超声联合微泡增效骨髓间充质干细胞归巢治疗缺血性脑卒中
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摘要
背景:国内外动物实验结果表明,超声微泡能够促进移植干细胞归巢到缺血区,增强血管新生和小动脉生成,改善缺血心肌局部血流量,恢复心肌收缩功能。目的:探讨超声联合微泡对静脉移植骨髓间充质干细胞归巢的影响及治疗缺血性脑卒中的有效性。方法:插线法建立大脑中动脉栓塞大鼠模型,建模72 h后,随机分为4组:PBS组(n=15),骨髓间充质干细胞组(n=18),超声+骨髓间充质干细胞组(n=18),超声+微泡+骨髓间充质干细胞组(n=18)。PBS组经尾静脉注射2 mL PBS;骨髓间充质干细胞组将骨髓间充质干细胞(约3×10~6)用2 mL PBS稀释后经尾静脉缓慢注射;超声+骨髓间充质干细胞组经颅骨超声(频率1 MHz,功率2 W/cm~2)辐照120 s,给予骨髓间充质干细胞;超声+微泡+骨髓间充质干细胞组在超声辐照前静注微泡对比剂0.1 mL/kg,再给予骨髓间充质干细胞。结果与结论:(1)移植后48 h,超声+微泡+骨髓间充质干细胞组细胞的脑内归巢率高于骨髓间充质干细胞组和超声+骨髓间充质干细胞组,差异均有显著性意义(P<0.05);(2)造模后28 d,超声+微泡+骨髓间充质干细胞组的神经损害程度较其余组轻,差异均有显著性意义(P<0.05);(3)移植后7 d,超声+微泡+骨髓间充质干细胞组的脑组织含水率较其余组低,差异均有显著性意义(P<0.05);(4)移植后7 d,超声+微泡+骨髓间充质干细胞组脑梗死体积较其余组小,差异有显著性意义(P<0.05);(5)结果表明,超声联合微泡可促进骨髓间充质干细胞的脑内靶向归巢,能够减少脑水肿和脑梗死体积,改善神经功能评分,增效骨髓间充质干细胞移植治疗缺血性脑卒中的效果。
BACKGROUND: In animal experiments, ultrasound-mediated microbubbles can promote the homing of transplanted stem cells to the ischemic area, enhance angiogenesis and small arterial formation, improve local blood flow in the ischemic myocardium and restore myocardial contractility.OBJECTIVE: To investigate the effect of ultrasound-mediated microbubbles on intravenously transplanted bone marrow mesenchymal stem cell(BMSC) homing and the therapeutic efficiency on ischemic stroke. METHODS: A middle cerebral artery occlusion(MCAO) model was induced by plug wire preparation. At 72 hours after MCAO, model rats were randomized into four groups: PBS group(n=15), BMSCs group(n=18), ultrasound+BMSCs group(n=18), ultrasound+microbubble+BMSCs group(n=18). Corresponding treatment was done in each group: 2 mL of PBS was injected via tail vein in the PBS group; about 3×10~6 BMSCs diluted by 2 mL of PBS were injected via tail vein slowly in the BMSCs group; after skull ultrasound radiation(1 MHz, 2 W/cm~2) for 120 seconds, BMSCs were injected via tail vein slowly in the ultrasound+BMSCs group; the same process as the ultrasound+BMSCs group was done following intravenous injection of 0.1 mL/kg microbubbles in the ultrasound+microbubble+BMSCs group. RESULTS AND CONCLUSION:(1) Forty-eight hours after BMSCs transplantation, the BMSCs homing rate in the brain was significantly higher in the ultrasound+microbubble+BMSCs group than the other two groups(P < 0.05).(2) Twenty-eight days after MCAO, nerve damage was significantly milder in the ultrasound+microbubble+BMSCs group than the other two groups(P < 0.05).(3) Seven days after transplantation, the water content in the brain tissue was significantly lower in the ultrasound+microbubble+BMSCs group than the other two groups(P < 0.05).(4) Seven days after transplantation, the cerebral infarction volume was significantly reduced in the ultrasound+microbubble+BMSCs group compared with the other two groups(P < 0.05). To conclude, ultrasound-mediated microbubbles can enhance the homing effect of intravenously transplanted BMSCs, reduce cerebral edema and cerebral infarction volume, improve the neurological function, and increase the therapeutic effect of BMSCs transplantation on ischemic stroke.
