The Study Of The Relation Between Triptolide Inhibits Peripheral Blood Mononuclear Cell To Secret IL-1β, IL-10 And Their Gene Polymorphism

(One)The correlation of the inhibitory effect of triptolide on interleukin—lβproduction from peripheral blood mononuclear cells with the gene polymorphisms of interleukin—lβObjective:To explore whether the inhibitory effect of triptolide(TP) on interleukin(IL)-1βproduction by peripheral blood mononuclear cells (PBMC) is associated with IL-1βgene polymorphisms. Methods:IL-1βgene polymorphism was analyzed in 31 healthy volunteers and 37 patients with rheumatoid arthritis(RA). From genomic DNA, the C-T polymorphism at IL-1β-511 was typed by PCR-RFLP. Meanwhile PBMC were cultured and stimulated by LPS and TP.Then IL-1βprotein was measured in the supernatants of the cultured and stimulated PBMC by ELISA, and IL-1βmRNA was detected in PBMC from 11 patients with RA by real-time fluorescent quantitative PCR. Results:After LPS stimulation in PBMC cultures of healthy volunteers and RA patients, the secretion levels of IL-1βin subjects who carried IL-1β-511 T/T genotype were higher than those in subjects who are not T/T genotype (P<0.05). In general, TP suppressed the production of IL-1βmRNA and protein significantly in LPS-treated human PBMC. In different genotype groups,low-dose TP suppressed the secretion of IL-1βsignificantly in LPS-treated human PBMC carried non-T/T genotype (P<0.05), but this significant inhibitory effect of TP was not seen in T/T genotype (P>0.05). Conclusion:The gene polymorphism at IL-1β-511 was related to the production of IL-1β, and the inhibitory effect of TP on the production of IL-1βwas different in non-T/T and T/T genotype of IL-1β-511, which may be one of the reasons that the therapy responses to Tripterygium wilfordii Hook.F.(TWHF) are different in different individuals.(Two) Inhibitory effect of triptolide on interleukin-10 production from peripheral blood mononuclear cells is associated with interleukin-10 gene–592A-C polymorphism Objective:To explore whether the inhibitory effect of triptolide(TP) on interleukin(IL)-10 secretion by peripheral blood mononuclear cells(PBMC)is associated with IL-10 gene–592 A-C polymorphism. Methods: IL-10 gene–592 A-C polymorphism was analyzed in 32 healthy volunteers by PCR-RFLP. Meanwhile IL-10 was also measured in the supernatant of the cultured and stimulated PBMC by ELISA. Results:In 32 subjects, 15 were A/A genotype, 17 subjects were non-A/A genotype. There was a tendency that the secretion level of IL-10 in non-A/A group was higher than in A/A group after LPS stimulation in PBMC cultures of healthy subjects,but there was not a statistical significance between the two groups. TP suppressed the production of IL-10 significantly in LPS-treated human PBMC (P<0.01). Furthermore, stronger suppressive effect was found in PBMC carried non-A/A genotype than in PBMC carried A/A genotype (P<0.05). Conclusion:The inhibitory effect of TP on the production of IL-10 is different between A/A and non-A/A genotype of IL-10–592 (stronger in non-A/A genotype), which may be one of the reasons for the phenomenon that patients respond to Tripterygium wilfordii Hook.F.(TWHF) differently. (Three)Influence of triptolide on proinflammatory and anti-inflammatory cytokine produced by peripheral blood mononuclear cellsObjective : To explore the effects of triptolide (TP) on the proinflammatory and anti-inflammatory cytokine produced by Peripheral blood mononuclear cells (PBMC) from healthy people. Methods:PBMC from healthy people were cultured and stimulated by lipopolysaccharide (LPS) or phytohemagglutinin (PHA), and were treated with or without TP simultaneously. The levels of proinflammatory cytokine (TNF-α) and anti-inflammatory cytokine (IL-4 , IL-10) were measured in the supernatants of cultured and stimulated PBMC by ELISA. Results:After LPS or LPS/PHA stimulation, the levels of TNF-αand IL-10 in PBMC cultures treated by TP significantly decreased than in those treated without TP (P<0.05). But TP had no effect on the secretion of IL-4 in PBMC cultures stimulated by LPS/PHA (P>0.05). Moreover the inhibitory effect of TP on IL-10 secretion was stronger than on TNF-αsecretion. Conclusion:TP can inhibit the secretion of both proinflammatory cytokine and anti-inflammatory cytokine directly, and have more powerful effect on the anti-inflammatory cytokine (IL-10), which may change the state of proinflammatory and anti-inflammatory cytokine network.