中医杂志

目的 评价黄芩清热除痹胶囊对类风湿关节炎（RA）患者感受及免疫炎症的综合干预效果，并探究其潜在作用机制。方法 回顾性收集452例RA患者的临床资料，分为对照组274例（采用常规西医治疗），观察组178例患者（在常规西医治疗基础上服用黄芩清热除痹胶囊且连续服用至少2周），疗程均为2周。采用倾向性评分匹配法（PSM）按1∶1比例将两组患者进行匹配，收集治疗前后患者感受指标[包括中文版简明健康调查量表（SF-36）、焦虑自评量表（SAS）、抑郁自评量表（SDS）、视觉模拟量表（VAS）评分及中国类风湿关节炎患者报告疾病活动指数（CPRI-RA）]和免疫炎症指标[包括红细胞沉降率（ESR）、超敏C反应蛋白（hs-CRP）、类风湿因子（RF）、抗环瓜氨酸肽抗体（anti-CCP）、白细胞介素-6(IL-6)、免疫球蛋白A(IgA)、免疫球蛋白G(IgG)、免疫球蛋白M(IgM)、补体C3、补体C4]。通过Spearman相关性分析评估患者感受指标与免疫炎症指标之间的关系，采用Logistic回归、关联规则及中介分析评估黄芩清热除痹胶囊对RA患者感受和免疫炎症指标的影响及作用路径。通过网络药理学研究方法筛选黄芩清热除痹胶囊治疗RA的药物活性成分及核心靶点，并进行分子对接验证。细胞实验中，分为正常组、模型组、20%含药血清组、80 nmol/L对照组，正常组予完全培养基常规培养人滑膜成纤维细胞（FLS），模型组予完全培养基常规培养人RA成纤维样滑膜细胞（RA-FLS）,20%含药血清组予含有20%黄芩清热除痹胶囊含药血清的培养基培养RA-FLS,80 nmol/L对照组予含有80 nmol/L甲氨蝶呤混悬液的完全培养基培养RA-FLS。各组细胞培养48 h后，CCK-8法检测细胞存活率，ELISA法检测细胞上清液白细胞介素-1β(IL-1β)、IL-6、白细胞介素-8(IL-8)、白细胞介素-10(IL-10)水平，Western Blot法检测细胞上清液基质金属蛋白酶9(MMP9)、转录因子AP-1亚基（JUN）、血管内皮生长因子A(VEGFA)、C-X-C基序趋化因子配体8(CXCL8)蛋白水平，Transwell实验检测细胞迁移能力。结果 PSM匹配后得到两组患者各178例，两组患者治疗后SF-36评分较治疗前升高，SAS评分、SDS评分、VAS评分、CPRI-RA、ESR、hs-CRP、IL-6、补体C3、补体C4均降低，观察组IgG、IgM亦降低（P<0.05）。SF-36中生理功能（相关系数-0.19,P<0.05）、社会功能（相关系数-0.18,P<0.05）评分与hs-CRP呈负相关，VAS评分与hs-CRP呈正相关（相关系数0.19,P<0.05）。黄芩清热除痹胶囊是多个患者感受指标改善的保护因素（P<0.05），其与多个患者感受指标及免疫炎症指标的改善具有较高关联度，且hs-CRP和ESR在黄芩清热除痹胶囊改善患者感受中发挥了部分中介作用。网络药理学研究筛选出黄芩清热除痹胶囊治疗RA的核心药物活性成分为汉黄芩素、槲皮素、α1-谷甾醇、β-谷甾醇、豆甾醇、黄芩素、藏红花酸，核心靶点包括JUN、IL-6、VEGFA、MMP9、IL-1β、CXCL8等。分子对接结果显示，槲皮素与VEGFA、JUN、MMP9、IL-6、IL-1β，黄芩素与VEGFA、MMP9，汉黄芩素与CXCL8结合能力较强。细胞实验结果显示，黄芩清热除痹胶囊和甲氨蝶呤均能抑制RA-FLS细胞存活率与细胞迁移能力，降低IL-1β、IL-6、IL-8水平以及MMP9、JUN、VEGFA、CXCL8蛋白水平，升高IL-10水平（P<0.05）。结论 黄芩清热除痹胶囊能通过调控免疫炎症反应来改善RA患者感受，其作用机制可能与该药多通路、多靶点抑制滑膜细胞炎症及细胞迁移有关。

Objective To evaluate the comprehensive intervention effects of Huangqin Qingre Chubi Capsule （黄芩清热除痹胶囊, HQC） on self-perception of patients(SPP) and immune inflammation in patients with rheumatoid arthritis(RA), and to explore its potential mechanisms. Methods Clinical data of 452 RA patients were retro??spectively collected. Patients were divided into a control group(274 cases), treated with conventional western medicine, and an observation group(178 cases), treated with HQC for at least 2 weeks in addition to conventional western medicine. The treatment duration was 2 weeks for both groups. Propensity score matching(PSM) was performed at a ratio of 1∶1 to match patients between groups. SPP including the Chinese version of the short form-36 health survey(SF-36), self-rating anxiety scale(SAS), self-rating depression scale(SDS), visual analog scale(VAS), and Chinese patient-reported index for rheumatoid arthritis(CPRI-RA), as well as immune inflammatory indicators, including erythrocyte sedimentation rate(ESR), high-sensitivity C-reactive protein(hs-CRP), rheumatoid factor(RF), anticyclic citrullinated peptide antibody(anti-CCP), interleukin-6(IL-6), immunoglobulin A(IgA), immunoglobulin G(IgG), immunoglobulin M(IgM), complement C3, and complement C4, were collected before and after treatment. Spearman correlation analysis was used to assess the relationships between SPP and immune inflammatory indicators. Logistic regression, association rule