川芎嗪对膜性肾病模型大鼠肾脏的保护作用
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摘要
背景:膜性肾病是成年人肾病综合征的常见病理类型,足细胞数目减少是膜性肾病发病的原因之一,而凋亡又是足细胞减少的原因,足细胞凋亡受到Bcl-2和Bax两种凋亡调控因子的调控,川芎嗪对于肾脏疾病有较好的治疗作用。目的:观察川芎嗪对阳离子化牛血清白蛋白诱导的膜性肾病大鼠的肾脏保护作用。方法:将西南医科大学实验动物中心提供的72只大鼠随机分为正常组、模型组和川芎嗪组,各24只,后2组采用改良Border法复制膜性肾病模型。川芎嗪组造模开始后每天腹腔注射川芎嗪注射液100mg/kg,连续4周。结果与结论:(1)与正常组相比,模型组可见肾小球系膜区及毛细血管壁Ig G强阳性颗粒样沉积;光镜下可见肾小球体积增大,马松染色见嗜复红蛋白沉积,六胺银染色见基底膜弥漫性增厚、"钉突"形成;电镜可见肾小球基底膜增厚,足细胞的足突融合,肾小球上皮电子致密物沉积。与模型组相比,川芎嗪组Ig G荧光强度及分布无明显差异,但肾小球基底膜增厚的程度及足突融合程度都有轻微改善;(2)与正常组相比,其他2组的血肌酐、24h尿蛋白升高,肌酐清除率降低,Bcl-2表达水平、WT-1阳性细胞比例减少,Bax表达水平、细胞凋亡数量增加;与模型组相比,川芎嗪组的血清白蛋白、肌酐清除率较高、血肌酐较低,Bcl-2表达水平和WT-1阳性细胞比例增加,Bax表达水平、细胞凋亡数量较少;(3)提示川芎嗪能改善阳离子牛血清白蛋白诱导膜性肾病大鼠的肾功能,升高血清白蛋白,具有一定疗效;川芎嗪可能通过调控Bcl-2和Bax途径减少膜性肾病大鼠肾固有细胞凋亡,抑制足细胞数量的减少,起到肾脏保护作用。
BACKGROUND: Membranous nephropathy, a common type of nephrotic syndrome in adults, is characterized by podocyte decrease caused by apoptosis. Podocyte apoptosis is regulated by Bcl-2 and Bax. Tetramethylpyrazine exerts good effectiveness in the treatment of renal diseases. OBJECTIVE: To observe the protective effect of tetramethylpyrazine on cationized bovine serum albumin-induced membranous nephropathy in rats. METHODS: Seventy-two rats provided by Laboratory Animal Center of Southwest University were randomly divided into normal, model and tetramethylpyrazine groups(n=24 per group). Rabbits in the latter two groups underwent membranous nephropathy modeling by modified Border method. The tetramethylpyrazine group received intraperitoneal injection of 100 mg/kg tetramethylpyrazine for four consecutive weeks. RESULTS AND CONCLUSION: Compared with the normal group, the model group showed strong IgG-like deposition in the glomerular mesangial area and capillary wall; glomerular volume increased, as shown by light microscopy. Masson staining showed chlorocruorin deposition. Gomori methenamine silver staining showed diffuse thickening of basement membrane and formation of "nail spikes". Electron microscope showed thickening of the glomerular basement membrane, fusion of foot process, and deposition of electron dense deposits on the glomerular epithelium. Compared with the model group, there was no significant difference in the fluorescence intensity and distribution of IgG in the tetramethylpyrazine group, but the degree of glomerular basement membrane thickening and the degree of foot process fusion were slightly improved. Compared with the normal group, the serum creatinine and 24-hour urinary protein levels were increased, creatinine clearance was decreased, expression level of Bcl-2 and the number of WT-1 positive cells were increased, and Bax expression level and the number of apoptotic cells were decreased in the other two groups. To conclude, tetramethylpyrazine can improve the renal function of rats with membranous nephropathy induced by cationized bovine serum albumin and increase serum albumin. It protects the kidney by reducing the cell apoptosis through regulating Bcl-2 and Bax pathways and inhibiting the reduction of podocytes.
