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目的研究毛果芸香碱对体外培养兔泪腺上皮细胞蛋白分泌与合成的影响。设计实验研究。研究对象培养的兔泪腺上皮细胞。方法对培养细胞进行广谱细胞角蛋白、波形蛋白免疫组化染色及电镜观察鉴定后,培养的兔泪腺上皮细胞中,分别加入700、70、7、0.7和0.07μmol/L的毛果芸香碱,培养24小时,分别测定培养液和细胞中的β-氨基已糖苷酶活性。在不同浓度毛果芸香碱的培养细胞中加入70μmol/L阿托品,培养24小时,分别测定培养液和细胞中的β-氨基已糖苷酶活性。主要指标β-氨基已糖苷酶活性(光密度值,OD)。结果毛果芸香碱在700、70、7、0.7和0.07μmol/L浓度时使泪腺上皮细胞培养液β-氨基已糖苷酶活性OD值分别增加17.7%(P=0.015)、14.7%(P=0.038)、5.7%(P=0.399)、4.3%(P=0.517)和2.0%(P=0.764),使泪腺上皮细胞冻融液β-氨基已糖苷酶活性OD值分别增加25.0%(P=0.000)、16.8%(P=0.001)、10.6%(P=0.023)、7.6%(P=0.089)和4.8% (P=0.271)。阿托品抑制毛果芸香碱的作用,使泪腺上皮细胞培养液β-氨基已糖苷酶活性OD值分别减少20.5%(P=0.018)、15.1%(P=0.029)、10.1%(P=0.097)、11.5%(P=0.118)和7.9%(P=0.085),使泪腺上皮细胞冻融液β-氨基已糖苷酶活性OD值分别减少10.0%(P=0.039)、4.8%(P=0.113)、6.2%(P=0.162)、2.9%(P=0.315)和0%(P=0.300)。结论毛果芸香碱可增加培养的泪腺上皮细胞的蛋白分泌和合成,阿托品可抑制毛果芸香碱的作用。
Objective To study the effect of pilocarpine on the secretion and synthesis of rabbit lacrimal gland epithelial cells cultured in vitro. Design experiment research. Rabbit lacrimal gland epithelial cells cultured in vitro Methods After the cultured cells were identified with broad-spectrum cytokeratin, vimentin immunohistochemistry and electron microscopy, the cultured rabbit lacrimal gland epithelial cells were treated with pilocarpine at 700, 70, 7, 0.7 and 0.07μmol / L, respectively. Hour, the β-aminohexosaminidase activity in the culture broth and in the cells was measured. The cells were cultured in different concentrations of pilocarpine 70μmol / L atropine, cultured for 24 hours, were measured in the culture fluid and cells β-aminohexosaminidase activity. The main index β-amino hexosaminidase activity (optical density value, OD). Results Pilocarpine increased the β-aminohexosaminidase activity OD by 17.7% (P = 0.015) and 14.7% (P = 0.038) at the concentrations of 700, 70, 7, 0.7 and 0.07 μmol / 5.7% (P = 0.399), 4.3% (P = 0.517) and 2.0% (P = 0.764), respectively, increased the OD value of β-aminohexosaminidase activity in lyophilized epithelial cells by 25.0% 16.8% (P = 0.001), 10.6% (P = 0.023), 7.6% (P = 0.089) and 4.8% (P = 0.271). The inhibitory effect of atropine on pilocarpine decreased the activity of β-aminohexosaminidase in cultured lacrimal gland epithelial cells by 20.5% (P = 0.018), 15.1% (P = 0.029), 10.1% (P = 0.097), 11.5% (P = 0.118) and 7.9% (P = 0.085) respectively. The OD values of β-aminohexosaminidase activity in the frozen-thawed lachrymal cells decreased 10.0%, 4.8% P = 0.162), 2.9% (P = 0.315) and 0% (P = 0.300). Conclusion Pilocarpine can increase the secretion and synthesis of cultured lacrimal gland epithelial cells, atropine can inhibit the role of pilocarpine.