analysis, and mediation analysis were performed to evaluate the effects and poten??tial pathways of HQC on SPP and immune inflammatory indicators. Network pharmacology was applied to identify the active components and core targets of HQC in the treatment of RA, followed by molecular docking verification. In cell experiments, cells were divided into normal group, model group, 20% medicated serum group, and 80 nmol/L control group. Human synovial fibroblasts(FLS) were cultured with complete medium in the normal group, while human rheumatoid arthritis fibroblast-like synoviocytes(RA-FLS) were cultured in the model group. In the 20% medicated serum group, RA-FLS were cultured with medium containing 20% HQC-medicated serum, and in the 80 nmol/L con??trol group, RA-FLS were cultured with complete medium containing 80 nmol/L methotrexate suspension. After 48 h of culture, cell viability was detected by cell counting kit-8(CCK-8) assay. Levels of interleukin-1β(IL-1β), interleukin-6(IL-6), interleukin-8(IL-8), and interleukin-10(IL-10) in the cell supernatant were measured by enzyme-linked immunosorbent assay(ELISA). Protein levels of matrix metalloproteinase 9(MMP9), transcrip??tion factor AP-1 subunit(JUN), vascular endothelial growth factor A(VEGFA), and C-X-C motif chemokine ligand 8(CXCL8) were detected by Western Blot, and cell migration ability was evaluated using Transwell assay. Results After PSM, 178 cases were included in each group. After treatment, SF-36 scores increased, while scores of SAS, SDS, VAS and CPRI-RA, levels of ESR, hs-CRP, IL-6, complement C3, and complement C4 levels decreased in both groups; IgG and IgM levels were also reduced in the observation group(P<0. 05). Physical functioning(corre??lation coefficient-0. 19, P<0. 05) and social functioning(correlation coefficient-0. 18, P<0. 05) of SF-36 were negatively correlated with hs-CRP, while VAS score was positively correlated with hs-CRP(correlation coefficient 0. 19, P<0. 05). HQC showed high associations with improvements in multiple indicators of SPP and immune inflammatory, and acted as a protective factor for the improvement of several SPP; hs-CRP and ESR played partial mediating roles in the improvement of SPP induced by HQC(P<0. 05). Network pharmacology analysis identified baicalein, quercetin, α1-sitosterol, β-sitosterol, stigmasterol, baicalin, and crocetin as the core active components, and JUN, IL-6, VEGFA, MMP9, IL-1β, and CXCL8 as the core targets. Molecular docking results showed strong binding affinities of quercetin with VEGFA, JUN, MMP9, IL-6, and IL-1β, of baicalin with VEGFA and MMP9, and of wogonin with CXCL8. Cell experiments demonstrated that HQC and methotrexate inhibited RA-FLS via??bility and migration, reduced levels of IL-1β, IL-6, and IL-8, decreased protein levels of MMP9, JUN, VEGFA, and CXCL8, and increased IL-10 levels(P<0. 05). Conclusion HQC can improve SPP in RA by regulating immune inflammatory responses. Its mechanism may be related to multi-pathway and multi-target inhibition of synovial cell inflammation and migration.