CAJ | 학술논문

目的探讨肿瘤坏死因子-α刺激基因-6蛋白(TSG-6)对脑梗死早期炎症反应和血脑屏障的影响.方法将90只雄性C57BL/6j小鼠按随机数字表法分为假手术组、溶剂治疗组、TSG-6治疗组,每组30只;后2组采用线栓法制备成大脑中动脉栓塞模型,造模成功后TSG-6治疗组小鼠立即通过尾静脉注射含有TSG-6(50 μg/只)PBS 200 μL,溶剂治疗组仅注射等体积PBS.脑梗死1、3d后采用改良神经功能缺损评分(mNSS)评价各组小鼠神经功能;脑梗死3d后采用氯化三苯四唑(TTC)染色评价脑梗死体积,干湿重法检测脑组织含水量,荧光分光光度法评估伊文氏蓝(EB)在梗死侧脑组织的渗出量,紫外分光光度法检测组过氧化物酶(MPO)活性,Western blotting检测基质金属蛋白酶-9(MMP-9)、高迁移率族蛋白B1(HMGB1)和Toll样受体4(TLR4)的表达量.结果脑梗死3d后,与溶剂治疗组mNSS评分(7.923 ±2.138)、脑梗死体积[(36.883±8.553)％]、脑组织EB含量[(11.867±4.135) μg/g]、脑组织含水量[(80.467±7.045)％]、MPO活性[(112.617±26.782)mU/g]以及梗死侧脑组织HMGB1 (0.861 ±0.137)、TLR4(0.833±0.193)和MMP-9(0.910±0.156)表达量比较,TSG-6治疗组小鼠mNSS评分(5.253±1.712)、脑梗死体积[(26.100±5.640)％]、脑组织EB含量[(7.233±3.434) μg/g]、脑组织含水量[(71.667±6.518)％]、MPO活性[(70.383±17.558) mU/g]以及梗死侧脑组织HMGB1 (0.503±0.230)、TLR4(0.482±0.159)和MMP-9(0.611±0.170)表达量明显降低或减小,差异均有统计学意义(P＜0.05).结论 TSG-6可显著减少HMGB1、TLR4的表达和中性粒细胞的浸润,进而减少MMP-9的表达发挥血脑屏障保护作用,减轻脑水肿和脑梗死体积,促进神经功能的恢复.
목적 탐토종류배사인자-α자격기인-6단백(TSG-6)대뇌경사조기염증반응화혈뇌병장적영향.방법 장90지웅성C57BL/6j소서안수궤수자표법분위가수술조、용제치료조、TSG-6치료조,매조30지;후2조채용선전법제비성대뇌중동맥전새모형,조모성공후TSG-6치료조소서립즉통과미정맥주사함유TSG-6(50 μg/지)PBS 200 μL,용제치료조부주사등체적PBS.뇌경사1、3d후채용개량신경공능결손평분(mNSS)평개각조소서신경공능;뇌경사3d후채용록화삼분사서(TTC)염색평개뇌경사체적,간습중법검측뇌조직함수량,형광분광광도법평고이문씨람(EB)재경사측뇌조직적삼출량,자외분광광도법검측조과양화물매(MPO)활성,Western blotting검측기질금속단백매-9(MMP-9)、고천이솔족단백B1(HMGB1)화Toll양수체4(TLR4)적표체량.결과 뇌경사3d후,여용제치료조mNSS평분(7.923 ±2.138)、뇌경사체적[(36.883±8.553)％]、뇌조직EB함량[(11.867±4.135) μg/g]、뇌조직함수량[(80.467±7.045)％]、MPO활성[(112.617±26.782)mU/g]이급경사측뇌조직HMGB1 (0.861 ±0.137)、TLR4(0.833±0.193)화MMP-9(0.910±0.156)표체량비교,TSG-6치료조소서mNSS평분(5.253±1.712)、뇌경사체적[(26.100±5.640)％]、뇌조직EB함량[(7.233±3.434) μg/g]、뇌조직함수량[(71.667±6.518)％]、MPO활성[(70.383±17.558) mU/g]이급경사측뇌조직HMGB1 (0.503±0.230)、TLR4(0.482±0.159)화MMP-9(0.611±0.170)표체량명현강저혹감소,차이균유통계학의의(P＜0.05).결론 TSG-6가현저감소HMGB1、TLR4적표체화중성립세포적침윤,진이감소MMP-9적표체발휘혈뇌병장보호작용,감경뇌수종화뇌경사체적,촉진신경공능적회복.
Objective To explore the effects of tumor necrosis factor-α stimulated gene-6 (TSG-6) on early inflammation and blood brain barrier (BBB) in mice after ischemic stroke (IS).Methods A total of 90 healthy male C57BL/6j mice were randomly divided into sham-operated group (n=30),vehicle group (n=30) and TSG-6 treatment group (n=30);the middle cerebral artery occlusion models (IS) in the later two groups were established with suture emboli method;50 μg TSG-6 or PBS was immediately injected into mice via tail vein after IS.Neurological function deficits were assessed by modified neurological severity scale (mNSS) one and three days after IS;infraction volume was examined by triphenyl tetrazolium chloride (TTC) staining,the changes of brain water content were examined by wet and dry weight method,evans blue (EB) extravasation was assessed by means of EB fluorescent quantitation,myeloperoxidase (MPO) activity was tested by ultraviolet specterphotometry,and Western blotting was used to detect the expressions of matrix metalloproteinase-9 (MMP-9),high-mobility group protein box 1 (HMGB1) and toll-like receptor 4 (TLR4) three days after IS.Results Three days after IS,the mNSS scores (5.253±1.712),infraction volume ([26.100±5.640]％),EB extravasation ([7.233± 3.434] μg/g),brainwater content ([71.667±6.518]％),MPO activity ([70.383±17.558] mU/g) and HMGB1,TLR4 and MMP-9 expressions (0.503±0.230,0.482±0.159 and 0.611±0.170) in the TSG-6 treatment group were significantly lower or smaller than those in the vehicle group ([7.923±2.138],([36.883±8.553]％),([11.867±4.135] μg/g),([80.467±7.045]％),([112.617±26.782] mU/g),[0.861± 0.137],[0.833 ±0.193] and [0.910 ±0.156]) three days after IS (P＜0.05);the expressions of MMP-9 (0.611±0.170),HMGB1 (0.503±0.230) and TLR4 (0.482±0.159) in mice of TSG-6 treatment group were significantly down-regulated as compared with those in vehicle group those days after IS (P＜0.05).Conclusion TSG-6 can exert neuroprotective effect by protecting BBB from damage and reducing brain edema by reduction of MMP-9 after stroke,which is mediated by downregulation of HMGB 1 and its receptor TLR4 and the infiltration ofneutrophile granulocytes.