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Research on Renal Expression of TSP-1 and Cytokines That Promote Interstitial Inflammation and Fibrosis in the Early Time of Rats with Protein-overload Proteinuria

Author: LiXiaoHui
Tutor: MaHong;LiuYuan
School: Shanxi Medical
Course: Pediatrics
Keywords: Kidney disease in young rats Thrombospondin Proteinuria Extracellular matrix Renal interstitial fibrosis
CLC: R692
Type: Master's thesis
Year: 2003
Downloads: 70
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Abstract


Objective To investigate the proteinuria caused by the molecular mechanism of the development of early renal interstitial fibrosis, tubular protein load condition, the renal tissue thrombospondin (TSP-1) expression trends and transforming growth factor (TGF-beta1), connective tissue growth factor (CTGF), urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor (PAI-1) expression, its role in the process in the early fibrosis formation. Explore the intervention of the cytokines. Methods the protein load nephropathy young rats for experimental kidney disease animal models, single stained with immunohistochemistry and double staining detection of TSP-1, TGF-beta1 and uPA expression at the protein level; tissue in situ hybridization of CTGF and PAI-1 expression at the mRNA level; analysis of the changes in the characteristics and correlation of the indicators in the process of lesion progression. Measured over the same period urine protein and blood biochemical indicators to observe the changes in the characteristics. I benzene that Plymouth and low molecular weight heparin treatment for 3 weeks, and changes in the indicators, to determine the effect of therapeutic interventions. Results in 3 days after the measured 24-hour urine protein given to rats by intraperitoneal injection of bovine serum albumin (BSA), you can see significantly higher (model group the 36.6 ± 15.19mg/24h control group 7.16 ± 1.42mg/24h The P = 0.000) . Reached the peak at 131.10 ± 18.3mg/24h (control group 7.31, ± 1.71mg/24h) on the 9th day, followed by a temporary decline, the first 30 days again appear the significantly increased 104.29 ± 21.77mg/24h (control group 6.51 ± 0.66mg/24h P = 0.000). General biopsy visible at all levels of tubular protein cast formation, but blood biochemical indicators does not appear to change renal failure. Semi-quantitative immunohistochemical analysis method measured TSP-1 protein expression positive or positive tubules staining integral value in the first three days was significantly higher for 2.42 ± 0.26 (control group, 2.12 ± 0.24, P = 0.04). Expression of the peak in the 9th day 3.39 ± 0.41 (control group, 2.06 ± 0.691, followed by the expression, although weakened, continued high level to 30 days was 2.71 ± 0.13 (control group 1.97 ± 0.39P = 0.001) with the progression immune group of semi-quantitative analysis method measured TGF-beta1 protein positive cells or positive tubules staining integral value on day 6 was significantly higher for 2.63 ± 0.14 (control group, 1.70 ± 0.56 P = 0.01), Shanxi Medical University, 2003 Master BU graduate degree Papers peak expression for the 9th day of 2.63 ± O.14 (control group, 1.70 ± 0.56), followed by the expression, although weakened, continued high levels exist to 30 days 1.65 ± 0.39, 2.75 ± 0.22 (control group). situ hybridization semi-quantitative Day 6 analysis method measured CTGF mRNA-positive cells or positive tubules staining integral value was significantly increased to 2.75 ± 0.13 (control group, 1.98 ± o.24p = 0.02) expression peak of 2.01 ± 0.16 to the 9th day 3.98 ± 0.35 (control group) , followed by expression although weakened, continued high levels exist to 30 days (control group 3.36 ± 0.12 1.94 ± 0.15P = 0.00). the TSP L, TGF December 1 protein expression and the expression of CTGFmRNA peak in the 9th day, with the progression of expression has decreased to 30 days continued high level of presence of the cytokine expression trends and proteinuria trends synchronization by correlation analysis showed TSP-1, TGF pl protein expression and CTGF mRNA expression the same trend. the three expression was a significant positive correlation with significant (rl = 0.971 0.842r2). semi-quantitative analysis of immunohistochemical measured uPA protein positive cells or positive tubules staining integral value from the first 9 days significantly decreased to 1.18 ± 0.12 (control group, 1.53 ± 0.30), with the progression of sustained low-level expression in situ hybridization semi-quantitative analysis method measured the PAI lmRNA expression of positive cells or positive tubules staining integral value starting from the first 3 days significantly increased to 2.33 ± 0.20 (control group 1 .75 ± 0.44) 1.55 persons peak of expression in the 9th day 3.76 ± 0.18 (control group, 64), the subsequent expression, although weakened, persistently high level present to 30 days to 2.76 ± 0.12 (control group, 1.47 ± 0.20) expression of TSP-1 protein expression and PAI lmRNA positive correlation. early in the disease TSP-1 protein expression and uPA protein expression negatively correlated with the course. progress this correlation is not obvious expression of uPA protein expression of PAI-lmRNA no obvious correlation TSP-1 protein expression and PAI lmRNA expression were consistent application benazepril and low molecular weight heparin combination therapy, the second week of treatment group and untreated group, 24-hour urinary protein was significantly decreased to 64.29 ± 4.44 mg (untreated group Shi-9.O6mg 89.00), this change is continuous to the first 3 weeks 46.71 of Guests, 7.93 mg (untreated group 1 04.29 ± 21 .77 mg). untreated group with the progression of urinary protein was gradually Bu rising trend in the treatment of urinary protein gradually Bu drop. the blood biochemistry __ between the two groups was no obvious different TSP-1 protein expression in the treatment group with the first three weeks of the untreated group was a decrease of 2.12 ± 0.14 (untreated group 2.71 Shi-0.13), with the progression of the TSP l eggs ease untreated group continued high expression in the treatment of Shanxi Medical University, 20 (3) Master Ren graduate degree thesis group was waning the TGF the pl protein expression treatment group and the untreated group compare the first 2 weeks significantly decreased to 2.14 ± 0.17 (untreated group, 2.78 ± 0.07), this change last until the 3rd week 1.7 ± 0.43 (untreated group. 5 ± 0.22) with the progression of TGF pl protein expression in the untreated group continued high expression in the treatment group gradually weakened CTGF mRNA expression in the treatment group and the untreated group 3 weeks uPA protein expression in the treatment group and the untreated group decreased to 2.07 ± 0. clever (untreated group 3.36 Shi-0.12). CTGF mRNA with the progression of the untreated group continued high expression, the treatment group CTGF mRNA expression decreased gradually. Compare two weeks significantly increased 1.92 ± 0.13 (untreated group, 1.47 ± 0.13) to 3 weeks 2.08 ± 0.17 (untreated group 1,41 ± 0.18) with the progression of the untreated group uPA protein expression decreased gradually, treatment sustained high expression group u Ensemble, and increased expression of PAI a lmRNA treatment group and the untreated group the first week decreased 2.39 Shi 0.65 (untreated group, 2.96 ± 0.19), this change is continuing to 3 weeks 1.71 Shi 0.02 (untreated group 2.76 Shi-0.12) with the progression of the team I a lmRNA in the untreated group, the persistently high expression, and the expression decreased gradually in the treatment group. Conclusions juvenile nephropathy in rats tubular egg?

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