BACKGROUND: In animal experiments, ultrasound-mediated microbubbles can promote the homing of transplanted stem cells to the ischemic area, enhance angiogenesis and small arterial formation, improve local blood flow in the ischemic myocardium and restore myocardial contractility.OBJECTIVE: To investigate the effect of ultrasound-mediated microbubbles on intravenously transplanted bone marrow mesenchymal stem cell(BMSC) homing and the therapeutic efficiency on ischemic stroke. METHODS: A middle cerebral artery occlusion(MCAO) model was induced by plug wire preparation. At 72 hours after MCAO, model rats were randomized into four groups: PBS group(n=15), BMSCs group(n=18), ultrasound+BMSCs group(n=18), ultrasound+microbubble+BMSCs group(n=18). Corresponding treatment was done in each group: 2 mL of PBS was injected via tail vein in the PBS group; about 3×10~6 BMSCs diluted by 2 mL of PBS were injected via tail vein slowly in the BMSCs group; after skull ultrasound radiation(1 MHz, 2 W/cm~2) for 120 seconds, BMSCs were injected via tail vein slowly in the ultrasound+BMSCs group; the same process as the ultrasound+BMSCs group was done following intravenous injection of 0.1 mL/kg microbubbles in the ultrasound+microbubble+BMSCs group. RESULTS AND CONCLUSION:(1) Forty-eight hours after BMSCs transplantation, the BMSCs homing rate in the brain was significantly higher in the ultrasound+microbubble+BMSCs group than the other two groups(P < 0.05).(2) Twenty-eight days after MCAO, nerve damage was significantly milder in the ultrasound+microbubble+BMSCs group than the other two groups(P < 0.05).(3) Seven days after transplantation, the water content in the brain tissue was significantly lower in the ultrasound+microbubble+BMSCs group than the other two groups(P < 0.05).(4) Seven days after transplantation, the cerebral infarction volume was significantly reduced in the ultrasound+microbubble+BMSCs group compared with the other two groups(P < 0.05). To conclude, ultrasound-mediated microbubbles can enhance the homing effect of intravenously transplanted BMSCs, reduce cerebral edema and cerebral infarction volume, improve the neurological function, and increase the therapeutic effect of BMSCs transplantation on ischemic stroke.
引文
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[8]童嘉毅,李金鹆,范国峰,等.超声波促骨髓间充质干细胞心肌内归巢的实验研究[J].中国心血管病研究杂志,2008,6(10):771-773.
[9]范国峰,冯毅,童嘉毅,等.超声联合微泡对内皮祖细胞体外增殖、凋亡及细胞周期的影响[J].东南大学学报:医学版,2009,28(2):109-112.
[10]Laing RJ,Jakubowski J,Laing RW.Middle cerebral artery occlusion without craniectomy in rats.Which method works best.Stroke.1993;24(2):294-297.
[11]Zhang ZH,Wang RZ,Wang RZ,et al.Transplantation of neural stem cells modified by human neurotrophin-3promotes functional recovery after transient focal cerebral ischemia in rats.Neurosci Lett.2008;444(3):227-230.
[12]Longa EZ,Weinstein PR,Carlson S,et al.Reversible middle cerebral artery occlusion without craniectomy in rats.Stroke.1989;20(1):84-91.
[13]Chen J,Li Y,Wang L,et al.Therapeutic benefit of intravenous administration of bone marrow stromal cells after cerebral ischemia in rats.Stroke.2001;32(4):1005-1011.
[14]Shahrokhi N,Haddad MK,Joukar S,et al.Neuroprotective antioxidant effect of sex steroid hormones in traumatic brain injury.Pak J Pharm Sci.2012;25(1):219-225.
[15]Luger D,Lipinski MJ,Westman PC,et al.IntravenouslyDelivered Mesenchymal Stem Cells:Systemic Anti-Inflammatory Effects Improve Left Ventricular Dysfunction in Acute Myocardial Infarction and Ischemic Cardiomyopathy.Circ Res.2017 Feb 23.[Epub ahead of print]
[16]Koizumi JI,Yoshida Y,Nakazawa T,et al.Experimental studies of ischemic brain edema:1.A new experimental model of cerebral embolism in rats in which recirculation can be introduced in the ischemic area.Jpn J Stroke.1986;8(1):1-8.
