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Effects of Phenylephrine on Transplanted Submandibular Gland

¹ Department of Oral and Maxillofacial Surgery, Peking University School and Hospital of Stomatology, Zhong Guan Cun South St. 22, 100081, Beijing, PRC;
² Department of Physiology and Pathophysiology, Peking University Health Science Center, 100083, Beijing;
³ Dalian Medical University School of Stomatology, 116011, Dalian; and
⁴ Institute of Vascular Medicine, Peking University Third Hospital and Key Laboratory of Molecular Cardiovascular Sciences, Ministry of Education, 100083, Beijing

View larger version (87K): [in this window] [in a new window]   Figure 1. Effects of phenylephrine on salivary secretion, mRNA expressions of 1-adrenoceptor subtypes, and localization of AQP5 in transplanted glands. Salivary flow (A), activity of -amylase (B), sodium concentration (C), and potassium concentration (D) of the saliva were measured as described in MATERIALS & METHODS. The transplanted glands were administrated phenylephrine from post-operative day 1. The expressions of 1A-, 1B-, and 1D-adrenoceptor mRNA were detected by RT-PCR in control, transplanted, and phenylephrine-treated glands (n = 6) (E,F,G). All values are given as means ± SEM. N, control submandibular gland; T, transplanted gland; T + PE, transplanted gland with phenylephrine treatment. *P < 0.05 and **P < 0.01 compared with N; #P < 0.05 compared with T. The reactivity to anti-AQP5 antibody was detected with FITC-conjugated secondary antibody. Sections were examined by confocal microscopy. (H) Control submandibular gland. AQP5 was located in both the apical membrane and the cytoplasm. (I) Transplanted gland. AQP5 was diffused in cytoplasm. (J) Translocation of AQP5 from cytoplasm to the apical membrane in phenylephrine-treated gland. Bars (H,I,J): 20 µm.

View larger version (204K): [in this window] [in a new window]   Figure 2. The histologic structure and ultrastructure of transplanted submandibular glands. All submandibular gland tissues were removed on post-operative day 7. (A) Histologic structure of a control submandibular gland. An acinar cell is indicated by an arrow. (B) Histologic structure of transplanted gland. Acinar cells show size reduction (arrow). (C) Histologic structure of phenylephrine-treated gland. Acinar cells show only mild size reduction (arrow). Light microscopy, magnification 400x. Bars (A,B,C): 20 µm. (D) Ultrastructure of control submandibular gland. Low-matrix-density secretory granule in acinar cell is indicated by an arrow. (E) Ultrastructure of transplanted submandibular gland. The low-matrix-density secretory granules (arrow) were decreased as compared with those in the control submandibular gland. (F) Ultrastructure of phenylephrine-treated gland. The low-density secretory granules (arrow) are similar to those in the control submandibular gland. TEM magnification, 5000x. Bars (D,E,F): 2 µm.

View larger version (83K): [in this window] [in a new window]   Figure 3. Alterations of PCNA and MAPK signal pathways in transplanted submandibular glands. Immunohistochemical localizations of PCNA in control submandibular gland (A), transplanted gland (B), and phenylephrine-treated gland (C) are shown. PCNA-positive cell is indicated by an arrow. Light microscopy, magnification 400x. Bars (A,B,C): 20 µm. (D) Semiquantitative scoring of PCNA-positive cells (n = 6). The PCNA-positive cells were counted in 10 different fields in each section (1 section/each animal) under 400x magnification. (E) Western blot analysis of phospho-PKC and PKC. An 80-µg quantity of protein was separated by 12% SDS-polyacrylamide gel electrophoresis and immunoblotted either with an antibody specific for phospho-PKC or for total PKC. (F) Western blot analysis of phospho-ERK1/2 and ERK1/2. An 80-µg quantity of protein was separated by 12% SDS-polyacrylamide gel electrophoresis and immunoblotted either with an antibody specific for phospho-ERK1/2 or for total ERK1/2. Filters were re-probed with antibody against actin to ensure equal loading of the lanes. The blot is a representative of 5 separated animals in each group, with similar results. All values are given as means ± SEM. N, control submandibular gland; T, transplanted gland; T + PE, transplanted gland with phenylephrine treatment. *P < 0.05 and **P < 0.01 compared with N; #P < 0.05 and ##P < 0.01 compared with T.

Journal of Dental Research, Vol. 85, No. 12, 1106-1111 (2006)
DOI: 10.1177/154405910608501208


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