目的 基于通过Toll样受体4（Toll-like receptor 4，TLR4）/核因子-κB（nuclear factor-κB，NF-κB）/NOD样受体热蛋白结构域相关蛋白3（NOD-like receptor thermal protein domain associated protein 3，NLRP3）通路探讨羟基红花黄色素A（hydroxysafflor yellow A，HSYA）抑制血管紧张素II（angiotensin II，ANG II）诱导的血管外膜成纤维细胞（vascular adventitial fibroblasts，VAFs）迁移作用。方法 组织贴块法培养大鼠胸主动脉VAFs，免疫荧光实验检测细胞中Vimentin和α-平滑肌肌动蛋白（α-smooth muscle actin，α-SMA）蛋白表达，鉴定VAFs。设置对照组、ANG II组、HSYA组、脂多糖（lipopolysaccharide，LPS）组和LPS＋HSYA组，对照组VAFs给予培养基正常培养，其余各组给予1×10^-7 mol/L ANG II处理24 h，然后更换为含100 nmol/L LPS或40 μmol/L HSYA的培养基，继续培养24 h，划痕实验检测VAFs的迁移能力；CCK-8法检测细胞活力；Western blotting检测TLR4、NF-κB和p-NF-κB蛋白表达；免疫荧光实验检测NLRP3炎性小体相关蛋白NLRP3、半胱氨酸天冬氨酸蛋白酶-1（cystein-asparate protease-1，Caspase-1）及凋亡斑点样蛋白（apoptotic spot-like protein，ASC）共表达情况。结果 免疫荧光结果证实VAFs培养成功。划痕实验结果显示，ANG II能够显著增加细胞的迁移率（P＜0.01），LPS刺激进一步提高VAFs的迁移率（P＜0.01），HSYA能够抑制ANG II及LPS对VAFs迁移的促进作用（P＜0.01）。Western blotting结果显示，ANG II显著提高细胞内TLR4和p-NF-κB的表达（P＜0.05、0.001），并促进NF-κB入核（P＜0.001）；HSYA显著抑制ANG II诱导的细胞内TLR4和p-NF-κB的表达（P＜0.05、0.01、0.001），并抑制NF-κB入核（P＜0.05、0.01、0.001）。免疫荧光实验结果表明，ANG II及LPS促进NLRP3炎症小体相关蛋白NLRP3、Caspase-1及ASC共表达，HSYA可减少ANG II及LPS诱导NLRP3、Caspase-1及ASC蛋白共表达，抑制NLRP3炎症小体的组装。结论 HSYA通过抑制TLR4/NF-κB/NLRP3通路减少ANG II诱导的VAFs迁移。

Objective To investigate the effect of hydroxysafflor yellow A (HSYA) on migration of advangiolar fibroblasts (VAFs) induced by angiotensin Ⅱ (ANG Ⅱ) based on Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB)/NOD-like receptor thermal protein domain associated protein 3 (NLRP3) pathway. Methods VAFs were cultured from rat thoracic aorta by tissue patch method, protein expressions of Vimentin and α-smooth muscle actin (α-SMA) was detected by laser confocal assay for identification of VAFs. Control group, ANG II group, HSYA group, lipopolysaccharide (LPS) group and LPS + HSYA group were set up. VAFs in control group were given normal culture medium, and the other groups were given 1×10^-7 mol/L ANG II for 24 h, and then changed to contain 100 nmol/L LPS or 40 μmol/L HSYA medium, continue to culture for 24 h, and migration ability of VAFs was tested by scratch test; CCK-8 method was used to detect cell viability; Western blotting was used to detect TLR4, NF-κB and p-NF-κB protein expression; The co-expression of NLRP3 inflammatory corpuscle-associated protein NLRP3, cystein-aspartate protease-1 (Caspase-1) and apoptotic spot-like protein (ASC) were detected by immunofluorescence assay. Results The results of immunofluorescence confirmed that VAFs were successfully cultured. The scratch test results showed that ANG II could significantly increase the cell migration rate (P < 0.01), LPS stimulation can further increase the migration of VAFs (P < 0.01), HSYA can inhibit the promotion of ANG II and LPS on VAFs migration (P < 0.01). Western blotting results showed that ANG II significantly increased intracellular TLR4 and p-NF-κB expressions (P < 0.05, 0.001), and promoted NF-κB entry (P < 0.001); HSYA significantly inhibited the intracellular TLR4 and p-NF-κB expressions induced by ANG II (P < 0.05, 0.01, 0.001), and inhibited NF-κB entered the nucleus (P < 0.05, 0.01, 0.001). The results of immunofluorescence test showed that ANG II and LPS promoted the co-expression of NLRP3, Caspase-1 and ASC proteins associated with NLRP3 inflammatory bodies, and HSYA could reduce the co-expression of NLRP3, Caspase-1 and ASC proteins induced by ANG II and LPS, and inhibited the assembly of NLRP3 inflammatory bodies. Conclusion HSYA can inhibit TLR4/NF-κB/NLRP3 pathway to reduce the migration of VAFs induced by ANG II.

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河北省自然科学基金资助项目（H2020423061）；河北省高等学校科学技术研究项目重点计划项目（ZD2017056）；河北省重点研发计划项目中医药创新专项（223777149D）；河北省中医药管理局科研计划项目（2017009，2022080）；河北省教育厅青年基金资助项目（QN2022092）