中国组织工程研究

中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (5): 743-747.doi: 10.3969/j.issn.2095-4344.2016.05.023

• 细胞损伤与修复动物模型 Animal models of cell injury and repair • 上一篇    下一篇

黄芪注射液干预人宫颈永生化上皮细胞体外实验模型的细胞凋亡变化

吕 玲1，肖晨光2，刘 青1，张 立3，李能莲3，舍雅莉3   

  1. 1甘肃省妇幼保健院，甘肃省兰州市 730050；2中核兰州铀浓缩有限公司职工医院，甘肃省兰州市 730065；3甘肃中医学院，甘肃省兰州市 730000
  • 收稿日期:2015-11-27 出版日期:2016-01-29 发布日期:2016-01-29
  • 通讯作者: 吕玲，硕士，主任医师，甘肃省妇幼保健院，甘肃省兰州市 730050
  • 作者简介:吕玲，女，1970年生，甘肃省庆阳市人，汉族，2008年兰州大学毕业，硕士，主任医师，从事妇科肿瘤研究。
  • 基金资助:
    甘肃省中医药管理局课题(GZK-2013-56)

Effects of astragalus injection on human immortalized cervical epithelial cell apoptosis in vitro 

Lv Ling1, Xiao Chen-guang2, Liu Qing1, Zhang Li3, Li Neng-lian3, She Ya-li3   

  1. 1Maternal and Child Health of Gansu Province, Lanzhou 730050, Gansu Province, China; 2Lanzhou Uranium Worker Hospital of China National Nuclear Corporation, Lanzhou 730065, Gansu Province, China; 3Traditional Chinese Medical College of Gansu, Lanzhou 730000, Gansu Province, China
  • Received:2015-11-27 Online:2016-01-29 Published:2016-01-29
  • Contact: Lv Ling, Master, Chief physician, Maternal and Child Health of Gansu Province, Lanzhou 730050, Gansu Province, China
  • About author:Lv Ling, Master, Chief physician, Maternal and Child Health of Gansu Province, Lanzhou 730050, Gansu Province, China
  • Supported by:

     the Traditional Chinese Medicine Administration Project of Gansu Province, No. GZK-2013-56

PDF

352

摘要：

文章快速阅读：

文题释义：

细胞永生化：是指体外培养的细胞经过自发的或受外界因素的影响从增殖衰老危机中逃离，从而具有无限增殖能力的过程。正常细胞的增殖能力有限，经过一定时间的增殖后，最终会进入一种生长抑制状态，而一些肿瘤细胞可以无限增殖。

黄芪：又名绵芪。多年生草本，高50-100 cm。主根肥厚，木质，常分枝，灰白色。茎直立，上部多分枝，有细棱，被白色柔毛。多年生草本，高50-100 cm。产于内蒙古、山西、甘肃、黑龙江等地。

 

背景：宫颈永生化上皮细胞 H8 在致癌因素诱导下可以发生癌变，当有协同因子共同作用时就会导致宫颈癌的发生。但临床上对于癌前病变的女性尚缺乏有效的干预治疗，此项治疗临床为空白。
目的：分析黄芪注射液对人宫颈永生化上皮细胞(H8细胞)凋亡的作用及机制。
方法：实验分2组，黄芪注射液药物组和空白对照组。①ELISA 法检测黄芪注射液作用后人宫颈永生化上皮细胞H8凋亡的 DNA 片断。②酶标仪分析黄芪注射液作用后人宫颈永生化上皮细胞H8中凋亡酶caspase-3、caspase-9 酶活性的变化。③Western blot 检测黄芪注射液作用后，人宫颈永生化上皮细胞H8中caspase-3、caspase-9、PARP 蛋白表达的变化。

结果与结论：①ELISA 法检测，20 g/L黄芪注射液分别作用0，6，12，24 h后，人宫颈永生化上皮细胞H8 DNA 片段随着药物作用时间的延长逐渐增多，且呈时间依赖性(P < 0.05)。②酶标仪检测，20 g/L黄芪注射液作用0，6，12，24 h后，人宫颈永生化上皮细胞H8 caspase-3和caspase -9 的活性增高，且呈时间依赖性(P < 0.05)。③20 g/L黄芪注射液分别作用0，6，12和24 h后，人宫颈永生化上皮细胞H8中cleaved caspase-3、cleaved caspase-9表达逐渐升高，差异有显著性意义(P < 0.05)；Cleaved PARP蛋白的表达逐渐下降，差异有显著性意义(P < 0.05)。结果说明，黄芪注射液对人宫颈永生化上皮细胞H8具有较显著的诱导凋亡作用，其机制可能与上调caspase-3和caspase-9蛋白表达有关。 

ORCID: 0000-0002-3808-126X(吕玲)

关键词: 实验动物, 细胞损伤与修复模型, 黄芪注射液, 宫颈永生化, 细胞凋亡

Abstract:

BACKGROUND: Immortalized cervical epithelial cells H8 can become cancerous under the induction of carcinogenic agent, and may cause cervical cancer when there is a cofactor interaction. However, there is still a lack of effective intervention for female patients with precancerous lesions, and this treatment is blank in the clinic. 
OBJECTIVE: To explore the effects and mechanism of astragalus injection on apoptosis of human immortalized cervical epithelial cells H8.
METHODS: This study contained two groups: astragalus drug group and the blank control group. (1) Enzyme linked immunosorbent assay (ELISA) was used to detect DNA fragments of apoptotic H8 after astragalus injection. (2) Enzyme-labeling instrument was used to analyze the changes in caspase-3 and caspase-9 activities a fter astragalus injection. (3) Western blot assay was used to detect the protein expression changes of caspase-3, caspase-9 and PARP in H8 cells after astragalus injection. 
RESULTS AND CONCLUSION: (1) ELISA results showed that at 0, 6, 12 and 24 hours after 20 g/L astragalus injection, DNA fragments were gradually increased with time prolonged in a time-dependent effect (P < 0.05). (2) Enzyme-labeling instrument demonstrated that at 0, 6, 12 and 24 hours after 20 g/L astragalus injection, caspase-3 and caspase-9 activities increased in a time-dependent manner (P < 0.05). (3) At 0, 6, 12 and 24 hours after 20 g/L astragalus injection, the expression of cleaved caspase-3 and cleaved caspase-9 were gradually increased in H8 cells (P < 0.05). Cleaved PARP protein expression was gradually decreased (P < 0.05). These findings indicate that astragalus injection could obviously induce H8 apoptosis, which may be associated with the upregulated protein expression of caspase-3 and caspase-9.