滋阴凉血汤对IgA肾病大鼠的肾脏保护机制研究
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摘要
【目的】观察滋阴凉血汤对IgA肾病大鼠蛋白尿、血尿、尿甘露糖结合凝集素(MBL)及肾小球、肾小管损害的影响,探讨其对肾脏的保护机制。【方法】将30只SD大鼠分为正常组、模型组、中药组、西药组及中西药结合组等5组,每组6只。模型组和3个治疗组采用牛血清白蛋白+脂多糖+四氯化碳的方法复制IgA肾病大鼠模型,在造模完成后1周,中药组给予滋阴凉血汤(剂量为11.5 g·kg~(-1)·d~(-1))灌胃,西药组给予贝那普利水溶液(剂量为2.5 mg·kg~(-1)·d~(-1))灌胃,中西药结合组用上述半剂量滋阴凉血汤水溶液与贝那普利水溶液灌胃,正常组与模型组均给予等体积生理盐水灌胃,每日1次。根据上述方案灌胃4周后,观察大鼠一般状况,采集大鼠尿液、静脉血,检测其尿红细胞数目、24 h尿蛋白定量、尿MBL及血生化指标。应用直接免疫荧光法测定其肾小球IgA沉积强度,苏木素—伊红(HE)染色法和高碘酸—无色品红(PAS)染色法观察肾小球及肾小管改变。应用电镜观察肾组织超微结构。【结果】中药组大鼠尿红细胞数目,尿蛋白、尿MBL含量较模型组显著减少,差异有统计学意义(P<0.05);中药组大鼠肾小球及肾小管病变较模型组明显改善。【结论】滋阴凉血汤能减少IgA肾病大鼠尿红细胞数目、尿蛋白、尿MBL,改善肾小球内系膜基质、细胞增生和肾小管萎缩及炎症反应等病理改变,对IgA肾病大鼠肾脏具有保护作用。
Objective To observe the effects of Ziyin Liangxue Decoction on proteinuria, hematuria, urinarymannose-binding lectin(MBL),and glomerular and renal tubular damages in IgA nephropathy(IgAN)rats,andto investigate its renal protective mechanism. Methods Thirty SD rats were divided into normal group, modelgroup,Chinese medicine group,western medicine group and integrated Chinese and western medicine group,6 rats in each group. The rats in the model group and the 3 medication groups were given bovine serum albumin,lipopolysaccharide and carbon tetrachloride to induce IgAN model. One week after establishment of the model,Chinese medicine group was given intragastric administration of Ziyin Liangxue Decoction(at the dosage of 11.5 g·kg~(-1)·d~(-1)),western medicine group was given intragastric administration of benazepril aqueous solution(at thedosage of 2.5 mg·kg~(-1)·d~(-1)),integrated Chinese and western medicine group was given intragastric administrationof Ziyin Liangxue Decoction and benazepril aqueous solution(at half dosage of the corresponding drugs),and thenormal group and the model group were given intragastric administration of the same volume of normal saline,oncea day. After medication for 4 weeks,the general situation of the rats in various groups were observed,urine andvenous blood were sampled for the examination of urinary erythrocytes,24-h urine protein,urinary MBL,andblood biochemical indexes. IgA deposition intensity of glomeruli was measured by direct immunofluorescenceassay, and the histological changes of glomeruli and renal tubules were observed by hematoxylin-eosin(HE)staining and periodic acid-Schiff(PAS)staining methods. The ultrastructure of rat renal tissues was observed underelectron microscope. Results The urinary erythrocyte count,and the contents of proteinuria and urinary MBL inChinese medicine group were significantly lower than those in the model group,the differences being statisticallysignificant(P < 0.05). The glomerular and renal tubular lesions of the rats in Chinese medicine group weresignificantly improved as compared with those of the model group. Conclusion Ziyin Liangxue Decoction canreduce urinary erythrocyte count and the contents of proteinuria and urinary MBL,and improving the pathologicalchanges involving glomerular mesangial matrix, cell proliferation, renal tubular atrophy, and inflammatoryresponse in IgAN rats,which results into the renal protective effects on IgAN.
