The effects of Forsythia suspense on the expression of Toll-like recepter 4, nuclear factor-κB and secretions of interleukin-6, interleukin-10 in the splenic lymphocytes of Wistar rats induced by endotoxin

Abstract

Objective To observe the effects of Forsythia suspense(FS) on the expression of Toll-like recepter 4(TLR4),nuclear factor-κB(NF-κB)and secretion of interleukin(IL)-6,IL-10 in the splenic lymphocytes of Wistar rats induced by Endotoxin(ET), to explore the anti-inflammatory and immunity modulation function of Forsythia suspense. Methods rats spleen and splenic lymphocytes were prepared by aseptic separation and the cells were suspended with DMEM and cultured in DMEM containing 20% fetal bovine serum.The experiment were divided into six groups:Control group: without any treatment; ET group:adding ET at final concentration of 10μg/L for 12 h; AFS1+ ET, AFS2+ ET, AFS3+ ET group:0. 5 h of AFS(50. 0, 25. 0, 12. 5 g/L) preconditioning before adding ET; Ploy B+ ET group:0. 5 h of Ploy B(10 mg/L in final concentration)preconditioning before adding ET.The expression of TLR4 and NF-κB m RNA on lymphocytes were measured by real-time quantitative polymerase chain reaction(Real-time PCR).The secretion levels of IL-6 and IL-10 in the lymphocytes supernatant were measured by enzyme linked immunosorbent assay (ELISA). Results Compared with Control group, the expression of TLR4, NF-κB m RNA and the secretion levels of IL-6, IL-10 were upregulated markedly in the ET group[0. 47±0. 01 vs.1. 38 ±0. 05, 0. 68±0. 09 vs.2. 37±0. 09,(16. 59±0. 23) vs.(65. 96±0. 26) ng/L,(9. 17±0. 14) vs. (21. 01±0. 16) ng/L, P 0. 05]. Conclusion Forsythia suspense preconditioning could inhibit the up-regulation the expression of TLR4, NF-κB mRNA and decrease the secretion levels of IL-6, IL-10 in the lymphocytes induced by ET, it was verified that Forsythia suspense has the function of anti-inflammatory and immunity modulation from the level of cells. Key words: Forsythia suspense; Endotoxin; Lymphocytes; Toll-like receptor 4; Cytokines