输尿管不全梗阻大鼠输尿管平滑肌损伤及肌球蛋白表达变化的临床意义
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摘要
目的观察大鼠输尿管不全梗阻3周内输尿管形态、平滑肌细胞结构及肌球蛋白表达变化,为临床判断输尿管不全梗阻患者的输尿管功能提供依据。方法选择SD大鼠120只,随机分为对照组30只、假手术组45只、模型组45只。模型组参照腰大肌隧道包埋法制备右侧输尿管不全梗阻模型,假手术组假手术处理,对照组不予处理。模型组造模1、2、3周,观察各组输尿管及肾脏形态,测量右侧输尿管直径,采用HE染色观察输尿管平滑肌组织细胞结构变化,采用免疫组化SP法检测输尿管平滑肌肌球蛋白表达,采用qRT-PCR法检测输尿管平滑肌肌球蛋白重链mRNA相对表达量。结果模型组造模1、2、3周近端输尿管逐渐增粗,肾脏体积逐渐增大、张力逐渐升高;右侧输尿管直径逐渐增加,且均大于对照组和假手术组(P均<0.01)。HE染色结果显示,模型组近端输尿管梗阻时间越长,平滑肌细胞肌层越薄、细胞越少、纤维增生越明显。模型组造模1、2周输尿管平滑肌肌球蛋白表达量及肌球蛋白重链mRNA相对表达量均高于对照组和假手术组,造模3周均低于对照组和假手术组(P均<0.01);造模1、2、3周,模型组平滑肌肌球蛋白表达量及肌球蛋白重链mRNA相对表达量各时间点两两比较P均<0.01。结论输尿管不全梗阻时间越长平滑肌损伤越重;梗阻超过2周将出现输尿管功能不可逆性损伤。
Objective To observe the changes of ureteral morphology,smooth muscle cell structure and myosin expression in rats with ureteral obstruction within 3 weeks,which provides a basis for clinical diagnosis of ureteral function in patients with ureteral obstruction.Methods Totally 120 SD rats were randomly divided into the control group(30 rats),sham operation group(45 rats),and model group(45 rats).The rats in the control group were fed normally,the rats in the sham operation group were sham operated,and the rats in the model group were prepared for right ureteral insufficiency obstruction models.The ureter and kidney morphology of the three groups were observed at 1,2,and 3 weeks after operation.The diameter of the right ureter was measured.The cell structure of ureter smooth muscle was observed by HE staining.The expression of myosin in the ureteral smooth muscle was detected by immunohistochemical SP method.The relative expression of ureter smooth muscle myosin heavy chain mRNA was detected by qRT-PCR.Results In the model group,the proximal ureter gradually thickened at 1,2,and 3 weeks after operation.The kidney volume gradually increased and the tension gradually increased.The diameter of the right ureter gradually increased,which was greater than that of the control group and the sham operation group(P<0.01).HE staining results showed that the longer the proximal ureteral obstruction in the model group,the thinner the muscle layer of smooth muscle cells,the fewer the cells,and the more obvious the fiber proliferation.The expression of myosin and the expression of myosin heavy chain mRNA in the model group was higher than that in the control group and the sham operation group at 1 and 2 weeks after operation,but at 3 weeks after operation,it was lower than that of the the control group and sham operation group(all P<0.01).The expression of myosin and the relative expression of myosin heavy chain mRNA in the model group decreased gradually at 2 weeks,1 week and 3 weeks after operation,and significant difference was found between every two groups at each time point(all P<0.01).Conclusion The longer the ureteral obstruction is,the more severe the smooth muscle injury is;when ureteral obstruction exceeds 2 weeks,the irreversible injury of ureteral function occurs.
