CAJ | 학술논문

目的分析基质金属蛋白酶-9(MMP-9)在小鼠结核性脑膜炎病理生理过程中的表达特点及其意义.方法侧脑室注射结核分枝杆菌H37RV悬液的16只小鼠作为模型组,注射等量0.9％氯化钠溶液的16只小鼠作为对照组,30 d后处死.分别进行脑组织结核分枝杆菌培养、病理学观察,明胶酶谱法检测MMP-9活性,检测血脑屏障通透性和脑组织含水量,免疫荧光染色脑组织MMP-9、胶原纤维酸性蛋白(GFAP)和整合素αM(OX-42).采用t检验比较两组间的差异.结果模型组小鼠注射菌量为(1.271±0.111)×106菌落形成单位(cfu)/只,感染30 d后脑组织匀浆培养出结核分枝杆菌,菌量为(4.900±1.407)×104 cfu/mL,光学显微镜下见蛛网膜下腔和脑室扩张,大量炎性细胞浸润.模型组小鼠脑组织MMP-9条带累积吸光度(A)值为47 821±19 932,对照组为10 082±3 544,差异有统计学意义(t=3.728,P=0.010).模型组小鼠脑组织伊文斯兰(EB)含量为(11.8±3.6) μg/g,对照组为(4.7±3.4)μg/g,差异有统计学意义(t=2.887,P=0.028).模型组小鼠脑组织含水量为0.849±0.035,对照组为0.775±0.037,差异有统计学意义(t=2.925,P=0.026).免疫荧光染色显示感染后小鼠脑组织高表达MMP-9、GFAP和OX-42,且MMP-9与GFAP和OX-42均明显重合.结论 MMP-9在结核性脑膜炎小鼠脑组织中活性增强,并参与破坏血脑屏障,导致组织水肿、炎性细胞渗出,小胶质细胞-星形胶质细胞网络参与MMP-9的分泌.
목적 분석기질금속단백매-9(MMP-9)재소서결핵성뇌막염병리생리과정중적표체특점급기의의.방법 측뇌실주사결핵분지간균H37RV현액적16지소서작위모형조,주사등량0.9％록화납용액적16지소서작위대조조,30 d후처사.분별진행뇌조직결핵분지간균배양、병이학관찰,명효매보법검측MMP-9활성,검측혈뇌병장통투성화뇌조직함수량,면역형광염색뇌조직MMP-9、효원섬유산성단백(GFAP)화정합소αM(OX-42).채용t검험비교량조간적차이.결과 모형조소서주사균량위(1.271±0.111)×106균락형성단위(cfu)/지,감염30 d후뇌조직균장배양출결핵분지간균,균량위(4.900±1.407)×104 cfu/mL,광학현미경하견주망막하강화뇌실확장,대량염성세포침윤.모형조소서뇌조직MMP-9조대루적흡광도(A)치위47 821±19 932,대조조위10 082±3 544,차이유통계학의의(t=3.728,P=0.010).모형조소서뇌조직이문사란(EB)함량위(11.8±3.6) μg/g,대조조위(4.7±3.4)μg/g,차이유통계학의의(t=2.887,P=0.028).모형조소서뇌조직함수량위0.849±0.035,대조조위0.775±0.037,차이유통계학의의(t=2.925,P=0.026).면역형광염색현시감염후소서뇌조직고표체MMP-9、GFAP화OX-42,차MMP-9여GFAP화OX-42균명현중합.결론 MMP-9재결핵성뇌막염소서뇌조직중활성증강,병삼여파배혈뇌병장,도치조직수종、염성세포삼출,소효질세포-성형효질세포망락삼여MMP-9적분비.
Objective To analyze the characteristics and significance of matrix metalloproteinase-9 (MMP-9) expression in the pathophysiological processes of tuberculous meningitis in mice.Methods Sixteen mice were intracerebroventricularly injected with H37RV suspension as the model group.Meanwhile,the other 16 mice were injected with 0.9％ sodium chloride solution as the control group.Thirty days later,all mice were decapitated and the brain tissue were respectively used to for Mycobacterium tuberculosis (M.tuberculosis) incubation,pathological changes observation,MMP-9 activity detection by zymography,blood-brain-barrier permeability and moisture content detection,and immunofluorescence stain of MMP-9,glial fibrillary acidic protein (GFAP) and integrin αM (OX-42).The t test was used to compare the differences between the two groups.Results Every experimental mouse was injected with (1.271±0.111) × 106 colony-forming units (cfu) M.tuberculosis.Thirty days later,the amount of M.tuberculosis in brain tissue homogenates was (4.900± 1.407) × 104 cfu/mL,and the hematoxylin and eosin staining showed dilatation of subarachnoid and ventricular and infiltration of a large number of inflammatory cells.The cumulative absorbance (A) of MMP-9 bands of brain tissue was 47 821 ± 19 932 in the model group and 10 082 ± 3 544 in the control group.The difference was statistically significant (t =3.728,P=0.010).The evans blue (EB) content of brain tissue was (11.8 ± 3.6) μg/g in model group and (4.7 ±3.4) μg/g in control group.The difference was statistically significant (t=2.887,P=0.028).The moisture of brain tissue was 0.849±0.035 in model group and 0.775±0.037 in control group.The difference was statistically significant (t=2.925,P=0.026).The immunofluorescence staining showed that the infected brain tissue expressed high degrees of MMP-9,GFAP and OX-42.And MMP-9 was overlapped with both GFAP and OX-42 obviously.Conclusions The activity of MMP-9 is significantly enhanced in brain tissue of mice suffering from tuberculous meningitis and participates in blood-brain barrier damage,tissue edema and inflammatory cells exudation.Microglia cells-astrocytes network is involved in the secretion of MMP-9.