蠲痹历节清方对改良痛风性关节模型大鼠滑膜的TLR4,NF-κB,PPARγ的影响

Effect of Juanbi Lijieqing Formula on TLR4,NF-κB and PPARγ in Synovial Membrane of Rats with Amelioration of Gouty Arthritis

目的：观察蠲痹历节清方对改良痛风性关节炎大鼠滑膜组织中Toll样受体4（Toll-like receptors 4,TLR4）,核转录因子kappa B（nuclear factor-kappa B,NF-κB）,过氧化物酶体增殖物激活受体γ（peroxisome proliferator-activated receptorγ,PPARγ）的影响,探讨蠲痹历节清方治疗急性痛风性关节炎局部组织炎症的可能作用机制。方法：将42只SD雄性大鼠,按随机数字表法选取6只为正常组,30只为造模组,造模组采用改良痛风性关节炎造模后随机分为模型组,蠲痹历节清方高、中、低剂量组（4 400,2 200,1 100 mg·kg^-1）,依托考昔组（11 mg·kg^-1）,吡格列酮组（20 mg·kg^-1）,每组6只。正常组及模型组的大鼠以生理盐水灌胃20 mL·kg^-1。每组每只大鼠每日灌胃给药2次,连续2 d后处死大鼠,取受试右踝关节,分离出滑膜组织,分为两部分,一部分用于病理形态学观察,一部分用逆转录聚合酶链式反应（RT-PCR）检测TLR4,NF-κB p65,PPARγmRNA的表达水平,蛋白质免疫印迹法（Western blot）检测TLR4,NF-κB p65,PPARγ蛋白表达水平。结果：与正常组比较,模型组中TLR4和NF-κB mRNA和蛋白表达显著升高（P〈0.01）,PPARγmRNA和蛋白表达升高（P〈0.05）;与模型组比较,吡格列酮组及蠲痹历节清方高、中、低剂量组TLR4和NF-κB mRNA和蛋白的表达显著降低（P〈0.01）,PPARγmRNA和蛋白的表达显著升高（P〈0.05）。结论：蠲痹历节清方可能通过上调PPARγ表达,抑制TLR4,NF-κB表达从而抑制急性痛风性关节炎局部组织炎症。

Objective： To observe the effect of Juanbi Lijieqing formula on Toll-like receptors 4（ TLR4）,nuclear factor-kappa B（ NF-κB）,peroxisome proliferator-activated receptor γ（ PPARγ） in synovial tissue of modified gouty arthritis rats,in order to explore the possible mechanism of Juanbi Lijieqing formula in the treatment of acute gouty arthritis with local tissue inflammation. Method： Among 42 male SD rats,6 were selected as blank group according to the random number table method,30 were included in the modeling group. The modeling group was randomly divided into model group, Juanbi Lijieqing formula group（ 4 400,2 200,1 100 mg·kg^-1）,celecoxib group（ 11 mg·kg^-1）,and pioglitazone group（ 20 mg·kg^-1）,with 6 mice in each group. Rats in normal control group and model group were given normal saline（ 20 mL·kg^-1）. Rats in each group were administrated with drugs for 2 times a day for 2 days; after that,the rats were put to death,the right ankle joints were collected,and synovial tissues were isolated. The tissues were divided into two parts,one part was used for the observation of pathomorphology,while the other part was used to detect mRNA expressions of TLR4,NF-κB p65 and PPARγ by reverse transcription-polymerase chain reaction（ RT-PCR）. Western blot was used to detect protein expressions of TLR4,NF-κB p65 and PPARγ. Result： Compared with the normal control group,mRNA and protein expressions of TLR4 and NF-κB increased significantly（ P〈0. 01） in the model group,and mRNA and protein expression of PPARγ in the model group increased（ P〈0. 05）. Compared with the model group,the expressions of TLR4 and NF-κB in the pioglitazone group and low,middle and high-dose Juanbi Lijieqing formula groups decreased significantly（ P〈0. 01）,and mRNA and protein expression of PPARγ increased significantly（ P〈0. 05）. Conclusion： Juanbi Lijieqing formula may inhibit local inflammation in acute gouty arthritis by upregulating the expression of PPARγ and inhibiting the expressions of TLR4 and NF-κB.