清络通痹方调控类风湿关节炎“免疫-骨侵蚀”的机制

目的观察清络通痹方对类风湿关节炎(RA)B细胞亚群、B细胞活化因子(BAFF)、骨侵蚀及骨形成指标以及核因子-κB受体活化因子配体(RANKL)/核因子-κB受体活化因子(RANK)/骨保护素(OPG)信号通路的影响,探讨本方调控RA“免疫-骨侵蚀”的机制。方法将64例RA患者随机分为对照组和研究组,32例/组,对照组口服甲氨蝶呤,研究组口服清络通痹方,两组疗程均为12周。采用流式细胞术检测RA患者外周血CD3-CD19+B细胞亚群、CD19+CD27+B细胞亚群和CD19+BAFFR+B细胞亚群百分比,ELISA法检测血清BAFF、RANKL、RANK、OPG水平,ELISA法检测RA患者治疗前后血清β胶原降解产物(β-CTX)、人抗酒石酸酸性磷酸酶5b(TRACP-5b)、血清骨钙素(BGP)、骨碱性磷酸酶(BALP)、Ⅰ型前胶原氨基端原肽(PINP)水平,DXEA双能X线骨密度仪测定骨密度。细胞实验:以RAW264.7细胞作为破骨细胞,分成空白组、模型组、清络通痹方低剂量组(125μg·mL^(-1))、中剂量组(250μg·mL^(-1))、高剂量组(500μg·mL^(-1)),甲氨蝶呤组(2μg·mL^(-1))。干预时间为48 h。采用免疫印迹法检测RANKL、RANK、OPG、Fos原癌基因(C-FOS)蛋白表达情况。结果RA患者B细胞与RANKL/RANK/OPG系统呈相关性,清络通痹方能下调RA患者外周血B细胞亚群百分比、血清BAFF水平,下调血清β-CTX及TRACP-5b水平,上调BGP、BALP、PINP水平,改善腰椎骨密度值(P<0.05),并与调控B细胞表达呈显著相关性(P<0.05);该处方能下调OC中RANKL、RANK、CFOS表达,上调OPG表达(P<0.05)。结论清络通痹方调控RA“免疫-骨侵蚀”的机制可能为通过调控B细胞亚群百分比表达及BAFF表达水平抑制骨破坏,并进一步干预RANKL/RANK/OPG通路抑制破骨细胞分化,从而抑制骨侵蚀的发生。

黄芩清热除痹胶囊配合中药外敷治疗急性痛风性关节炎的效果及对细胞因子的影响

目的观察黄芩清热除痹胶囊(Huangqin Qingre Chubi capsule,HQC)口服配合芙蓉膏(Furong Ointment,FRO)外敷治疗急性痛风性关节炎(AGA)患者的临床疗效及对细胞因子的影响。方法选取2017年1月至2019年3月安徽中医药大学第一附属医院风湿免疫科92例AGA患者,按随机数字表法分为治疗组与对照组各46例,对照组使用非甾类消炎止痛药,治疗组在此基础上加用HQC口服配合FRO外敷肿痛关节区,疗程为7 d,同时选择46名健康组。应用ELISA法检测血清白细胞介素-1β(IL-1β)、肿瘤坏死因子α(TNF-α)、白细胞介素-4(IL-4)、白细胞介素-10(IL-10),仪器法检测红细胞沉降率(ESR),生化仪检测超敏C反应蛋白(hs-CRP),并观察患者视觉模拟评分法(VAS)评分、关节肿胀评分、中医证候积分。结果(1)治疗组总有效率93.48%,对照组为76.09%,治疗组明显优于对照组(P<0.05);(2)两组患者治疗后VAS评分、关节肿胀、中医证候积分、ESR、hs-CRP均明显降低(P<0.05或P<0.01);与对照组相比,治疗组VAS评分、关节肿胀、ESR、hs-CRP均明显降低(P<0.05或P<0.01);(3)与健康组相比,AGA患者TNF-a、IL-1β显著升高,IL-4、IL-10显著降低(P<0.01);(4)两组患者治疗后TNF-a、IL-1β显著降低,IL-4、IL-10显著升高(P<0.01);与对照组相比,治疗组TNF-a显著降低,IL-10显著升高(P<0.05或P<0.01)。结论HQC口服配合FRO外敷治疗AGA疗效显著,不仅能显著改善患者临床症状,而且能明显降低炎性反应。

“从脾治痹”治疗膝骨关节炎疗效观察

目的观察以“从脾治痹”理论为指导,采用健脾祛湿中成药新风胶囊治疗膝骨关节炎(KOA),观察其临床疗效。方法 60例KOA患者按随机数字表法分为观察组和对照组,观察组口服新风胶囊及硫酸氨基葡萄糖,对照组给予硫酸氨基葡萄糖口服,疗程均为4周。观察两组治疗前后HAQ、SF-36评分,VAS评分、WOMAC评分、LequesneMG评分,以及中医证候积分变化、中医总体疗效,实验室指标[血沉(ESR)、C反应蛋白(CRP)、血小板计数(PLT)、甘油三酯(TG)、总胆固醇(TC)]变化,并评估其安全性。结果观察组改善HAQ评分较对照组差异有统计学意义(P<0.01),改善SF-36评分中8个维度治疗后较治疗前差异有统计学意义(P<0.01或P<0.05);两组治疗后比较,其中在改善生理功能、躯体疼痛、一般健康状况、生命活力、社会职能、精神健康方面差异有统计学意义(P<0.05);观察组改善VAS评分、WOMAC评分、LequesneMG评分较对照组差异有统计学意义(P<0.01或P<0.05);观察组在降低中医证候积分变化方面较对照组差异有统计学意义(P<0.05);观察组在降低ESR、PLT、TC、TG方面治疗前后差异有统计学意义(P<0.01或P<0.05),治疗后观察组与对照组比较,在降低ESR、PLT方面差异有统计学意义(P<0.05)。结论新风胶囊能提高膝骨关节炎患者生活质量和疗效,改善其关节症状、证候积分及相关指标,临床使用安全且值得长期推广。

