中华器官移植杂志
中華器官移植雜誌
중화기관이식잡지
CHINESE JOURNAL OF ORGAN TRANSPLANTATION
2011年
8期
502-505
,共4页
孙煦勇%秦科%农江%文宁%赖彦华%董建辉%聂峰%蔡文娥%覃音红%黄晨
孫煦勇%秦科%農江%文寧%賴彥華%董建輝%聶峰%蔡文娥%覃音紅%黃晨
손후용%진과%농강%문저%뢰언화%동건휘%섭봉%채문아%담음홍%황신
胰岛%移植%细胞凋亡%氧化损伤
胰島%移植%細胞凋亡%氧化損傷
이도%이식%세포조망%양화손상
Islet%Transplantation%Apoptosis%Injury of oxidation
目的 探讨胰岛在获取、分离及纯化等过程中出现细胞凋亡的相关因素及应对策略.方法 经供者腹主动脉灌注4℃的UW液后,切取供者胰腺,并进行胶原酶灌注、消化及梯度离心,分离、提取胰岛.分别于胶原酶灌注前、灌注后、消化后及胰岛分离后等时点,采用原位末端标记法(TUNEL法)检测胰岛细胞凋亡情况,采用比色法检测超氧化物歧化酶(SOD)与丙二醛(MDA)水平,采用HE染色和双硫腙染色观察胰岛的形态学改变.结果 在分离、纯化胰岛的各个过程中,均出现了胰岛细胞凋亡的现象,其中在胶原酶灌注后和消化后最为明显,并伴随MDA的高表达,表达水平分别为(6.18±2.38)nmol/mgprot和(9.21±2.75)nmol/mgprot,均明显高于灌注前的(4.21±1.83)nmol/mgprot(P＜0.05和P＜0.01);而此时SOD水平则显著下降,至胰岛分离后仍处于较低水平,与灌注前相比,差异均有统计学意义(P＜0.05和P＜0.01).胶原酶灌注前,胰岛结构完整;胶原酶灌注后,胰岛周围组织结构膨胀;消化后,胰岛膜性结构被破坏,但能勉强维持岛状结构;胰岛分离后结构基本完整.结论 在提取胰岛的过程中,胶原酶灌注及消化可引起胰岛细胞凋亡,其机制可能与氧化损伤有关,抗氧化措施可作为移植前保护胰岛的手段.
目的 探討胰島在穫取、分離及純化等過程中齣現細胞凋亡的相關因素及應對策略.方法 經供者腹主動脈灌註4℃的UW液後,切取供者胰腺,併進行膠原酶灌註、消化及梯度離心,分離、提取胰島.分彆于膠原酶灌註前、灌註後、消化後及胰島分離後等時點,採用原位末耑標記法(TUNEL法)檢測胰島細胞凋亡情況,採用比色法檢測超氧化物歧化酶(SOD)與丙二醛(MDA)水平,採用HE染色和雙硫腙染色觀察胰島的形態學改變.結果 在分離、純化胰島的各箇過程中,均齣現瞭胰島細胞凋亡的現象,其中在膠原酶灌註後和消化後最為明顯,併伴隨MDA的高錶達,錶達水平分彆為(6.18±2.38)nmol/mgprot和(9.21±2.75)nmol/mgprot,均明顯高于灌註前的(4.21±1.83)nmol/mgprot(P＜0.05和P＜0.01);而此時SOD水平則顯著下降,至胰島分離後仍處于較低水平,與灌註前相比,差異均有統計學意義(P＜0.05和P＜0.01).膠原酶灌註前,胰島結構完整;膠原酶灌註後,胰島週圍組織結構膨脹;消化後,胰島膜性結構被破壞,但能勉彊維持島狀結構;胰島分離後結構基本完整.結論 在提取胰島的過程中,膠原酶灌註及消化可引起胰島細胞凋亡,其機製可能與氧化損傷有關,抗氧化措施可作為移植前保護胰島的手段.
목적 탐토이도재획취、분리급순화등과정중출현세포조망적상관인소급응대책략.방법 경공자복주동맥관주4℃적UW액후,절취공자이선,병진행효원매관주、소화급제도리심,분리、제취이도.분별우효원매관주전、관주후、소화후급이도분리후등시점,채용원위말단표기법(TUNEL법)검측이도세포조망정황,채용비색법검측초양화물기화매(SOD)여병이철(MDA)수평,채용HE염색화쌍류종염색관찰이도적형태학개변.결과 재분리、순화이도적각개과정중,균출현료이도세포조망적현상,기중재효원매관주후화소화후최위명현,병반수MDA적고표체,표체수평분별위(6.18±2.38)nmol/mgprot화(9.21±2.75)nmol/mgprot,균명현고우관주전적(4.21±1.83)nmol/mgprot(P＜0.05화P＜0.01);이차시SOD수평칙현저하강,지이도분리후잉처우교저수평,여관주전상비,차이균유통계학의의(P＜0.05화P＜0.01).효원매관주전,이도결구완정;효원매관주후,이도주위조직결구팽창;소화후,이도막성결구피파배,단능면강유지도상결구;이도분리후결구기본완정.결론 재제취이도적과정중,효원매관주급소화가인기이도세포조망,기궤제가능여양화손상유관,항양화조시가작위이식전보호이도적수단.
Objective To observe the changes of islet cell apoptosis and oxidation-antioxidation before the transplantation, and to explore the pathways of islet protection. Methods Fifteen human pancreases were perfused with the Hanks solution containing collagenase, then digested and isolated. During the procedure, islet cell apoptosis was detected by TUNEL, SOD and MDA in the pancreas were measured by colorimetric method, and the morphologic changes were observed by H-E staining and dithizone staining. Results In the procedure of human islet isolation, especially in the stage of digestion, the apoptosis of human islet cells occurred. In the stages of perfusion and digestion, the MDA contents reached the high levels (6. 18 ± 2. 38 and 9. 21 ± 2. 75 umol/mg protein respectively),and the structures of the islets and tissues around the islets were damaged. Conclusion In the stages of perfusion and digestion, apoptosis of islet cells can be caused by oxidation. It suggests that antioxidation is a pathway for protection of islets before transplantation.
