference to Katafuchi and other methods,the glomerular,tubulointerstitial were received semi-quantitative integration.The relationship between clinical parameters and different pathological types and pathological changes were analyzed.According to Lee’s renal tissue changes,they are divided into I ~ V level,and grade I ~ III level for a group,meanwhile IV and V level for another group.The patients’ clinical data of general condition were recorded,including urine protein,serum albumin,serum creatinine,uric acid,β2-microglobulin,total cholesterol,triglyceride,fasting blood glucose,urinary osmotic pressure.The simplified MDRD formula was applied to evaluate glomerular filtration rate.40 cases of normal healthy adults were enrolled as the control group during the same time in our hospital examination center.10 cases of renal tissue from the lesion site were taken as normal tissue control after renal resection of urology in thesecond hospital of Hebei Medical University.The levels of urinary TNF-α and IL-6 were measured by enzyme-linked immunosorbent assay(ELISA).The results of urinary L-6 and urinary TNF-α had to be divided by urinary creatinine.Immunohistochemical method was used to stain TNF-α,IL-6 and WT1 in renal tissue,and the results were analyzed by image analysis software.Statistical analysis was performed by using IBM spss21.0 statistical software.Results:1 The general data:(1)Compared with control group,the systolic blood pressure,diastolic blood pressure,serum creatinine,uric acid,β2-microglobulin,total cholesterol,triglycerides in IgA nephropathy patients were significantly increased,while the serum albumin,eGFR were decreased,The difference was statistically significant(P <0.05).There was no significant difference in gender,age and fasting blood glucose between the two groups(P>0.05).(2)Between the experiment groups: Compared with patients of grade I~III,the levels of systolic blood pressure,diastolic blood pressure,urinary protein,serum creatinine,uric acid,β2-microglobulin,total cholesterol,triglycerides were increased(P<0.05).Serum albumin,eGFR,urine osmolality were significantly lower,the difference was statistically significant(P <0.05).There was no significant difference in gender,age and fasting blood glucose between the two groups(P> 0.05).2 The changes of TNF-α:(1)The expression of TNF-α in renal tissue:TNF-α is mainly expressed in the cytoplasm of proximal tubule epithelial cells,and the proximal tubules are the most obvious,which appeared yellow or brownish.Among the experimental groups,the average optical density of TNF-α in renal tissue of patients with grade I ~ III Ig A nephropathy was(17.12 ± 5.22),and the average optical density of TNF-α in renal tissue of patients with grade IV ~ V IgA nephropathy was(35.15 ± 9.42),the difference was statistically significant(P<0.05).(2)The urinary TNF-α of Ig A nephropathy patients was(23.18 ± 10.09)ng / mg · Cr,the urinary TNF-αof normal control group is(11.92 ± 2.53)ng / mg · Cr.Comparing the two groups,the experimental group was significantly increased,the difference wasstatistically significant(P <0.05).Comparing the experimental groups,the urinary TNF-α of IgA nephropathy patients with grade I ~ III was(15.70 ±4.98)ng / mg · Cr,the urinary TNF-α of IgA nephropathy patients with grade IV ~ V was(29.95 ± 8.49)ng / mg · Cr.The heavier the pathological type,the higher the level of TNF-α,the difference was statistically significant(P<0.05).3 The changes of IL-6:(1)The expression of IL-6 in renal tissue: IL-6 is mainly expressed in the cytoplasm of proximal tubule epithelial cells,and the proximal tubules are the most obvious,which was yellow or brownish.Compared with the two groups,the average optical density of IL-6 in renal tissue of patients with IgA nephropathy with grade I ~ III was(24.25 ± 11.98),and the average optical density of IL-6 in renal tissue of patients with grade IV~ V Ig A nephropathy was(52.57 ± 20.15),the difference was statistically significant(P <0.05).