骨髓间充质干细胞穴位移植对痴呆大鼠内源性神经干细胞增殖分化的影响

Effect of Bone Marrow Mesenchymal Stem Cell Transplantation at Acupoint on the Proliferation and Differentiation of Endogenous Neural Stem Cells in Dementia Rats

目的观察骨髓间充质干细胞(BMSC)风府穴穴位移植对痴呆大鼠海马区内源性神经干细胞增殖、分化的影响。方法SD大鼠50只,随机抽取10只作为对照组,其余采用Aβ1-42寡聚体侧脑室灌注建立大鼠阿尔茨海默病模型,然后再随机分为模型组、风府穴组、足三里穴组、尾静脉组,每组10只。水迷宫检测造模成功进行相应干预,风府穴组及足三里组分别以大鼠BMSC穴位移植,尾静脉组移植等量BMSC,模型组于风府穴注射等量的生理盐水,对照组作为空白对照。4周后,水迷宫实验检测大鼠的学习记忆能力,免疫荧光检测大鼠海马区内源性Nestin﹢、β-Tubulin-Ⅲ﹢表达。结果造模后水迷宫检测大鼠逃避潜伏期(EL)较对照组延长(P<0.01),穿越平台次数显著减少(P<0.001)。干预后,与模型组比较,风府穴组EL无明显缩短(P>0.05),穿越平台次数无明显增加(P>0.05)。Nestin﹢细胞数升高(P<0.05),β-Tubulin-Ⅲ﹢细胞数无明显增加(P>0.05)。结论大鼠BMSC风府穴穴位移植能诱导AD大鼠海马区内源性神经干细胞增殖,但尚未证实向成熟神经细胞定向分化。

Objective To observe the effect of bone marrow mesenchymal stem cell(BMSC) transplantation at Fengfu(GV16) acupoint on the proliferation and differentiation of endogenous neural stem cells in the hippocampus of dementia rats. Method Of 50 Sprague Dawley(SD) rats, 10 were randomly selected as a control group, and the rest received lateral ventricular perfusion of Aβ1-42 oligomer to develop Alzheimer disease(AD) rat models and then were randomized into a model group, a Fengfu point group, a Zusanli(ST36) point group and a tail vein group. When the model was confirmed a success by water maze test, the corresponding interventions were carried out. The Fengfu point group and Zusanli point group received transplantation of BMSCs at the corresponding acupoints;the same amount of BMSCs was transplanted in the tail vein group;the model group was injected with the same dose of normal saline at Fengfu point;the control group was taken as the blank control. Four weeks later, the learning and memory abilities of rats were examined by water maze test, and the expressions of nestin﹢ and b-Tubulin-Ⅲ﹢ in rat’s hippocampus were detected using immunofluorescence. Result The water maze test showed that the model rats had prolonged escape latency(EL)(P<0.01) and reduced number of platform crossings(P<0.001) compared with the control group. After intervention, the EL was not significantly shortened(P>0.05) and the number of platform crossings was not notably increased(P>0.05) in the Fengfu point group compared with the model group;the number of Nestin﹢ cells was larger in the Fengfu point group than in the model group(P<0.05);compared with the model group, the number of b-Tubulin-Ⅲ﹢ cells was not notably increased in the Fengfu point group(P>0.05). Conclusion BMSC transplantation at Fengfu acupoint can induce the proliferation of endogenous neural stem cells in hippocampus of AD rats, but there is no evidence fount to support the differentiation towards mature nerve cells.