基于名老中医经验挖掘的卓越中医风湿医师培养模式和培养体系研究

探讨“名老中医经验数据挖掘培养模式”结合了中医师承和信息时代数据挖掘方法的优势,借鉴传统中医人才培养模式,培养具有良好医德、注重知识交融、具有思辨能力、符合个性发展的卓越中医师,真正做好中医药的传承和创新创新,建立痹病学科卓越中医医师教育体系和培养保障体系,走好中医痹病学科卓越医师教学发展的创新之路。为中医药高等教育的传承与创新提供旺盛的生命力。

Drug-containing serum of Xinfeng capsules protect against H9C2 from death by enhancing miRNA-21 and inhibiting toll-like receptor 4/phosphorylated p-38(p-p38)/p-p65 signaling pathway and proinflammatory cytokines expression

OBJECTIVE: To investigate effect of drug-containing serum of Xinfeng capsules on myocardial cell growth.METHODS: Drug-containing serum of Xinfeng capsules rat models were established by intragastricly administrated Xinfeng capsules. MTT assay wasused to evaluated H9C2 cells viability. H9C2 cells were divided into normal control group, triptolide group, lipopolysaccharide(LPS) group, drug-containing serum group and mi RNA-21 inhibitor group. micro RNA-21(mi RNA-21) inhibitor was structured and transfected into H9C2 cells. Western blot and immunofluorescence assay were applied to examine toll-like receptor 4(TLR4), phosphorylated p-38(p-p38) and p-p65 expression. Quantitative real-time PCR(q RT-PCR) was used to evaluated m RNA levels of mi RNA-21. Enzyme linked immunosorbent(ELISA) was used to measure tumor necrosis factor α(TNF-α), IL-6 and IL-17 levels.RESULTS: Drug-containing serum treatment significantly increased cell viability compared to LPS treated group. q RT-PCR results indicated that mi RNA-21 levels were significantly decreased in drugcontaining serum group compared to LPS group.Early and late apoptosis in drug-containing serum group were significantly decreased compared to LPS group. Western blot and immunofluorescence assay results showed that TLR4, p-p38 and p-p65 levels in drug-containing serum group were significantly decreased compared to LPS group. ELISA findings indicated that drug-containing serum significantly decreased inflammatory cytokine levels of TNF-α, IL-6 and IL-17.CONCLUSION: Drug-containing serum of Xinfeng capsules protect against lipopolysaccharide instructed H9C2 cells from death by enhancing mi RNA-21 and inhibiting TLR4/p-p38/p-p65 signaling pathway and proinflammatory cytokines expression.

Mechanism studies of Xinfeng capsule on improving cardiovascular function though toll-like receptor 4/nuclear factor kappa B pathway in a rat model of adjuvant arthritis

OBJECTIVE: To observe the impact of Xinfeng capsule(XFC) on cardiovascular function in adjuvant arthritis(AA) model rats and investigate the mechanism though toll-like receptor 4(TLR4)/nuclear factor kappa B(NF-κB) signaling pathway.METHODS: Seventy rats were randomly divided into seven groups: normal control(NC), model control(MC), tripterygium glycosides tablet(TPT),methotrexate(MTX), high, moderate and low dose XFC group. The administration began from day 19 after modeling for 30 day. Paw swelling, arthritic in-dex(AI), cardiac function indexes and myocardial pathological pattern were detected. The expression of TLR4, myeloid differentiation factor(My D) 88, interleukin-1 receptor-associated kinase(IRAK) 1, tumor necrosis factor receptor associated factor(TRAF) 6, NF-κB, tumor necrosis factor-alpha(TNF-α) proteins in myocardial tissue were determined by western blot method.RESULTS: Paw swelling and AI in MC group increased in MC group(P < 0.01), and decreased in high and moderate dose XFC groups(P < 0.01 or P > 0.05). Left ventricular systolic pressure(LVSP),left ventricular end-diastolic pressure(LVEDP),heart rate(HR) were elevated in MC group(P <0.01), and ± dp/dtmax and CI were reduced(P <0.01); while LVSP, LVEDP and HR declined and ±dp/dtmax, CI improved in high dose XFC group(P <0.05 or P < 0.01). LVSP in high dose XFC group were reduced more than other treatment groups(P <0.05 or P < 0.01). The improvements on LVEDP, dp/dt-max were superior to MTX and low dose XFC group, and the improvement on CI was better than low dose XFC group(P < 0.05 or P < 0.01). Myocardial fibers arranged irregular in MC group with intracellular edema and mitochondria damage. The modifications on myocardial structural were shown in each treatment group, but more prominent in TPT, high and moderate dose XFC group. The proteins of TLR4, My D88, IRAK1, TRAF6, NF-κB, TNF-αwere highly expressed in MC group, and those proteins declined in high and moderate dose XFC group(P < 0.05 or P < 0.01). High dose XFC group was superior to MTX and low dose XFC group on reducing TLR4, NF-κB, TNF-α(P < 0.05).CONCLUSION: XFC can not only inhibit the excessive activation of TLR4/NF-κB signaling pathway and the increased inflammatory mediators, but also reduce the damage of myocardial tissue and cells.