BACKGROUND: Membranous nephropathy, a common type of nephrotic syndrome in adults, is characterized by podocyte decrease caused by apoptosis. Podocyte apoptosis is regulated by Bcl-2 and Bax. Tetramethylpyrazine exerts good effectiveness in the treatment of renal diseases. OBJECTIVE: To observe the protective effect of tetramethylpyrazine on cationized bovine serum albumin-induced membranous nephropathy in rats. METHODS: Seventy-two rats provided by Laboratory Animal Center of Southwest University were randomly divided into normal, model and tetramethylpyrazine groups(n=24 per group). Rabbits in the latter two groups underwent membranous nephropathy modeling by modified Border method. The tetramethylpyrazine group received intraperitoneal injection of 100 mg/kg tetramethylpyrazine for four consecutive weeks. RESULTS AND CONCLUSION: Compared with the normal group, the model group showed strong IgG-like deposition in the glomerular mesangial area and capillary wall; glomerular volume increased, as shown by light microscopy. Masson staining showed chlorocruorin deposition. Gomori methenamine silver staining showed diffuse thickening of basement membrane and formation of "nail spikes". Electron microscope showed thickening of the glomerular basement membrane, fusion of foot process, and deposition of electron dense deposits on the glomerular epithelium. Compared with the model group, there was no significant difference in the fluorescence intensity and distribution of IgG in the tetramethylpyrazine group, but the degree of glomerular basement membrane thickening and the degree of foot process fusion were slightly improved. Compared with the normal group, the serum creatinine and 24-hour urinary protein levels were increased, creatinine clearance was decreased, expression level of Bcl-2 and the number of WT-1 positive cells were increased, and Bax expression level and the number of apoptotic cells were decreased in the other two groups. To conclude, tetramethylpyrazine can improve the renal function of rats with membranous nephropathy induced by cationized bovine serum albumin and increase serum albumin. It protects the kidney by reducing the cell apoptosis through regulating Bcl-2 and Bax pathways and inhibiting the reduction of podocytes.
引文
[1]Rozenberg I,Kotliroff A,Zahavi T,et al.Outcome of Idiopathic Membranous Nephropathy:A Retrospective Study.Isr Med Assoc J.2018;20(3):186-189.
[2]Ronco P,Debiec H.Advances in membranous nephropathy:success stories of a long journey.Clin Exp Pharmacol Physiol.2011;38(7):460-466.
[3]Akiyama S,Akiyama M,Imai E,et al.Prevalence of anti-phospholipase A2 receptor antibodies in Japanese patients with membranous nephropathy.Clin Exp Nephrol.2015;19(4):653-660.
[4]洪虹,李秀珍,徐文莲.膜性肾病临床流行病学分析[J].中国全科医学,2017,20(S2):16-18.
[5]Improving Global Outcomes(KDIGO)Glomerulonephritis Work Group.KDIGO Clinical Practice Guidelinne for Glomerulonephritis.Kidney Inter.2012;Suppl 2:139-274.
[6]Xu J,Zhang W,Xu Y,et al.Tacrolimus combined with corticosteroids in idiopathic membranous nephropathy:a randomized,prospective,controlled trial.Contrib Nephrol.2013;181:152-162.
[7]Choi JY,Kim DK,Kim YW,et al.The Effect of Mycophenolate Mofetil versus Cyclosporine as Combination Therapy with Low Dose Corticosteroids in High-risk Patients with Idiopathic Membranous Nephropathy:a Multicenter Randomized Trial.JKorean Med Sci.2018;33(9):e74.
[8]黄智敏,陈科杰.环孢素A和环磷酰胺治疗特发性膜性肾病的疗效、复发率及副作用研究[J].北方药学,2016,13(9):66.
[9]金佳丽,苏行,姚馨怡,等.炙黄芪对阿霉素所致小鼠蛋白尿的预防作用[J].中华中医药学刊,2013,31(3):504-506,708.
[10]Wang Z,Liu J,Sun W.Effects of asiaticoside on levels of podocyte cytoskeletal proteins and renal slit diaphragm proteins in adriamycin-induced rat nephropathy.Life Sci.2013;93(8):352-358.
[11]赵建荣,屈磊,李晓玫.黄芪当归合剂对梗阻性肾病大鼠肾间质纤维化的防治作用[J].北京大学学报(医学版),2004,36(2):119-123.
[12]李晓玫,王海燕,蔡琪.黄芪当归合剂对大鼠急性缺血/再灌注肾损伤的保护及加速修复作用[J].中华肾脏病杂志,2000,16(6):387-391.
[13]李彪,唐嘉薇,蔡少青,等.黄芪当归合剂抑制马兜铃酸Ⅰ导致的肾小管上皮细胞损伤[J].北京大学学报(医学版),2006,38(4):381-384.
