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目的 研究转化生长因子β3(transforming growth factorβ3,TGF-β3)对鼠颌下腺细胞 (ratsubmandibular gland cells,RSGCs)分泌淀粉酶功能的影响。 方法 原代及传代培养 Wistar大鼠 RSGCs,免疫组织化学鉴定细胞来源 ,按加入不同浓度 TGF-β3(0 .5、1.0、5 .0、10 .0、2 5 .0 ng/ ml)分组 ,未加入 TGF-β3的为对照组。采用噻唑蓝 (MTT)比色法、自动生化分析仪和免疫印迹法检测 2 4、4 8、72和 96小时后 TGF-β3对细胞增殖及淀粉酶分泌功能的影响。 结果 培养的 RSGCs免疫组织化学鉴定 CK 8.13、S- 10 0蛋白染色呈阳性 ,波形丝蛋白染色呈阴性。 MTT法检测各组细胞吸光度 A值与对照组比较 ,差异无统计学意义 (P>0 .0 5 )。 72和 96小时后 ,0 .5～ 10 .0 ng/ ml TGF-β3组与对照组比较 ,细胞分泌的淀粉酶明显增加 ,差异有统计学意义 (P<0 .0 1)。免疫印迹法检测颌下腺组织及培养细胞的淀粉酶提取物形成蛋白表达一致的电泳带。 结论 TGF-β3能促进 RSGCs的分化和淀粉酶的分泌功能 ,但对细胞的增殖无明显影响
Objective To investigate the effect of transforming growth factorβ3 (TGF-β3) on amylase secretion in rat submandibular gland cells (RSGCs). Methods Primary and subculture Wistar rat RSGCs were harvested and cells were identified by immunohistochemistry. The cells were divided into groups according to the different concentrations of TGF-β3 (0 .5,1.0,5 .0,10 .0,25 .0 ng / ml) The control group did not join TGF-β3. The effects of TGF-β3 on proliferation and amylase secretion function were detected by MTT colorimetric assay, automated biochemical analyzer and Western blotting after 24, 48, 72 and 96 hours. Results The cultured RSGCs were immunohistochemically identified positive for CK 8.13 and S-10 0 protein and negative for silk fibroin staining. MTT assay for each group of cells absorbance A value compared with the control group, the difference was not statistically significant (P> 0.05). After 72 and 96 hours, compared with the control group, the amount of amylase secreted by the cells in the 0.5 ~ 10 .0 ng / ml TGF-β3 group was significantly increased (P <0.01). Immunoblotting was used to detect the amylase in the submandibular gland tissues and cultured cells to form the electrophoresis bands with the same protein expression. Conclusion TGF-β3 can promote the differentiation of RSGCs and the secretion of amylase, but have no significant effect on the proliferation of the cells