黄芩清热除痹胶囊通过调控LncRNA NONHSAT118801.2抑制NF-κB通路改善强直性脊柱炎湿热痹阻证患者免疫炎症反应OA
Huangqin Qingre Chubi Capsules Improve Immune Inflammatory Response in Patients with Damp-Heat Bi Syndrome of Ankylosing Spondylitis by Regulating LncRNA NONHSAT118801.2 to Inhibit the NF-κB Pathway
目的:探讨黄芩清热除痹胶囊含药血清改善强直性脊柱炎湿热痹阻证患者免疫炎症反应的机制.方法:构建 NONHSAT118801.2 功能敲减及过表达的巨噬细胞模型;细胞计数试剂盒 8 法(CCK-8)筛选最佳 HQC 含药血清浓度和作用时间;采用流式细胞术检测巨噬细胞标志物;Elisa 试剂盒检测 TNF-α、IL-1β、IL-6、IL-10、Argl、CXCR5、BLC-1/CXCL13 的表达;RT-qPCR 检测各组 NONHSAT118801.2、IKBα、NF-κB p65 mRNA 的表达;免疫荧光和 Western Blot 技术检测巨噬细胞中 NKRF、p-IKBα、及 p-NF-κB p65 蛋白的表达.结果:CCK-8 法结果显示,含 10%HQC 的血清干预 24h 时对巨噬细胞抑制作用最强.HQC 干预后 CD86 表达显著下调,CD163 表达明显上调,转染 pc DNA3.1-NONHSAT118801.2 可促进 CD86 表达,抑制 CD163 表达,功能回复实验中,HQC 含药血清可逆转过表达 NONHSAT118801.2 对 CD86 表达升高,和 CD163 表达降低的影响;HQC 干预后 IL-10、ARG1 的表达明显升高,BLC-1/CXCL13、CXCR5、IL-6、IL-1β、TNF-α 表达明显下降,转染 pcDNA3.1-NONHSAT118801.2 后,显著促进 BLC-1/CXCL13、CXCR5、IL-6、IL-1β、TNF-α 表达,抑制 IL-10、ARG1 的表达,功能回复实验中,HQC 含药血清可逆转过表达 NONHSAT118801.2 对 BLC-1/CXCL13、CXCR5、IL-6、IL-1β、TNF-α 表达升高和 IL-10、ARG1 表达下降的影响;HQC 干预后 NONHSAT118801.2、IKBα、NF-κB p65 mRNA 及 p-IKBα 及 p-NF-κB p65蛋白表达明显降低,NKRF 蛋白表达明显升高,转染 pcDNA3.1-NONHSAT118801.2 后,NONHSAT118801.2、IKBα、NF-κB p65 mRNA 及 p-IKBα 及 p-NF-κB p65 蛋白表达明显升高,NKRF 蛋白表达明显降低,功能回复实验中,HQC 含药血清可逆转过表达 NONHSAT118801.2 对 NONHSAT118801.2、IKBα、NF-κB p65 mRNA 及 p-IKBα 及p-NF-κB p65、NKRF 蛋白表达影响.结论:HQC 含药血清可能通过下调 NONHSAT118801.2 表达,抑制 NF-κB信号通路活化,调控巨噬细胞极化和炎性因子表达,从而抑制 AS 巨噬细胞炎症反应.
