鳖甲煎丸含药血清通过调控TLR4-NF-κB通路抑制血小板衍生生长因子诱导的肝星状细胞激活  

Biejiajian containing serum inhibits platelet derived growth factor induced hepatic stellate cells activation by regulating TLR4-NF-κB pathway

作　　者：庞杰[1] 贺松其[1] 李健薇 杨惠琳 林佳鑫 冯明辉 PANG Jie;HE Songqi;LI Jianwei;YANG Huilin;LIN Jiaxin;FENG Minghui(School of Chinese Medicine,Southern Medical University,Guangzhou 510515,China;Department of Infertility and Sexual Medicine,The Third Affilicatal Hospital,SunYat-Sen University,Guangzhou 510630,China;Department of Oncology,Third Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine,Guangzhou 515000,China)

机构地区：[1]南方医科大学中医药学院,广东广州510515 [2]中山大学附属第三医院不育与性医学科,广东广州510630 [3]广州中医药大学第三附属医院肿瘤科,广东广州510378

出　　处：《沈阳药科大学学报》2022年第4期440-446,共7页Journal of Shenyang Pharmaceutical University

基　　金：国家自然科学基金资助项目(81704016);广东省自然科学基金资助项目(2017A030313687);广东省中医药管理局项目(20171163)。

摘　　要：目的研究鳖甲煎丸(Biejiajian Pill,BJJ)含药血清对血小板衍生生长因子(platelet derived growth factor,PDGF)诱导肝星状细胞(hepatic stellate cells,HSC)活化的抑制作用及其机制。方法通过PDGF诱导HSC激活并给予鳖甲煎丸含药血清处理,MTT实验检测PDGF处理后HSC细胞增殖活性,EdU染色检测HSC细胞增殖。ELISA实验检测HSC激活相关细胞因子,最后通过Western blotting实验研究Toll样受体-4(Toll-like receptor 4,TLR 4)/核因子κB(nuclear factor kappa-B,NF-κB)信号通路相关蛋白表达情况。结果MTT和EdU实验结果显示鳖甲煎丸含药血清可显著抑制PDGF引起的HSC细胞增殖活性;ELISA实验结果显示鳖甲煎丸含药血清可显著降低PDGF处理后HSC中Ⅰ型胶原(collagen typeⅠ,Col-Ⅰ)、白介素-6(interleukin 6,IL-6)、PDGF-AA和肿瘤坏死因子-α(tumor necrosis factor alpha,TNF-α)含量;并可显著抑制PDGF处理后TGF-β1和α-SMA表达;免疫印迹实验显示鳖甲煎丸含药血清可抑制TLR4、p-IKK表达并升高p-IκBα表达。结论鳖甲煎丸含药血清可通过作用于TLR4-NF-κB信号通路抑制HSC细胞增殖和激活。Objective To study the inhibitory effect of Biejiajian Pill(BJJ)on the activation of hepatic stellate cells(HSC)induced by platelet derived growth factor(PDGF)and its mechanism.Methods HSC activation was induced by PDGF and treated with BJJ containing serum.HSC cell activity was detected by MTT assay and HSC cell proliferation was detected by EdU staining.HSC activation related cytokines was detected by ELISA.The expression of Toll like receptor 4(TLR 4)/nuclear factor kappa-B(NF-κB)signaling pathway related proteins was studied by Western blotting.Results The results of MTT and EdU staining showed that BJJ containing serum could significantly inhibit the cell viability and proliferation of HSC cells;the results of ELISA showed that BJJ containing serum could significantly reduce the content of collagen typeⅠ(Col-Ⅰ),interleukin-6(IL-6),PDGF-AA and tumor necrosis factor alpha(TNF-α)in HSC after PDGF treatment.BJJ containing serum could significantly inhibit the content of TGF-β1 andα-SMA after PDGF treatment,and inhibit the expression of TLR4,p-IKK and increase the expression of p-IκBα.Conclusion BJJ containing serum can inhibit the proliferation and activation of HSC cells by acting on TLR4-NF-κB pathway.

关 键 词：肝星状细胞 鳖甲煎丸 血小板衍生生长因子 α-平滑肌肌动蛋白 Toll样受体-4/核因子κB 

分 类 号：R96[医药卫生—药理学]