[17]Shen LH,Li Y,Chen J,et al.Intracarotid transplantation of bone marrow stromal cells increases axon-myelin remodeling after stroke.Neuroscience.2006;137(2):393-399.
[18]Shen LH,Li Y,Chen J,et al.One-year follow-up after bone marrow stromal cell treatment in middle-aged female rats with stroke.Stroke.2007;38(7):2150-2156.
[19]Iihoshi S,Honmou O,Houkin K,et al.A therapeutic window for intravenous administration of autologous bone marrow after cerebral ischemia in adult rats.Brain Res.2004;1007(1-2):1-9.
[20]Ruan GP,Han YB,Wang TH,et al.Comparative study among three different methods of bone marrow mesenchymal stem cell transplantation following cerebral infarction in rats.Neurol Res.2013;35(2):212-220.
[21]Jiang Y,Zhu W,Zhu J,et al.Feasibility of delivering mesenchymal stem cells via catheter to the proximal end of the lesion artery in patients with stroke in the territory of the middle cerebral artery.Cell Transplant.2013;22(12):2291-2298.
[22]Imada T,Tatsumi T,Mori Y,et al.Targeted delivery of bone marrow mononuclear cells by ultrasound destruction of microbubbles induces both angiogenesis and arteriogenesis response.Arterioscler Thromb Vasc Biol.2005;25(10):2128-2134.
[23]Zen K,Okigaki M,Hosokawa Y,et al.Myocardium-targeted delivery of endothelial progenitor cells by ultrasound-mediated microbubble destruction improves cardiac function via an angiogenic response.J Mol Cell Cardiol.2006n;40(6):799-809.
[24]Tung YS,Marquet F,Teichert T,et al.Feasibility of noninvasive cavitation-guided blood-brain barrier opening using focused ultrasound and microbubbles in nonhuman primates.Appl Phys Lett.2011;98(16):163704.
[25]Tung YS,Vlachos F,Feshitan JA,et al.The mechanism of interaction between focused ultrasound and microbubbles in blood-brain barrier opening in mice.J Acoust Soc Am.2011;130(5):3059-3067.
[26]Lv W,Li WY,Xu XY,et al.Bone marrow mesenchymal stem cells transplantation promotes the release of endogenous erythropoietin after ischemic stroke.Neural Regen Res.2015;10(8):1265-1270.
[27]魏红,童嘉毅,范国峰,等.超声介导微泡对内皮祖细胞一氧化氮及一氧化氮合酶活性的影响[J].中国医科大学学报,2010,39(10):806-808.
[28]卞叶萍,童嘉毅,沈祥波,等.超声联合一氧化氮微泡介导间充质干细胞移植对心肌梗死大鼠心功能的影响[J].中华物理医学与康复杂志,2013,35(7):523-527.
[2]Go AS,Mozaffarian D,Roger VL,et al.Heart disease and stroke statistics--2013 update:a report from the American Heart Association.Circulation.2013;127(1):e6-e245.
[3]Dharmasaroja P.Bone marrow-derived mesenchymal stem cells for the treatment of ischemic stroke.J Clin Neurosci.2009;16(1):12-20.
[4]Bao X,Wei J,Feng M,et al.Transplantation of human bone marrow-derived mesenchymal stem cells promotes behavioral recovery and endogenous neurogenesis after cerebral ischemia in rats.Brain Res.2011;1367:103-113.
[5]Wakabayashi K,Nagai A,Sheikh AM,et al.Transplantation of human mesenchymal stem cells promotes functional improvement and increased expression of neurotrophic factors in a rat focal cerebral ischemia model.J Neurosci Res.2010;88(5):1017-1025.
[6]Chen J,Li Y,Wang L,et al.Therapeutic benefit of intravenous administration of bone marrow stromal cells after cerebral ischemia in rats.Stroke.2001;32(4):1005-1011.
[7]Chen J,Sanberg PR,Li Y,et al.Intravenous administration of human umbilical cord blood reduces behavioral deficits after stroke in rats.Stroke.2001;32(11):2682-2688.
[8]童嘉毅,李金鹆,范国峰,等.超声波促骨髓间充质干细胞心肌内归巢的实验研究[J].中国心血管病研究杂志,2008,6(10):771-773.
[9]范国峰,冯毅,童嘉毅,等.超声联合微泡对内皮祖细胞体外增殖、凋亡及细胞周期的影响[J].东南大学学报:医学版,2009,28(2):109-112.
