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
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摘要
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 m L·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 m L·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.
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 m L·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.
引文
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[18]吴玉乱,许雅清,李海龙,等.久泻灵颗粒对脾肾阳虚型溃疡性结肠炎大鼠TLR4及NF-κB p65表达的影响[J].中国实验动物学报,2016,24(1):47-52.
[19]WANG R C,JIANG D M.PPAR-γagonist pioglitazone affects rat gouty arthritisby regulating cytokines[J].Genet Mol Res,2014,13(3):6577-6581.
[20]黄火高,韩星海,尚健,等.PPARγ激动刑吡格列酮对尿酸钠晶体诱导炎症的防治作用[J].中华风湿病学杂志,2005,9(8):463-468.
[21]黄火高,胡明,王建和,等.吡格列酮对大鼠尿酸钠晶体诱导的炎症组织细胞中S100A8和S100A9的mRNA表达的影响[J].中国药理学与毒理学杂志,2007,21(2):113-117.
[22]周子朋,郑福增,孟庆良,等.基于NALP3炎性体信号通路研究栀黄止痛颗粒治疗大鼠痛风性关节炎的作用机制[J].中国实验方剂学杂志,2018,24(8):140-147.
[2]Singh J A.Racial and gender disparities among patients with gout[J].Curr Rheumatol Rep,2013,15(2):307.
[3]匡红,曾琳,刘书蓉,等.成都地区28097例健康体检者血尿酸水平分析[J].国际检验医学杂志,2013,34(16):2121-2122.
[4]李建,张洁瑛,孙鹏,等.平胃散合桂枝芍药知母汤加减治疗慢性痛风性关节炎的疗效机制[J].中国实验方剂学杂志,2018,24(1):180-185.
[5]陆小龙,郭玉星,熊辉,等.蠲痹历节清方治疗湿热蕴结型痛风性关节炎的临床观察[J].中医药导报,2015,21(5):23-25.
[6]郭玉星,熊辉,陆小龙,等.蠲痹历节清方治疗急性痛风性关节炎的临床研究[J].云南中医学院学报,2016,39(1):81-84.
[7]周彪,郭玉星,陆小龙,等.蠲痹历节清方对大鼠痛风性关节炎关节肿胀指数和滑膜组织中炎症因子的影响[J].云南中医学院学报,2017,40(3):15-18.
[8]尤卓,熊辉,吴海金,等.蠲痹历节清方对急性痛风性关节炎大鼠的影响及其作用机制[J].中医正骨,2018,30(2):1-6.
[9]齐新宇,熊辉,周彪,等.蠲痹历节清方干预鸡急性痛风性关节炎模型的实验研究[J].中医正骨,2015,27(3):5-11.
[10]Bolzetta F,Veronese N,Manzato E,et al.Chronic gout in the elderly[J].Aging Clin Exp Res,2013,25(2):129-137.
[11]贾萍,陈刚.豨桐丸对大鼠痛风性关节炎的影响及机制[J].中国实验方剂学杂志,2018,24(1):96-101.
[12]Scott P,MA H,Viriyakosol S,et al.Engagment of CD14mediates the inflamm-atory potential of monosodiun urate crystals[J].J Immunol,2006,177(9):6370-6378.
[13]邵明伟.痛风宁颗粒对急性痛风性关节炎模型大鼠TNF-α,IL-6,COX-2及NF-κB影响的实验研究[D].福州:福建中医药大学,2011.
[14]傅玉辉.痛风宁颗粒对SD大鼠急性痛风性关节炎模型TLR/My D88影响的研究[D].福州:福建中医药大学,2011.
[15]QING Y F,ZHANG Q B,ZHOU J G,et al.Changes in toll-like receptor TLR4-NF-κB-IL-1βsignaling in male gout patients might be involved in the patho-genesis of primary gouty arthritis[J].Rheumatol Int,2013,34(2):213-220.
[16]YIN Y,HOU G,LI E R,et al.Regulation of cigarette smoke-induced toll like receptor 4 expression by peroxisome proliferator-activated receptor-gammaagonists in bronchial epithelial cells[J].Respirology,2013,18(S3):30-39.
[17]郭玉星,熊辉,陆小龙,等.改良痛风性关节炎大鼠模型的复制[J].云南中医学院学报,2017,40(2):18-23.
[18]吴玉乱,许雅清,李海龙,等.久泻灵颗粒对脾肾阳虚型溃疡性结肠炎大鼠TLR4及NF-κB p65表达的影响[J].中国实验动物学报,2016,24(1):47-52.
[19]WANG R C,JIANG D M.PPAR-γagonist pioglitazone affects rat gouty arthritisby regulating cytokines[J].Genet Mol Res,2014,13(3):6577-6581.
[20]黄火高,韩星海,尚健,等.PPARγ激动刑吡格列酮对尿酸钠晶体诱导炎症的防治作用[J].中华风湿病学杂志,2005,9(8):463-468.
[21]黄火高,胡明,王建和,等.吡格列酮对大鼠尿酸钠晶体诱导的炎症组织细胞中S100A8和S100A9的mRNA表达的影响[J].中国药理学与毒理学杂志,2007,21(2):113-117.
[22]周子朋,郑福增,孟庆良,等.基于NALP3炎性体信号通路研究栀黄止痛颗粒治疗大鼠痛风性关节炎的作用机制[J].中国实验方剂学杂志,2018,24(8):140-147.