Objective:To investigate the mechanism by which serum containing Scutellaria baicalensis Qingre Chubi capsules improves the immune inflammatory response in patients with ankylosing spondylitis and damp-heat obstruction syndrome.Methods:A macrophage model with knockdown and over-expression of NONHSAT118801.2 was construct-ed;the optimal serum concentration and treatment time of HQC were screened using the CCK-8 assay;macrophage markers were detected by flow cytometry;the expression of TNF-α,IL-1β,IL-6,IL-10,Arg1,CXCR5,and BLC-1/CXCL13 were detected by ELISA;the expression of NONHSAT118801.2,IKBα,and NF-κB p65 mRNA in each group was detected by RT-qPCR;and the expression of NKRF,p-IKBα,and p-NF-κB p65 proteins in macrophages were detected by immunofluorescence and Western blotting.Results:CCK-8 assay showed that serum containing 10%HQC had the strongest inhibitory effect on macrophages after 24 hours of intervention.After HQC intervention,CD86 expression was significantly down-regulated and CD163 expression was significantly up-regulated.Transfection with pc DNA3.1-NONHSAT118801.2 promoted CD86 expression and inhibited CD163 expression.In the functional recovery experiment,HQC-containing serum could reverse the effect of over-expression of NONHSAT118801.2 on the increase in CD86 expression and decrease in CD163 expression.After HQC intervention,the expression of IL-10 and ARG1 was significantly increased,while the expression of BLC-1/CXCL13,CXCR5,IL-6,IL-1β,and TNF-α was signifi-cantly decreased.Transfection with pcDNA3.1-NONHSAT118801.2 significantly promoted the expression of BLC-1/CXCL13,CXCR5,IL-6,IL-1β,and TNF-α,and inhibited the expression of IL-10 and ARG1.In the functional recovery experiment,HQC.The effects of reversible over-expression of NONHSAT118801.2 in HQC-containing serum on increased expression of BLC-1/CXCL13,CXCR5,IL-6,IL-1β,and TNF-α,and decreased expression of IL-10 and ARG1;after HQC intervention,the expression of NONHSAT118801.2,IKBα,NF-κB p65 mRNA,and p-IKBα and p-NF-κB p65 proteins were significantly decreased,while NKRF protein expression was significantly increased.After transfection with pcDNA3.1-NONHSAT118801.2,the expression of NONHSAT118801.2,IKBα,NF-κB p65 mRNA,and p-IKBα and p-NF-κB p65 proteins were significantly increased,while NKRF protein expression was significantly decreased.In the functional recovery experiment,reversible overexpression of NONHSAT118801.2 in HQC-containing serum had a significant effect on...The drug HQC affected the expression of NONHSAT118801.2,IKBα,NF-κB p65 mRNA,and p-IKBα,p-NF-κB p65,and NKRF proteins.Conclusion:HQC-containing serum may inhibit the inflammatory response of AS macrophages by down-regulating NONHSAT118801.2 expression,inhib-iting NF-κB signaling pathway activation,and regulating macrophage polarization and inflammatory factor expression.
黄旦;刘健;虞佳琪;汪元;万磊;张皖东;方妍妍
安徽中医药大学第一附属医院 安徽 合肥 230031||现代中医内科应用基础与开发研究安徽省重点实验室 安徽 合肥 230031安徽中医药大学第一附属医院 安徽 合肥 230031||现代中医内科应用基础与开发研究安徽省重点实验室 安徽 合肥 230031安徽中医药大学第一附属医院 安徽 合肥 230031安徽中医药大学第一附属医院 安徽 合肥 230031||现代中医内科应用基础与开发研究安徽省重点实验室 安徽 合肥 230031安徽中医药大学第一附属医院 安徽 合肥 230031||现代中医内科应用基础与开发研究安徽省重点实验室 安徽 合肥 230031安徽中医药大学第一附属医院 安徽 合肥 230031||现代中医内科应用基础与开发研究安徽省重点实验室 安徽 合肥 230031安徽中医药大学第一附属医院 安徽 合肥 230031
医药卫生
强直性脊柱炎湿热痹阻证黄芩清热除痹胶囊炎症反应
Ankylosing spondylitisDamp-heat obstruction syndromeScutellaria baicalensis clearing heat and re-lieving pain capsuleInflammatory response
《中医药临床杂志》 2026 (6)
1234-1242,9
国家自然科学基金青年基金(82205054)安徽省高校自然科学基金重点项目(2024AH0551005)安徽中医药大学2024年临床研究项目(2024YFYLCZX02)
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