Objective To observe the changes of ureteral morphology,smooth muscle cell structure and myosin expression in rats with ureteral obstruction within 3 weeks,which provides a basis for clinical diagnosis of ureteral function in patients with ureteral obstruction.Methods Totally 120 SD rats were randomly divided into the control group(30 rats),sham operation group(45 rats),and model group(45 rats).The rats in the control group were fed normally,the rats in the sham operation group were sham operated,and the rats in the model group were prepared for right ureteral insufficiency obstruction models.The ureter and kidney morphology of the three groups were observed at 1,2,and 3 weeks after operation.The diameter of the right ureter was measured.The cell structure of ureter smooth muscle was observed by HE staining.The expression of myosin in the ureteral smooth muscle was detected by immunohistochemical SP method.The relative expression of ureter smooth muscle myosin heavy chain mRNA was detected by qRT-PCR.Results In the model group,the proximal ureter gradually thickened at 1,2,and 3 weeks after operation.The kidney volume gradually increased and the tension gradually increased.The diameter of the right ureter gradually increased,which was greater than that of the control group and the sham operation group(P<0.01).HE staining results showed that the longer the proximal ureteral obstruction in the model group,the thinner the muscle layer of smooth muscle cells,the fewer the cells,and the more obvious the fiber proliferation.The expression of myosin and the expression of myosin heavy chain mRNA in the model group was higher than that in the control group and the sham operation group at 1 and 2 weeks after operation,but at 3 weeks after operation,it was lower than that of the the control group and sham operation group(all P<0.01).The expression of myosin and the relative expression of myosin heavy chain mRNA in the model group decreased gradually at 2 weeks,1 week and 3 weeks after operation,and significant difference was found between every two groups at each time point(all P<0.01).Conclusion The longer the ureteral obstruction is,the more severe the smooth muscle injury is;when ureteral obstruction exceeds 2 weeks,the irreversible injury of ureteral function occurs.
引文
[1]王浩宇,代林勇,李前伟,等.上尿路梗阻解除后患侧肾功能恢复情况的回顾性研究[J].中华泌尿外科杂志,2017,38(3):170-173.
[2]敖劲松,邱承俊,姚小兵,等.压力灌注对不同程度积水肾氧化损伤的影响[J].中华实验外科杂志,2015,32(6):1267-1269.
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[5]李时军,曾晓勇.手术治疗上尿路结石梗阻肾积水患者肾功能的恢复情况[J].中国临床研究,2017,30(3):379-381.
[6]Aboumarzouk OM,Monga M,Kata SG,et al.Flexible ureteroscopy and laser lithotripsy for stones>2 cm:a systematic review and meta-analysis[J].J Endourol,2012,26(10):1257-1263.
[7]Sergio M,Galarreta CI,Thornhill BA,et al.The fate of nephrons in congenital obstructive nephropathy:adult recovery is limited by nephron number despite early release of Obstruction[J].J Urol,2015,194(5):1463-1472.
[8]谭付清,沈柏华,谢立平,等.一侧输尿管结石梗阻致急性肾功能衰竭的治疗经验[J].中华医学杂志,2013,93(20):1580-1582.
[9]吴小芬,陈挺,马登扬,等.单侧上尿路梗阻解除前后肾皮质厚度与肾小球滤过率的关系[J].中国内镜杂志,2017,23)11):14-18.
[10]王庆伟,陶德赏,王晓飞,等.神经源性尿路功能障碍大鼠输尿管平滑肌细胞Ca2+变化的意义[J].中华实用儿科临床杂志,2016,31(12):943-946.
[11]Lin S,Mequanint K.Bioreactor-induced mesenchymal progenitor cell differentiation and elastic fiber assembly in engineered vascular tissues[J].Acta Biomater,2017(59):200-209.
[12]Moore JR,Leinwand L,Warshaw DM.Understanding cardiomyopathy phenotypes basedon the functional impact of mutations in the myosin motor[J].Circ Res,2012,111(3):375-385.
[13]尹楠,茹峰,王东文.输尿管部分梗阻后平滑肌细胞中游离Ca2+浓度改变[J].中国医药导报,2011,8(8):6-8.
[14]He YM,Gu MM.Research progress of myosin heavy chain genes in human genetic diseases[J].Yi Chuan,2017,39(10):877-887.
