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糖尿病脑血管并发症是其致死致残的常见原因。鉴于内皮祖细胞(EPCs)在血管形成中的重要作用,近年来在心脑血管方面的研究越来越受到重视。多项研究证实EPCs移植治疗对脑缺血性疾病有改善作用[1-2]。然而糖尿病可影响EPCs功能[3],单纯EPCs移植治疗的效果并不理想。前期研究证实雌激素体外孵育可改善糖尿病大鼠的EPCs的功能。因此,本研究通过分离培养糖尿病大鼠EPCs,经雌激素孵育后对糖尿病缺血性脑卒中大鼠进行移植治疗并评价治疗的效果,为临床治疗糖尿病脑血管并发症提供新的思路。
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与CI组比较,DM+CI组脑梗死体积明显增加(P<0.01);经EPCs移植治疗后糖尿病大鼠的脑缺血体积明显减小且经雌激素体外干预后的EPCs移植能进一步减少脑缺血梗死体积,说明雌激素干预可能加强了EPCs对脑卒中大鼠脑部缺血损伤的治疗作用(P<0.05, P<0.01,与 DM+CI组比较; P<0.05, 与 CI组比较),见图1。
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水迷宫实验结果显示, DM+CI组大鼠游泳路程和总时间相较于CI组明显增加(P<0.05),平均速度和潜伏时间则无明显变化(P>0.05);与DM+CI组比较,DM+CI+EPCs组以及DM+CI+EPCs+ estrogen组游泳总时间明显缩短(P<0.05),平均速度均明显提高(P<0.05),游泳总路程则无明显变化(P>0.05);但只有DM+CI+EPCs+ estrogen组潜伏期明显降低(P<0.01),见图2和表1。
表 1 不同处理组大鼠Morris迷宫实验指标对比(
$\bar x $ +s)组别(n=8) 总时间(s) 总路程(mm) 平均速度(mm/s) 潜伏期(s) CI 30.71±18.28 6721.56±4838.88 198.36±78.74 26.34±21.20 DM+CI 35.15±15.29* 7855.55±4262.89* 197.49±59.9 27.93±20.36 DM+CI+EPCs 29.49±13.46# 7459.41±4829.74 236.85±56.49# 25.14±16.04 DM+CI+EPCs+estrogen 28.80±13.75## 8477.04±5226.29 269.68±94.16## 22.41±14.16## *P<0.05,与CI组比较;#P<0.01,##P<0.01,与DM+CI组比较。 -
免疫荧光结果显示,与CI组比较,DM+CI组大鼠脑梗死区域新生血管数量明显降低(P<0.01);而DM+CI+EPCs组以及DM+CI+EPCs+ estrogen组大鼠脑梗死区域新生血管数量明显增加(P<0.01),且DM+CI+EPCs+ estrogen新生血管数量高于CI组(P<0.05),见图3。
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EPCs移植后第3天,取大鼠脑组织,切片后观察EPCs归巢情况。与CI组比较,DM+CI组大鼠脑组织缺血区域EPCs数量明显降低;而EPCs移植组大鼠脑组织梗死区域EPCs数量均明显增加,其中经过雌激素干预的EPCs移植组显示出梗死区域有更多数量的EPCs(图4)。
Therapeutic effects of estrogen-intervened EPCs transplantation on diabetic ischemic stroke rats
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摘要:
目的 探讨雌激素干预的EPCs移植对糖尿病缺血性脑卒中的治疗作用。 方法 制备大鼠糖尿病缺血性脑卒中模型,24 h后通过尾静脉移植经PKH26标记的糖尿病EPCs和雌激素干预的糖尿病EPCs。移植3 d后测定各组大鼠脑缺血体积、大鼠行为学变化、缺血部位血管新生及EPCs的归巢情况。 结果 与糖尿病脑缺血大鼠相比,雌激素干预的糖尿病EPCs移植能降低脑缺血体积、改善行为学评分和缺血部位的血管新生及促进EPCs归巢到损伤部位(P<0.05)。 结论 雌激素干预的糖尿病EPCs移植,通过促进EPCs归巢和血管新生对糖尿病缺血性脑卒中有较好的治疗作用。 Abstract:Objective To explore the therapeutic effects of estrogen-intervened endothelial progenitor cells( EPCs) transplantation on diabetic ischemic stroke rats. Methods PKH26-labeled diabetic EPCs and estrogen-intervened diabetic EPCs were injected into rats via the tail vein 24 h after cerebral ischemia. Cerebral ischemic volume, behavioral changes, ischemic site vascularization and homing of EPCs were measured 3 d after EPCs injection. Results Compared with diabetic ischemic rats, estrogen-intervened EPCs transplantation had reduced infarct volumes, improved behavioral scores and ischemic site revascularization and promoted homing of EPCs to sites of injury(P<0.05). Conclusion Estrogen-intervened EPCs transplantation had a better therapeutic effect on diabetic ischemic stroke by promoting EPCs homing to injury site and EPCs-medicated neovascularization . -
Key words:
- estrogen /
- EPCs transplantation /
- diabetic ischemic stroke /
- neovascularization /
- homing
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表 1 不同处理组大鼠Morris迷宫实验指标对比(
$\bar x $ +s)组别(n=8) 总时间(s) 总路程(mm) 平均速度(mm/s) 潜伏期(s) CI 30.71±18.28 6721.56±4838.88 198.36±78.74 26.34±21.20 DM+CI 35.15±15.29* 7855.55±4262.89* 197.49±59.9 27.93±20.36 DM+CI+EPCs 29.49±13.46# 7459.41±4829.74 236.85±56.49# 25.14±16.04 DM+CI+EPCs+estrogen 28.80±13.75## 8477.04±5226.29 269.68±94.16## 22.41±14.16## *P<0.05,与CI组比较;#P<0.01,##P<0.01,与DM+CI组比较。 -
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