中华糖尿病杂志
中華糖尿病雜誌
중화당뇨병잡지
CHINES JOURNAL OF DLABETES MELLITUS
2014年
1期
37-41
,共5页
魏玲玲%邓绍平%杨卯竹%钟明%黄孝伦
魏玲玲%鄧紹平%楊卯竹%鐘明%黃孝倫
위령령%산소평%양묘죽%종명%황효륜
糖尿病%异种移植%微囊化%成体猪胰岛分离%腹腔移植
糖尿病%異種移植%微囊化%成體豬胰島分離%腹腔移植
당뇨병%이충이식%미낭화%성체저이도분리%복강이식
Diabetes%Xenotransplantation%Encapsulation%Adult pig islet isolation%Peritoneal cavity transplantation
目的 探讨猪胰岛细胞微囊化后腹腔移植治疗小鼠糖尿病的效果.方法 采用胶原酶灌注消化法及不连续密度梯度离心法分离并纯化成体猪胰岛,并用海藻酸钠、多聚赖氨酸对猪胰岛进行微囊化.双硫腙(DTZ)染色观察分离胰岛纯度,葡萄糖刺激胰岛素释放实验检测纯化胰岛和微囊化胰岛体外功能.利用链脲佐菌素(STZ)200 mg/kg腹腔内注射,制作BALB/c小鼠1型糖尿病动物模型.成模后动物随机分为3组:猪胰岛微囊组、空微囊组和生理盐水对照组.每只小鼠腹腔内分别移植500个相应胰岛,检测移植前后小鼠的血糖变化.采用单因素方差分析法进行统计分析.结果 不连续密度梯度离心纯化后,得到胰岛的纯度为＞90％;碘化丙啶/双醋酸荧光素(PI/FDA)染色后,测定胰岛的活力为＞90％;葡萄糖刺激胰岛素分泌检测,分离纯化的胰岛和微囊化胰岛的刺激指数均＞2.0;将微囊化胰岛移植到STZ诱导的糖尿病小鼠腹腔内,能够使小鼠血糖恢复并维持正常超过60 d(＜9.3 mmol/L).与空囊移植组和生理盐水对照组比较,血糖为正常水平的控制时间明显延长(F=13.587,P＜0.05).病理学检查发现,微囊内胰岛形态基本完整,胰岛素染色为阳性.结论 微囊化胰岛腹腔移植后,能有效降低小鼠血糖,微囊具有较好的免疫隔离作用.
目的 探討豬胰島細胞微囊化後腹腔移植治療小鼠糖尿病的效果.方法 採用膠原酶灌註消化法及不連續密度梯度離心法分離併純化成體豬胰島,併用海藻痠鈉、多聚賴氨痠對豬胰島進行微囊化.雙硫腙(DTZ)染色觀察分離胰島純度,葡萄糖刺激胰島素釋放實驗檢測純化胰島和微囊化胰島體外功能.利用鏈脲佐菌素(STZ)200 mg/kg腹腔內註射,製作BALB/c小鼠1型糖尿病動物模型.成模後動物隨機分為3組:豬胰島微囊組、空微囊組和生理鹽水對照組.每隻小鼠腹腔內分彆移植500箇相應胰島,檢測移植前後小鼠的血糖變化.採用單因素方差分析法進行統計分析.結果 不連續密度梯度離心純化後,得到胰島的純度為＞90％;碘化丙啶/雙醋痠熒光素(PI/FDA)染色後,測定胰島的活力為＞90％;葡萄糖刺激胰島素分泌檢測,分離純化的胰島和微囊化胰島的刺激指數均＞2.0;將微囊化胰島移植到STZ誘導的糖尿病小鼠腹腔內,能夠使小鼠血糖恢複併維持正常超過60 d(＜9.3 mmol/L).與空囊移植組和生理鹽水對照組比較,血糖為正常水平的控製時間明顯延長(F=13.587,P＜0.05).病理學檢查髮現,微囊內胰島形態基本完整,胰島素染色為暘性.結論 微囊化胰島腹腔移植後,能有效降低小鼠血糖,微囊具有較好的免疫隔離作用.
목적 탐토저이도세포미낭화후복강이식치료소서당뇨병적효과.방법 채용효원매관주소화법급불련속밀도제도리심법분리병순화성체저이도,병용해조산납、다취뢰안산대저이도진행미낭화.쌍류종(DTZ)염색관찰분리이도순도,포도당자격이도소석방실험검측순화이도화미낭화이도체외공능.이용련뇨좌균소(STZ)200 mg/kg복강내주사,제작BALB/c소서1형당뇨병동물모형.성모후동물수궤분위3조:저이도미낭조、공미낭조화생리염수대조조.매지소서복강내분별이식500개상응이도,검측이식전후소서적혈당변화.채용단인소방차분석법진행통계분석.결과 불련속밀도제도리심순화후,득도이도적순도위＞90％;전화병정/쌍작산형광소(PI/FDA)염색후,측정이도적활력위＞90％;포도당자격이도소분비검측,분리순화적이도화미낭화이도적자격지수균＞2.0;장미낭화이도이식도STZ유도적당뇨병소서복강내,능구사소서혈당회복병유지정상초과60 d(＜9.3 mmol/L).여공낭이식조화생리염수대조조비교,혈당위정상수평적공제시간명현연장(F=13.587,P＜0.05).병이학검사발현,미낭내이도형태기본완정,이도소염색위양성.결론 미낭화이도복강이식후,능유효강저소서혈당,미낭구유교호적면역격리작용.
Objective To evaluate the effects of intraperitoneal transplantation of encapsulated adult pit islets on blood glucose of diabetic mice.Methods Pancreata were digested with collagenase P using Ricordi chamber and purified with discontinued density gradient centrifugation and subsequently cultured.Islets were encapsulated using alginate-polylysine polymers.The quantity and purity of islets was determined by manual and counter machine after dithizone (DTZ) staining.Glucose stimulated insulin secretion test was performed to test the function of purified islets and encapsulated islets in vitro.BALB/c mice were intraperitoneal injected with streptozocin (STZ) 200 mg/kg body weight to induce type 1 diabetes model.Diabetic mice were randomly divided into 3 groups:encapsulated pig islet group,empty capsule group and saline control group.Encapsulated capsules (equivilent to 500 pig islets),empty caplules and normal saline were respectively transplanted into peritoneal cavity of three groups.The blood glucose of all the mice were regularly monitored before and after transplantation.Results After purification of discontinued density gradient centrifugation,the mean purity of the islets was more than 90％.The viability of the islets was more than 90％ by propidium iodide (PI) and fluorescein diacetate (FDA) staining.Glucose-stimulated insulin secretion (GSIS) of purified islet and encapsulated islet was more than 2.0 in vitro.In vivo,after transplantation,encapsulated pig islets group had a significantly improved survival and biochemical profile,compared to mice transplanted with empty capsule group and saline control group.Blood glucose of encapsulated pig islets group significantly reduced more than 60 days (＜ 9.3 mmol/L),Duration time of normal blood glucose in encapsulated pig islets group was significantly longer,compared with empty capsules group and saline control group (F =13.587,P ＜ 0.05).The pathologic structure of encapsulated islets was normal,which of insulin staining was positive.Conclusions Encapsulated pig islet transplantation exerts good effects of reduction of blood glucose on diabetic mice and the capsules have good immuno-isolating function.
