目的揭示自体动静脉内瘘失功血管内膜病理改变和血管剪切应力的变化规律。方法研究对象:选取C57BL/6小鼠,分假手术组(n=20)和手术组(n=20)。手术组应用显微外科技术建立小鼠颈动静脉内瘘模型,假手术组不建立动静脉内瘘。检测指标:①血流动力学:测定动脉压、心率、血流速度、剪切应力值;②组织病理学:HE染色观察静脉壁厚度;PAS染色观察血管壁蛋白粘多糖;Gomori银染色观察血管壁网状纤维组织;醛品红弹力纤维染色观察血管壁弹力纤维组织;CD31、CD34免疫染色观察血管壁血管内皮细胞变化。结果①血流动力学检查:与假手术组比,手术组颈动脉内径变细,血管剪切应力明显增加(t=-6.840,P<0,001)。②组织病理学检查:HE染色显示手术组静脉血管壁明显增厚,管腔狭窄;PAS染色显示手术组中性粘多糖含量明显增多;Gomori银染色显示手术组纤维增生明显;醛品红弹力纤维染色显示手术组弹力纤维增生明显,排列紊乱;CD31、CD34免疫染色显示手术组血管内皮细胞膜缺乏连续性,被纤维组织增生破坏。结论动静脉内瘘手术后血管剪切应力增加,血管内膜异常增生,伴纤维增生紊乱、炎性细胞浸润增殖,进而血管壁增厚、管腔狭窄,是导致维持性血液透析患者自体动静脉内瘘功能障碍的重要机制。
Objective To reveal the rules of vascular intima and shear force changes in the autologous arteriovenous fistula failure. Methods C57BL/6 mice were divided into sham operation group (n=20) and surgical group (n=20). The operation group were subjected to a microsurgery to establish an internal arteriovenous fistula between carotid artery and jugular vein, while the sham group did not have the fistula. The examination indicators after the surgery included ①hemodynamics: arterial pressure, heart rate, blood flow velocity and shear stress value; ②histopathology: thickness of the venous wall by HE staining, mucopolysaccharide in vascular wall by PAS staining, damage of reticular fibers in vascular wall by Gomori silver staining, damage of elastic fibers in vascular wall by aldehyde-fuchsin elastic fiber staining, and changes of vascular endothelial cells by CD31 and CD34 immunostaining. Results ①Hemodynamics: Compared with the sham group, the inner diameter of the carotid artery shrank, and the vascular shear stress increased significantly (t=-6.840, P<0.001) in the operation group. ②Histopathology: In the operation group, pathological findings included thickened wall and narrowed lumen of the vein by HE staining, increased neutral mucopolysaccharide by PAS staining, evident fibrosis by Gomori silver staining, proliferation and disarrangement of elastic fibers by aldehyde-fuchsin elastic fiber staining, and interrupted endothelial cell layer by proliferated fibrous tissues in CD31 and CD34 immunostaining. Conclusion After arteriovenous fistula surgery, the increased shear stress in the fistula, the intimal hyperplasia, the proliferation and disarrangement of fibrous tissues and the infiltration of inflammatory cells lead to the thickness of the vessel wall and the stenosis of the vessel lumen, which account for the failure of autologous arteriovenous fistula in maintenance hemodialysis patients.
[1] Lee, T. Novel paradigms for dialysis vascular access: downstream vascular biology--is there a final common pathway?, Clin J Am Soc Nephrol,2013, 8 (12):2194-2201.
[2] 王萌萌, 鄢艳. 动静脉内瘘功能障碍机制及防治进展. In: 第九届中国血液净化论坛暨2017年中国医院协会血液净化中心管理分会年会论文集. 北京; 2017,1-13.
[3] 靖永胜, 王小平, 刘芳, 等. 动静脉内瘘功能障碍病理及发病机制的研究进展, 中华肾脏病杂志. 2017, 4 (33):318-320.
[4] Stolic R. Most Important Chronic Complications of Arteriovenous Fistulas for Hemodialysis, Medical Principles and Practice,2012, 22 (3):220-228.
[5] W?gs?ter Dick. MMP-2 and MMP-9 are prominent matrix metalloproteinases during atherosclerosis development in the Ldlr(-/-)Apob(100/100) mouse, International Journal of Molecular Medicine. 2011, 28 (2):247-253.
[6] Joerg H, Dallit M, Christine W, Daniele V, Meyer F, Mcconnell JP, Mario G, Lerman LO, Amir L. Expression of lipoprotein-associated phospholipase A2 in carotid artery plaques predicts long-term cardiac outcome, European Heart Journal. 2009, 23:2930-2938.
[7] 于海波. 剪切力对动静脉内瘘血管内皮细胞的影响. 2016.
[8] Davies PF. Flow-mediated endothelial mechanotransduction, Physiological Reviews. 75 (3):519-560.
[9] Tarbell JM, Simon SI, Curry F. Mechanosensing at the vascular interface, Annual Review of Biomedical Engineering. 2014, 16 (1):505-532.
[10] Piezol integration of vascular architecture with physiological force, Nature. 2014, 515(13): 279-282.
[11] Brahmbhatt A, Remuzzi A, Franzoni M, Misra S. The molecular mechanisms of hemodialysis vascular access failure, other. 2016, 89 (2):303-316.
[12] 陈荣荣, 俞燕. 动静脉内瘘血管内膜增生的细胞和分子病理机制, 肾脏病与透析肾移植杂志. 2016, 25 (005):475-478,455.
[13] Prabir RC, Wang Y, Mahesh K, Zhang J, Rupak B, Rino M, Sue H, Lois A. Cellular phenotypes in human stenotic lesions from haemodialysis vascular access, Nephrol Dial Transplant. 2009, 9:2786-2791.
[14] Michael, Allon. Novel paradigms for dialysis vascular access: Introduction, Clinical journal of the American Society of Nephrology : CJASN. 2013, 8(12):2183-2185.
[15] Owens CD, Nicole W, Min KJ, Dirk H, Conte MS, Andres S. Endothelial function predicts positive arterial-venous fistula remodeling in subjects with stage IV and V chronic kidney disease, Journal of Vascular Access. 2010, 11 (4):329-334.
[16] Brahmbhatt A, Misra S. The Biology of Hemodialysis Vascular Access Failure, Seminars in Interventional Radiology. 2016, 33 (01):015-020.
[17] Kang L, Grande JP, Farrugia G, Croatt AJ, Katusic ZS, Nath KA. Functioning of an arteriovenous fistula requires heme oxygenase-2, American journal of physiology Renal physiology. 2013, 305 (4): F545-F552.
[18] 董建华, 葛永纯. 自体动静脉内瘘功能障碍的机制, 肾脏病与透析肾移植杂志. 2019, v.28;No.162 (04):85-90.
[19] Kang L, Grande JP, Hillestad ML, Croatt AJ, Barry MA, Katusic ZS, Nath KA. A New Model of an Arteriovenous Fistula in Chronic Kidney Disease in the Mouse: Beneficial Effects of Upregulated Heme Oxygenase-1, Am J Physiol Renal Physiol. 2016, 310 (6): F466.
[20] Nath, A K. Heme oxygenase-1: a provenance for cytoprotective pathways in the kidney and other tissues, Kidney International. 2006, 70 (3):432-443.