水通道基因治疗小型猪腮腺放射损伤的研究  

Adenoviral-mediated transfer of the human aquaporin-1 cDNA to irradiated minipig parotid glands

作　　者：单兆臣[1] 李钧[1] 刘晓勇[2] 张春梅[1] Baum BJ 王松灵[1] 

机构地区：[1]首都医科大学口腔医学院基因治疗分子生物实验室,北京100050 [2]首都医科大学口腔医学院病理科,北京100050 [3]美国国立卫生研究院

出　　处：《北京口腔医学》2005年第3期141-144,共4页Beijing Journal of Stomatology

基　　金：国家自然科学基金资助项目(30271400);国家杰出青年科学基金资助项目(30125042)

摘　　要：目的研究腺病毒介导水通道基因对小型猪腮腺放射损伤的治疗效果及其相应表达关系。方法19只实验用小型猪一侧腮腺给予20Gy放射剂量照射,照射后17周,转导不同浓度的水通道蛋白基因、荧光素酶基因和病毒缓冲液,分析唾液流率变化和目的基因表达。结果109pfu AdCMVhAQP1转导后3天和7天腮腺唾液流率明显增加,分别恢复到放射前腮腺唾液流率81±18%(P=0.024)和69±20%(P=0.058),14天下降到转导前水平,对照组均未见腮腺唾液流率明显变化。转导109pfu AdCMVhAQP1组免疫组织化学显示,AQP1主要表达在导管上皮,Western blotting分析有外源性水通道蛋白表达,而对照组未见AQP1表达。结论109pfu AdCMVhAQP1明显增加小型猪腮腺放射损伤唾液流率,转导的水通道基因具有潜在治疗涎腺放射损伤的临床应用价值。Objective To evaluate the effect of adenoviral-mediated transfer of human aquaporin-1 cDNA to irradiated miniature pig （minipig）parotid glands on parotid saliva output and the expression of aquaporin-1 cDNA. Methods Nineteen minipigs were subjected to 20Gy IR directed at one parotid gland. At 17 weeks after irradiation, minipigs were administered either infusate buffer, or infusate buffer containing either the AdS vectors AdAQP1 or AdLuc. Parotid flow rate was measured, immunohistochemical and Western blot assessment were performed on day 3 and 14 after vector delivery. Results Three days following administration of 10^9 pfu of AdAQP1, a highly significant increase in parotid saliva output was observed （ P = 0. 024）. On average, the level of saliva output achieved following AdAQP1 was 81 ± 18% of pre-IR values. By day 7 following AdAQP1 delivery, parotid saliva output had decreased to 69 ± 20% of pre-IR values （P = 0. 058） , and by day 14 was further reduced. Conversely, there was no significant change in saliva output in other groups. Immunohistochemica] detection showed that 3 days following administration of 10^9 pfu AdAQP1, aquaporin-1 protein was expressed mainly in ductal cells. Membranes from AdCMVluc-infected tissue had a background level of immunoreactive AQP1, with a major band of -28 Kda, likely representing the endogenous porcine AQP1 present in capillaries and venules, while membranes from tissue administered with 10^9 pfu AdhAQPI showed a higher level of this immunopositive band, indicating hAQP1 transgene expression in the transduced salivary glands. Conclusion Localized delivery of AdAQP1 to IR-damaged salivary glands can significantly increase parotid flow rate and may be clinically used for patients with IR-induced salivary hypofunction.

关 键 词：基因转导 放射损伤 小型猪 水通道蛋白 

分 类 号：R818[医药卫生—放射医学]