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小鼠肺移植术后早期肺呼吸阻力与γδT淋巴细胞变化规律的实验研究

The early changes of respiratory system resistance andγδT lymphocytes infiltrated in graft after lung transplantation of mouse

摘要目的:观察实验动物肺移植术后早期肺呼吸阻力和γδT淋巴细胞的变化规律。方法研究实施时间为2014年3月至2015年5月。雄性C57BL/6小鼠35只,BALB/c 小鼠10只;随机分为对照组(5只), C57BL/6至 C57BL/6同型移植组(10只), BALB/c 至 C57BL/6异型移植组(10只);移植组再随机分为3 d和7 d亚组(各5只)。移植组进行小鼠原位左肺移植手术。术后第3、7天对各组小鼠进行肺功能检查、移植肺病理学检查,计数移植肺γδT淋巴细胞及其亚型的变化,检测移植肺白细胞介素17A的mRNA表达水平。计量资料组间比较采用方差分析,两两比较采用LSD法检验。结果肺呼吸阻力术后3 d 同型移植组为(2?61±0?59)cmH2O·s/ml(1 cmH2O=0?098 kPa),异型移植组为(2?84±0?31)cmH2O·s/ml,均高于对照组(1?39±0?17)cmH2O·s/ml(P=0?001,0?000);术后7 d异型移植组为(4?33±0?67)cmH2O·s/ml,高于同型移植组的(1?87±0?27) cmH2O·s/ml和对照组的(1?39±0?17)cmH2O·s/ml(P=0?000,0?000)。异型移植组术后肺内炎症细胞浸润较同型移植组更明显,其中7d亚组移植肺表现为严重排斥反应。移植肺浸润γδT淋巴细胞术后3 d同型移植组为3?90%±0?86%,异型移植组为4?40%±0?57%,均高于对照组的2?00%±0?23%( P=0?000,0?000);术后7 d异型移植组为5?40%±0?98%,高于同型移植组的2?60%±0?54%和对照组的2?00%±0?23%(P=0?000,0?000)。移植肺白细胞介素17A mRNA表达水平术后3 d和7 d 同型移植组分别为3?37±0?55和5?23±1?50,异型移植组分别为6?77±0?93和27?32±4?20,均高于对照组的0?99±0?08(P均=0?000),异型移植组高于同型移植组(P=0?000,0?000)。结论实验动物肺移植术后早期移植肺呼吸阻力升高,可能与局部募集的高表达白细胞介素17A的γδT淋巴细胞有关。

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abstractsObjectives To generate an orthotopic left lung transplantation model in mice, and to observe the early changes of respiratory system resistance andγδT lymphocytes infiltrated in grafts? Methods The research time was from March 2014 to May 2015? The male C57BL/6 mice ( n=35) and BALB/c mice (syngenic group,n=10) were randomly divided into five groups. Control group (n=5): wild C57BL/6 mice; syngenic transplant group ( n=10 ): C57BL/6→C57BL/6; allogenic transplant group ( allogenic group,n=10): BALB/c→C57BL/6; each transplant group was randomly divided into 3?day and 7?day subgroups ( n=5 )? Respiratory system resistance and histological features of grafts were assessed, and differences in graft infiltrating γδT lymphocytes and mRNA expression of interleukin ( IL )?17A were quantified on 3 and 7 days after transplantation? Multiple comparisons were performed using one?way analysis of variance and least significant difference analysis? Results ( 1 ) The respiratory system resistance of syngenic group and allogenic group were (2?61±0?59) cmH2O·s/ml and (2?84±0?31) cmH2O·s/ml 3 days post?operation, both of them increased compared to control group (1?39±0?17) cmH2O·s/ml (1 cmH2O=0?098 kPa) (P=0?001, 0?000). The respiratory system resistance of allogenic group were (4?33±0?67) cmH2 O·s/ml 7 days post?operation, which was significantly higher than that of syngenic 7?day subgroup (1?87±0?27) cmH2O·s/ml and control group (1?39±0?17) cmH2O·s/ml (P=0?000, 0?000)?(2) The isografts of syngenic group showed a relatively normal histological appearance with minimal infiltration of inflammatory cells, and the allografts of allogenic group infiltrated apparently by inflammatory cells, especially 7?day subgroup showed acute cellular rejection? ( 3) The percentage of γδT lymphocytes infiltrated in isografts and allografts were 3?90%± 0?86% and 4?40%± 0?57%, respectively, which were significantly increased compared to that of control lungs 2?00%±0?23% 3 days post?operation(P=0?000, 0?000);The percentage ofγδT lymphocytes infiltrated in 7 days allografts was 5?40%±0?98% , which was higher compared to that of 7 days isografts 2?60%± 0?54% and control lungs 2?00%± 0?23% ( P=0?000, 0?000)? (4) IL?17A mRNA expression levels were 3?37±0?55 and 5?23±1?50 in isografts and 6?77± 0?93 and 27?32±4?20 in allografts, on postoperative day 3 and 7 respectively? All of them were significantly upregulated compared to that of control lungs 0?99±0?08 (P=0?000, 0?000), and allografts exhibited significantly greater IL?17A transcript levels compared to isografts on postoperative day 3 and 7 ( P=0?000, 0?000) . Conclusion The rise of respiratory system resistance of lung grafts after transplantation may relate to the increased IL?17A?producing γδT lymphocytes infiltrated in the grafts.

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中华外科杂志

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2016年54卷12期

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