大鼠颌下腺腺病毒介导荧光素酶基因转导及组织学变化

Gene expression and pathological changes of rat submandibular glands after adenovirus-mediated gene transfer

目的 观察大白鼠颌下腺腺病毒介导的荧光素酶基因转导及其组织学变化。方法 将4 0只Wistar大鼠随机分为 5组 ,经颌下腺导管转导腺病毒介导荧光素酶基因重组体即AdCMVLuc ,3d及 1、2、4、8周后观察其基因表达及病理变化。结果 颌下腺转基因表达 3d时最高 ,以后逐渐下降 ,至 4周、8周时仍可测到表达。组织病理学变化 :3d～ 1周时可见腺泡受挤压 ,腺导管扩张 ;2周时部分腺泡轻度萎缩 ,腺泡间距离加大 ;小叶内及小叶间导管周围淋巴细胞呈灶状浸润 ;4周时可见有腺泡及腺泡腔结构变清晰 ;8周时腺泡、导管基本恢复正常 ,炎症不明显。超微结构变化 :注入基因后 3d ,腺泡和导管细胞内可见大量粗面内质网 ,粘液腺泡中粘液滴增多 ,融合成片 ;1周后 ,导管腔的微绒毛突起减少 ,胞质内可见线粒体显著增多 ;2周及 4周 ,腺泡腔可见少量的微绒毛突起及细丝网状物。粘液腺泡中可见粘液滴。基底部可见粗面内质网 ,基底膜增厚。结论 本研究报道了腺病毒介导的大鼠颌下腺基因转导后的超微结构变化 ,提示腺体蛋白合成体系功能活跃。大鼠涎腺基因转导能产生生物活性蛋白 。

Objective To investigate luciferase gene expression and pathological changes of submandibular glands (SMG)of rats after adenovirus mediated gene transfer Methods Adenovirus mediated luciferase gene (AdCMVLuc, 10 8pfu in 50 μl) was injected into SMG of forty wistar rats The SMGs were harvested for gene expression measurement and pathological study after 3 days,1?2?4 and 8 weeks Results Peak expression was observed in three days following administation of the vector However, gene expression in submandibular glands decreased rapidly The pathological changes induced by retrograde injection of AdCMVLuc included: after 3 days to one week, compression of acini, dilation of terminal ducts; after two weeks, slight atrophy of a part of acini, increase of interacinar distance and focal lymphocyte infiltration in lobules and interlobular ducts; after 4 weeks, recovery evidence was found in acini; after 8 weeks, normal acini and ducts were found The ultrastructural changes included: 3 days, much more rough endoplasmic reticulum was found both in acini and duct epithelial cells; a lot of mucus drops were found in acini; after 1 week, microvillus decreased in duct epithelial cells, mitochondria increased significantly in acini ; intercellular space was enlarged Conclusions Adenovirus mediated gene transfer can produce biological proteins and induce marked inflammatory changes in rat SMG The ultrastructural changes suggest high protein synthesis activity in the acinar cells after gene transfer.