(2)Urinary IL-6 in patients with IgA nephropathy was(20.45 ± 10.34)pg / mg · Cr,in normal control group,urinary IL-6 was(5.90± 2.31)pg / mg · Cr,the difference was statistically significant(P <0.05).Comparing two experimental groups,the urinary IL-6 of IgA nephropathy patients with grade I ~ III was(13.18 ± 5.30)ng / mg · Cr,the urinary IL-6 of IgA nephropathy patients with grade IV ~ V was(27.01 ± 9.36)ng / mg · Cr.The heavier the pathological type,the higher the level of IL-6,the difference was statistically significant(P <0.05).4 Expression of WT1 in renal tissue: WT1 is mainly expressed in normal podocyte nuclei,yellow or brownish granules.Comparing the experimental groups,the average optical density of WT1 in renal tissue of patients with grade I ~ III Ig A nephropathy was(14.15 ± 5.30),and the average optical density of WT1 in renal tissue of patients with grade IV ~ V IgA nephropathy was(9.44±3.51),which was statistically significant(P <0.05).5 Correlation analysis:(1)the correlation analysis of TNF-α in renal tissue with other indexes: there was a positive correlation between the expression of TNF-α in renal tissue of IgA nephropathy and systolic blood pressure,diastolic blood pressure,urine protein,serum creatinine,uric acid,β2-microglobulin,total cholesterol,triglyceride,mesangial hyperplasia,glomerular sclerosis,interstitial fibrosis,tubular atrophy,IL-6 positive expression rate,urinary TNF-α and urinary IL-6;there was a negtive correlation between the expression of TNF-α in renal tissue and serum albumin,eGFR,urinary osmotic pressure,WT1 positive expression rate;age,fasting blood glucose were not correlated with TNF-α level in renal tissue.(2)the correlation analysis of urinary TNF-α with other indexes: there was a positive correlation between urinary TNF-α in patients with IgA nephropathy and systolic blood pressure,diastolic blood pressure,urinary protein,serum creatinine,uric acid,β2-microglobulin,total cholesterol,mesangial hyperplasia,the degree of sclerosis,interstitial fibrosis,tubular atrophy,TNF-α positive expression rate,IL-6 positive expression rate and urinary IL-6;there was a negtive correlation between urinary TNF-α and serum albumin,eGFR,urinary osmotic pressure and wT1 positive expression rate;age,fasting blood glucose and triglyceride were not correlated with urinary TNF-α level.(3)the correlation analysis of IL-6 in renal tissue with other indexes: there was a positive correlation between the expression of IL-6 in renal tissue of Ig A nephropathy and systolic blood pressure,diastolic blood pressure,urinary protein,serum creatinine,uric acid,β2-microglobulin,total cholesterol,mesangial hyperplasia,glomerular sclerosis,interstitial fibrosis,tubular atrophy,TNF-α positive expression rate,urinary TNF-α and urinary IL-6;there was a negtive correlation between the expression of IL-6 in renal tissue and serum albumin,eGFR,urinary osmotic pressure,WT1 positive expression rate;age,fasting blood glucose and triglyceride were not correlated with IL-6level in renal tissue.(4)the correlation analysis of urinary IL-6 with other indexes: there was a positive correlation between urinary IL-6 in patients with IgA nephropathy and systolic blood pressure,diastolic blood pressure,urinary protein,serum creatinine,uric acid,β2-microglobulin,total cholesterol,mesangial hyperplasia,the degree of sclerosis,interstitial fibrosis,tubular atrophy,TNF-α positive expression rate,IL-6 positive expression rate and urinary TNF-α;there was a negtive correlation between urinary IL-6 andserum albumin,eGFR,urinary osmotic pressure and wT1 positive expression rate;age,fasting blood glucose and triglyceride were not correlated with urinary IL-6 level.Conclusions:1 TNF-α,IL-6 aggravates Ig AN podocyte injury and promotes disease progression.2 The levels of TNF-α and IL-6 in urine are expected to be a reliable indicators of local TNF-α and IL-6 metabolism in renal tissue,which provides a noninvasive method for clinical monitoring the progression of Ig A nephropathy. |
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