摘要
目的研究酸枣仁汤(SZRD)含药血清对NG2细胞分泌IL-1β、TNF-α和BDNF的影响及其机制。方法制备SZRD含药血清,免疫荧光鉴定NG2细胞;CCK-8法筛选SZRD含药血清作用浓度和时间,将NG2细胞分为正常血清组,SZRD含药血清组和艾司唑仑组。ELISA法检测细胞上清中IL-1β、TNF-α和BDNF的蛋白含量,RT-PCR法检测细胞IL-1β、TNF-α、BDNF、cAMP和EpacmRNA的表达,WB法检测细胞IL-1β、TNF-α、BDNF、cAMP和Epac蛋白的表达。结果与同体积分数的正常血清组比较,10%SZRD含药血清组细胞IL-1β mRNA和蛋白的表达增加(P<0.05),BDNF mRNA和蛋白的表达降低(P<0.05);20%SZRD含药血清组细胞IL-1β、TNF-α、BDNFmRNA和蛋白的表达增加(P<0.05);30%SZRD含药血清组细胞IL-1β、TNF-α、BDNF、cAMP和Epac mRNA和蛋白的表达增加(P<0.05);艾司唑仑组细胞IL-1β、TNF-α、BDNF、cAMP、Epac mRNA和蛋白表达均显著增加(P<0.01)。结论SZRD可影响NG2细胞分泌和表达IL-1β、TNF-α和BDNF活性物质及上调cAMP/Epac信号通路mRNA和蛋白的表达。
Objective To study the effect of sour date seed soup(SZRD)containing serum on the secretion of IL-1β、TNF-α and BDNF by NG2 cells and its mechanism.Methods SZRD-containing serum was prepared and NG2 cells were identified by immunofluorescence;the concentration and duration of action of SZRD-containing serum were screened by CCK-8 method,and NG2 cells were divided into normal serum group,SZRD-containing serum group and eszopiclone group.The protein content of IL-1β、TNF-α and BDNF in cell supernatant was detected by ELISA,and the expression of IL-1β、TNF-α、BDNF、cAMP and Epac mRNA in cells was detected by RT-PCR.TNF-α、BDNF、cAMP and Epac mRNA expression by RT-PCR,and IL-1β、TNF-α、BDNF、cAMP and Epac protein expression by WB method.Results Compared with the normal serum group with the same volume fraction,the expression of IL-1β mRNA and protein in the 10%SZRD-containing serum group increased(P<0.05)and BDNF mRNA and protein decreased(P<0.05);the expression of IL-1β、TNF-α、BDNF in the 20% SZRD-Containingserum groups increased(P<0.05),the expression of IL-1β、TNF-α、BDNF、cAMP and Epac in the 30% SZRD-containing serum groups increased(P<0.05);the expression of IL-1β、TNF-α、BDNF、cAMP and Epac in the eszopiclone group increased significantly(P<0.05).mRNA and protein expression were increased(P<0.05);IL-1β、TNF-α、BDNF、cAMP、Epac mRNA and protein expression were significantly increased in the eszopiclone group(P<0.01).Conclusion SZRD can affected the secretion and expression of IL-1β、TNF-α and BDNF active substances and upregulated the expression of mRNA and protein of cAMP/Epac signaling pathway in NG2 cells.
作者
杨婷婷
王慧
石鹏
滕柳
YANG Ting-ting;WANG Hui;SHI Peng;TENG Liu(Guizhou University of Traditional Chinese Medicine,Guiyang 550025,China)
出处
《时珍国医国药》
CAS
CSCD
北大核心
2022年第1期16-20,共5页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金(81760816)。
