CAJ | 학술논문

目的：通过大鼠皮肤移植急性排斥模型与正常生理稳态大鼠比较，探讨TLR4、9、10在脾脏中的表达特点及意义。方法：供体分别为6只Wistar和SD大鼠，24只受体SD大鼠随机分为四组，每组6只，取供体大鼠背部皮片移植到受体，A组：为同种异基因组，Wistar大鼠皮肤移植到SD大鼠；B组：为同种同基因组，SD大鼠皮肤移植到SD大鼠；C组：为假手术组，行皮片自体移植；D组：为正常大鼠空白对照组。在术后第1、4、7、10、14、21天时间点取各组大鼠脾脏，对脾脏单个核细胞行实时荧光定量（ real-time PCR）检测TLR4、9、10的mRNA表达以及脾脏组织免疫组化检测TLR4、9、10的蛋白表达情况。结果：A组TLR4、9、10的mRNA表达明显高于B、C组，差异有统计学意义（ P值＜0.05）；B组与C组之间的mRNA表达水平无统计学意义（ P值＞0.05）。 TLR4、9、10蛋白在A组中的表达强呈中等阳性（＋＋）到强阳性（＋＋＋），而在B、C、D三组中的表达基本呈现弱阳性（＋）。结论：通过对大鼠皮肤移植急性排斥模型中脾脏TLR4、9、10的mRNA和蛋白表达特点进行检测，发现TLR4、9、10参与了急性排斥反应；三者之间的表达可能存在协同效应；通过抑制Toll样受体的激活可能为移植术后防治急性排斥反应发生提供治疗靶点。
목적：통과대서피부이식급성배척모형여정상생리은태대서비교，탐토TLR4、9、10재비장중적표체특점급의의。방법：공체분별위6지Wistar화SD대서，24지수체SD대서수궤분위사조，매조6지，취공체대서배부피편이식도수체，A조：위동충이기인조，Wistar대서피부이식도SD대서；B조：위동충동기인조，SD대서피부이식도SD대서；C조：위가수술조，행피편자체이식；D조：위정상대서공백대조조。재술후제1、4、7、10、14、21천시간점취각조대서비장，대비장단개핵세포행실시형광정량（ real-time PCR）검측TLR4、9、10적mRNA표체이급비장조직면역조화검측TLR4、9、10적단백표체정황。결과：A조TLR4、9、10적mRNA표체명현고우B、C조，차이유통계학의의（ P치＜0.05）；B조여C조지간적mRNA표체수평무통계학의의（ P치＞0.05）。 TLR4、9、10단백재A조중적표체강정중등양성（＋＋）도강양성（＋＋＋），이재B、C、D삼조중적표체기본정현약양성（＋）。결론：통과대대서피부이식급성배척모형중비장TLR4、9、10적mRNA화단백표체특점진행검측，발현TLR4、9、10삼여료급성배척반응；삼자지간적표체가능존재협동효응；통과억제Toll양수체적격활가능위이식술후방치급성배척반응발생제공치료파점。
Objective:The expression patterns of TLR4,9,10 in the rat spleen were compared between the rat model of acute skin-transplant rejection and physiologically normal rats ,and their physiological significance were accessed.Methods:6 Wistar and 6 SD rats were used as donors ,and another 24 SD rats were used as recipients which were divided into 4 groups of 6.The same back skin slices were transplanted from donor to recipient , Group A:allograft ( Wistar to SD );Group B:homograft ( SD to SD );Group C:autograft;and Group D:normal SD rats as control.Rat spleens were collected at 1,4,7,10,14,and 21 days after surgery ,and mRNA levels of TLR4,9,10 in splenic mononuclear cells were determined by real-time PCR and corresponding protein expression was accessed by immunohistochemistry.Results:TLR4,9,10 mRNA levels of Group A were significantly higher than that of Group B and C ( P<0.05);TLR4,9,10 mRNA levels showed no significant difference between Group B and C (P>0.05).TLR4,9,10 expressions were moderate (++) to strong (+++) in Group A,and generally weak (+) in Group B,C and D.Conclusion:The patterns of TLR4,9,10 mRNA and protein expression in the rat model of acute skin-transplant rejection indicate that the three genes are involved in acute transplant rejection;the three genes may be expressed coordinately;inhibition of the Toll-like receptors may provide therapeutic targets for acute transplant rejection.