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1. 西安交通大学医学院第二附属医院:内分泌科,传染科
2. 第四军医大学基础医学部
纸质出版日期:2013,
网络出版日期:2013-09-15,
移动端阅览
王俊红, 王枫, 董鹏, 等. 双链腺相关病毒介导表达Exendin-4治疗糖尿病大鼠的研究[J]. 陆军军医大学学报, 2013,35(17):1831-1835.
WANG JUNHONG, WANG FENG, DONG PENG, et al. Efficiency of Exendin-4 expression mediated by a recombinant double-stranded adeno-associated virus vector in treatment of diabetic rats. [J]. Journal of army medical university, 2013, 35(17): 1831-1835.
王俊红, 王枫, 董鹏, 等. 双链腺相关病毒介导表达Exendin-4治疗糖尿病大鼠的研究[J]. 陆军军医大学学报, 2013,35(17):1831-1835. DOI:
WANG JUNHONG, WANG FENG, DONG PENG, et al. Efficiency of Exendin-4 expression mediated by a recombinant double-stranded adeno-associated virus vector in treatment of diabetic rats. [J]. Journal of army medical university, 2013, 35(17): 1831-1835. DOI:
目的 构建 、鉴定自身互补双链DNA的腺相关病毒(scAAV)重组载体,使其分泌表达Exendin-4,检测其转导效率并观察在糖尿病大鼠模型中的治疗作用。 方法 应用基因工程方法改建穿梭质粒pSSHG-CMV,插入外源性基因Exendin-4,构建重组scAAV载体,感染HEK293细胞,ELISA检测转染NIH3T3细胞上清Exendin-4滴度,链佐霉素诱导20只6周龄体质量180~220 g SD成年雄性大鼠为糖尿病鼠模型,逆向注射重组scAAV于糖尿病大鼠颌下腺,检测其血糖及胰岛素分泌水平。 结果 重组scpSSHG/exn4可有效包装和复制,病毒滴度为2.5×10
11
pfu/mL,转染细胞上清分泌Exendin-4浓度可达到4.53 ng/mL,scAAV治疗组血糖浓度在2、4周及8周均低于对照组[分别为(639.17±27.89)
vs
(396.00±34.00),(657.02±39.87)
vs
(315.62±42.56),(215.6±24.7)
vs
(458.6±19.7) mg/dL
]
,胰岛素浓度均高于对照组[分别为(156.8±24.5)
vs
(535.9±35.6),(236.5±12.3)
vs
(495.3±18.6),(620.43±46.90)
vs
(381.56±21.78) pg/mL
]
,二者比较有显著统计学差异(
P
<
0.05)。 结论 成功构建重组双链腺伴病毒scAAV-Ex-4,具有高效转导能力,对糖尿病大鼠模型具有控制血糖及增加胰岛素分泌作用。
Objective To construct and identity a self complementary double-stranded recombinant adeno-associated virus vector (scAAV) to secrete Exendin-4 (Ex-4). The transduction efficiency of the recombinant scAAV-Ex-4 was evaluated and its efficacy in tre
atment of diabetic rats was observed. Methods The shuttle vector pSSHG-CMV inserted with Ex-4 gene was constructed by genetic engineering method. The scAAV-Ex-4 vector was co-transferred into HEK-293 cells. Ex-4 level in the supernatants from transfected NIH3T3 cells was determined by an enzyme immunoassay kit. The rat model of type 2 diabetes (T2DM) was successfully induced by streptozotocin in twenty six-week-old Sprague-Dawley (SD) male rats (160-180 g). The recombinant scAAV was injected into the salivary gland of T2DM model rats. The levels of blood glucose and insulin of the rats were assessed. Results The recombinant scAAV-Ex-4 was successfully constructed and packaged with a high titer of virus (2.5×10
11
pfu/L). The Ex-4 concentration of the supernatants from transfected cells could be 4.53 ng/mL. The mean levels of blood glucose and insulin at 2
4 and 8 weeks were significantly different between scAAV treatment groups and control groups (
P
<
0.05). The mean level of blood glucose in scAAV treated rats was lower than that in the control group (639.17±27.89
vs
396.00±34.00
657.02±39.87
vs
315.62±42.56
215.6±24.7
vs
458.6±19.7 mg/dL)
but the mean level of insulin was higher (156.8±24.5
vs
535.9±35.6
236.5±12.3
vs
495.3±18.6
620.43±46.90
vs
381.56±21.78 pg/mL). Conclusion The scAAV-Ex-4 can improve glycemic control and increase insulinotropic effect in T2DM model rats.
双链AAVExendin-42型糖尿病大鼠
double-stranded AAVExendin-4type 2 diabetic rats
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