中华器官移植杂志
中華器官移植雜誌
중화기관이식잡지
CHINESE JOURNAL OF ORGAN TRANSPLANTATION
2010年
12期
728-732
,共5页
田磊%周燕%贾新干%顾永耀%吴向华%李杰华
田磊%週燕%賈新榦%顧永耀%吳嚮華%李傑華
전뢰%주연%가신간%고영요%오향화%리걸화
胰岛移植%外分泌细胞%淀粉酶%α-1抗胰蛋白酶
胰島移植%外分泌細胞%澱粉酶%α-1抗胰蛋白酶
이도이식%외분비세포%정분매%α-1항이단백매
Islet transplantation%exocrine cells%amylase%α-1 antitrypsin
目的 探讨胰腺外分泌细胞对胰岛移植物的损伤作用及α-1抗胰蛋白酶(A1AT)对胰岛β细胞的保护作用.方法 (1)体内实验:消化成人尸体部分胰腺,人工挑选胰岛,并收集胰腺外分泌细胞.链脲霉素腹腔注射诱导Balb/c-Nu裸小鼠成为糖尿病小鼠.对照组(n=6)小鼠每只于左肾包膜下移植成人胰岛250个;共同移植组(n=7)小鼠每只分别于左肾包膜上、下极同时植入胰岛250个和等体积的胰腺外分泌细胞.持续检测受鼠血糖,28d后切取左肾,以免疫组织化学染色(SP法)观察左肾包膜下抗淀粉酶抗体的表达,并继续检测血糖.(2)体外实验.纯化胰岛组(n=6),每孔250个胰岛进行培养;非纯化胰岛组(n=6),每孔250个胰岛和等体积外分泌细胞同时培养;非纯化胰岛+A1AT组(n=6),每孔250个胰岛和等体积外分泌细胞同时培养,并加入A1AT0.5 mg/ml.48 h后,测定各孔胰岛中胰岛素含量和上清液中胰蛋白酶浓度.结果 (1)胰岛移植后,对照组和共同移植组受鼠血糖均逐步恢复正常.共同移植组较对照组血糖恢复正常时间延迟,组间比较差异有统计学意义(P＜0.01).切取受鼠左肾后5 d(移植后33 d),两组受鼠血糖均＞21 mmol/L.共同移植组可见肾包膜下有大量抗淀粉酶抗体阳性细胞,对照组少见.(2)纯化胰岛组胰岛素含量为(2.02±0.33)μg/孔,非纯化胰岛组为(1.13±0.27)μg/孔,非纯化胰岛+A1AT组为(1.68±0.17)μg/孔,各组间差异均有统计学意义(P＜0.01).纯化胰岛组胰蛋白酶浓度为(1.03±0.25)ng/ml,非纯化胰岛组为(6.92±1.21)ng/ml,非纯化胰岛+A1AT组为(3.23±0.55)ng/ml,各组间差异均有统计学意义(P＜0.01).结论 胰腺外分泌细胞与胰岛同时移植会延迟植入胰岛功能恢复正常的时间,这与腺泡细胞内胰蛋白酶被激活、释放有关;在胰岛、胰腺外分泌细胞共同培养时加入A1AT,可以缓解腺泡细胞分泌的蛋白酶对胰岛的破坏.
目的 探討胰腺外分泌細胞對胰島移植物的損傷作用及α-1抗胰蛋白酶(A1AT)對胰島β細胞的保護作用.方法 (1)體內實驗:消化成人尸體部分胰腺,人工挑選胰島,併收集胰腺外分泌細胞.鏈脲黴素腹腔註射誘導Balb/c-Nu裸小鼠成為糖尿病小鼠.對照組(n=6)小鼠每隻于左腎包膜下移植成人胰島250箇;共同移植組(n=7)小鼠每隻分彆于左腎包膜上、下極同時植入胰島250箇和等體積的胰腺外分泌細胞.持續檢測受鼠血糖,28d後切取左腎,以免疫組織化學染色(SP法)觀察左腎包膜下抗澱粉酶抗體的錶達,併繼續檢測血糖.(2)體外實驗.純化胰島組(n=6),每孔250箇胰島進行培養;非純化胰島組(n=6),每孔250箇胰島和等體積外分泌細胞同時培養;非純化胰島+A1AT組(n=6),每孔250箇胰島和等體積外分泌細胞同時培養,併加入A1AT0.5 mg/ml.48 h後,測定各孔胰島中胰島素含量和上清液中胰蛋白酶濃度.結果 (1)胰島移植後,對照組和共同移植組受鼠血糖均逐步恢複正常.共同移植組較對照組血糖恢複正常時間延遲,組間比較差異有統計學意義(P＜0.01).切取受鼠左腎後5 d(移植後33 d),兩組受鼠血糖均＞21 mmol/L.共同移植組可見腎包膜下有大量抗澱粉酶抗體暘性細胞,對照組少見.(2)純化胰島組胰島素含量為(2.02±0.33)μg/孔,非純化胰島組為(1.13±0.27)μg/孔,非純化胰島+A1AT組為(1.68±0.17)μg/孔,各組間差異均有統計學意義(P＜0.01).純化胰島組胰蛋白酶濃度為(1.03±0.25)ng/ml,非純化胰島組為(6.92±1.21)ng/ml,非純化胰島+A1AT組為(3.23±0.55)ng/ml,各組間差異均有統計學意義(P＜0.01).結論 胰腺外分泌細胞與胰島同時移植會延遲植入胰島功能恢複正常的時間,這與腺泡細胞內胰蛋白酶被激活、釋放有關;在胰島、胰腺外分泌細胞共同培養時加入A1AT,可以緩解腺泡細胞分泌的蛋白酶對胰島的破壞.
