| PrefaceAristolochic Acid Nephropathy (AAN) is a kind of tubulointerstitial injury caused by Aristolochic Acid (AA),which can be expressed as acute renal failure,chronic renal falure and tubules acidosis. There were several lesions under light microscope could be observed in renal pathological slices, including tubules epithelial cells necrosis and/or loss, tubules base membrance hypoxia, tubulointerstitial fibrosis, microcapillary damage, Vascular wall Thickening, Vascular Cavity narrow, even to close totally. Recently, studies focused on AAN. Few therapeutic methods on AAN has been proved effectively。The purpose of this work is to observe the effect of calcium dobesilate on the urine protein, the blood urea nitrogen(BUN),Serum creatinie (Scr),and.expression of CD34 and vWF in peritubular capillary (PTC) in Chronic arisolochic acid nephrapathy (CAAN) rats'renal tissues and to explore the probable mechanism of calcium dobesilate improving microcirculation of kidney in rats with CAAN.Materials and Methods 1 Materials1.1 Animals:60 female wistar rats, weighting 190-220g, aged 7-8 weeks, were bought from China medical university animal center, the rats were divided randomly into three groups.1.2 Major reagent and physics: manshuriensis, calcium dobesilate, CD34 test kits, vWF test kits.1.3 Apparatus: spectrophotometer, auto-biochemical-analyzer, centrifugal machine, microcamera, microimage analyzer, optical microscope.2.Methods2.1 Manshuriensis Dutchmanspipe solution was made with intensity of 2g/ml.2.2 The division of animals: 60 rats were divided into three groups randomly:①calcium dobesilate treatment group: 20 rats were infused into stomachs with Aristolochia manshuriensis decoction 10ml/kg.d for 12 weeks, then turned to be treated by calcium dobesilate at the following 4 weeks solely.②NX group: 20 rats were gavaged with Aristolochia manshuriensis decoction 10ml/kg/d for 12 weeks first, and infused with distilled water for 4 weeks later.③SHAM group: 20 rats in the SHAM were infused with distilled water althrough the 16 weeks. All 60 rats feed by enough water and food.2.3 Biochemical detection8 rats were killed in which three groups respectively at 12 weeks. The others sacrificed at 16 week, the end of this study. After being anaesthesized introperitoneally with 10% chloral hydrate, the kidney and blood of rats were collected, kidney was perfused with saline until pale before it was removed quickly and packed. Routine biochemical examination included serum creatine(Scr), blood urea nitrogen (BUN), 24 hours urinary protein were assayed.2.4 Pathology detectionThe kidney tissue was fixed with 4%neutral formaldehyde and embeded in paraffin. Section of 3μm was made, which was then stained with Masson for the evaluation of renal mesenchymal lesions. The sections were observed under light microscope.2.5 Immnutohisto-chemical analysisExpression of CD34 and vWF (Von Willebrand factor) were examined by ABC immunohistochemistry. Five high power(400×) were chose randomly in every slice and measured the value of Intergrated optional density(IOD) using microimage analyzer under light microscope.2.6 Statistical analysisAll data were handled by SPSS 13.0. Metering data between two groups were tested by t test.Results1.Urinary protein, serum creatinine and BUN:the levels of 24 hours urinary protein, serum creatinine and BUN were increased markedly in rats after infused with Aristolochia manshuriensis decoction for 12th weeks comparing to those gavaged by distilled water(P＜0.01).At the end of study, 16th weeks, we found that Urinary protein, serum creatinine and BUN in calcium dobesilate treated group were much lower than those of NX group (P＜0.01).There was no change in CON group throughout the 16 weeks study. 2.Renal histologyTubulointerstitial fibrosis was observed in both of NX and calcium dobesilate treatment groups at 12th weeks, since the lesion in the later is much attenuate than the former at 16 weeks(P＜0.01). No significant pathologic changes were found in SHAM group at either 12 or 16 weeks.3.Level of expression of CD34 measured by immunohistochemistry:Expression of CD34 measured by immunohistochemistry in calcium dobesilate treated group and NX group evaluated significantly than that of CON at 12th weeks and 16th weeks(P＜0.01).The IOD value of CD34~+ increased significantly in calcium dobesilate treated group ((16.7±4.2)×10~3) comparing to NX group ((3.19±1.4)×10~3) at 16th weeks (P＜0.01).4. Level of expression of vWF measured by immunohistochemistry:Expression of vWF measured by immunohistochemistry in calcium dobesilate treated group and NX group evaluated signficiantly than that of SHAM at 12th weeks and 16th weeks (P＜0.01).The IOD value of vWF~+ decreased in calcium dobesilate treated group((10.16±1.68)×10~3) comparing to NX group ((18.66±4.65)×10~3)at 16th weeks (P＜0.01).Conclusions1.Calcium dobesilate could decreased the levels of urinary protein, and improved the renal function in aristolochic acid-induced rats.2.Ccalcium dobesilate could attenuate the damage of interstitial peritubules capillaries.3. Calcium dobesilate could upregulate the expression of CD34~+ and density of peritubular capillary (PTC) in CAAN rats'renal tissues. Calcium dobesilate could deduce the level of vWF~+ and decreased thrombosis。...
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