鸡豆黄素A通过雌激素受体抑制LPS诱导的小胶质细胞活化及其机制

Bioch A inhibits LPS-induced microglial activation throughestrogen receptors and its mechanism

目的探讨鸡豆黄素A(biochanin,Bioch A)能否通过雌激素受体抑制LPS诱导的小胶质细胞活化及其机制的研究。方法BV2细胞分为:Control组、LPS(10 mg·L-1)组、Bioch A(5μmol·L-1)组、Bioch A(5μmol·L-1)+ICI(0.1μmol·L-1)组、ER(0.1μmol·L-1)组、ER(0.1μmol·L-1)+ICI(0.1μmol·L-1)组、ICI(0.1μmol·L-1)组、Rosup(0.1μmol·L-1)组、MAPK通路特异性抑制剂组。加入上述相应药物后,进行2 h预处理,除control组外,其余各组加LPS(10 mg·L-1)进行36 h孵育。测定NO的含量通过Griess法;检测ROS通过DCFH-DA探针法;ELISA法测定IL-1β、TNF-α、PGE2含量;Western blot法测定P47phox、gp91phox、iNOS、NF-κB、p-NF-κB以及p-ERK、p-JNK、p-p38表达水平。结果与LPS组相比,Bioch A组ROS、NO、IL-1β、TNF-α和PGE2含量下降,iNOS、p-ERK、P47phox、p-p38、p-JNK、p-NF-κB和gp91phox蛋白表达减少;Bioch A+ICI组上述指标未出现明显变化。结论通过雌激素受体,LPS诱导小胶质细胞的活化受到Bioch A抑制,其机制可能与抑制促分裂原活化蛋白激酶(MAPK)信号通路有关。

Aim To investigate whether the effect of biochanin A(biochanin,Bioch A)on LPS-induced microglia activation can be inhibited by estrogen receptor.Methods The BV2 cells were divided into:Control group,LPS group(10 mg·L-1),Bioch A group(5μmol·L-1),Bioch A(5μmol·L-1)+ICI group(0.1μmol·L-1),ER group(0.1μmol·L-1),ER(0.1μmol·L-1)+ICI group(0.1μmol·L-1),Rosup group(0.1μmol·L-1),and MAPK pathway specific inhibitor group.After adding the above drugs,2 h pretreatment was performed,and except the control group,all the other groups were added LPS(10 mg·L-1),incubated together for 36 h.NO content was determined using Griess reaction;ROS content was detected using DCFH-DA probe method;The content of IL-1β,TNF-αand PGE2 was detected by ELISA method.p47phox,gp91phox,iNOS,NF-κB,p-NF-κB and p-ERK,p-JNK,p-p38 expression levels were assessed by Western blot.Results Compared to LPS group,ROS,NO,IL-1β,TNF-αand PGE2 content and iNOS,p-ERK,P47phox,gp91phox,p-NF-κB,p-JNK and p-p38 of Bioch A group decreased;the above indicators showed no significant change in Bioch A+ICI group.Conclusions Bioch A inhibits the activation of LPS-induced microglial through estrogen receptors,and its mechanism may be related to the inhibition of MAPK signaling pathway.