| Objective: To confirm the existence of Telocytes(TCs)in the submandibular gland of mice and observe distribution and morphological,preliminary discussion the effects of Telocytes on salivary secretion of submandibular gland by imatinib mesylate's regulation,it lays a foundation for the study of the biological function of TCs in the submandibular gland and its clinical application.Methods: 1)We let the submandibular glands of healthy Kunming mice as the research object,the TCs immune-labellings C-kit/CD117 and CD34 were double labelled with immunofluorescence staining technology to localize TCs in the submandibular gland of mice.2)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.3)We established the imatinib mesylate drug model in 48 male Kunming mice of healthy and clean adult.The mice were divided into 5 groups:normal group(24),1week-,2week-,3week-and 4 week-medicine groups(6 in each group,24).Medicine groups were administered medicine(imatinib mesylate)by gavage at 80mg/(kg·d)daily,normal group were insteaded of same volume of PBS.In per week we take fresh submandibular glands from each group Kunming mice according to the time required for the experiment.Immunofluorescence staining technology was used to observe the change of network-like structure of TCs.Transmission Electron Microscopy was used to observe the change of the ultrastructure of TCs and Telopodes(TPs).Western blotting was used to evaluated the change of level of CD117/CD34 and?-Amy protein expression in submandibular gland.Results: 1)The immunofluorescence staining showed that TCs were widely distributed in connective tissues in submandibular gland and cell nucleus was large,cellular body was small with protrusions,or TPs.TPs were connected each other and formed network-like structure,surrounding acinus and ducts.Compared with the normal group,the network structure consisting of TCs protrusions in the medication groups were gradually reduced with the time of experiment.2)The electron microscopy observation showed TCs with small cellular body,few cytoplasm and larger nucleus,in which there are many organelles such as mitochondria and endoplasmic reticulum,ect.TPs with like-moniliform structure,which are alternately composed of podmers and podoms.These TPs connected with adjacent tissues and surrounded by extracellular vesicles.Compared with the normal group,the number of TPs and the intracellular organelles decreased but the intracellular vesicles increased with the time of experiment.3)Western blotting showed that within the experimental intervention time of imatinib mesylate,the level of CD117/CD34 protein expression were also declined.Compared with the normal group,the difference was statistically significant(p<0.01).The expression of ?-Amy protein represented by salivary amylase decreased with the prolongation of treatment time.Compared with the normal group,the difference was statistically significant(p<0.01).The level of?-Amy and CD117/CD34 protein expression positively correlated with each other.Conclusion: There are TCs in the submandibular gland of mice.Intervention of TCs with imatinib mesylate may suppress the salivary function of submandibular gland by affecting the structure,immunolabel and intercellular communication of TCs. |
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