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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 role of TLR4 in hBD-2 mRNA expression by interfering with 16HBE on behalf of Fangshen Suyin (“Taiping Huiminheji Decoction”) and its disassembled serum containing Yiqikang Table. Methods: The 16HBE cells were subcultured and divided into 5 groups. The model of cell inflammation was established by LPS. Anti-Human CD284, a TLR4 inhibitor, was used to replace the culture medium containing the advanced preparation of Shen Su Yin and its disassembled drug-containing serum The medium was taken out from the corresponding groups at different time points after administration (1 / 2h, 1h, 3h, 5h, 8h) to extract the total RNA of the cells. Real time PCR was used to detect the NF in the 16HBE inflammation model -κBp65 mRNA, hBD-2 mRNA expression. Results: Compared with the blank group, the relative expression of NF-κBp65 mRNA in model group was significantly increased at 1h and 5h (P <0.05), while the relative expression of hBD-2 mRNA was significantly increased at 1h (P <0.05). Compared with the model group, the expression of NF-κBp65 mRNA and hBD-2 mRNA in ShenShenYuQuanFang group increased significantly only 5h (P <0.05), while the expression of NF-κBp65 mRNA and hBD- The expression level increased significantly at 3h (P <0.05). Conclusion: Anti-Human CD284 blocks the expression of NF-κBp65 mRNA and hBD-2mRNA in 16HBE induced by ShenShenYin, indicating that ShenShuYin up-regulates NF-κBp65 mRNA and hBD-2 mRNA.