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目的研究痛风泰颗粒对TLRs/NF-κB信号通路中关键基因的调控作用,探究该药物抗炎的作用机制。方法将24只大鼠随机分为正常对照组、模型对照组、阳性对照组、痛风泰颗粒低剂量组、中剂量组及高剂量组。除正常对照组外,均向每组大鼠踝关节腔注射尿酸钠混悬液,造成急性痛风大鼠模型,随后按照分组和剂量灌胃给药5 d。末次灌服后分别采用ELISA及免疫组化法测定大鼠血清中TNF-α的含量和关节滑膜组织中NF-κB p65的表达。另将15只大鼠随机分为A、B、C、D、E组,分别灌服生理盐水、戴芬溶液、低剂量痛风泰颗粒、中剂量痛风泰颗粒及高剂量痛风泰颗粒,得到5种含药血清。取8只大鼠,构建痛风大鼠模型后取关节滑膜组织培养炎性的原代滑膜细胞,分别用A、B、C、D、E组血清刺激相应的A、B、C、D、E组细胞,利用RT-PCR检测不同组别血清刺激8 h后TLR2、TLR3及TLR4基因mRNA的表达。结果与正常对照组相比,模型对照组血清中TNF-α含量、关节滑膜组织中NF-κB p65表达量均升高(P<0.01);与模型对照组相比,阳性对照组、低、中、高剂量组的大鼠血清中TNF-α含量、关节滑膜组织中NF-κB p65表达量都有所下降(P<0.05,P<0.01,P<0.001);血清刺激后,与A组相比,B、C、D、E组细胞TLR4 mRNA的相对表达量均有所降低,呈现出一定变化趋势(P<0.05,P<0.001)。但TLR2和TLR3基因的mRNA相对表达量无任何趋势及表达差异。结论痛风泰颗粒能够降低TLRs/NF-κB信号通路中关键基因TLR4、TNF-α及NF-κB的表达,作用效果与戴芬相近。该药物可能通过这一通路发挥抗炎作用,达到治疗痛风的效果。
Objective To study the regulatory effect of gao fengtai granule on key genes in TLRs / NF-κB signaling pathway and explore the anti-inflammatory mechanism of this drug. Methods Twenty-four rats were randomly divided into normal control group, model control group, positive control group, Tongfengtai Granules low dose group, middle dose group and high dose group. In addition to the normal control group, each group of rats were injected sodium hyaluronate suspension into the ankle joint cavity, causing acute gout rat model, followed by grouping and dose gavage for 5 d. Serum levels of TNF-α and NF-κB p65 in synovial tissues were measured by ELISA and immunohistochemistry respectively after the last oral administration. Another 15 rats were randomly divided into groups A, B, C, D, E, were drenched with saline, Dai Fen solution, low dose of goutytai particles, the amount of goutytai particles and high dose of goutytai particles, get 5 Species containing serum. Eight rats were selected to establish synovial tissue of primary gout model. Primary synovial cells were cultured and cultured in vitro. Serum A, B, C, D and E were used to stimulate the corresponding synovial cells of A, B, C, D , E group. RT-PCR was used to detect the mRNA expression of TLR2, TLR3 and TLR4 in different groups of serum for 8 h. Results Compared with the normal control group, the levels of TNF-α in serum and the expression of NF-κB p65 in synovial tissue of model control group were significantly increased (P <0.01). Compared with the model control group, the positive control group (P <0.05, P <0.01, P <0.001). The levels of TNF-α and the expression of NF-κB p65 in synovial tissues of rats in middle, Compared with group A, the relative expression of TLR4 mRNA in group B, C, D, E decreased, showing a trend of change (P <0.05, P <0.001). However, TLR2 and TLR3 gene mRNA relative expression of any trend and expression differences. CONCLUSION: Tongfutai Granules can reduce the expression of TLR4, TNF-α and NF-κB in TLRs / NF-κB signaling pathway with similar effect to Dai Fen. The drug may exert anti-inflammatory effect through this pathway to achieve the effect of treating gout.