목적 탐토이선외분비세포대이도이식물적손상작용급α-1항이단백매(A1AT)대이도β세포적보호작용.방법 (1)체내실험:소화성인시체부분이선,인공도선이도,병수집이선외분비세포.련뇨매소복강주사유도Balb/c-Nu라소서성위당뇨병소서.대조조(n=6)소서매지우좌신포막하이식성인이도250개;공동이식조(n=7)소서매지분별우좌신포막상、하겁동시식입이도250개화등체적적이선외분비세포.지속검측수서혈당,28d후절취좌신,이면역조직화학염색(SP법)관찰좌신포막하항정분매항체적표체,병계속검측혈당.(2)체외실험.순화이도조(n=6),매공250개이도진행배양;비순화이도조(n=6),매공250개이도화등체적외분비세포동시배양;비순화이도+A1AT조(n=6),매공250개이도화등체적외분비세포동시배양,병가입A1AT0.5 mg/ml.48 h후,측정각공이도중이도소함량화상청액중이단백매농도.결과 (1)이도이식후,대조조화공동이식조수서혈당균축보회복정상.공동이식조교대조조혈당회복정상시간연지,조간비교차이유통계학의의(P＜0.01).절취수서좌신후5 d(이식후33 d),량조수서혈당균＞21 mmol/L.공동이식조가견신포막하유대량항정분매항체양성세포,대조조소견.(2)순화이도조이도소함량위(2.02±0.33)μg/공,비순화이도조위(1.13±0.27)μg/공,비순화이도+A1AT조위(1.68±0.17)μg/공,각조간차이균유통계학의의(P＜0.01).순화이도조이단백매농도위(1.03±0.25)ng/ml,비순화이도조위(6.92±1.21)ng/ml,비순화이도+A1AT조위(3.23±0.55)ng/ml,각조간차이균유통계학의의(P＜0.01).결론 이선외분비세포여이도동시이식회연지식입이도공능회복정상적시간,저여선포세포내이단백매피격활、석방유관;재이도、이선외분비세포공동배양시가입A1AT,가이완해선포세포분비적단백매대이도적파배.
Objective To investigate the protective effects of α-1 antitrypsin on human islets injured by protease released from pancreas exocrine cells. Methods ( 1 ) in vivo experiment. Parts of the cadaveric pancreas was digested with collagenase, islets were selected artificially, and pancreatic exocrine cells were collected. 8-9 weeks olds male BALB/c-Nu nude mice were induced into diabetic mice with STZ (240 mg/kg body weight, i. p) and randomly divided into two groups: the control group (n = 6), 250 islets were transplanted into left kidney subcapsule of diabetic nude mice; cotransplant group (n = 7), 250 islets and the equal volume of pancreatic exocrine cells were transplanted into different regions of left kidney subcapsule. Blood glucose level was monitored. Nephrectomies were performed after 28 days. The expression of anti-amylase antibodies in subcapsule was detected by using immunohistochemical staining. (2) Islets culture: Three groups were randomly set up. Group 1: purified islet group, 250 islets were incubated into a 6-well culture plate; Group 2: non-purified islet group, 250 purified islets and equal volume of exocrine cells were incubated; Group 3: nonpurified islet + Al AT group, 250 purified islets and equal volume of exocrine cells were incubated with α-1 antitrypsin added (0. 5 mg/ml). After 48 h, insulin content of islets in each well and trypsin concentration in the supernatant of each well were measured. Results 10000 islets were collected.After islets transplantation, the blood glucose levels in control and co-transplant groups were normal,but a delayed islet function in reversing diabetes was in the co-transplant group, and ehe mice in both groups became hyperglycemic after nephrectomy. A large number of anti-amylase antibody-positive cells were found in renal subcapsule in the co-transplant group while little seen in the control group.Insulin levels in the non-purified islet group were decreased as compared with purified islet group,those in the non-purified islet group + A1AT group were higher than in the non-purified islet group,but lower than in the purified islet groups. Trypsin concentration in the non-purified islet group was increased as compared with purified group, that in the non-purified islet group + A1AT group was lower than the non-purified islet group, but higher than in the purified islets group (all P＜0. 01).Conclusion Protease released from acinar cells during pancreatic digestion has detrimental effect on islet function after transplantation. Co-cultivation of islets and pancreatic exocrine cells with A1AT added can prevent islet cell damage caused by trypsin.
