摘要
目的观察黄芪多糖(APS)在骨髓间充质干细胞(BMSCs)向肿瘤相关成纤维细胞(TAFs)分化中对IL-6/STAT3、TNF-α/NF-κB通路的影响,探讨其可能的调控机制。方法以白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)诱导BMSCs,建立向TAFs方向分化的炎性微环境模型;采用CCK-8法分别检测不同浓度IL-6/STAT3通路阻断剂AG490和TNF-α/NF-κB通路阻断剂BMS345541对BMSCs和诱导后BMSCs增殖能力的影响,Western blot检测阻断剂所对应的靶向蛋白JAK2和IKKβ的表达。将BMSCs分为空白组、模型组、模型+AG490组、模型+BMS345541组、模型+APS组,检测各组TAFs标记分子α-平滑肌肌动蛋白(α-SMA)、成纤维细胞活化蛋白(FAP)和IL-6/STAT3、TNF-α/NF-κB通路关键蛋白STAT3、p65的表达。结果与空白组比较,模型组BMSCs增殖水平明显增强(P<0.05);10μmol/L AG490和5μmol/L BMS345541较模型组BMSCs抑制作用明显增强(P<0.05)。10μmol/L AG490对应的靶蛋白JAK2较其他各组表达减弱,5μmol/L BMS345541对应的靶蛋白IKKβ较其他各组表达减弱,故选为最佳作用浓度。通路阻断剂干预结果表明,与空白组比较,模型组TAFs标记分子α-SMA、FAP和通路关键蛋白STAT3、p65表达均明显升高(P<0.05);与模型组比较,各干预组TAFs标记分子α-SMA、FAP和通路蛋白STAT3、p65表达均明显下降(P<0.05)。结论 APS对IL-6、TNF-α诱导的BMSCs向TAFs方向分化有抑制作用,其机制可能与调控IL-6/STAT3、TNF-α/NF-κB通路有关。
Objective To observe the effects of Astragalus polysaccharides（APS） on pathways of IL-6/STAT3 and TNF-α/NF-κB in the process of differentiation from bone marrow mesenchymal stem cells（BMSCs） to tumor-associated fibroblasts（TAFs）; To discuss the possible regulatory mechanism. Methods BMSCs was induced by IL-6 and TNF-α, and an inflammatory microenvironment model which differentiated into TAFs direction was established. CCK-8 method was used to detect the effects of different concentrations of IL-6/STAT3 pathway blocker AG490 and TNF-α/NF-κB pathway blocker BMS345541 on BMSCs and its proliferation after induction. Target protein expressions of JAK2 and IKKβ corresponding to blocker were detected by Western blot. BMSCs were divided into blank group, model group, model＋AG490 group, model＋BMS345541 group, and model＋APS group; expressions of α-SMA and FAP of TAFs marker molecules and the key proteins of STAT3 and p65 of IL-6/STAT3 and TNF-α/NF-κB pathway were detected. Results Compared with the blank group, the proliferation ability of the model group increased significantly（P〈0.05）; Compared with the model group, 10 μmol/L AG490 and 5 μmol/L BMS345541 had a strong inhibitory effect on the proliferation ability（P〈0.05）. The expression of JAK2, a target protein corresponding to 10 μmol/L AG490, was weaker than that of other groups. The target protein IKKβcorresponding to 5 μmol/L BMS345541 was weaker than the other groups, so it was selected as the best concentration. The results of pathway intervening blockers showed that compared with the blank group, the expressions of TAFs makers α-SMA and FAP, and key proteins STAT3 and p65 of pathway in the model group increased significantly（P〈0.05）; Compared with model group, the expressions of TAFs makers α-SMA and FAP, and key proteins STAT3 and p65 of pathway in other groups decreased significantly（P〈0.05）. Conclusion APS has inhibitory effects on the differentiation of BMSCs into TAFs induced by IL-6 and TNF-α, and its mechanism may be related to the regulation of pathways of IL-6/STAT3 and TNF-α/NF-κB.
作者
张艳辉
骆亚莉
刘永琦
王磊
许小敏
冯彩琴
李研
ZHANG Yan-hui;LUO Ya-li;LIU Yong-qi;WANG Lei;XU Xiao-min;FENG Cai-qin;LI Yan(Gansu University of Chinese Medicine,Provincial Key Laboratory of Major Disease Prevention and Control of Molecular Medicine and Traditional Chinese Medicine Research in Gansu Province,Key Laboratory of Dunhuang Medicine and Transformation at Provincial and Ministerial Level,Lanzhou 730000,China)
出处
《中国中医药信息杂志》
CAS
CSCD
2018年第10期54-59,共6页
Chinese Journal of Information on Traditional Chinese Medicine
基金
国家自然科学基金(817614217)
作者简介
通讯作者：刘永琦,E-mail：liuyongqi73@163.com
