Chinese Journal of Tissue Engineering Research
Zhang Sheng-ning, Ran Jiang-hua, Liu Jing, Li Zhu, Li Lai-bang, Gao Yang, Zhang Xi-bing, Wu Shu-yuan, Li Li
Received:
2013-03-22
Revised:
2013-03-27
Online:
2013-07-30
Published:
2013-07-30
Contact:
Ran Jiang-hua, M.D., Professor, Chief physician, First Department of Hepatopancreatobiliary Surgery, the Affiliated Ganmei Hospital of Kunming Medical University and the First People’s Hospital of Kunming City, Liver Transplantation Center of Organ Transplantation Institute of Yunnan Province, Kunming 650011, Yunnan Province, China
rjh2u@163.com
About author:
Zhang Sheng-ning☆, M.D., Associate chief physician, Master’s supervisor, First Department of Hepatopancreatobiliary Surgery, the Affiliated Ganmei Hospital of Kunming Medical University and the First People’s Hospital of Kunming City, Liver Transplantation Center of Organ Transplantation Institute of Yunnan Province, Kunming 650011, Yunnan Province, China
zsn813@163.com
CLC Number:
Zhang Sheng-ning, Ran Jiang-hua, Liu Jing, Li Zhu, Li Lai-bang, Gao Yang, Zhang Xi-bing, Wu Shu-yuan, Li Li. Expression of nuclear factor-kappa B p65 in acute rejection of liver transplantation in rhesus monkey[J]. Chinese Journal of Tissue Engineering Research, doi: 10.3969/j.issn.2095-4344.2013.31.001.
2.1 实验动物数量分析 肝移植后受体动物共8只,急性排斥反应组和对照组各4只,急性排斥反应组中途丢失2只,对照组中途丢失1只,丢失原因为移植后腹腔出血及呼吸循环功能衰竭,丢失动物均及时补齐,计4个时点,每个时点获取标本4个,每组共获取肝脏标本16个。 2.2 肝移植后血清丙氨酸氨基转移酶活性 2组恒河猴肝移植后血清丙氨酸氨基转移酶活性同期比较,急性排斥反应组由于没有使用抗免疫排斥药物,移植后未使用激素等原因导致肝细胞受到机体炎性细胞的攻击损伤较对照组严重。2组移植后6 h和12 h的丙氨酸氨基转移酶活性比较接近,组间比较差异无显著性意义(P > 0.05)。恒河猴肝移植后,随着时间的延长,肝移植后6,12,24和72 h的肝功能丙氨酸氨基转移酶活性呈逐渐上升趋势,因而2组组内比较,差异有显著性意义(P < 0.05);并且急性排斥反应组比对照组变化大,肝移植后24 h到72 h的变化趋势最明显,见表1。"
表2可见,2组恒河猴肝移植后血清总胆红素同期比较,急性排斥反应组由于没有使用抗免疫排斥药物,移植后未使用激素等原因导致肝细胞受到机体炎性细胞的攻击损伤较对照组严重。2组随着移植后时间的延长总胆红素呈逐渐上升的趋势,2组移植后6 h和12 h的总胆红素水平比较接近,组间比较差异无显著性意义(P > 0.05)。恒河猴肝移植后,随着时间的延长,肝移植后6,12,24和72 h的肝功能总胆红素逐渐上升趋势,因而2组组内比较差异有显著性意义(P < 0.05);并且急性排斥反应组比对照组变化大,肝移植后24 h到72 h的变化趋势最明显。 2.4 肝移植后白细胞介素2水平 2组随着移植后时间延的长白细胞介素2呈逐渐上升的趋势,2组移植后6 h白细胞介素2水平比较接近,组间比较差异无显著性意义(P > 0.05)。恒河猴肝移植后,随着时间的延长,肝移植后6,12,24和72 h的血清白细胞介素2呈逐渐上升趋势,因而2组组内比较,差异有显著性意义(P < 0.05);并且急性排斥反应组比对照组变化大,肝移植后24 h到72 h的变化趋势最明显,见表3。"
2.5 肝移植后肝脏组织病理学变化 见图1。 移植后6 h:2组的肝脏组织基本上正常,没有排斥反应的镜下特点,可见肝小叶结构正常,汇管区有少量以中性粒细胞为主的炎性细胞浸润,未见淋巴细胞浸润或者仅有极少量的淋巴细胞,小叶间静脉内膜基本正常,但小叶间静脉内膜水肿,内膜下可见少量中性粒细胞侵润,汇管区可见有少量的混合炎症细胞侵润,胆管、血管均存在,但胆管上皮未受损伤。 移植后12 h:急性排斥反应组肝脏苏木精-伊红染色显示,肝小叶结构基本正常,汇管区有中等量以中性粒细胞为主的炎性细胞侵润,其中可以看到较多的淋巴细胞侵润,小叶间静脉内膜水肿明显增厚,内膜下可见中等量中性粒细胞侵润(淋巴细胞占的比例增多),胆管、血管均存在,但胆管上皮未明显受损伤,Banff评分Ⅰ-Ⅱ级。 对照组可见肝脏实质内有中等量的中性粒细胞侵润,汇管区仅见有少量中性粒细胞侵润(其中的淋巴细胞极少),排斥反应的分级不明确,Banff评分0-Ⅰ级。 移植后24 h:急性排斥反应组肝脏苏木精-伊红染色,中到重度排斥反应,汇管区大量单个核细胞浸润,并累及胆管、血管,肝小叶结构消失,伴有局灶性的出血、坏死。Banff评分Ⅱ-Ⅲ级。 对照组汇管区出现少量的单核细胞浸润,但侵及肝实质、胆管和血管的炎细胞浸润程度达到轻度急性排斥的诊断标准,Banff评分0-Ⅰ级。"
移植后72 h:急性排斥反应组肝脏苏木精-伊红染色,重度排斥反应,汇管区大量单个核细胞浸润,并累及胆管、血管,肝小叶结构消失明显,且伴有明显的局灶性出血、坏死,Banff评分Ⅲ级。 对照组汇管区出现少到中等量的单核细胞浸润,侵及肝实质程度较急性排斥反应组轻,Banff评分0-Ⅰ级。 2.6 肝移植后核因子κB p65蛋白表达变化 核因子κB p65条带(Mr 65 000)与相应β-actin(Mr 42 000)做比值后得到的数据进行统计学分析,对不同组采用单因素方差分析,组间对比采用t 检验。 整体变化趋势:急性排斥反应组6 h开始明显增高;对照组呈现下降趋势,12,24和72 h 3个时间点组急性排斥反应组明显高于对照组,差异有显著性意义(P=0.023,0.002,0.001,均< 0.05);6 h 2组之间差异无显著性意义(P=0.364,> 0.05),见表4及图2。"
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