清热除痹汤含药血清对尿酸钠结晶刺激下THP-1细胞活性及分泌白细胞介素-1β的影响

Influence of Serum Containing Qingre Chubi Decoction on THP-1 Cell Viability and Interleukin-1β Release Stimulated by Monosodium Urate Crystals

【目的】探讨清热除痹汤含药血清对尿酸钠结晶刺激下THP-1细胞的增殖活性及分泌白细胞介素-1β(IL-1β)功能的影响。【方法】体外培养人单核细胞THP-1细胞,分为5组,空白血清组,模型对照组,中药血清高、中、低浓度组(浓度分别为体积分数20%、10%、5%),除空白血清组外,其他各组均加入浓度为500 mg/L的尿酸钠结晶,于培养0、12、24、48 h时间点采用四甲基偶氮唑盐比色(MTS)法检测细胞的增殖活性,培养48 h后采用酶联免疫吸附(ELISA)法检测细胞上清液IL-1β含量。【结果】各组THP-1细胞活性均随着时间的延长而增加,模型对照组各时间点细胞活性均较空白血清组显著增高(P<0.05或P<0.01),48 h模型对照组的IL-1β水平较空白血清组显著增高(P<0.01)。12、24 h中药血清各浓度组,48 h中药血清高、中浓度组细胞活性均较模型对照组显著下降(P<0.05或P<0.01),48 h中药血清各浓度组IL-1β水平均较模型对照组显著下降(P<0.01)。【结论】清热除痹汤含药血清对尿酸钠结晶刺激下THP-1细胞的活性有抑制作用,机制与其可抑制IL-1分泌有关。

Objective To investigate the influence of serum containing Qingre Chubi Decoction （ QCD） on the THP-1 cell viability and the release of interleukin 1 beta （ IL-1β） stimulated by monosodium urate crystals in vitro. Methods The cultured human monocyte THP-1 strain were divided into blank serum group, model control group, and high-, middle- and low-concentration （ volume fraction being 20%, 10%, 5%） QCD-containing serum groups. Except for the blank serum group , the other groups were all given 500 mg/L of monosodium urate crystals. On culturing hour 0, 12, 24 and 48, THP-1 cell viability was tested by methy1 thiazolyl tetrazolium celorimetry （ MTS） method. On culturing hour 48, the content of IL-1β in the supernatant of THP-1 cells was detected by enzyme-linked immunosorbent assay （ ELISA） . Results The THP-1 cell viability in various groups was increased along with the prolongation of culturing time. The THP-1 cell viability in the model control group was increased as compared with that in the blank serum group at＆amp;nbsp;different time points （P〈0.05 or P〈0.01） . And the content of IL-1β in the model control group was increased significantly as compared with that in the blank serum group on culturing hour 48 （P〈0.01） . The THP-1 cell viability in various QCD-containing serum groups on culturing hour 12 and 24, and in high- and middle-concentration QCD-containing serum group on culturing hour 48 was decreased significantly as compared with that of the model control group at the same time point （ P〈0.05 or P〈0.01） . The content of IL-1β in various serum containing QCD groups was markedly decreased as compared with that in the model control group on culturing hour 48 （ P〈0.01） . Conclusion Serum containing QCD can inhibit the viability of THP-1 cells stimulated by monosodium urate crystals, and the possible mechanism is related with the inhibition of IL-1 release.