高压氧治疗对早期兔动静脉瘘模型中静脉血管平滑肌细胞增殖和迁移的影响
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摘要
该文探讨了高压氧治疗对早期兔动静脉瘘模型中静脉血管平滑肌细胞增殖和迁移的影响。通过腺嘌呤诱导36只成年新西兰兔建立慢性肾衰模型,然后将其随机分为3组(每组12只):假手术组;单纯动静脉瘘组;高压氧治疗和动静脉瘘组。应用Masson染色检测静脉内膜增生和血管重塑的情况;免疫组化染色和Western blot检测α-SMA、PCNA、PDGF-BB、VEGF-A、TNF-α和MMP-9表达情况; RT-PCR检测α-SMA、PDGF-BB和VEGF-A表达水平。结果显示,相较于假手术组,单纯动静脉瘘组的静脉管壁弹力纤维明显增生,中内膜增厚,血管明显重塑。与单纯动静脉瘘组相比较,高压氧治疗和动静脉瘘组的弹力纤维增生受到抑制,以及相关的各目的蛋白和mRNA的表达水平下降。这说明,动静脉瘘术后,高压氧治疗可明显抑制动静脉瘘静脉血管平滑肌细胞的增殖和迁移,改善血管重塑。
The aim of this study was to develop a rabbit model of arteriovenous fistula(AVF) and then use it to evaluate the effects of Hyperbaric oxygen(HBO) therapy on proliferation and migration of venous vascular smooth muscle cells in early stage. A total of 36 New Zealand rabbits were fed with adenine diet in order to establish a chronic renal failure model. The rabbits were randomly divided into three groups(n=12 for each group):the sham group underwent sham operation; the AVF alone group underwent fistulization, but did not receive HBOT therapy; the AVF+HBO group underwent fistulization and received HBO therapy. We used Masson staining to detect the vein intimal hyperplasia and vascular remodeling. The expression levels of α-SMA, PCNA, PDGF-BB,VEGF-A, TNF-α and MMP-9 were analyzed by Immunohistochemical staining and Western blot. The expression levels of α-SMA, PDGF-BB and VEGF-A were assessed by RT-PCR assay. Compared with the sham group, the elastic fibers of the venous wall increased significantly, the venous intima media became thicker, and the blood vessels remodeled significantly in the AVF alone group. Compared with the AVF alone group, the proliferation of the venous-wall elastic fibers were inhibited and the expression of related proteins and mRNAs were decreased in the AVF+HBO group. Our study suggested that continuous HBO therapy following AVF could not only significantly inhibit the proliferation and migration of vascular smooth muscle cells in AVF-vein tissue, but also improve vascular remodeling.
The aim of this study was to develop a rabbit model of arteriovenous fistula(AVF) and then use it to evaluate the effects of Hyperbaric oxygen(HBO) therapy on proliferation and migration of venous vascular smooth muscle cells in early stage. A total of 36 New Zealand rabbits were fed with adenine diet in order to establish a chronic renal failure model. The rabbits were randomly divided into three groups(n=12 for each group):the sham group underwent sham operation; the AVF alone group underwent fistulization, but did not receive HBOT therapy; the AVF+HBO group underwent fistulization and received HBO therapy. We used Masson staining to detect the vein intimal hyperplasia and vascular remodeling. The expression levels of α-SMA, PCNA, PDGF-BB,VEGF-A, TNF-α and MMP-9 were analyzed by Immunohistochemical staining and Western blot. The expression levels of α-SMA, PDGF-BB and VEGF-A were assessed by RT-PCR assay. Compared with the sham group, the elastic fibers of the venous wall increased significantly, the venous intima media became thicker, and the blood vessels remodeled significantly in the AVF alone group. Compared with the AVF alone group, the proliferation of the venous-wall elastic fibers were inhibited and the expression of related proteins and mRNAs were decreased in the AVF+HBO group. Our study suggested that continuous HBO therapy following AVF could not only significantly inhibit the proliferation and migration of vascular smooth muscle cells in AVF-vein tissue, but also improve vascular remodeling.
引文
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2 Cheung AK,Imrey PB,Alpers CE,Robbin ML,Radeva M,Larive B,et al.Intimal hyperplasia,stenosis,and arteriovenous fistula maturation failure in the hemodialysis fistula maturation study.J Am Soc Nephrol 2017;28(10):3005-13.
3 Woodside KJ,Bell S,Mukhopadhyay P,Repeck KJ,Robinson IT,Eckard AR,et al.Arteriovenous fistula maturation in prevalent hemodialysis patients in the United States:A national study.Am J Kidney Dis 2018;71(6):793-801.
