中药药理与临床

目的:探讨穿山龙复方对痛风性关节炎(GA)合并高尿酸血症(HUA)模型大鼠Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核转录因子-κB(NF-κB)(TLR4/MyD88/NF-κB)信号通路的影响。方法:先运用分子对接技术预测穿山龙复方中主要活性成分与TLR4/MyD88/NF-κB信号通路相关靶蛋白的分子间相互作用，再建立痛风性关节炎合并高尿酸血症大鼠复合模型进行体内实验验证。将48只雄性SD大鼠分为正常对照组、模型对照组、阳性对照秋水仙碱0.3 mg/kg组及穿山龙复方0.2、0.4、0.8 g/kg组。用尿酸钠、氧嗪酸钾和10%酵母饲料造模的同时，分别给予相应的药物灌胃。7 d后取材，测定踝关节肿胀度；HE染色观察踝关节病理学变化，血清白细胞介素-8(IL-8)、IL-1β、肿瘤坏死因子-α(TNF-α)含量；以Real-timePCR法测定大鼠踝关节中Tlr4、Myd88、Nfkb mRNA表达。结果:分子对接结果表明复方的主要活性成分与TLR4/MyD88/NF-κB信号通路的靶蛋白均有良好的结合活性，其中薯蓣皂苷与TLR4结合活性最强。体内实验结果表明，与正常对照组相比，模型对照组大鼠血尿酸含量以及踝关节损伤程度均显著增加，血清IL-1、IL-8、TNF-α的含量显著升高(P<0.01),踝关节中Tlr4、Myd88、Nfkb mRNA表达显著上调(P<0.01);与模型对照组比较，穿山龙复方各剂量组和秋水仙碱组对GA&HUA复合模型大鼠血尿酸含量以及踝关节损伤程度显著减轻，血清中IL-1β、IL-8、TNF-α的含量显著降低(P<0.01),踝关节中Tlr4、Myd88、Nfkb mRNA表达显著下调(P<0.01)。结论:穿山龙复方可以通过调节TLR4/MyD88/NF-κB信号通路达到抗痛风性关节炎和高尿酸血症的作用。

Objective:To explore the effect of Chuanshanlong(穿山龙) Compound Prescription on the Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB) signaling pathway in a rat model of gouty arthritis(GA) with hyperuricemia(HUA). Methods:Molecular docking technology was first used to predict the molecular interactions between the main active components of Chuanshanlong Compound Prescription and the target proteins related to the TLR4/MyD88/NF-κB signaling pathway. Subsequently, an in vivo experiment was conducted to validate the findings in the rat model of GA with HUA. Forty-eight male SD rats were divided into a normal control group, a model control group, a positive control group(colchicine),and high-,medium-,and low-dose Chuanshanlong Compound Prescription groups(0.8,0.4,and 0.2 g/kg). Sodium urate, oteracil potassium, and 10% yeast feed were used to induce the model, and corresponding drugs were orally administered. After 7 days, rats were sampled and ankle joint swelling was measured. Histopathological changes in the ankle joint were observed by HE staining, and the levels of interleukin-8(IL-8),interleukin-1β(IL-1β),and tumor necrosis factor-α(TNF-α) in serum were determined. Real-time PCR was used to measure the mRNA expression of Tlr4,Myd88 and Nf-kb in the rat ankle joint. Results:The molecular docking results showed that the main active components of Chuanshanlong Compound Prescription exhibited good binding affinity to the target proteins of the TLR4/MyD88/NF-κB signaling pathway, with dioscin showing the strongest binding affinity with TLR4. The in vivo experimental results demonstrated that compared with the normal control group, the model control group had significantly increased blood uric acid levels and ankle joint injury, significantly elevated levels of serum IL-1,IL-8,and TNF-α(P<0.01),and significantly up-regulated mRNA expression of Tlr4,Myd88 and Nf-kb in the ankle joint(P<0.01). Compared with the model control group, Chuanshanlong Compound Prescription at different doses and colchicine significantly reduced blood uric acid levels and ankle joint injury, significantly decreased the levels of IL-1β,IL-8 and TNF-α in serum(P<0.01),and significantly down-regulated the mRNA expression of Tlr4,Myd88 and Nf-kb in the ankle joint(P<0.01). Conclusion:Chuanshanlong Compound Prescription can exert its effects against GA and HUA by regulating the TLR4/MyD88/NF-κB signaling pathway.