[10]Laing RJ,Jakubowski J,Laing RW.Middle cerebral artery occlusion without craniectomy in rats.Which method works best.Stroke.1993;24(2):294-297.
[11]Zhang ZH,Wang RZ,Wang RZ,et al.Transplantation of neural stem cells modified by human neurotrophin-3promotes functional recovery after transient focal cerebral ischemia in rats.Neurosci Lett.2008;444(3):227-230.
[12]Longa EZ,Weinstein PR,Carlson S,et al.Reversible middle cerebral artery occlusion without craniectomy in rats.Stroke.1989;20(1):84-91.
[13]Chen J,Li Y,Wang L,et al.Therapeutic benefit of intravenous administration of bone marrow stromal cells after cerebral ischemia in rats.Stroke.2001;32(4):1005-1011.
[14]Shahrokhi N,Haddad MK,Joukar S,et al.Neuroprotective antioxidant effect of sex steroid hormones in traumatic brain injury.Pak J Pharm Sci.2012;25(1):219-225.
[15]Luger D,Lipinski MJ,Westman PC,et al.IntravenouslyDelivered Mesenchymal Stem Cells:Systemic Anti-Inflammatory Effects Improve Left Ventricular Dysfunction in Acute Myocardial Infarction and Ischemic Cardiomyopathy.Circ Res.2017 Feb 23.[Epub ahead of print]
[16]Koizumi JI,Yoshida Y,Nakazawa T,et al.Experimental studies of ischemic brain edema:1.A new experimental model of cerebral embolism in rats in which recirculation can be introduced in the ischemic area.Jpn J Stroke.1986;8(1):1-8.
[17]Shen LH,Li Y,Chen J,et al.Intracarotid transplantation of bone marrow stromal cells increases axon-myelin remodeling after stroke.Neuroscience.2006;137(2):393-399.
[18]Shen LH,Li Y,Chen J,et al.One-year follow-up after bone marrow stromal cell treatment in middle-aged female rats with stroke.Stroke.2007;38(7):2150-2156.
[19]Iihoshi S,Honmou O,Houkin K,et al.A therapeutic window for intravenous administration of autologous bone marrow after cerebral ischemia in adult rats.Brain Res.2004;1007(1-2):1-9.
[20]Ruan GP,Han YB,Wang TH,et al.Comparative study among three different methods of bone marrow mesenchymal stem cell transplantation following cerebral infarction in rats.Neurol Res.2013;35(2):212-220.
[21]Jiang Y,Zhu W,Zhu J,et al.Feasibility of delivering mesenchymal stem cells via catheter to the proximal end of the lesion artery in patients with stroke in the territory of the middle cerebral artery.Cell Transplant.2013;22(12):2291-2298.
[22]Imada T,Tatsumi T,Mori Y,et al.Targeted delivery of bone marrow mononuclear cells by ultrasound destruction of microbubbles induces both angiogenesis and arteriogenesis response.Arterioscler Thromb Vasc Biol.2005;25(10):2128-2134.
[23]Zen K,Okigaki M,Hosokawa Y,et al.Myocardium-targeted delivery of endothelial progenitor cells by ultrasound-mediated microbubble destruction improves cardiac function via an angiogenic response.J Mol Cell Cardiol.2006n;40(6):799-809.
[24]Tung YS,Marquet F,Teichert T,et al.Feasibility of noninvasive cavitation-guided blood-brain barrier opening using focused ultrasound and microbubbles in nonhuman primates.Appl Phys Lett.2011;98(16):163704.
[25]Tung YS,Vlachos F,Feshitan JA,et al.The mechanism of interaction between focused ultrasound and microbubbles in blood-brain barrier opening in mice.J Acoust Soc Am.2011;130(5):3059-3067.
[26]Lv W,Li WY,Xu XY,et al.Bone marrow mesenchymal stem cells transplantation promotes the release of endogenous erythropoietin after ischemic stroke.Neural Regen Res.2015;10(8):1265-1270.
[27]魏红,童嘉毅,范国峰,等.超声介导微泡对内皮祖细胞一氧化氮及一氧化氮合酶活性的影响[J].中国医科大学学报,2010,39(10):806-808.
[28]卞叶萍,童嘉毅,沈祥波,等.超声联合一氧化氮微泡介导间充质干细胞移植对心肌梗死大鼠心功能的影响[J].中华物理医学与康复杂志,2013,35(7):523-527.
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