润喉开音颗粒对体外培养人声带小结成纤维细胞细胞周期的影响(1)
第1页 |
参见附件。
摘 要: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细胞周期的影响,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人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、金嗓散结丸组及空白对照组,流式细胞仪检测各组细胞细胞周期。结果:润喉开音颗粒各浓度组细胞处于G0/G1期及G2/M期细胞比例均较空白对照组升高,处于S期细胞比例较空白对照组明显降低。结论:润喉开音颗粒可以将人声带小结成纤维细胞阻滞于G0/G1期及G2/M期,使DNA合成受阻,阻抑细胞的有丝分裂。
关键词:润喉开音颗粒;声带小结;成纤维细胞;细胞周期
中图分类号:R76
文献标识码:A
文章编号:1673-7717(2012)04-0786-03
Effects of Runhou Kaiyin Granule on Proliferation And Apoptosis of Human Vocal Nodules Fibroblast Cultured in Vitro
HUANG Lei1,ZHU Xuan-xuan1,SHEN Feng1,WANG Jin1,ZHU Wu-yuan2
(1.Jiangsu Hospital of TCM,Nanjiang 210029,Jiangsu,China;
2.Nanjing University of TCM,210029,Jiangsu,China)
Abstract:Objective:To observe the effects of Runhou Kaiyin Granule on cell cycle of Human vocal nodules fibroblast cultured in vitro and discuss the mechanisms of action. Methods:Human vocal nodules tissue obtained from patients with vocal nodules was rinsed by PBS-antibiotic solution,cut into 1-mm3 pieces,and incubated in 5% carbon dioxide at 37℃.A confluent monolayer was established within 14 days.For this experiment, fibroblast cells from three generations were used. The cell cycle of fibroblast cells was assessed by flow cytometry with PI staining.Results: The ratios of G0/G1 phase and G2 phase were higher than control group,the ratio of S phase was lower than control group.Conclusions: Runhou Kaiyin Granule can inhibit the proliferation and DNA synthesis of fibroblast cultured in vitro, which may be accomplished by blocking the cell cycle.
Key words:Runhou Kaiyin Granule;human vocal nodules; fibroblast; cell cycle
声带小结是耳鼻喉科常见病、多发病,为慢性喉炎的一种常见类型,以长期声音嘶哑伴喉部不适、干燥感,说话时有喉痛感为特征。其发病病因复杂,目前公认用声不当或过度用声是其发病的主要原因[1]。当各种病因刺激喉部后,声带发生血管扩张,经久可以发生黏膜下渗血积聚、间质性水肿及单核细胞为主的炎性渗出物。黏膜上皮可有散在的部分表皮剥脱,继之,病变部位有成纤维细胞侵入,致纤维组织增生和玻璃样变性,并伴有组织结构增厚和畸形,黏膜肿胀。其病理组织变化多以声带水肿、玻璃样变性或纤维化为主[2]。江苏省中医院耳鼻喉科在中医理论的指导下,使用润喉开音颗粒治疗声带小结,疗效显著。为探讨润喉开音颗粒治疗声带小结的作用机理,现观察润喉开音颗粒对体外培养人声带小结成纤维细胞细胞周期的影响。
1 实验材料及方法
1.1 实验材料
1.1.1 实验药物及试剂 DMED细胞培养液:GIBCO公司提供,批号:727635。胎牛血清:杭州四季青生物工程材料有限公司,批号:090512。青霉素:山东鲁抗医药股份有限公司,B081107。
链霉素:山东鲁抗医药股份有限公司, 081103。胰蛋白酶:美国Gibco公司,批号:2011/08。润喉开音颗粒:江苏省中医院制剂部提供,批号:20100308。金嗓散结丸:西安碑林药业股份有限公司,批号:200908226。细胞凋亡PI染色试剂盒:南京凯基生物,批号:KGA214。RNaseA:美国Sigma公司,2012/08。
1.1.2 实验仪器 HF151UV CO2培养箱:上海力申。倒置显微镜(XDS-1B):重庆光学仪器厂。SW-CJ-1F净化工作台:苏州安泰空气技术有限公司 ......
您现在查看是摘要介绍页,详见PDF附件(3841kb)。