小檗碱对小鼠痛风性关节炎模型中NLRP3/TLRs的调控作用

Regulatory effect of berberine on NLRP3/TLRs in mice with gouty arthritis

摘要:

目的：探究小檗碱对痛风性关节炎小鼠NLRP3/TLRs信号通路的调控作用。方法：C57BL/6小鼠随机均分为6组，分别为正常对照组，模型组，秋水仙碱组（阳性对照，0.65 mg/kg），小檗碱高、中、低剂量组（150、100、50 mg/kg），造模给药7 d，每天测定小鼠踝关节肿胀度，观察治疗作用；同时，采用ELISA法测定血清白细胞介素-2（interleukin-2，IL-2）、白细胞介素-6（inter－leukin-6，IL-6）、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）及干扰素-α（interferon-α，IFN-α）表达水平。进一步采用RT-PCR、Western blot分别检测关节滑膜组织中NLRP3/TLRs关键靶点基因及蛋白的表达。结果：关节肿胀度结果显示，小檗碱高、中、低剂量组（分别为18.35%、21.35%、25.35%）和秋水仙碱组（15.35%）与模型组（31.34%）比较明显降低，差异具有统计学意义（F=11.240，P=0.005）。且各剂量小檗碱治疗后，小鼠踝关节滑膜组织中炎性细胞浸润及水肿现象明显减轻。ELISA结果显示，与模型组比较，小檗碱各组小鼠血清IL-2、IL-6、TNF-α及IFN-α表达水平均明显下降（IL-2：F=5.690，P=0.005；IL-6：F=10.370，P=0.009；TNF-α：F=15.240，P=0.002；IFN-α：F=4.520，P=0.02）。RT-PCR及Western blot结果显示，小檗碱高、中、低剂量组的小鼠滑膜组织NLRP3、TLR3、TLR7、MyD88及NF-?资B p65的基因和蛋白表达均明显降低（P<0.05）。结论：小檗碱能缓解小鼠痛风性关节炎，其机制可能与下调炎症因子水平，抑制NLRP3/TLRs信号通路有关。

Abstract:

Objective：To investigate the regulatory effect of berberine on NLRP3/TLRs signal pathway in mice with gouty arthritis. Methods：C57BL/6 mice were randomly divided into 6 groups：normal control group，model group，colchicine group（positive control，0.65 mg/kg），and berberine groups with high，middle and low doses（150，100，50 mg/kg），respectively. Administration for 7 days after modeled，and the swelling degree of the ankle joint was measured every day to observe therapeutic effects. Meanwhile，serum inter－leukin-2（IL-2），interleukin-6（IL-6），tumor necrosis factor-α（TNF-α） and interferon-α（IFN-α） expression levels were measured by ELISA. And the expressions of key target genes and proteins of NLRP3/TLRs in the mice's ankle synovial tissue were detected by RT-PCR and Western blot assays. Results：Compared with the model group（31.34%），the swelling degrees of the ankle joint of the berberine groups with high，medium and low doses（18.35%，21.35%，25.35%，respectively） and the colchicine group（15.35%） were significantly reduced，and the difference was statistically significant（F=11.240，P=0.005）. After the berberine treatment with different doses，inflammatory cell infiltration and edema in the ankle synovial tissue were significantly alleviated. The ELISA results showed that compared with the model group，the serum levels of IL-2，IL-6，TNF-α and IFN-α in the berberine groups were significantly decreased（IL-2：F=5.690，P=0.005；IL-6：F=10.370，P=0.009；TNF-α：F=15.240，P=0.002；IFN-α：F=4.520，P=0.020）. RT-PCR and Western blot results showed that the expression levels of NLRP3，TLR3，TLR7，MyD88 and NF-?资B p65 genes and proteins were obviously decreased in the mice's synovial tissue of the berberine groups with high，medium and low doses（P<0.05）. Conclusion：Berberine can alleviate gouty arthritis in mice，and its mechanism may be related to down-regulating inflammatory cytokine levels and inhibiting NLRP3/TLRs signal pathway.