CAJ | 학술논문

目的研究脓毒症大鼠肺组织中组织抑制因子-1(TIMP-1)的表达变化及其在脓毒症导致的肺损伤中的意义.方法实验在三峡大学医学院实验中心完成.雄性SD大鼠40只,随机(随机数字法)分为假手术(Sham)组(8只)和模型组(32只),模型组分6h、12 h、24 h、48 h4个时相组,每组8只.脓毒症模型制备采用盲肠结扎穿孔法(CLP).造模后在不同时相取血、处死并摘取肺组织.用光镜观察肺组织结构变化.用RT-PCR法检测TIMP-1 mRNA在不同时相肺组织中的表达,并同时检测凋亡因子Bax及凋亡抑制因子Bcl-2 mRNA的表达,采用免疫组织化学法以CD18标记不同时相肺组织中的炎症细胞并计数.计量资料以均数±标准差((x)±s)表示,组间比较采用t检验.结果与Sham组相比,CLP后各时相肺组织均有不同程度的损伤；CLP后各时相肺组织中TIMP-1 mRNA表达量均显著升高(P＜0.01),其峰值见于CLP后24h (P＜0.01)；CLP后12 ～48 h Bax mRNA表达量降低(P＜0.05),CLP后48 h最低(P＜0.01)；CLP后Bcl-2 mRNA表达量无明显改变.免疫组化染色表明CLP后各时相肺组织中CD18阳性细胞数均显著高于Sham组(P＜0.01),24 h达高峰(P＜0.01),且与TIMP-1 mRNA的表达量差异具有统计学意义,相关系数r0.426 (P＜0.01).结论 TIMP-1与脓毒症所导致的肺损伤关系密切,其机制可能是抑制了炎症细胞的凋亡,持续的炎症反应造成组织损伤,最终导致器官功能障碍.
목적 연구농독증대서폐조직중조직억제인자-1(TIMP-1)적표체변화급기재농독증도치적폐손상중적의의.방법 실험재삼협대학의학원실험중심완성.웅성SD대서40지,수궤(수궤수자법)분위가수술(Sham)조(8지)화모형조(32지),모형조분6h、12 h、24 h、48 h4개시상조,매조8지.농독증모형제비채용맹장결찰천공법(CLP).조모후재불동시상취혈、처사병적취폐조직.용광경관찰폐조직결구변화.용RT-PCR법검측TIMP-1 mRNA재불동시상폐조직중적표체,병동시검측조망인자Bax급조망억제인자Bcl-2 mRNA적표체,채용면역조직화학법이CD18표기불동시상폐조직중적염증세포병계수.계량자료이균수±표준차((x)±s)표시,조간비교채용t검험.결과 여Sham조상비,CLP후각시상폐조직균유불동정도적손상；CLP후각시상폐조직중TIMP-1 mRNA표체량균현저승고(P＜0.01),기봉치견우CLP후24h (P＜0.01)；CLP후12 ～48 h Bax mRNA표체량강저(P＜0.05),CLP후48 h최저(P＜0.01)；CLP후Bcl-2 mRNA표체량무명현개변.면역조화염색표명CLP후각시상폐조직중CD18양성세포수균현저고우Sham조(P＜0.01),24 h체고봉(P＜0.01),차여TIMP-1 mRNA적표체량차이구유통계학의의,상관계수r0.426 (P＜0.01).결론 TIMP-1여농독증소도치적폐손상관계밀절,기궤제가능시억제료염증세포적조망,지속적염증반응조성조직손상,최종도치기관공능장애.
Objective To investigate the expression and the effects of tissue inhibitor of metalloproteinases-1 (TIMP-1) on lungs of rats with sepsis.Methods Forty Sprague-Dawley (SD) rats were randomly divided into two groups,namely sham group (n =8) and sepsis model group (n =32).The rats of model group were modeled by cecal ligation and puncture (CLP),and were further divided into four subgroups as per the time after modeling,namely 6 h (n =8),12 h (n =8),24 h (n =8),48 h (n =8)subgroups.Blood and lung samples were taken 6 h,12 h,24 h and 48 h after modeling.The histological changes in lungs of the rats were observed under light microscope.Expressions of TIMP-1 mRNA,Bax mRNA and Bcl-2 mRNA in lungs were measured by RT-PCR.The immunohistochemistry was used to label the CD18 in lungs during different phases of sepsis.The data were processed by t test.Results Compared with sham group,the lung tissues of rats in model group were injured to a certain extent after CLP.The expression of TIMP-1 mRNA and the number of CD18 positive cells increased at the same time (P ＜ 0.01),and peaked 24 hours later (P ＜ 0.01).While the expression of Bax mRNA in model group decreased markedly 12-48 hours after modeling (P ＜ 0.01-0.05),and reached minimum 48 hours later (P ＜ 0.01).The expression of Bcl-2 mRNA in model group changed unnoticeable.The positive correlation between variations in number of CD18 positive cells and expression of TIMP-1 mRNA was found in model group (r =0.426,P ＜ 0.01).Conclusions The increase in expression of TIMP-1 mRNA in lungs is closely associated with the lung injury of sepsis.The mechanism of lung injury is likely attributed to the preservation of inflammatory cells from apoptosis,and the persistent inflammation response causes tissue damage,leading to organ dysfunction.