TLR4在参苏饮及其拆方含药血清诱导炎症16HBE hBD-2mRNA表达中的作用机制研究  

Mechanism of action of Toll-like receptor 4 in mRNA expression of human β-defensin-2 in inflammatory 16HBE cells treated with drug-containing serum of Shensu drink and its separated recipes

作　　者：陈瑶[1] 马萍[1] 胡晋婷 徐春肖[1] 张发君 

机构地区：[1]成都中医药大学,四川成都610075

出　　处：《湖南中医杂志》2017年第2期123-125,共3页Hunan Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(编号:81173365)

摘　　要：目的:使用益气解表代表方参苏饮(《太平惠民和剂局方》)及其拆方的含药血清对炎症16HBE进行干预,从中探讨TLR4在hBD-2mRNA表达中的作用。方法:传代培养16HBE细胞后将其分为5组,用LPS建立细胞炎症模型,加入TLR4抑制剂Anti-Human CD284后分别将培养液替换为含有提前制备的参苏饮及其拆方含药血清培养液,在给药后不同时间点(1/2h,1h,3h,5h,8h)分别取出相应组别细胞培养板,提取细胞总RNA,使用real time PCR检测16HBE炎症模型中以上时间点NF-κBp65 mRNA、hBD-2 mRNA表达量。结果:与空白组比较,模型组NF-κBp65 mRNA相对表达量在1h、5h显著升高(P<0.05),而hBD-2 mRNA相对表达量仅在1h显著升高(P<0.05)。与模型组比较,参苏饮全方组NF-κBp65 mRNA、hBD-2 mRNA相对表达量仅在5h显著升高(P<0.05);益气解表组NF-κBp65 mRNA、hBD-2 mRNA相对表达量仅在3h显著升高(P<0.05)。结论:Anti-Human CD284阻断TLR4后,参苏饮诱导炎症16HBE表达NF-κBp65 mRNA、hBD-2mRNA明显受抑,表明参苏饮上调NF-κBp65 mRNA、hBD-2 mRNA需要TLR4参与。Objective： To investigate the effect of drug- containing serum of the representative qi- tonifying and exterior syndrome- relieving prescription Shensu drink（ Prescriptions of Peaceful Benevolent Dispensary） and its separated recipes on inflammatory 16 HBE cells and the role of Toll- like receptor 4（ TLR4）in the mRNA expres sion of human β- defensin- 2（ hBD- 2）. Methods： Subcultured 16 HBE cells were divided into five groups,and lipopolysaccharide was used to establish a cell inflammation model. After the TLR4 inhibitor anti- human CD284 was added,the culture solution was changed to the culture solution containing the drug- containing serum of Shensu drink or its separated. The cell culture plate was removed at different time points（ 1 /2,1,3,5,and 8 hours） after administration,total RNA was extracted,and real- time PCR was used to measure the mRNA expression of NF-κBp65 and hBD- 2 in the 16 HBE inflammation model. Results： Compared with the blank group,the model group had significant increases in the relative mRNA expression of NF- κBp65 at 1 and 5 hours（ P 0. 05） and a significant increase in the relative mRNA expression of hBD- 2 at 1 hour（ P 0. 05）. Compared with the model group,the Shensu drink whole prescription group had significant increases in the relative mRNA expression of NF- κBp65and hBD- 2 at 5 hours（ P 0. 05）,and the qi- tonifying exterior syndrome- relieving group had significant increases in the relative mRNA expression of NF- κBp65 and hBD- 2 at 3 hours（ P 0. 05）. Conclusion： After TLR4 is inhibited by anti- human CD284,Shensu drink significantly inhibits the mRNA expression of NF- κBp65and hBD- 2 in inflammatory 16 HBE cells,indicating that TLR4 is involved in the upregulation of the mRNA expression of NF- κBp65 and hBD- 2 by Shensu drink.

关 键 词：参苏饮 16HBE TLR4 HBD-2 mRNA 实验研究 

分 类 号：R285.5[医药卫生—中药学]