4 Allon M,Lockhart ME,Lilly RZ,Gallichio MH,Young CJ,Barker J,et al.Effect of preoperative sonographic mapping on vascular access outcomes in hemodialysis patients.Kidney Int2001;60(5):2013-20.
5 Allon M,Robbin ML,Young CJ,Deierhoi MH,Goodman J,Hanaway M,et al.Preoperative venous intimal hyperplasia,postoperative arteriovenous fistula stenosis,and clinical fistula outcomes.Clin J Am Soc Nephrol 2013;8(10):1750-5.
6 Davies MG,Hagen PO.Pathobiology of intimal hyperplasia.Br J Surg 1994;81(9):1254-69.
7 Janardhanan R,Yang B,Vohra P,Roy B,Withers S,Bhattacharya S,et al.Simvastatin reduces venous stenosis formation in a murine hemodialysis vascular access model.Kidney Int 2013;84(2):338-52.
8 Brahmbhatt A,Misra S.The biology of hemodialysis vascular access failure.Semin Intervent Radiol 2016;33(1):15-20.
9 Choudhry H,Harris AL.Advances in hypoxia-inducible factor biology.Cell Metabolism 2018;27(2):281-98.
10 Lim CS,Kiriakidis S,Sandison A,Paleolog EM,Davies AH.Hypoxia-inducible factor pathway and diseases of the vascular wall.J Vasc Surg 2013;58(1):219-30.
11 Muto A,Fitzgerald TN,Pimiento JM,Maloney SP,Teso D,Paszkowiak JJ,et al.Smooth muscle cell signal transduction:implications of vascular biology for vascular surgeons.J Vasc Surg,2007;45(Suppl A):A15-24.
12 Chanakira A,Dutta R,Charboneau R,Barke R,Santilli SM,Roy S.Hypoxia differentially regulates arterial and venous smooth muscle cell proliferation via PDGFR-βand VEGFR-2 expression.Am J Physiol Heart Circ Physiol 2011;302(5):H1173-84.
13 Misra S,Fu AA,Rajan DK,Juncos LA,McKusick MA,Bjarnason H,et al.Expression of hypoxia inducible factor-1alpha,macrophage migration inhibition factor,matrix metalloproteinase-2 and-9,and their inhibitors in hemodialysis grafts and arteriovenous fistulas.J Vasc Interv Radiol 2008;19(2Pt 1):252-9.
14 Misra S,Shergill U,Yang B,Janardhanan R,Misra KD.Increased expression of HIF-1alpha,VEGF-A and its receptors,MMP-2,TIMP-1,and ADAMTS-1 at the venous stenosis of arteriovenous fistula in a mouse model with renal insufficiency.JVasc Interv Radiol 2010;21(8):1255-61.
15 Yan L,Liang T,Cheng O.Hyperbaric oxygen therapy in China.Med Gas Res 2015;5(3):1-6.
16 Li Y,Hao Y,Wang T,Wei L,Wang W,Liang Y,et al.The effect of hyperbaric oxygen therapy on myocardial perfusion after the implantation of drug-eluting stents.Ann Clin Lab Sci 2018;48(2):158-63.
17 Rits Y,Uzieblo M,Shanley CJ.Hyperbaric oxygen decreases intimal thickness and area after carotid artery balloon injury in a rat model.Ann Vasc Surg 2013;27(6):785-90.
18 Li Z,Li M,Li X,Zhang M,Zhao Y,Ren W,et al.Hyperbaric oxygen inhibits venous neointimal hyperplasia following arteriovenous fistulization.Int J Mol Med 2017;39(5):1299-306.
19 Duque J,Martinez L,Mesa A,Wei Y,Tabbara M,Salman L,et al.CD4(+)lymphocytes improve venous blood flow in experimental arteriovenous fistulae.Surg Annu 2015;158(2):529-36.
20 Chen S,Qin S,Wang M,Zhang S.Expression and significance of NELIN and SM22αin varicose vein tissue.Exp Ther Med 2015;9(3):845-49.
21 Sezer S,Ozdemir FN,Arat Z,Turan M,Haberal M.Triad of malnutrition,inflammation,and atherosclerosis in hemodialysis patients.Nephron 2002;91(3):456-62.
22 Kokubo T,Ishikawa N,Uchida H,Chasnoff SE,Xie X,Mathew S,et al.CKD accelerates development of neointimal hyperplasia in arteriovenous fistulas.J Am Soc Nephrol 2009;20(6):1236-45.
2 Cheung AK,Imrey PB,Alpers CE,Robbin ML,Radeva M,Larive B,et al.Intimal hyperplasia,stenosis,and arteriovenous fistula maturation failure in the hemodialysis fistula maturation study.J Am Soc Nephrol 2017;28(10):3005-13.
