CAJ | 학술논문

目的研究蛋白酪氨酸磷酸酶SHP2在细菌性脑膜炎中的表达及意义.方法脑池内注射肺炎链球菌诱导幼鼠脑膜炎模型,对照组脑池内注射0.9%氯化钠灭菌溶液.应用反转录(RT)-PCR、免疫印迹、免疫组织化学技术检测不同时间点脑组织酪氨酸磷酸酶SHP2的表达,观察SHP2蛋白表达与脑脊液TNF-α、WBC计数的关系.正态分布的两组数据比较采用t检验,多组数据的两两比较采用q检验.非正态分布的数据采用Kruskal-Wallis检验.相关性分析用Pearson检验.结果与健康对照组相比,细菌性脑膜炎引起皮层SHP2在mRNA和蛋白水平表达均明显升高(F=12.74,P<0.01;F=198,P<0.01).SHP2 mRNA在脑膜炎第3天达高峰,约为健康对照的5倍,第7天仍维持在较高水平.SHP2蛋白表达在脑膜炎第3天开始升高,第7天达高峰,约为健康对照的9倍,第14天时仍维持较高水平.免疫组织化学显示,SHP2在皮层神经元和胶质细胞广泛表达,脑膜炎组第三脑室周围免疫染色阳性细胞数明显增多,从形态上看主要为胶质细胞.皮层SHP2蛋白表达与脑脊液WBC计数呈显著正相关(r=0.77,P<0.01),而与脑脊液TNF-α浓度无明显相关性.结论 SHP2参与了细菌性脑膜炎的病理生理过程,可能发挥抑制炎性反应、促进炎性反应修复的作用,可作为病情变化的参考指标.
목적 연구단백락안산린산매SHP2재세균성뇌막염중적표체급의의.방법 뇌지내주사폐염련구균유도유서뇌막염모형,대조조뇌지내주사0.9%록화납멸균용액.응용반전록(RT)-PCR、면역인적、면역조직화학기술검측불동시간점뇌조직락안산린산매SHP2적표체,관찰SHP2단백표체여뇌척액TNF-α、WBC계수적관계.정태분포적량조수거비교채용t검험,다조수거적량량비교채용q검험.비정태분포적수거채용Kruskal-Wallis검험.상관성분석용Pearson검험.결과 여건강대조조상비,세균성뇌막염인기피층SHP2재mRNA화단백수평표체균명현승고(F=12.74,P<0.01;F=198,P<0.01).SHP2 mRNA재뇌막염제3천체고봉,약위건강대조적5배,제7천잉유지재교고수평.SHP2단백표체재뇌막염제3천개시승고,제7천체고봉,약위건강대조적9배,제14천시잉유지교고수평.면역조직화학현시,SHP2재피층신경원화효질세포엄범표체,뇌막염조제삼뇌실주위면역염색양성세포수명현증다,종형태상간주요위효질세포.피층SHP2단백표체여뇌척액WBC계수정현저정상관(r=0.77,P<0.01),이여뇌척액TNF-α농도무명현상관성.결론 SHP2삼여료세균성뇌막염적병리생리과정,가능발휘억제염성반응、촉진염성반응수복적작용,가작위병정변화적삼고지표.
Objective To explore the expression and significance of protein tyrosine phosphatase SHP2 in experimental bacterial meningitis.Methods A juvenile rat bacterial meningitis model was established by direct intraeisternal injection with Streptococcus pneumoniae. Uninfected control animals were mock-infected with sterile saline.The transcription and expression of SHP2 were detected by reverse transcription-polymerase chain reaction(RT-PCR),Western blot and immunohistochemistry techniques respectively at 1,3,7 and 14 days after infection.White blood cell(WBC)count,concentration of tumor necrosis factor-alpha(TNF-α)in cerebrospinal fluid(CSF)were also measured.Variables that were not normally distributed were compared by the Kruskal-Wallis test.Multiple comparison used Student-Newman-Keuls(SNK).The association between variables was assessed by using the Pearson correlation coefficient.Results Compared with uninfected controls,rats with bacterial meningitis showed a significant upregulation of SHP2 at both mRNA and protein levels(F=12.74,P<0.01;F=198,P<0.01).S HP2 mRNA levels peaked at 3 days after infection increasing more than five fold and remained at high levels at 7 days.In parallel,SHP2 protein levels began to increase at 3 days after infection,reaching a maximal increase of nearly nine fold at 7 days and remained at high levels at 14 days. Immunohistochemical analysis of SHP2 expression in the juvenile rat brain demonstrated that SHP2 labelling cells,identified as neuronal and glial cells,widely distributed in the cerebral cortex and the increased immunoreactive cells around the third ventricle were mainly glial cells.In addition,the protein levels of SHP2 and WBC counts were significantly correlated with each other(r=0.77,P<0.01),while there were no significant correlations between levels of SHP2 and TNF-a (r=0.08,P>0.05).Conclusions SHP2 may participate the pathological progress of the bacterial meningitis,restrating the inflammation and accelerating the renovation,so it can be regard as an index to measure the state of the illness.