目的 观察血必净对内毒素刺激的THP-1细胞的影响,并探究其是否具有诱导内毒素耐受的作用。方法 THP-1细胞先以不同浓度的血必净(10,25,50 mg/mL)作用不同时间(4,12,24 h),再用内毒素进行刺激。用ELISA法测定上清液中TNF-α水平,Real time-PCR技术检测TLR4和IRAK-M mRNA的表达差异。结果 不同浓度血必净不同时间作用下,各实验组细胞TNF-α分泌水平均无显著变化(P>0.05)。在高浓度(50 mg/mL)血必净组,作用24 h时细胞TLR4表达上调,是4 h时的1.547倍(P<0.05);在作用24 h后,50 mg/mL血必净组IRAK-M表达上调,是对照组的1.349倍(P<0.05);但其余各浓度及时间组细胞的TLR4和IRAK-M mRNA表达均无显著差异(P>0.05)。结论 血必净不能阻断内毒素刺激的THP-1细胞TNF-α的分泌,不能诱导内毒素耐受;高浓度血必净长时间作用后虽可能上调THP-1细胞TLR4和IRAK-M mRNA表达,但确切含义仍不清楚。
Objective To study the effect of Xuebijing, a Chinese traditional medicine injection, on the THP-1 cells challenged by endotoxin, and to explore whether it induces endotoxin tolerance. Methods The THP-1 cells were pretreated with Xuebijing in different concentrations (10, 25, 50 mg/mL) and times (4, 12, 24 hours), and then challenged by endotoxin. The level of TNF-α in culture supernatant was detected by ELISA assay, and the expression of TLR4 and IRAK-M mRNA were detected by Real time-PCR technique. Results There was no significant difference in TNF-α level among all the groups (pretreated with different concentrations of Xuebijing for different time) (P>0.05). Only in 50 mg/mL Xuebijing group, TLR4 mRNA was 1.547-fold increase in 24 h than in 4 h group (P<0.05). Only when pretreated for 24 h, IRAK-M mRNA was 1.349-fold increase in 50 mg/mL Xuebijing group than in control group (P<0.05). However,there was no significant difference among other groups (P>0.05). Conclusions Xuebijing can not block the release of TNF-α from the THP-1 cells challenged by endotoxin; and it does not induce endotoxin tolerance. When pretreated with high concentration of Xuebijing for long time, the expression of both TLR4 and IRAK-M mRNA is up-regulated, but its significance is not yet clear.