中华器官移植杂志
中華器官移植雜誌
중화기관이식잡지
CHINESE JOURNAL OF ORGAN TRANSPLANTATION
2008年
11期
645-648
,共4页
谢林%魏庆信%李文鑫%王树森%李荣%曾梦华%朱珉%郭晖%陈实
謝林%魏慶信%李文鑫%王樹森%李榮%曾夢華%硃珉%郭暉%陳實
사림%위경신%리문흠%왕수삼%리영%증몽화%주민%곽휘%진실
转基因%移植,异种%补体%小鼠
轉基因%移植,異種%補體%小鼠
전기인%이식,이충%보체%소서
Transgenes%Transplantation,heterologous%Complement%Mice
目的 观察转人源性膜辅助蛋白(CD46)基因对转基因小鼠心脏中人补体沉积的抑制作用.方法 以近交系昆明小鼠为对象,采用显微注射法制备转人CD46基因小鼠,用逆转录聚合酶链法(RT-PCR法)检测外源基因的整合情况.以Fo代转基因小鼠11只为实验组,同窝非转基因小鼠10只为对照,快速切取小鼠心脏,用改良的Langendorff法经小鼠主动脉逆行灌注预先制备的含补体的B型血人血浆.观察并记录小鼠心脏搏动时间;采用免疫荧光和免疫组织化学法检测小鼠心脏组织中补体C3c及C9的沉积情况.结果 Fo代转基因小鼠外源基因的整合率为33.3%.实验组小鼠心脏搏动时间为(42.6±20.6)min(15～77 min),长于对照组的(20.2±12.5)min(7～40 min),差异有统计学意义(P＜0.01).实验组小鼠心脏组织中补体C3c及C9的沉积均少于对照组.结论 通过显微注射法制备的转人CD46基因小鼠,其外源基因可稳定表达;转人CD46基因可以抑制人补体C3c及C9在转基因小鼠心脏组织中的沉积.
目的 觀察轉人源性膜輔助蛋白(CD46)基因對轉基因小鼠心髒中人補體沉積的抑製作用.方法 以近交繫昆明小鼠為對象,採用顯微註射法製備轉人CD46基因小鼠,用逆轉錄聚閤酶鏈法(RT-PCR法)檢測外源基因的整閤情況.以Fo代轉基因小鼠11隻為實驗組,同窩非轉基因小鼠10隻為對照,快速切取小鼠心髒,用改良的Langendorff法經小鼠主動脈逆行灌註預先製備的含補體的B型血人血漿.觀察併記錄小鼠心髒搏動時間;採用免疫熒光和免疫組織化學法檢測小鼠心髒組織中補體C3c及C9的沉積情況.結果 Fo代轉基因小鼠外源基因的整閤率為33.3%.實驗組小鼠心髒搏動時間為(42.6±20.6)min(15～77 min),長于對照組的(20.2±12.5)min(7～40 min),差異有統計學意義(P＜0.01).實驗組小鼠心髒組織中補體C3c及C9的沉積均少于對照組.結論 通過顯微註射法製備的轉人CD46基因小鼠,其外源基因可穩定錶達;轉人CD46基因可以抑製人補體C3c及C9在轉基因小鼠心髒組織中的沉積.
목적 관찰전인원성막보조단백(CD46)기인대전기인소서심장중인보체침적적억제작용.방법 이근교계곤명소서위대상,채용현미주사법제비전인CD46기인소서,용역전록취합매련법(RT-PCR법)검측외원기인적정합정황.이Fo대전기인소서11지위실험조,동와비전기인소서10지위대조,쾌속절취소서심장,용개량적Langendorff법경소서주동맥역행관주예선제비적함보체적B형혈인혈장.관찰병기록소서심장박동시간;채용면역형광화면역조직화학법검측소서심장조직중보체C3c급C9적침적정황.결과 Fo대전기인소서외원기인적정합솔위33.3%.실험조소서심장박동시간위(42.6±20.6)min(15～77 min),장우대조조적(20.2±12.5)min(7～40 min),차이유통계학의의(P＜0.01).실험조소서심장조직중보체C3c급C9적침적균소우대조조.결론 통과현미주사법제비적전인CD46기인소서,기외원기인가은정표체;전인CD46기인가이억제인보체C3c급C9재전기인소서심장조직중적침적.
Objective To investigate the inhibition of human complements deposition by human CD46 expression on transgenic mouse hearts.Methods The whole length cDNA encoding human CD46 was amplified by PCR using human cDNA library.The human CD46 gene was introduced into fertilized mice eggs by microinjection to produce transgenic mice expressing human CD46.Isolated hearts from two groups of mice(wild type and hCD46-expressing)were perfused ex vivo with 10% pooled human plasma by modified Langendorff profusion system.Heart function was evaluated by mean heart beating time.Deposition of complements C3c and C9 in the perfused muffne hearts was examined by immunofluorescence and immunohistochemistry respectively.Results Mean heart beating time was prolonged to 42.6±20.6 min in the hCD46 expression group(P＜0.01)in contrast to 20.2 ±12.5 min in the wild type group.The expression of hCD46 in transgenic mice was detected by RTPCR analysis.In immunohistological examination,hCD46-expressing hearts exhibited a reduction in deposition of complements C9 and C3 compared with nontransgenic mouse hearts.Conclusion Human CD46 transgenic mice have been obtained by microinjection.Expression of human CD46 gene in xenografts could inhibit deposition of human complements C3c and C9.
