益气解毒方对转基因小鼠上皮细胞及基因影响  

Effects of Yiqijiedu formula on cell proliferation and genes expressive levels of epithelia in mice

作　　者：何迎春[1] 田道法[1] 卢芳国[1] 江洁琼[1] 贺安意[1] 

机构地区：[1]湖南中医药大学中西医结合学院,长沙410007

出　　处：《中国公共卫生》2009年第4期419-421,共3页Chinese Journal of Public Health

基　　金：国家自然科学基金(30672738;30572408);湖南省杰出青年基金(03JJY1006);湖南省科技厅国际合作项目(06WK3016);湖南省教育厅优秀青年基金(06B070);一般项目(07C473)

摘　　要：目的研究益气解毒方对TgN(p53mt-LMP1)/HT转基因小鼠的鼻腔和鼻咽粘膜上皮细胞增殖及肿瘤坏死因子受体相关因子2(TRAF2)和p16基因表达的影响。方法苏木素-伊红(H-E)染色法观察小鼠鼻腔和鼻咽粘膜上皮细胞增殖特征;免疫组织化学染色法检测小鼠鼻腔和鼻咽粘膜上皮细胞TRAF2、p16基因的表达水平,并采用蛋白印迹(Westernblot)对基因表达进行验证。结果与TgN(p53mt-LMP1)/HT小鼠诱癌生理盐水对照组(TIC组)比较,TgN(p53mt-LMP1)/HT小鼠诱癌益气解毒方干预组(TIM组鼻腔和鼻咽粘膜上皮增生不明显,癌前病变率显著下降(P<0.01);其TRAF2基因表达水平(TIC组、TIM组鼻腔的分别为129.45±9.53和160.96±11.04,鼻咽的分别为126.38±12.69和160.88±6.84)显著降低(P<0.01);p16基因表达水平(TIC组、TIM组鼻腔的分别为157.21±10.44和131.72±10.06,鼻咽的分别为155.41±11.77和132.63±10.22)显著增高(P<0.01)。TIM组的TRAF2和p16基因表达水平接近野生型C57BL/6J小鼠对照组(鼻腔和鼻咽的TRAF2基因表达水平分别为165.07±10.58和164.39±7.88,p16基因表达水平分别为129.62±12.16和129.12±11.67),差异无统计学意义。结论中药益气解毒方可阻逆二亚硝基哌嗪(DNP)对TgN(p53mt-LMP1)/HT小鼠鼻腔和鼻咽粘膜上皮细胞异常增殖的诱导效应,其阻逆作用的发挥与下调TRAF2基因表达和上调p16基因表达密切相关。Objective To investigate the effects of Yiqijiedu formula on cell proliferation of nasal and nasopharyngeal epithelia and the expressive levels of TRAF2 and p16 in TgN（ p53mt-LMP1 ）/HT mice. Methods The cell proliferative characteristics of nasal mucosa and nasopharyngeal epithelia was evaluated by H-E staining procedures and the expressive levels of TRAF2 and p16 genes were assayed by immunohistochemical methods. At the same time,the genes expressive levels were also identified by western blot. Results Compared with those of TgN（ p53mt-LMP1 ）/HT mice of normal saline （NS） group（ TIC group）, hyperplasia of nasal and/or nasopharyngeal epithelia was not obvious in TgN（p53mt-LMP1 ）/HT cancerous lesion-inducing mice with Yiqijiedu fromula（ TIM group） and the occurring rate of precancerous lesions in nasal and/or nasopharyngeal epithelia was reduced significantly （ P 〈 0. 01 ）. The TRAF2 expressive levels （ 129. 45 ± 9. 53, 160. 96 ± 11.04 of nasal cavity epithelia of TIC group, and TIM group; 126. 36 ± 12. 69,160. 88 ± 6. 84 in nasopharygeal epithelia） were decreased（ P 〈 0. 01 ） and p16 expressive levels（ 157. 21 ± 10. 44,131.72 ± 10. 06 in nasal epithelia of TIC and TIM group; 155.41 ±11.77,132.63 ± 10.22 in nasopharyngeal epithelia of TIC and TIM group） were increased markedly（ P 〈0.01 ）. TRAF2 and p16 expressive levels were similar to those of C57 BL/6J wild-type mice（ TRAF2 gene： 165.07 ± 10. 58 in nasal epithelia, 164. 39 ± 7.88 in nasopharygeal epithelia;p16 gene： 129. 62 ± 12. 16 in nasal epithelia and 129. 12 ± 11.67 in nasopharygeal epithelia） with no significant differece. Conclusion The effects of N,N＇-dinitrosopiperazinc（DNP） on nasal/nasopharyngeal epithelial abnormal proliferation of TgN（p53mt- LMP）/HT mice can be inhibited by Yiqijiedu formula herbal medicine. The effects of Yiqijiedu fromula is closely related with TRAF2 gene downregulation expression and p16 gene expression upregulation in nasal/nasopharyngeal epithelia

关 键 词：癌前病变 益气解毒方 He瘤坏死因子受体相关因子2(TRAF2) P16 

分 类 号：Q786[生物学—分子生物学]