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目的 探究白头翁皂苷A(PSA)对肿瘤坏死因子（TNF-α）诱导的滑膜细胞异常增殖、迁移和侵袭的作用及机制。方法 使用TNF-α诱导滑膜炎模型，并使用LRP1的抑制剂RAP和LRP1-siRNA构建LRP1沉默模型。采用不同浓度的PSA处理SW982滑膜细胞，CCK-8法检测细胞活力。Western blot检测低密度脂蛋白受体相关蛋白1(LRP1)、基质金属蛋白酶3(MMP3)、神经钙黏蛋白（N-cadherin）的表达水平。EdU增殖染色试剂盒检测滑膜细胞的异常增殖能力。Transwell和划痕实验检测滑膜细胞迁移和侵袭的情况。逆转录实时荧光定量PCR(RT-qPCR)检测滑膜细胞LRP1、MMP3、N-cadherin、白细胞介素（IL）-1β、IL-6和TNF-α的mRNA表达水平。免疫荧光染色观察滑膜细胞迁移和侵袭相关蛋白MMP3、N-cadherin的表达情况。同时使用LRP1的抑制剂受体相关蛋白（RAP）验证MMP3、N-cadherin的表达。结果 与TNF-α诱导的滑膜细胞炎症模型相比，PSA组SW982滑膜细胞活性降低（P<0.05）,MMP3、N-cadherin的表达降低（均P<0.01）;PSA组细胞在EdU染色后，阳性细胞比例减少（P<0.05）。此外，与正常滑膜细胞相比，TNF-α诱导的滑膜细胞炎症模型中LRP1的表达上升（P<0.01），而加入PSA后LRP1的表达下调（P<0.05）。同时，与模型组相比，敲除LRP1后迁移侵袭相关蛋白MMP3、N-cadherin的表达降低（均P<0.01）。与模型组相比，PSA+RAP组滑膜细胞的MMP3、N-cadherin表达下降（均P<0.01）。结论 PSA可以通过下调LRP1的表达抑制TNF-α诱导的滑膜异常增殖、迁移和侵袭。
Abstract：Objective To investigate the effects and mechanisms of pulchinenoside A(PSA) on tumor necrosis factor-α(TNF-α)-induced abnormal proliferation, migration, and invasion of synovial fibroblasts.Methods A synovitis model was established by stimulating SW982 synovial cells with TNF-α. The low-density lipoprotein receptor-related protein 1(LRP1) was knocked down using LRP1-specific siRNA or inhibited by the receptor-associated protein(RAP). Cells were treated with various concentrations of PSA, and cell viability was assessed by CCK-8 assay. Protein levels of LRP1, matrix metalloproteinase-3(MMP3), and N-cadherin were measured by Western blotting. Proliferation was evaluated using an EdU incorporation assay. Migration and invasion capacities were examined by Transwell and wound-healing assays. mRNA expression of LRP1, MMP3, N-cadherin, IL-1β, IL-6, and TNF-α was detected by reverse transcription quantitative real-time PCR(RT-qPCR). Immunofluorescence staining was used to visualize the expression of MMP3 and N-cadherin. The involvement of LRP1 was further verified with the inhibitor RAP.Results Compared with the TNF-α-treated model group, PSA treatment significantly reduced SW982 cell viability(P<0. 05), downregulated MMP3 and N-cadherin expression(both P<0. 01), and decreased the proportion of EdU-positive cells(P<0. 05). Moreover, TNF-α stimulation upregulated LRP1 expression compared with normal controls(P<0. 01), whereas PSA treatment reversed this increase(P<0. 05). LRP1 knockdown or inhibition markedly decreased MMP3 and N-cadherin levels compared with the model group(both P<0. 01). Similarly, the combination of PSA and RAP led to significant reductions in MMP3 and Ncadherin expression compared with the model group(both P<0. 01).Conclusion PSA inhibits TNF-α-induced abnormal proliferation, migration, and invasion of synovial, at least in part, by downregulating LRP1 expression.
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基本信息:
DOI：10.19405/j.cnki.issn1000-1492.2026.09.010
中图分类号:R285
引用信息:
[1]毛丽芳,陈天宇,王金虹,等.白头翁皂苷A下调LRP1抑制滑膜炎性细胞的增殖、迁移和侵袭[J].安徽医科大学学报,2026(09):1584-1593.DOI:10.19405/j.cnki.issn1000-1492.2026.09.010.
基金信息:
安徽省自然科学基金项目(编号:2208085MH215); 安徽省卫生健康科研项目(编号:AHWJ2024Aa40016)
2026-08-11
2026-08-11
2026-08-11