Forskolin inhibited the secretion of interleukin－1β in activated human THP－1 macrophages
10.3760/cma.j.issn.2095-428X.2018.23.017
- VernacularTitle:佛司可林对活化的人单核巨噬细胞分泌白细胞介素-1β的抑制作用
- Author:
Yan CHEN
1
;
Jinjin FENG
;
Shaohua YAN
;
Yidong LI
;
Yunlong LI
;
Jianguo WEN
Author Information
1. 450052 郑州大学第一附属医院临床医学重点学科开放实验室;450052 郑州大学第一附属医院小儿尿动力学中心&泌尿外科
- Keywords:
Inflammation;
Forskolin;
Macrophage;
Inflammasome;
Interleukin－1β
- From:
Chinese Journal of Applied Clinical Pediatrics
2018;33(23):1823-1827
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of Forskolin on the activation of nucleotide－binding oligome-rization domain like receptor family,pyrin domain－containing 3(NLRP－3)inflammasome and the secretion of inter-leukin－1β(IL－1β)in activated human THP－1 macrophages,which can provide evidence for clinical treatment of inflammatory diseases in children.Methods Human monocyte cell line THP－1 cells were induced to differentiate into macrophages by Phorbol－12－myristate－13－acetate(PMA),and Forskolin(10,50,100 μmol/L)stimulated activa-ted macrophages by nigericin.The mRNA of the inflammasome markers NLRP－3,IL－1β and Caspase－1 were detec-ted by real－time quantitative polymerase chain reaction(qPCR)method.The protein of NLRP －3,pro －IL －1β, pro－Caspase－1 and Caspase－1 were detected by Western blot.The secretion of inflammatory cytokines IL－1β was detected by enzyme－linked immuno－sorbent assay(ELISA).Results Nigericin activated the cells and the mRNA expression of NLRP－3,IL－1β and Caspase－1 increased by 7.59 times(P＜0.001),579.10 times(P＜0.001)and 3. 59 times (P ＜0. 001 ),compared to non － activated cells;Forskolin had no effect on the mRNA expression of NLRP－3,Caspase－1 and IL－1β on activated THP－1 macrophages(P＞0.05).Western blot showed that Forskolin also had no effect on the protein expression of pro－Caspase－1 and pro－IL－1β in activated THP－1 macrophages (P＞0. 05),but the expression of NLRP －3 and Caspase －1 decreased significantly (P ＜0.001). The results of ELISA showed that the IL －1β secretion increased from basal 584. 0 nmol/L to activated 2 695. 6 nmol/L (t ＝16.031 1,P＜0. 001)on THP －1 macrophages by nigericin,but Forskolin(10,50,100 μmol/L)reduced it to 1 858. 4 nmol/L(t＝5.365 5,P ＜0. 001),1 467.9 nmol/L(t ＝8.047 5,P ＜0.001)and 1 246.7 nmol/L(t ＝10.199 0,P＜0.001)on the activated THP－1 macrophages.Conclusions Forskolin can not affect the expression of pro－cytokines at the gene transcription and protein translation levels in activated THP－1 cells,but inhibit the secre-tion of inflammatory cytokines IL－1β,so as to inhibit inflammatory response,which can be used to treat pediatric in-flammatory diseases caused by IL－1β.
