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甲磺酸伊马替尼调控Telocytes对下颌下腺唾液分泌功能的影响
摘要
目的探讨甲磺酸伊马替尼调控Telocytes(TCs)对下颌下腺唾液分泌功能的影响。方法采用免疫荧光染色技术对TCs免疫标记物(C-kit/CD117、CD34)进行双标记染色,进而对小鼠下颌下腺内TCs进行定位。采用透射电子显微镜(TEM)显示小鼠下颌下腺内TCs的超微结构及其与周围细胞之间的关系。建立甲磺酸伊马替尼干预模型,实验小鼠分5组:正常组(共24只),用药1周、2周、3周、4周组(共24只),药物以80 mg/(kg·d)每日灌胃,免疫荧光显示,用药前后下颌下腺内TCs的变化;免疫印迹法(Western blotting)观察用药前后CD117、CD34及下颌下腺内α唾液淀粉酶(α-Amy)蛋白表达水平的变化。结果免疫荧光染色显示,TCs广泛分布于下颌下腺结缔组织内,胞体较小有突起(Tps),Tps相互连接成网络样结构,包绕着腺泡及导管,随甲磺酸伊马替尼干预时间延长,用药组Tps构成的网络样结构变稀疏。超微结构显示,Tps呈念珠状,与邻近组织紧密相连,周围可见胞外囊泡。随用药时间的增加Tps减少,TCs胞内囊泡增多,胞内细胞器减少。免疫印迹法显示,用药后CD117、CD34、α-Amy蛋白表达水平相应减少且互为正相关。结论小鼠下颌下腺内存在TCs,甲磺酸伊马替尼对TCs的干预可能会通过影响TCs的结构、免疫表型和细胞间通讯降低下颌下腺唾液分泌功能。
Objective To explore the effect of Telocytes(TCs) on salivary secretion of submandibular gland by imatinib mesylate's regulation.Methods The TCs immune-labellings C-kit/CD117 and CD34 were double labelled with immunofluorescent staining technology to localize TCs in the submandibular gland of mice.The transmission electron microscopy(TEM) method was used in the present study to observe the ultrastructure of TCs and their relationship with neighboring cells in the submandibular gland of mice.The mice were divided into 5 groups:normal group(24),1-week,2-week,3-week and 4-week medicine groups(24),which were administered medicine at 80 mg/(kg·d) daily in order to study the effect of imatinib mesylate treatment on TCs function.Immunofluorescent staining technology was used to observe the change of TCs.Western blotting was used to evaluated the change of level of CD117/CD34/α-amylase(α-Amy) protein expression.Results The immunofluorescent staining showed that TCs were widely distributed in connective tissues in submandibular gland and cellular body was small with protrusions,or telopodes(Tps).Tps were connected each other and formed network-like structure,surrounding acinus and ducts.The network structure consisting of TCs protrusions in the medication groups were gradually reduced with the time of experiment.The electron microscopy observation showed that the TPs possess like-moniliform structure,connecting with adjacent tissues and being surrounded by extracellular vesicles.The number of TPs and the intracellular organelles decreased but the intracellular vesicles increased with the time of experiment.Western blotting showed that the level of CD117/CD34/α-Amy protein expression were also declined,and positively correlated with each other.Conclusion There are TCs in the submandibular gland of mice.Intervention of TCs withimatinib mesylate may reduce the salivary function of submandibular gland by affecting the structure,immunolabel and intercellular communication of TCs.
引文
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