Apoptosis and Revascularization of Rat Islet Grafts Transfected by Adenovirus-Mediated Constitutively Active Akt1 Gene_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

LIU Fangfeng , CHENG Ying , LIU Yongfeng , SHI Rui , ZHAO Ning

* Department of General Surgery, Division of Transplantation, The First Hospital of China Medical University, Shengyang 110001, ChinaCorresponding Author: LIU Yong-feng, E-mail： yfliu@mail.cum.edu.cn;

Corresponding author：

Keywords：

Islet transplantation; Rat; Gene transfection; Diabetes【Foundation item】 Foundation of Department of Education of Liaoning Province （No.20061002）

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Objective To investigate the effect of constitutively active Akt1 gene on rat engrafted islets in apoptosis and revascularization, and to explore potential method of gene therapy in the islet transplantation. Methods Rat islet which was transfected constitutively actived Akt1 gene via adenovirus vector using MOI=500. Thirty-six streptozotocin induced diabetic Wistar rats were divided into 3 groups complete randomly: Adv-CA-Akt1 group, Adv-LacZ group and simple transplantation group. Blood glucose and insulin were determined after operation. TUNEL was used to detect the apoptotic islet cells. HE and immunohistochemical staining of insulin were used to evaluate the histology of the islet grafts. The microvessel density (MVD) was determined by CD31 immunohistochemical staining. Results The fasting glucose level in Adv-CA-Akt1 group restored to normal 2 days after transplantation. However, in Adv-LacZ group and simple transplantation group, it reduced but still kept being hyperglycemia. And the serum insulin level was higher than other two groups ( P ＜ 0.05). Compared to simple transplantation group and Adv-LacZ group, apoptotic rate decreased 25% in Adv-CA-Akt1 group, a large number of islet grafts were seen under the capsule of the kidney, which were positively stained by insulin antibody. In the other two groups, the islet groups mass were lighter, and few positively stained by insulin antibody. MVD showed lighter positive endothelial cells stained by CD31 antibody in the other two groups than Adv-CA-Akt1 group ( P ＜ 0.05). Conclusion Constitutively activate Akt1 gene can prolong graft survival during early posttransplant period, and can accelerate the revascularization of islet grafts effectively.

Citation： LIU Fangfeng,CHENG Ying,LIU Yongfeng,SHI Rui,ZHAO Ning,.. Apoptosis and Revascularization of Rat Islet Grafts Transfected by Adenovirus-Mediated Constitutively Active Akt1 Gene. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2009, 16(1): 28-31. doi: Copy

1.	徐泽宽，苗毅. 胰腺移植和胰岛移植的现状 [Ｊ] . 中国普外基础与临床杂志， 2003； 10(6)： 532.
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7.	霍鑫，冯金炜，刘方峰，等. pEGFP-C1/Akt体外转染骨髓间充质干细胞对后肢缺血大鼠血管生成的影响 [Ｊ] . 中国普外基础与临床杂志， 2008； 15(6)： 402.
8.	程颖，刘永锋，张佳林，等. 血管内皮生长因子促进大鼠的移植胰岛再血管化并提高其存活率 [Ｊ] . 中华器官移植杂志， 2007； 28(6)： 332.
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10.	Li Y, Li G, Dong W, et al . Transplantation of rat islets transduced with human heme oxygenase-1 gene using adenovirus vector [Ｊ] . Pancreas, 2006; 33(3)： 280.
11.	Bernal-Mizrachi E, Wen W, Stahlhut S, et al . Islet β cell expression of constitutively active Akt1/PKB α induces striking hypertrophy, hyperplasia, and hyperinsulinemia [Ｊ] . J Clin Invest, 2001; 108 (11)： 1631.
12.	Bernal-Mizrachi E, Fatrai S, Johnson JD, et al . Defective insulin secretion and increased susceptibility to experimental diabetes are induced by reduced Akt activity in pancreatic islet β cells [Ｊ] . J Clin Invest, 2004; 114(7)： 928.
13.	Tuttle RL, Gill NS, Pugh W, et al . Regulation of pancreatic beta-cell growth and survival by the serine/threonine protein kinase Akt1/PKBalpha [Ｊ] . Nat Med, 2001; 7(10)： 1133.
14.	Garcia-Ocana A, Takane KK, Reddy VT, et al . Adenovirus-mediated hepatocyte growth factor expression in mouse islets improves pancreatic islet transplant performance and reduces beta cell death [Ｊ] . J Biol Chem, 2003; 278(1)： 343.
15.	Richards SK, Parton LE, Leclerc I, et al . Over-expression of AMP-activated protein kinase impairs pancreatic beta-cell function in vivo [Ｊ] . J Endocrinol, 2005; 187(2)： 225.