Objective To observe the effects of Ziyin Liangxue Decoction on proteinuria, hematuria, urinarymannose-binding lectin(MBL),and glomerular and renal tubular damages in IgA nephropathy(IgAN)rats,andto investigate its renal protective mechanism. Methods Thirty SD rats were divided into normal group, modelgroup,Chinese medicine group,western medicine group and integrated Chinese and western medicine group,6 rats in each group. The rats in the model group and the 3 medication groups were given bovine serum albumin,lipopolysaccharide and carbon tetrachloride to induce IgAN model. One week after establishment of the model,Chinese medicine group was given intragastric administration of Ziyin Liangxue Decoction(at the dosage of 11.5 g·kg~(-1)·d~(-1)),western medicine group was given intragastric administration of benazepril aqueous solution(at thedosage of 2.5 mg·kg~(-1)·d~(-1)),integrated Chinese and western medicine group was given intragastric administrationof Ziyin Liangxue Decoction and benazepril aqueous solution(at half dosage of the corresponding drugs),and thenormal group and the model group were given intragastric administration of the same volume of normal saline,oncea day. After medication for 4 weeks,the general situation of the rats in various groups were observed,urine andvenous blood were sampled for the examination of urinary erythrocytes,24-h urine protein,urinary MBL,andblood biochemical indexes. IgA deposition intensity of glomeruli was measured by direct immunofluorescenceassay, and the histological changes of glomeruli and renal tubules were observed by hematoxylin-eosin(HE)staining and periodic acid-Schiff(PAS)staining methods. The ultrastructure of rat renal tissues was observed underelectron microscope. Results The urinary erythrocyte count,and the contents of proteinuria and urinary MBL inChinese medicine group were significantly lower than those in the model group,the differences being statisticallysignificant(P < 0.05). The glomerular and renal tubular lesions of the rats in Chinese medicine group weresignificantly improved as compared with those of the model group. Conclusion Ziyin Liangxue Decoction canreduce urinary erythrocyte count and the contents of proteinuria and urinary MBL,and improving the pathologicalchanges involving glomerular mesangial matrix, cell proliferation, renal tubular atrophy, and inflammatoryresponse in IgAN rats,which results into the renal protective effects on IgAN.
引文
[1]Wyatt R J,Julian B A.Ig A nephropathy[J].N Eng1 J Med,2013,368(25):2402.
[2]Li L S,Liu Z H.Epidemiologic data of renal diseases from a single unit in China:analysis based on 13519 renal biopsies[J].Kidney Int,2004,66(3):920.
[3]王海燕.肾脏病学[M].3版.北京:人民卫生出版社,2008:993.
[4]Pozzi C,Andrulli S,Pani A,et al.Addition of azathioprine to corticosteroids does not benefit patients with Ig A nephropathy[J].J Am Soc Nephrol,2010,21(10):1783.
[5]汤颖,娄探奇,成彩联,等.实验性Ig A肾病模型的改进[J].中山大学学报,2006,27(2):184.
[6]Barratt J,Eitner F,Feehally J,et al.Immune complex formation in Ig A nephropathy:a case of the‘right’antibodies in the‘wrong’place at the‘wrong’time[J].Nephrol Dial Transplant,2009,24(12):3620.
[7]Onda K,Ohi H M,Ohsawal I.Hypercomplementemia in adult patients with Ig A nephropathy[J].J Clin Lab Anal,2007,21(2):77.
[8]Zwirner J,Burg M,Schulze M.Activated complement C3:a potentially novel predictor of progressive Ig A nephropathy[J].Kidney Int,1997,51(4):1257.
[9]Wyatt R J,Julian B A.Ig A nephropathy[J].N Engl J Med,2013,368(25):2402.
[10]De Vries B,Walter SJ,Peutz-Kootstra C J,et al.The mannose-binding lectin-pathway is involved in complement activation in the course of renal ischemia-reperfusion injury[J].Am J Pathol,2004,165(5):1677.
[11]Hovind P,Hansen T K,Tarnow L,et al.Mannose-binding lectin as a predictor of microalbuminuria in type 1 diabetes:an inception cohort study[J].Diabetes,2005,54(5):1523.
[12]Sato N,Ohsawa I,Nagamachi S,et al.Significance of glomerular activation of the alternative pathway and lectin pathway in lupus nephritis[J].Lupus,2011,20(13):1378.
[13]Goeldner I,Skare T L,Utiyama S R,et al.Mannose binding lectin and susceptibility to rheumatoid arthritis in brazilian patients and their relatives[J].PLo S One,2014,9(4):e95519.
[14]Koch A,Melbye M,Sorensen P,et al.Acute respiratory tract infections and mannose-binding lectin insufficiency during early childhood[J].JAMA,2001,285(10):1316.
[15]Hegele R A,Ban M R,Anderson C M,et al.Infectionsusceptibility alleles of mannose-binding lectin are associated with increased carotid plaque area[J].J Investig Med,2000,48(3):198.
[16]Endo M,Ohi H,Ohsawa I,et al.Glomerular deposition of mannose-binding lectin(MBL)indicates a novel mechanism of complement activation in Ig A nephropathy[J].Nephrol Dial Transplant,1998,13(8):1984.
[17]Matsuda M,Shikata K,Wada J,et al.Deposition of mannan binding protein and mannan binding protein-mediated complement activation in the glomeruli of patients with Ig A nephropathy[J].Nephron,1998,80(4):408.
[18]KDIGO clinical practice guidelines for glomerulonephritis(2012)Chapter 10:Immunoglobulin A nephropathy[J].Kidney Int Suppl(2011),2012,2(2):209.