[14]郭晓媛,蔡月茹,孙广宇,等.扶正祛风方对膜性肾病大鼠蛋白尿及肾脏病理改变的影响[J].中国实验方剂学杂志,2018,24(1):136-141.
[15]张春兵,丁兴,詹臻.川芎嗪通过干预Bax/Bcl-2平衡调控Caspase9途径抑制谷氨酸诱导的PC-12细胞凋亡[J].河南中医学院学报,2008,23(2):24-27.
[16]王汉民,谭华,赵洪雯,等.川芎嗪对大鼠肾脏缺血/再灌注损伤保护作用及抗细胞凋亡作用[J].第四军医大学学报,2008,29(9):833-836.
[17]刘晓华,黎七雄.川芎嗪对顺铂肾损伤大鼠肾细胞凋亡及凋亡相关蛋白表达的影响[J].中国药理学与毒理学杂志,2005,19(5):352-356.
[18]Langer S,Kreutz R,Eisenreich A.Metformin modulates apoptosis and cell signaling of human podocytes under high glucose conditions.J Nephrol.2016;29(6):765-773.
[19]宣柳,顾娟,王学东.川芎嗪对佐剂性关节炎大鼠滑膜细胞凋亡的诱导作用[J].中国临床药理学杂志,2015,17(6):1766-1768.
[20]冷辉,孙海波,马贤德,等.川芎嗪对顺铂诱导的豚鼠耳聋模型耳蜗组织Bax及Bcl-2表达的影响[J].中华中医药学刊,2017,35(7):1827-1830,1936.
[21]Debiec H,Lefeu F,Kemper MJ,et al.Early-childhood membranous nephropathy due to cationic bovine serum albumin.N Engl J Med.2011;364(22):2101-2110.
[22]Beck LH Jr,Salant DJ.Membranous nephropathy:from models to man.J Clin Invest.2014;124(6):2307-2314.
[23]王慧,王净,李静,等.膜性肾病小鼠模型的建立与鉴定[J].细胞与分子免疫学杂志,2013,29(2):89-91.
[24]母传贤,刘国玲,田华,等.川芎嗪对胶原性关节炎大鼠血清IL-1、IL-6、IL-2水平及关节浸液NO、PGE2的影响[J].中国中西医结合杂志,2014,34(2):214-217.
[25]黄海泉,刘必成,罗东东,等.STZ诱导糖尿病大鼠肾脏CTGF表达改变及意义探讨[J].中国病理生理杂志,2007,23(3):553-558.
[26]金周慧,陈以平,邓跃毅,等.蝉花菌丝延缓肾小球硬化的作用机制研究[J].中国中西医结合肾病杂志,2005,6(3):132-136.
[27]刘晶晶,赵砚丽,程会平,等.缺血后处理对肾缺血再灌注损伤大鼠肾组织细胞凋亡及bcl-2和bax基因表达的影响[J].第四军医大学学报,2007,28(21):1960-1963.
[28]保莉,毕逢辰,郑亚莉.nephrin及WT-1在嘌呤霉素大鼠肾病模型足细胞中的表达及意义[J].宁夏医学杂志,2014,36(3):201-203.
[29]殷玉红,李英.α-actinin-4在糖尿病肾病大鼠的表达及其与足细胞数目的相关性[J].中国中西医结合肾病杂志,2009,10(4):299-302,378.
[30]Lai WL,Yeh TH,Chen PM,et al.Membranous nephropathy:a review on the pathogenesis,diagnosis,and treatment.JFormos Med Assoc.2015;114(2):102-111.
[31]Levandoski KA,Girardi NA,Loss MJ.Eruptive sebaceous hyperplasia as a side effect of oral tacrolimus in a renal transplant recipient.Dermatol Online J.2017;23(5):13030/qt7x0125gz.
[32]Sendeyo K,Audard V,Zhang SY,et al.Upregulation of c-mip is closely related to podocyte dysfunction in membranous nephropathy.Kidney Int.2013;83(3):414-425.
[33]Asanuma K.The role of podocyte injury in chronic kidney disease.Nihon Rinsho Meneki Gakkai Kaishi.2015;38(1):26-36.
[34]Burlaka I,Nilsson LM,Scott L,et al.Prevention of apoptosis averts glomerular tubular disconnection and podocyte loss in proteinuric kidney disease.Kidney Int.2016;90(1):135-148.
