CAJ | 학술논문

目的:观察脂肪来源的干细胞(ADSC)移植对脑缺血大鼠脑组织IL-10及TNF含量变化, 探讨ADSC对脑缺血损伤的保护机制.方法:将72只清洁级成年雄性Sprague-Darley大鼠随机分为假手术组(Sham组)、脑缺血组(MCAO组)、溶剂对照组(Vehicle组)和ADSC治疗组(ADSC组)四组, 每组18 只, 采用线栓法制作大脑中动脉栓塞(MCAO)模型, ADSC移植前用DAPI标记, ADSC 组于造模成功1 d后经侧脑室注射入30 μL的经DAPI标记的ADSC细胞悬液, 内含1×10~6细胞, Vehicle组则注射同等剂量的PBS, 各组分别于术后4 d、 7 d和14 d分批处死后取出脑脑织, 采用ELISA法、免疫组织化学法和半定量RT-PCR法, 检测大鼠脑组织中IL-10和TNF-α表达的变化.结果:与假手术组相比较, MCAO组大鼠4 d和7 d时脑组织中TNF-α和IL-10的表达明显上调(P<0.05).MCAO组与vehicle组各相应时间点IL-10和TNF-α表达均无统计学差异(P>0.05).与vehicle组相比, ADSC组4 d、 7 d和14 d时脑组织中IL-10的表达显著上调, TNF-α的表达明显下调(P<0.05).结论:ADSC移植可上调脑缺血大鼠IL-10的表达, 并下调TNF-α的表达, 这可能是ADSC具有脑缺血保护作用的机制之一.
목적:관찰지방래원적간세포(ADSC)이식대뇌결혈대서뇌조직IL-10급TNF함량변화, 탐토ADSC대뇌결혈손상적보호궤제.방법:장72지청길급성년웅성Sprague-Darley대서수궤분위가수술조(Sham조)、뇌결혈조(MCAO조)、용제대조조(Vehicle조)화ADSC치료조(ADSC조)사조, 매조18 지, 채용선전법제작대뇌중동맥전새(MCAO)모형, ADSC이식전용DAPI표기, ADSC 조우조모성공1 d후경측뇌실주사입30 μL적경DAPI표기적ADSC세포현액, 내함1×10~6세포, Vehicle조칙주사동등제량적PBS, 각조분별우술후4 d、 7 d화14 d분비처사후취출뇌뇌직, 채용ELISA법、면역조직화학법화반정량RT-PCR법, 검측대서뇌조직중IL-10화TNF-α표체적변화.결과:여가수술조상비교, MCAO조대서4 d화7 d시뇌조직중TNF-α화IL-10적표체명현상조(P<0.05).MCAO조여vehicle조각상응시간점IL-10화TNF-α표체균무통계학차이(P>0.05).여vehicle조상비, ADSC조4 d、 7 d화14 d시뇌조직중IL-10적표체현저상조, TNF-α적표체명현하조(P<0.05).결론:ADSC이식가상조뇌결혈대서IL-10적표체, 병하조TNF-α적표체, 저가능시ADSC구유뇌결혈보호작용적궤제지일.
AIM: To explore the effects of adipose-derived stem cell (ADSC) transplantation on the expression of IL-10 amd TNF-α after cerebral ischaemia in Middle cerebral artery occlusion (MCAO) rats. METHODS: 72 male adult Sprague-Dawley rats were randomly divided into 4 groups: sham group, MCAO group, Vehicle group and ADSC group (n=18). Rat's cerebral ischemia model was established by MCAO with Longa' s method. ADSC were labeled by DAPI before transplantation. One day after MCAO, 30 μL of cell suspension containing 1×10~6 ADSCs were injected into the lateral ventricle of ADSC group and the same dose of PBS was given to the Vehicle group. At day 4, day 7 and day 14 after MCAO, the rats were decapitated to detect the expression of IL-10 and TNF-α in ischaemic rat' s brain by ELISA, immunohistochemistry and RT-PCR. RESULTS: Compared with sham group, the expression of IL-10 and TNF-α significantly up-regulated at 4 d, 7 d of MCAO group(P<0.05). There was no statistical difference of IL-10 and TNF-α expression between MCAO and vehicle group ant any time point(P>0.05). Compared with Vehicle group, the expression of IL-10 significantly up-regulated while TNF-α expression significantly decreased of ADSC-treated group at any timepoint post MCAO(P<0.05). CONCLUSION: The transplantation of ADSC could up-regulate the expression of IL-10 and down-regulate the expression of TNF-α in MCAO rat' s brain, which might contribute to its protective role upon cerebral ischaemia.