Chelluboina Bharath, Nalamolu Koteswara Rao, Mendez Gustavo G, Klopfenstein Jeffrey D, Pinson David M, Wang David Z, Veeravalli Krishna Kumar
Departments of Cancer Biology and Pharmacology, University of Illinois College of Medicine at Peoria, Peoria, Illinois, USA.
Departments of Neurosurgery, University of Illinois College of Medicine at Peoria, Peoria, Illinois, USA.
Cell Physiol Biochem. 2017;44(4):1360-1369. doi: 10.1159/000485533. Epub 2017 Nov 30.
BACKGROUND/AIMS: Stem cell treatment is one of the potential treatment options for ischemic stroke. We recently demonstrated a protective effect of human umbilical cord blood-derived mesenchymal stem cells (HUCB-MSCs) in a rat model of ischemic stroke. The treatment attenuated apoptosis and prevented DNA damage. A collection of published studies, including several from our laboratory, indicated the induction and detrimental role for several matrix metalloproteinases (MMPs) in post-stroke brain injury. We hypothesized that the HUCB-MSCs treatment after focal cerebral ischemia prevents the dysregulation of MMPs and induces the expression of endogenous tissue inhibitors of metalloproteinases (TIMPs) to neutralize the elevated activity of MMPs.
To test our hypothesis, we administered HUCB-MSCs (0.25 million cells/animal and 1 million cells/animal) intravenously via tail vein to male Sprague-Dawley rats that were subjected to a transient (two-hour) right middle cerebral artery occlusion (MCAO) and one-day reperfusion. Ischemic brain tissues obtained from various groups of rats seven days after reperfusion were subjected to real-time PCR, immunoblot, and immunofluorescence analysis.
HUCB-MSCs treatment prevented the induction of MMPs, which were upregulated in ischemia-induced rats that received no treatment. HUCB-MSCs treatment also prevented the induction of TIMPs expression. The extent of prevention of MMPs and TIMPs induction by HUCB-MSCs treatment is similar at both the doses tested.
Prevention of stroke-induced MMPs upregulation after HUCB-MSCs treatment is not mediated through TIMPs upregulation.
背景/目的：干细胞治疗是缺血性中风潜在的治疗选择之一。我们最近在缺血性中风大鼠模型中证实了人脐带血间充质干细胞（HUCB-MSCs）的保护作用。该治疗减轻了细胞凋亡并防止了DNA损伤。一系列已发表的研究，包括我们实验室的几项研究，表明几种基质金属蛋白酶（MMPs）在中风后脑损伤中具有诱导作用且发挥有害影响。我们假设局灶性脑缺血后给予HUCB-MSCs治疗可防止MMPs失调，并诱导内源性金属蛋白酶组织抑制剂（TIMPs）表达，以中和升高的MMPs活性。
为验证我们的假设，我们通过尾静脉向雄性Sprague-Dawley大鼠静脉注射HUCB-MSCs（每只动物0.25×10⁶个细胞和1×10⁶个细胞），这些大鼠经历了短暂（两小时）的右侧大脑中动脉闭塞（MCAO）并再灌注一天。在再灌注七天后从各组大鼠获取的缺血脑组织进行实时PCR、免疫印迹和免疫荧光分析。
HUCB-MSCs治疗可防止MMPs的诱导，MMPs在未接受治疗的缺血诱导大鼠中上调。HUCB-MSCs治疗还可防止TIMP表达的诱导。在测试的两种剂量下，HUCB-MSCs治疗对MMPs和TIMP诱导的预防程度相似。
HUCB-MSCs治疗后对中风诱导的MMPs上调的预防不是通过TIMP上调介导的。