[35]Macconi D,Bonomelli M,Benigni A,et al.Pathophysiologic implications of reduced podocyte number in a rat model of progressive glomerular injury.Am J Pathol.2006;168(1):42-54.
[36]姜宇懋,王丹巧.川芎嗪药理作用研究进展[J].中国现代中药,2016,18(10):1364-1370.
[37]任玉伟,宿华威.Bcl-2基因家族研究进展[J].大连医科大学学报,2015,37(2):202-205.
[38]Hassan M,Watari H,AbuAlmaaty A,et al.Apoptosis and molecular targeting therapy in cancer.Biomed Res Int.2014;2014:150845.
[39]Schenk RL,Strasser A,Dewson G.BCL-2:Long and winding path from discovery to therapeutic target.Biochem Biophys Res Commun.2017;482(3):459-469.
[40]Brown LM,Hanna DT,Khaw SL,et al.Dysregulation of BCL-2family proteins by leukemia fusion genes.J Biol Chem.2017;292(35):14325-14333.
[2]Ronco P,Debiec H.Advances in membranous nephropathy:success stories of a long journey.Clin Exp Pharmacol Physiol.2011;38(7):460-466.
[3]Akiyama S,Akiyama M,Imai E,et al.Prevalence of anti-phospholipase A2 receptor antibodies in Japanese patients with membranous nephropathy.Clin Exp Nephrol.2015;19(4):653-660.
[4]洪虹,李秀珍,徐文莲.膜性肾病临床流行病学分析[J].中国全科医学,2017,20(S2):16-18.
[5]Improving Global Outcomes(KDIGO)Glomerulonephritis Work Group.KDIGO Clinical Practice Guidelinne for Glomerulonephritis.Kidney Inter.2012;Suppl 2:139-274.
[6]Xu J,Zhang W,Xu Y,et al.Tacrolimus combined with corticosteroids in idiopathic membranous nephropathy:a randomized,prospective,controlled trial.Contrib Nephrol.2013;181:152-162.
[7]Choi JY,Kim DK,Kim YW,et al.The Effect of Mycophenolate Mofetil versus Cyclosporine as Combination Therapy with Low Dose Corticosteroids in High-risk Patients with Idiopathic Membranous Nephropathy:a Multicenter Randomized Trial.JKorean Med Sci.2018;33(9):e74.
[8]黄智敏,陈科杰.环孢素A和环磷酰胺治疗特发性膜性肾病的疗效、复发率及副作用研究[J].北方药学,2016,13(9):66.
[9]金佳丽,苏行,姚馨怡,等.炙黄芪对阿霉素所致小鼠蛋白尿的预防作用[J].中华中医药学刊,2013,31(3):504-506,708.
[10]Wang Z,Liu J,Sun W.Effects of asiaticoside on levels of podocyte cytoskeletal proteins and renal slit diaphragm proteins in adriamycin-induced rat nephropathy.Life Sci.2013;93(8):352-358.
[11]赵建荣,屈磊,李晓玫.黄芪当归合剂对梗阻性肾病大鼠肾间质纤维化的防治作用[J].北京大学学报(医学版),2004,36(2):119-123.
[12]李晓玫,王海燕,蔡琪.黄芪当归合剂对大鼠急性缺血/再灌注肾损伤的保护及加速修复作用[J].中华肾脏病杂志,2000,16(6):387-391.
[13]李彪,唐嘉薇,蔡少青,等.黄芪当归合剂抑制马兜铃酸Ⅰ导致的肾小管上皮细胞损伤[J].北京大学学报(医学版),2006,38(4):381-384.
[14]郭晓媛,蔡月茹,孙广宇,等.扶正祛风方对膜性肾病大鼠蛋白尿及肾脏病理改变的影响[J].中国实验方剂学杂志,2018,24(1):136-141.
[15]张春兵,丁兴,詹臻.川芎嗪通过干预Bax/Bcl-2平衡调控Caspase9途径抑制谷氨酸诱导的PC-12细胞凋亡[J].河南中医学院学报,2008,23(2):24-27.
[16]王汉民,谭华,赵洪雯,等.川芎嗪对大鼠肾脏缺血/再灌注损伤保护作用及抗细胞凋亡作用[J].第四军医大学学报,2008,29(9):833-836.
[17]刘晓华,黎七雄.川芎嗪对顺铂肾损伤大鼠肾细胞凋亡及凋亡相关蛋白表达的影响[J].中国药理学与毒理学杂志,2005,19(5):352-356.
