基于血管内皮生长因子及其受体途径的局灶节段性肾小球硬化大鼠的艾灸干预随机对照实验
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摘要
目的:观察在艾灸干预下局灶节段性肾小球硬化(FSGS)大鼠肾组织中血管内皮生长因子(VEGF)及其受体(VEGFR-2)的表达。方法:102只健康雄性SD大鼠,按体质量随机分为正常组7只、假手术组7只、造模组88只。采取间隔2周2次尾静脉注射阿霉素并单侧肾切除的方法建立FSGS大鼠模型。造模成功后大鼠根据尿蛋白定量随机分为模型组、氯沙坦组、肾俞灸法(短时、中时、长时)组和膈俞灸法(短时、中时、长时)组,每组11只,各组采用相应干预。12周后运用免疫组化法检测VEGF、VEGFR-2、ET-1、AngⅡ、AT1R在肾组织的蛋白表达;硝酸还原酶法检测NO在肾组织的含量。结果:与假手术组比较,模型组VEGF、VEGFR-2、ET-1、AngⅡ、AT1R蛋白表达显著升高,NO含量显著降低(P<0.01);与模型组比较,氯沙坦组、肾俞长时组和膈俞长时组VEGF、VEGFR-2、ET-1、AngⅡ、AT1R表达降低,NO含量升高(P<0.05,P<0.01)。结论:艾灸可能通过下调VEGF及其受体VEGFR-2的蛋白表达起到保护肾组织的作用。
Objective:To observe the effects of moxibustion intervention on treating focal segmental glomerulosclerosis(FSGS) rat model by observing the expression of vascular endothelial growth factor(VEGF) and its receptor(VEGFR-2) in renal tissue.Methods:A total of 102 SD rats were randomly divided into the normal group(n=7), sham operation group(n=7) and model group(n=88).The FSGS rat model was established by tail intravenous injection of adriamycin twice a week for two times and unilateral renal resection.After making a successful model, the rats were randomly divided into model group, losartan group, Shenshu(BL23) moxibustion group(short-term, medium-term and long-term) and Geshu(BL17) moxibustion group(shortterm, medium-term and long-term), with 11 rats in each group.Rats in each group were given corresponding intervention.After 12 weeks, immunohistochemical method was used to detect the protein expression of VEGF, VEGFR-2, ET-1, Ang II and AT1 R in renal tissue; nitrate reductase method was used to measure the content of NO in renal tissue.Results:Compared with the sham operation group, the protein expressions of VEGF, VEGFR-2, ET-1, Ang II and AT1 R in the model group were significantly increased and the contents of NO were significantly decreased(P<0.01); compared with the model group, the expressions of VEGF, VEGFR-2, ET-1, Ang II and AT1 R in losartan group, long-term Shenshu(BL23) moxibustion group and long-term Geshu(BL17) moxibustion group were decreased and the contents of NO were increased(P<0.05, P<0.01).Conclusion:Moxibustion may protect the renal tissue by down-regulating the protein expression of VEGF and its receptor VEGFR-2.
Objective:To observe the effects of moxibustion intervention on treating focal segmental glomerulosclerosis(FSGS) rat model by observing the expression of vascular endothelial growth factor(VEGF) and its receptor(VEGFR-2) in renal tissue.Methods:A total of 102 SD rats were randomly divided into the normal group(n=7), sham operation group(n=7) and model group(n=88).The FSGS rat model was established by tail intravenous injection of adriamycin twice a week for two times and unilateral renal resection.After making a successful model, the rats were randomly divided into model group, losartan group, Shenshu(BL23) moxibustion group(short-term, medium-term and long-term) and Geshu(BL17) moxibustion group(shortterm, medium-term and long-term), with 11 rats in each group.Rats in each group were given corresponding intervention.After 12 weeks, immunohistochemical method was used to detect the protein expression of VEGF, VEGFR-2, ET-1, Ang II and AT1 R in renal tissue; nitrate reductase method was used to measure the content of NO in renal tissue.Results:Compared with the sham operation group, the protein expressions of VEGF, VEGFR-2, ET-1, Ang II and AT1 R in the model group were significantly increased and the contents of NO were significantly decreased(P<0.01); compared with the model group, the expressions of VEGF, VEGFR-2, ET-1, Ang II and AT1 R in losartan group, long-term Shenshu(BL23) moxibustion group and long-term Geshu(BL17) moxibustion group were decreased and the contents of NO were increased(P<0.05, P<0.01).Conclusion:Moxibustion may protect the renal tissue by down-regulating the protein expression of VEGF and its receptor VEGFR-2.