3 Woodside KJ,Bell S,Mukhopadhyay P,Repeck KJ,Robinson IT,Eckard AR,et al.Arteriovenous fistula maturation in prevalent hemodialysis patients in the United States:A national study.Am J Kidney Dis 2018;71(6):793-801.
4 Allon M,Lockhart ME,Lilly RZ,Gallichio MH,Young CJ,Barker J,et al.Effect of preoperative sonographic mapping on vascular access outcomes in hemodialysis patients.Kidney Int2001;60(5):2013-20.
5 Allon M,Robbin ML,Young CJ,Deierhoi MH,Goodman J,Hanaway M,et al.Preoperative venous intimal hyperplasia,postoperative arteriovenous fistula stenosis,and clinical fistula outcomes.Clin J Am Soc Nephrol 2013;8(10):1750-5.
6 Davies MG,Hagen PO.Pathobiology of intimal hyperplasia.Br J Surg 1994;81(9):1254-69.
7 Janardhanan R,Yang B,Vohra P,Roy B,Withers S,Bhattacharya S,et al.Simvastatin reduces venous stenosis formation in a murine hemodialysis vascular access model.Kidney Int 2013;84(2):338-52.
8 Brahmbhatt A,Misra S.The biology of hemodialysis vascular access failure.Semin Intervent Radiol 2016;33(1):15-20.
9 Choudhry H,Harris AL.Advances in hypoxia-inducible factor biology.Cell Metabolism 2018;27(2):281-98.
10 Lim CS,Kiriakidis S,Sandison A,Paleolog EM,Davies AH.Hypoxia-inducible factor pathway and diseases of the vascular wall.J Vasc Surg 2013;58(1):219-30.
11 Muto A,Fitzgerald TN,Pimiento JM,Maloney SP,Teso D,Paszkowiak JJ,et al.Smooth muscle cell signal transduction:implications of vascular biology for vascular surgeons.J Vasc Surg,2007;45(Suppl A):A15-24.
12 Chanakira A,Dutta R,Charboneau R,Barke R,Santilli SM,Roy S.Hypoxia differentially regulates arterial and venous smooth muscle cell proliferation via PDGFR-βand VEGFR-2 expression.Am J Physiol Heart Circ Physiol 2011;302(5):H1173-84.
13 Misra S,Fu AA,Rajan DK,Juncos LA,McKusick MA,Bjarnason H,et al.Expression of hypoxia inducible factor-1alpha,macrophage migration inhibition factor,matrix metalloproteinase-2 and-9,and their inhibitors in hemodialysis grafts and arteriovenous fistulas.J Vasc Interv Radiol 2008;19(2Pt 1):252-9.
14 Misra S,Shergill U,Yang B,Janardhanan R,Misra KD.Increased expression of HIF-1alpha,VEGF-A and its receptors,MMP-2,TIMP-1,and ADAMTS-1 at the venous stenosis of arteriovenous fistula in a mouse model with renal insufficiency.JVasc Interv Radiol 2010;21(8):1255-61.
15 Yan L,Liang T,Cheng O.Hyperbaric oxygen therapy in China.Med Gas Res 2015;5(3):1-6.
16 Li Y,Hao Y,Wang T,Wei L,Wang W,Liang Y,et al.The effect of hyperbaric oxygen therapy on myocardial perfusion after the implantation of drug-eluting stents.Ann Clin Lab Sci 2018;48(2):158-63.
17 Rits Y,Uzieblo M,Shanley CJ.Hyperbaric oxygen decreases intimal thickness and area after carotid artery balloon injury in a rat model.Ann Vasc Surg 2013;27(6):785-90.
18 Li Z,Li M,Li X,Zhang M,Zhao Y,Ren W,et al.Hyperbaric oxygen inhibits venous neointimal hyperplasia following arteriovenous fistulization.Int J Mol Med 2017;39(5):1299-306.
19 Duque J,Martinez L,Mesa A,Wei Y,Tabbara M,Salman L,et al.CD4(+)lymphocytes improve venous blood flow in experimental arteriovenous fistulae.Surg Annu 2015;158(2):529-36.
20 Chen S,Qin S,Wang M,Zhang S.Expression and significance of NELIN and SM22αin varicose vein tissue.Exp Ther Med 2015;9(3):845-49.
21 Sezer S,Ozdemir FN,Arat Z,Turan M,Haberal M.Triad of malnutrition,inflammation,and atherosclerosis in hemodialysis patients.Nephron 2002;91(3):456-62.
22 Kokubo T,Ishikawa N,Uchida H,Chasnoff SE,Xie X,Mathew S,et al.CKD accelerates development of neointimal hyperplasia in arteriovenous fistulas.J Am Soc Nephrol 2009;20(6):1236-45.