1. 徐泽宽，苗毅. 胰腺移植和胰岛移植的现状 [Ｊ] . 中国普外基础与临床杂志， 2003； 10(6)： 532.
2. Woodgett JR. Recent advances in the protein kinase B signaling pathway [Ｊ] . Curr Opin Cell Biol, 2005; 17(2)： 150.
3. Martínez-Gac L, Alvarez B, García Z, et al . Phosphoinositide 3-kinase and Forkhead, a switch for cell division [Ｊ] . Biochem Soc Trans, 2005; 32(Pt 2)： 360.
4. Amaravadi R, Thompson CB. The survival kinases Akt and Pim as potential pharmacological targets [Ｊ] . J Clin Invest, 2005; 115 (10)： 2618.
5. Wideman RD, Yu IL, Webber TD, et al . Improving function and survival of pancreatic islets by endogenous production of glucagon-like peptide 1 (GLP-1) [Ｊ] . Proc Natl Acad Sci USA, 2006; 103 (36)： 13468.
6. Shiojima I, Walsh K. Role of Akt signaling in vascular homeostasis and angiogenesis [Ｊ] . Circ Res, 2002; 90(12)： 1243.
7. 霍鑫，冯金炜，刘方峰，等. pEGFP-C1/Akt体外转染骨髓间充质干细胞对后肢缺血大鼠血管生成的影响 [Ｊ] . 中国普外基础与临床杂志， 2008； 15(6)： 402.
8. 程颖，刘永锋，张佳林，等. 血管内皮生长因子促进大鼠的移植胰岛再血管化并提高其存活率 [Ｊ] . 中华器官移植杂志， 2007； 28(6)： 332.
9. Emamaullee JA, Shapiro AM. Factors influencing the loss of beta-cell mass in islet transplantation [Ｊ] . Cell Transplant, 2007; 16(1)： 1.
10. Li Y, Li G, Dong W, et al . Transplantation of rat islets transduced with human heme oxygenase-1 gene using adenovirus vector [Ｊ] . Pancreas, 2006; 33(3)： 280.
11. Bernal-Mizrachi E, Wen W, Stahlhut S, et al . Islet β cell expression of constitutively active Akt1/PKB α induces striking hypertrophy, hyperplasia, and hyperinsulinemia [Ｊ] . J Clin Invest, 2001; 108 (11)： 1631.
12. Bernal-Mizrachi E, Fatrai S, Johnson JD, et al . Defective insulin secretion and increased susceptibility to experimental diabetes are induced by reduced Akt activity in pancreatic islet β cells [Ｊ] . J Clin Invest, 2004; 114(7)： 928.
13. Tuttle RL, Gill NS, Pugh W, et al . Regulation of pancreatic beta-cell growth and survival by the serine/threonine protein kinase Akt1/PKBalpha [Ｊ] . Nat Med, 2001; 7(10)： 1133.
14. Garcia-Ocana A, Takane KK, Reddy VT, et al . Adenovirus-mediated hepatocyte growth factor expression in mouse islets improves pancreatic islet transplant performance and reduces beta cell death [Ｊ] . J Biol Chem, 2003; 278(1)： 343.
15. Richards SK, Parton LE, Leclerc I, et al . Over-expression of AMP-activated protein kinase impairs pancreatic beta-cell function in vivo [Ｊ] . J Endocrinol, 2005; 187(2)： 225.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Apoptosis and Revascularization of Rat Islet Grafts Transfected by Adenovirus-Mediated Constitutively Active Akt1 Gene

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