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目的:观察润喉开音颗粒对体外培养人声带小结成纤维细胞增殖抑制及凋亡的影响,探讨润喉开音颗粒治疗声带小结的作用机制。方法:采用组织块贴壁法对临床摘取声带小结标本进行体外培养,有成纤维细胞游出并顺利传代者视为培养成功,将成纤维细胞传至第3代后用于正式实验。体外培养人声带小结成纤维细胞随即分为润喉开音颗粒组、金嗓散结丸组、曲安奈德组,倒置显微镜观察各组细胞生长情况并拍照记录,MTT法检测各组细胞增殖状况;Hoechst33258荧光染色法观察各组细胞凋亡情况,荧光倒置显微镜观察并拍照记录。结果:润喉开音颗粒能浓度依赖性地抑制体外培养人声带小结成纤维细胞的增殖,并能促进其凋亡。结论:润喉开音颗粒对声带小结的治疗作用与其抑制成纤维细胞的增殖及促进成纤维细胞凋亡有关。
Objective: To observe the effect of Ruhuan Kaiyin Granules on the proliferation inhibition and apoptosis of cultured human vocal nodule fibroblasts in vitro, and to explore the mechanism of action of Ruhuan Kaiyin Granule on the treatment of vocal nodules. Methods: The specimens of vocal nodules in clinical samples were cultured in vitro with tissue block adherent method. Fibroblasts were successfully aspirated and successfully passaged. The fibroblasts were passaged to the third generation and used for formal experiments. In vitro culture of vocal nodules fibroblasts then divided into Run Throat open sound particle group, Jinsang Sanjie pill group, triamcinolone acetonide group, inverted microscope to observe the growth of the cells in each group and take pictures recorded, MTT assay of cell proliferation Hoechst33258 fluorescence staining was used to observe the apoptosis of the cells in each group. Fluorescence inverted microscope was used to observe and take pictures. Results: Run-throat open-tone particles can inhibit the proliferation of vocal nodules fibroblasts in vitro and promote their apoptosis in a concentration-dependent manner. Conclusion: The therapeutic effect of “Throat Openings” particles on the vocal nodules is related to inhibiting the proliferation of fibroblasts and promoting fibroblast apoptosis.