[15]Chitano P,Wang L,Tin GYY,et al.Smooth muscle function and myosin polymerization[J].J Cell Sci,2017,130(15):2438-2480.
[16]Ming HU,Hong L.The accommodative function of myosin on cell biomechanics effect[J].Med Recapitul,2015,21(3):387-390.
[17]韩庆,沈德良,王勃,等.瑞舒伐他汀抑制血管平滑肌细胞表型转化的机制研究[J].中国现代医学杂志,2017,27(3):27-33.
[18]白志明,刘振湘,张冲,等.膀胱出口梗阻后逼尿肌收缩蛋白表达和膀胱重量的改变[J].中国男科学杂志,2007,21(8):14-17.
[2]敖劲松,邱承俊,姚小兵,等.压力灌注对不同程度积水肾氧化损伤的影响[J].中华实验外科杂志,2015,32(6):1267-1269.
[3]Wen J,Ringgaard S,Jorgensen T,et al.Long-term effects of partial unilateral ureteral obstruction on renal hemodynamics and morphology in newborn rats:a magnetic resonance imaging study[J].Urol Res,2002,30(4):205-212.
[4]李泸平,范应中,张谦,等.小儿泌尿系结石并急性肾功能衰竭41例临床分析[J].中华儿科杂志,2013,51(4):295-297.
[5]李时军,曾晓勇.手术治疗上尿路结石梗阻肾积水患者肾功能的恢复情况[J].中国临床研究,2017,30(3):379-381.
[6]Aboumarzouk OM,Monga M,Kata SG,et al.Flexible ureteroscopy and laser lithotripsy for stones>2 cm:a systematic review and meta-analysis[J].J Endourol,2012,26(10):1257-1263.
[7]Sergio M,Galarreta CI,Thornhill BA,et al.The fate of nephrons in congenital obstructive nephropathy:adult recovery is limited by nephron number despite early release of Obstruction[J].J Urol,2015,194(5):1463-1472.
[8]谭付清,沈柏华,谢立平,等.一侧输尿管结石梗阻致急性肾功能衰竭的治疗经验[J].中华医学杂志,2013,93(20):1580-1582.
[9]吴小芬,陈挺,马登扬,等.单侧上尿路梗阻解除前后肾皮质厚度与肾小球滤过率的关系[J].中国内镜杂志,2017,23)11):14-18.
[10]王庆伟,陶德赏,王晓飞,等.神经源性尿路功能障碍大鼠输尿管平滑肌细胞Ca2+变化的意义[J].中华实用儿科临床杂志,2016,31(12):943-946.
[11]Lin S,Mequanint K.Bioreactor-induced mesenchymal progenitor cell differentiation and elastic fiber assembly in engineered vascular tissues[J].Acta Biomater,2017(59):200-209.
[12]Moore JR,Leinwand L,Warshaw DM.Understanding cardiomyopathy phenotypes basedon the functional impact of mutations in the myosin motor[J].Circ Res,2012,111(3):375-385.
[13]尹楠,茹峰,王东文.输尿管部分梗阻后平滑肌细胞中游离Ca2+浓度改变[J].中国医药导报,2011,8(8):6-8.
[14]He YM,Gu MM.Research progress of myosin heavy chain genes in human genetic diseases[J].Yi Chuan,2017,39(10):877-887.
[15]Chitano P,Wang L,Tin GYY,et al.Smooth muscle function and myosin polymerization[J].J Cell Sci,2017,130(15):2438-2480.
[16]Ming HU,Hong L.The accommodative function of myosin on cell biomechanics effect[J].Med Recapitul,2015,21(3):387-390.
[17]韩庆,沈德良,王勃,等.瑞舒伐他汀抑制血管平滑肌细胞表型转化的机制研究[J].中国现代医学杂志,2017,27(3):27-33.
[18]白志明,刘振湘,张冲,等.膀胱出口梗阻后逼尿肌收缩蛋白表达和膀胱重量的改变[J].中国男科学杂志,2007,21(8):14-17.