CAJ | 학술논문

目的探讨腺病毒载体介导激活性Akt1基因(Adv-CA-Akt1)转染大鼠胰岛对异种移植胰岛功能和存活的影响.方法以BALB/C糖尿病小鼠为受体,分离纯化雄性Wistar大鼠胰岛,体外培养,Adv-CA-Akt1转染后异种胰岛移植.受体小鼠分3组,实验组:Adv-CA-Akt1转染的大鼠胰岛体外培养24 h,小鼠肾被膜下移植,并口服环孢素A(CsA)30 mg·kg-1·d-1;CsA组:未转染胰岛移植,同剂量环孢素口服;对照组:单纯胰岛移植.每只接受300胰岛当量(IEQs)移植.检测术后血糖,移植物存活时间及组织病理学.结果实验组和CsA组术后2 d血糖即降至正常,胰岛功能存活时间分别为(21.0±3.65)d和(9.0±2.54)d,而对照组血糖短暂下降后再次升高,胰岛功能存活时间(4.2±2.6)d.实验组小鼠生存时间为(31.0±5.67)d比CsA组(17.0±3.35)d和对照组(10.0±1.52)d明显延长,三组比较差异有统计学意义(P<0.05);胰岛素免疫组化染色实验组.肾被膜下见较多有功能胰岛细胞团,而CsA组和对照组胰岛素染色阳性细胞数减少.结论 Adv-CA-Akt1转染大鼠胰岛联合应用免疫抑制剂,可提高胰岛功能,延长异种胰岛移植物存活时间.
목적 탐토선병독재체개도격활성Akt1기인(Adv-CA-Akt1)전염대서이도대이충이식이도공능화존활적영향.방법 이BALB/C당뇨병소서위수체,분리순화웅성Wistar대서이도,체외배양,Adv-CA-Akt1전염후이충이도이식.수체소서분3조,실험조:Adv-CA-Akt1전염적대서이도체외배양24 h,소서신피막하이식,병구복배포소A(CsA)30 mg·kg-1·d-1;CsA조:미전염이도이식,동제량배포소구복;대조조:단순이도이식.매지접수300이도당량(IEQs)이식.검측술후혈당,이식물존활시간급조직병이학.결과 실험조화CsA조술후2 d혈당즉강지정상,이도공능존활시간분별위(21.0±3.65)d화(9.0±2.54)d,이대조조혈당단잠하강후재차승고,이도공능존활시간(4.2±2.6)d.실험조소서생존시간위(31.0±5.67)d비CsA조(17.0±3.35)d화대조조(10.0±1.52)d명현연장,삼조비교차이유통계학의의(P<0.05);이도소면역조화염색실험조.신피막하견교다유공능이도세포단,이CsA조화대조조이도소염색양성세포수감소.결론 Adv-CA-Akt1전염대서이도연합응용면역억제제,가제고이도공능,연장이충이도이식물존활시간.
Objective To investigate the effect of constitutively active Akt1/PKB gene for rat islets transplantation on the function of the grafts and the life span of the mice. Methods Wistar islet was transfected with constitutively active Akt1 gene via adenovirus vector using MOI = 500. Each BALB/C streptozotucin induced diabetic mouse was transplanted with 300 IEQs rat islets under the capsule of kidney. Cyclosporin A (CsA) was administered at the same time in experimental group and CsA group, but not in control group. Blood glucose and life span were determined after operation. HE and immunohistochemical staining of insulin was used to evaluate the histology of the grafts. Results Glucose level in the experimental and CsA group returned to normal in 2 days after transplantation, and maintained for 21.0 ± 3.65 d and 9. 0 ± 2. 54 d respectively. The life span of mice in experimental group was longer than that of the other 2 groups (31.0 ± 5. 67 d and 17. 0 ± 3.35 d, 10. 0 ± 1.52 d, respectively) ( P < 0. 05 ). In control group, the blood glucose level was kept in normal in only 4. 2 ± 2. 6 d (P < 0. 05 ). Large amount of islet grafts were seen under the capsule of the kidney, which were positively stained by insulin-Ab. In the other two groups, the islet groups mass were lower, and few positively stained by insulin-Ab. Conclusion Constitutively active Akt1 gene can increase the function of the graft and prolong the life span effectively in xeon-genic islet transplantation.