引文
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[3]李屹,黄迪,何立群.灸药结合对脾肾气虚型慢性肾脏病患者肾功能及尿β-痕迹蛋白的影响.上海中医药杂志,2012,46(9):41-43
[4]徐海山,庄永泽,曾俊婷.单侧肾切除+股静脉注射阿霉素制作进展性肾病模型.当代医学,2009,15(34):41-42
[5]孟凡谋,肖晓辉,王月飞,等.VEGF受体Fn-1,FLK-1在大鼠过度刺激卵巢的表达研究.齐齐哈尔医学院学报,2007,28(9):1045-1046
[6]Ljungberg B J,Jacobsen J,Rudolfsson S H,et al.Different vascular endothelial growth factor(VEGF),VEGF-receptor l and 2 m RNA expression profiles between clear cell and papillary renal cell carcinoma.BJU Int,2006,98(3):661-667
[7]闫滨,贾成友,王锋,等.康脑灵冲剂对高脂合并脑缺血大鼠自由基代谢的影响.山东中医药大学学报,2003,27(4):304-306
[8]Neuhofer W,Pittrow D.Role of endothelin and endothelin receptor antagonists in renal disease.Eur J Clin Invest,2006,36(3):78-88
[9]程虹.内皮素-1与肾素-血管紧张素-醛固酮系统的相互作用.肾脏病与透析肾移植杂志,2003,12(4):371-374
[10]K Nitta,K Uchida,N kimata,et al.Endothelin-1 mediates erythropoietin-stimulated glomerular endothelial celldependent proliferation of mesangial cells.Eur J Pharmacol,1995,293(4):491-494
[11]Korbet S M.Treatment of primary FSGS in adults.J Am Soc Nephrol,2012,23(11):1769-1776
[12]涂远茂,李世军.局灶节段性肾小球硬化的治疗现状.肾脏病与透析肾移植杂志,2014,23(4):371-376
[2]王玉梅,邓安国.血管内皮生长因子在阿霉素肾病大鼠中的表达及其意义.中华肾病杂志,2002,18(5):374-375
[3]李屹,黄迪,何立群.灸药结合对脾肾气虚型慢性肾脏病患者肾功能及尿β-痕迹蛋白的影响.上海中医药杂志,2012,46(9):41-43
[4]徐海山,庄永泽,曾俊婷.单侧肾切除+股静脉注射阿霉素制作进展性肾病模型.当代医学,2009,15(34):41-42
[5]孟凡谋,肖晓辉,王月飞,等.VEGF受体Fn-1,FLK-1在大鼠过度刺激卵巢的表达研究.齐齐哈尔医学院学报,2007,28(9):1045-1046
[6]Ljungberg B J,Jacobsen J,Rudolfsson S H,et al.Different vascular endothelial growth factor(VEGF),VEGF-receptor l and 2 m RNA expression profiles between clear cell and papillary renal cell carcinoma.BJU Int,2006,98(3):661-667
[7]闫滨,贾成友,王锋,等.康脑灵冲剂对高脂合并脑缺血大鼠自由基代谢的影响.山东中医药大学学报,2003,27(4):304-306
[8]Neuhofer W,Pittrow D.Role of endothelin and endothelin receptor antagonists in renal disease.Eur J Clin Invest,2006,36(3):78-88
[9]程虹.内皮素-1与肾素-血管紧张素-醛固酮系统的相互作用.肾脏病与透析肾移植杂志,2003,12(4):371-374
[10]K Nitta,K Uchida,N kimata,et al.Endothelin-1 mediates erythropoietin-stimulated glomerular endothelial celldependent proliferation of mesangial cells.Eur J Pharmacol,1995,293(4):491-494
[11]Korbet S M.Treatment of primary FSGS in adults.J Am Soc Nephrol,2012,23(11):1769-1776
[12]涂远茂,李世军.局灶节段性肾小球硬化的治疗现状.肾脏病与透析肾移植杂志,2014,23(4):371-376