CAJ | 학술논문

目的观察自然杀伤(NK)细胞在异纲(罗非鱼/小鼠)胰岛移植免疫排斥中的作用.方法胶原酶消化分离罗非鱼胰岛.链脲霉素小鼠腹腔内注射,建立药物性糖尿病小鼠模型.消化分离罗非鱼胰岛后,移植入糖尿病小鼠肝脏内,作为移植组(18只)；设立生理盐水注射组(12只)(简称盐水组)为对照组.动态观察移植后两组小鼠血糖的变化.并对移植物进行病理检查和对NK细胞免疫组织化学检测.结果移植后1、12h时,移植组的血糖水平明显下降,浓度分别为(9.34±3.05) mmol/L及(7.80±2.10) mmol/L,与对照组比较差异有统计学意义(P＜0.05).但在移植后24h时血糖浓度为(28.13±1.56) mmol/L,与对照组比较差异无统计学意义(P＞0.05).实验组在移植后12h,病理检查可见移植物周围有轻度的炎性反应,各级血管处可见明显的凝血反应,有炎性细胞的聚集,胰岛结构仍完整；在移植后24 h,可见移植物周围有明显的炎性细胞浸润.免疫组织化学检查显示在移植物周围有少量的NK细胞浸润.移植后3d,病理检查可见移植物周围可见大量炎性细胞浸润及纤维化表现.对照组在移植后的病理检查始终未发现移植部位周围有任何炎性细胞聚集.结论罗非鱼胰岛肝脏内移植能短暂的降低糖尿病小鼠血糖,血液介导的即刻炎性反应是导致胰岛早期死亡的原因,NK细胞参与了这个排斥反应.
목적 관찰자연살상(NK)세포재이강(라비어/소서)이도이식면역배척중적작용.방법 효원매소화분리라비어이도.련뇨매소소서복강내주사,건립약물성당뇨병소서모형.소화분리라비어이도후,이식입당뇨병소서간장내,작위이식조(18지)；설립생리염수주사조(12지)(간칭염수조)위대조조.동태관찰이식후량조소서혈당적변화.병대이식물진행병리검사화대NK세포면역조직화학검측.결과 이식후1、12h시,이식조적혈당수평명현하강,농도분별위(9.34±3.05) mmol/L급(7.80±2.10) mmol/L,여대조조비교차이유통계학의의(P＜0.05).단재이식후24h시혈당농도위(28.13±1.56) mmol/L,여대조조비교차이무통계학의의(P＞0.05).실험조재이식후12h,병리검사가견이식물주위유경도적염성반응,각급혈관처가견명현적응혈반응,유염성세포적취집,이도결구잉완정；재이식후24 h,가견이식물주위유명현적염성세포침윤.면역조직화학검사현시재이식물주위유소량적NK세포침윤.이식후3d,병리검사가견이식물주위가견대량염성세포침윤급섬유화표현.대조조재이식후적병리검사시종미발현이식부위주위유임하염성세포취집.결론 라비어이도간장내이식능단잠적강저당뇨병소서혈당,혈액개도적즉각염성반응시도치이도조기사망적원인,NK세포삼여료저개배척반응.
Objective To observe the roles of natural killer (NK) cells in fish-to-mice islet xenograft rejection.Methods Tilapia islets were harvested,and isolated with the collagenase enzyme.The BALB/c mouse diabetic model was made by intraperitoneal injection of stretopzin (STZ).Diabetic mice were divided randomly into two groups:islets transplantation (islets) group (n =18),and control (NS)group (n =12).The blood glucose levels were tested and compared after transplantation.The samples were evaluated and examined histologically by HE and immunohistochemical staining.Results Mean blood glucose levels were reduced to (9.34±3.05) and (7.80 ±2.10) mmol/L at 1 h and 12 h post-transplantation in islets transplantation group respectively,which were significantly lower than in control group (P ＜0.05).At 24 h post-transplantation,the mean blood glucose levels in islets transplantation group were increased to (28.13 ± 1.56) mmol/L,showing no significant difference in comparison to control group (P ＞0.05).At 12 h post-transplantation,the mild inflammatory response was found in islets transplantation group.The graft showed mild inflammatory cells infiltration.Only partially necrotic islet tissue was observed,whereas the majority of the islet tissue was spared.At 24 h post-transplantation,globally necrotic islet was replaced by dense eosinophilic fibrotic tissue and neutrophils.Immunocytochemical staining revealed the grafts were surrounded by fewer NKp46 immunopositive cells.The islet tissue remained intact and no inflammation process was found at anytime post-transplantation in control group.Conclusion Tilapia islet grafts transplanted into diabetic mice could decrease temporarily the blood glucose levels.The instant blood-mediated inflammatory response is the main reason to cause early islet death.NK cells seem to be involved in the rejection of islets transplantation in discordant tilapia-to-mice model.