[18]Langer S,Kreutz R,Eisenreich A.Metformin modulates apoptosis and cell signaling of human podocytes under high glucose conditions.J Nephrol.2016;29(6):765-773.
[19]宣柳,顾娟,王学东.川芎嗪对佐剂性关节炎大鼠滑膜细胞凋亡的诱导作用[J].中国临床药理学杂志,2015,17(6):1766-1768.
[20]冷辉,孙海波,马贤德,等.川芎嗪对顺铂诱导的豚鼠耳聋模型耳蜗组织Bax及Bcl-2表达的影响[J].中华中医药学刊,2017,35(7):1827-1830,1936.
[21]Debiec H,Lefeu F,Kemper MJ,et al.Early-childhood membranous nephropathy due to cationic bovine serum albumin.N Engl J Med.2011;364(22):2101-2110.
[22]Beck LH Jr,Salant DJ.Membranous nephropathy:from models to man.J Clin Invest.2014;124(6):2307-2314.
[23]王慧,王净,李静,等.膜性肾病小鼠模型的建立与鉴定[J].细胞与分子免疫学杂志,2013,29(2):89-91.
[24]母传贤,刘国玲,田华,等.川芎嗪对胶原性关节炎大鼠血清IL-1、IL-6、IL-2水平及关节浸液NO、PGE2的影响[J].中国中西医结合杂志,2014,34(2):214-217.
[25]黄海泉,刘必成,罗东东,等.STZ诱导糖尿病大鼠肾脏CTGF表达改变及意义探讨[J].中国病理生理杂志,2007,23(3):553-558.
[26]金周慧,陈以平,邓跃毅,等.蝉花菌丝延缓肾小球硬化的作用机制研究[J].中国中西医结合肾病杂志,2005,6(3):132-136.
[27]刘晶晶,赵砚丽,程会平,等.缺血后处理对肾缺血再灌注损伤大鼠肾组织细胞凋亡及bcl-2和bax基因表达的影响[J].第四军医大学学报,2007,28(21):1960-1963.
[28]保莉,毕逢辰,郑亚莉.nephrin及WT-1在嘌呤霉素大鼠肾病模型足细胞中的表达及意义[J].宁夏医学杂志,2014,36(3):201-203.
[29]殷玉红,李英.α-actinin-4在糖尿病肾病大鼠的表达及其与足细胞数目的相关性[J].中国中西医结合肾病杂志,2009,10(4):299-302,378.
[30]Lai WL,Yeh TH,Chen PM,et al.Membranous nephropathy:a review on the pathogenesis,diagnosis,and treatment.JFormos Med Assoc.2015;114(2):102-111.
[31]Levandoski KA,Girardi NA,Loss MJ.Eruptive sebaceous hyperplasia as a side effect of oral tacrolimus in a renal transplant recipient.Dermatol Online J.2017;23(5):13030/qt7x0125gz.
[32]Sendeyo K,Audard V,Zhang SY,et al.Upregulation of c-mip is closely related to podocyte dysfunction in membranous nephropathy.Kidney Int.2013;83(3):414-425.
[33]Asanuma K.The role of podocyte injury in chronic kidney disease.Nihon Rinsho Meneki Gakkai Kaishi.2015;38(1):26-36.
[34]Burlaka I,Nilsson LM,Scott L,et al.Prevention of apoptosis averts glomerular tubular disconnection and podocyte loss in proteinuric kidney disease.Kidney Int.2016;90(1):135-148.
[35]Macconi D,Bonomelli M,Benigni A,et al.Pathophysiologic implications of reduced podocyte number in a rat model of progressive glomerular injury.Am J Pathol.2006;168(1):42-54.
[36]姜宇懋,王丹巧.川芎嗪药理作用研究进展[J].中国现代中药,2016,18(10):1364-1370.
[37]任玉伟,宿华威.Bcl-2基因家族研究进展[J].大连医科大学学报,2015,37(2):202-205.
[38]Hassan M,Watari H,AbuAlmaaty A,et al.Apoptosis and molecular targeting therapy in cancer.Biomed Res Int.2014;2014:150845.
[39]Schenk RL,Strasser A,Dewson G.BCL-2:Long and winding path from discovery to therapeutic target.Biochem Biophys Res Commun.2017;482(3):459-469.
[40]Brown LM,Hanna DT,Khaw SL,et al.Dysregulation of BCL-2family proteins by leukemia fusion genes.J Biol Chem.2017;292(35):14325-14333.