[19]Yuzawa Y,Yamamoto R,Takahashi K,et al.Evidence-based clinical practice guidelines for Ig A nephropathy 2014[J].Clin Exp Nephrol,2016,20(4):511.
[20]Coppo R,Peruzzi L,Amore A,et al.Ig ACE:a placebocontrolled,randomized trial of angiotensin-converting enzyme inhibitors in children and young people with Ig A nephropathy and moderate proteinuria[J].J Am Soc Nephrol,2007,18(6):1880.
[21]Li P K,Leung C B,Chow K M,et al.Hong Kong Study Using Valsartan in Ig A Nephropathy(HKVIN):a double-blind,randomized,placebo-controlled study[J].Am J Kidney Dis,2006,47:751.
[2]Li L S,Liu Z H.Epidemiologic data of renal diseases from a single unit in China:analysis based on 13519 renal biopsies[J].Kidney Int,2004,66(3):920.
[3]王海燕.肾脏病学[M].3版.北京:人民卫生出版社,2008:993.
[4]Pozzi C,Andrulli S,Pani A,et al.Addition of azathioprine to corticosteroids does not benefit patients with Ig A nephropathy[J].J Am Soc Nephrol,2010,21(10):1783.
[5]汤颖,娄探奇,成彩联,等.实验性Ig A肾病模型的改进[J].中山大学学报,2006,27(2):184.
[6]Barratt J,Eitner F,Feehally J,et al.Immune complex formation in Ig A nephropathy:a case of the‘right’antibodies in the‘wrong’place at the‘wrong’time[J].Nephrol Dial Transplant,2009,24(12):3620.
[7]Onda K,Ohi H M,Ohsawal I.Hypercomplementemia in adult patients with Ig A nephropathy[J].J Clin Lab Anal,2007,21(2):77.
[8]Zwirner J,Burg M,Schulze M.Activated complement C3:a potentially novel predictor of progressive Ig A nephropathy[J].Kidney Int,1997,51(4):1257.
[9]Wyatt R J,Julian B A.Ig A nephropathy[J].N Engl J Med,2013,368(25):2402.
[10]De Vries B,Walter SJ,Peutz-Kootstra C J,et al.The mannose-binding lectin-pathway is involved in complement activation in the course of renal ischemia-reperfusion injury[J].Am J Pathol,2004,165(5):1677.
[11]Hovind P,Hansen T K,Tarnow L,et al.Mannose-binding lectin as a predictor of microalbuminuria in type 1 diabetes:an inception cohort study[J].Diabetes,2005,54(5):1523.
[12]Sato N,Ohsawa I,Nagamachi S,et al.Significance of glomerular activation of the alternative pathway and lectin pathway in lupus nephritis[J].Lupus,2011,20(13):1378.
[13]Goeldner I,Skare T L,Utiyama S R,et al.Mannose binding lectin and susceptibility to rheumatoid arthritis in brazilian patients and their relatives[J].PLo S One,2014,9(4):e95519.
[14]Koch A,Melbye M,Sorensen P,et al.Acute respiratory tract infections and mannose-binding lectin insufficiency during early childhood[J].JAMA,2001,285(10):1316.
[15]Hegele R A,Ban M R,Anderson C M,et al.Infectionsusceptibility alleles of mannose-binding lectin are associated with increased carotid plaque area[J].J Investig Med,2000,48(3):198.
[16]Endo M,Ohi H,Ohsawa I,et al.Glomerular deposition of mannose-binding lectin(MBL)indicates a novel mechanism of complement activation in Ig A nephropathy[J].Nephrol Dial Transplant,1998,13(8):1984.
[17]Matsuda M,Shikata K,Wada J,et al.Deposition of mannan binding protein and mannan binding protein-mediated complement activation in the glomeruli of patients with Ig A nephropathy[J].Nephron,1998,80(4):408.
[18]KDIGO clinical practice guidelines for glomerulonephritis(2012)Chapter 10:Immunoglobulin A nephropathy[J].Kidney Int Suppl(2011),2012,2(2):209.
[19]Yuzawa Y,Yamamoto R,Takahashi K,et al.Evidence-based clinical practice guidelines for Ig A nephropathy 2014[J].Clin Exp Nephrol,2016,20(4):511.
[20]Coppo R,Peruzzi L,Amore A,et al.Ig ACE:a placebocontrolled,randomized trial of angiotensin-converting enzyme inhibitors in children and young people with Ig A nephropathy and moderate proteinuria[J].J Am Soc Nephrol,2007,18(6):1880.
[21]Li P K,Leung C B,Chow K M,et al.Hong Kong Study Using Valsartan in Ig A Nephropathy(HKVIN):a double-blind,randomized,placebo-controlled study[J].Am J Kidney Dis,2006,47:751.