虎杖对胶原诱导性关节炎大鼠滑膜PPARγ/ NF-κB信号途径的影响
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摘要
目的通过观察虎杖对胶原诱导性关节炎(CIA)大鼠踝关节病理及滑膜组织、血清中过氧化物酶体增殖物激活受体-γ(PPARγ)、p65、白细胞介素17(IL-17)表达的影响,探讨虎杖抑制类风湿关节炎(RA)滑膜免疫炎性损伤的作用机制。方法采用大鼠尾根部注射牛CⅡ方法制备CIA模型,将成模大鼠按随机数字表法分为模型组、阳性对照组、虎杖2倍剂量组、虎杖常量组,另设正常对照组,每组6只。正常对照组与模型组每天予去离子水10 mL/kg灌胃,阳性对照组每天予来氟米特1.87 mg/kg灌胃,虎杖2倍剂量组、常量组每天分别以8、4 g/kg灌胃,连续干预12周。采用光镜观察各组大鼠踝关节病理改变。运用RT-PCR、Western Blot及ELISA检测技术,观察各组大鼠滑膜组织、血清中PPARγ、p65、IL-17 mRNA及蛋白表达情况。结果与正常对照组比较,模型组大鼠踝关节关节周围纤维组织增生,炎性细胞浸润;与模型组比较,虎杖常量组大鼠关节软骨、关节腔未见明显病变。与正常对照组比较,模型组PPARγ、p65、IL-17 mRNA及蛋白表达上调(P<0.01)。与模型组比较,虎杖2倍剂量组及常量组p65、IL-17 mRNA及蛋白表达均下调(P<0.01);PPARγmRNA蛋白表达水平上调(P<0.01)。结论虎杖改善CIA模型大鼠关节滑膜免疫炎性损伤的机制,可能与调节PPARγ/NF-κB信号途径相关。
Objective To observe the effects of Polygonum cuspidatum on ankle joint pathology, synovial tissue, and serum peroxisome proliferator activated receptor gamma(PPAR gamma),p65, IL-17 expression in rats with collagen-induced arthritis(CIA), and to study its mechanism for inhibiting synovial immune inflammatory injury in rheumatoid arthritis(RA). Methods CIA model was established by injecting bovine CⅡ into the tail root of rats. The rats were divided into model group, positive control group, 2 times dosage group of Polygonum cuspidatum, and constant dosage group of Polygonum cuspidatum by random digit table, 6 in each group. Another normal control group was set up(n=6). Deionized water 10 mL·kg~(-1)·d~(-1) was administered to rats in the normal control group and the model group by gastrogavage. Leflunomide 1.87 mg·kg~(-1)·d~(-1) was administered to rats in the positive control group by gastrogavage. Eight and 4 g·kg~(-1)·d~(-1) Polygonum cuspidatum was respectively administered to rats in the 2 times dosage group of Polygonum cuspidatum, and constant dosage group by gastrogavage. All medication lasted for 12 successive weeks. The pathological changes of ankle joint of rats in each group were observed by light microscopy. RT-PCR, Western Blot and ELISA were used to detect the expressions of PPAR-gamma, p65 and IL-17 in synovial tissue and serum of rats in each group. ResultsCompared with the normal control group, the fibrous tissue around the ankle joint in the model group were proliferated and inflammatory cells infiltrated. Compared with the model group, there were no obvious pathological changes in the articular cartilage and articular cavity in the constant dosage group of Polygonum cuspidatum. Compared with the normal control group, mRNA and protein expressions of PPAR gamma, p65, and IL-17 in the model group were up-regulated(P<0.01). Compared with the model group, mRNA and protein expressions of p65 and IL-17 in the two Polygonum cuspidatum groups were down-regulated(P<0.01); the expressions of PPAR gamma in the two Polygonum cuspidatum groups were up-regulated(P<0.01). Conclusion The mechanism of Polygonum cuspidatum for improving synovial immune inflammatory injury in CIA rats might be related to regulating PPARgamma/NF-kappa B signaling pathway.
Objective To observe the effects of Polygonum cuspidatum on ankle joint pathology, synovial tissue, and serum peroxisome proliferator activated receptor gamma(PPAR gamma),p65, IL-17 expression in rats with collagen-induced arthritis(CIA), and to study its mechanism for inhibiting synovial immune inflammatory injury in rheumatoid arthritis(RA). Methods CIA model was established by injecting bovine CⅡ into the tail root of rats. The rats were divided into model group, positive control group, 2 times dosage group of Polygonum cuspidatum, and constant dosage group of Polygonum cuspidatum by random digit table, 6 in each group. Another normal control group was set up(n=6). Deionized water 10 mL·kg~(-1)·d~(-1) was administered to rats in the normal control group and the model group by gastrogavage. Leflunomide 1.87 mg·kg~(-1)·d~(-1) was administered to rats in the positive control group by gastrogavage. Eight and 4 g·kg~(-1)·d~(-1) Polygonum cuspidatum was respectively administered to rats in the 2 times dosage group of Polygonum cuspidatum, and constant dosage group by gastrogavage. All medication lasted for 12 successive weeks. The pathological changes of ankle joint of rats in each group were observed by light microscopy. RT-PCR, Western Blot and ELISA were used to detect the expressions of PPAR-gamma, p65 and IL-17 in synovial tissue and serum of rats in each group. ResultsCompared with the normal control group, the fibrous tissue around the ankle joint in the model group were proliferated and inflammatory cells infiltrated. Compared with the model group, there were no obvious pathological changes in the articular cartilage and articular cavity in the constant dosage group of Polygonum cuspidatum. Compared with the normal control group, mRNA and protein expressions of PPAR gamma, p65, and IL-17 in the model group were up-regulated(P<0.01). Compared with the model group, mRNA and protein expressions of p65 and IL-17 in the two Polygonum cuspidatum groups were down-regulated(P<0.01); the expressions of PPAR gamma in the two Polygonum cuspidatum groups were up-regulated(P<0.01). Conclusion The mechanism of Polygonum cuspidatum for improving synovial immune inflammatory injury in CIA rats might be related to regulating PPARgamma/NF-kappa B signaling pathway.
引文
[1] 李雪,栗占国.2015年亚洲太平洋地区风湿病学学会联盟类风湿关节炎治疗建议[J].中华风湿病学杂志,2016,20(4):286-288.
[2] 陆妍,王亚南,刘慧,等.中药藤莓汤对Ⅱ型胶原诱导性大鼠关节炎性病理损伤的影响[J].中国比较医学杂志,2015,25(4):48-57.
[3] 王洁,王亚南,陆妍,等.藤莓汤对胶原诱导性关节炎大鼠滑膜PPARγ/NF-κB信号途径的影响[J].中国中西医结合杂志,2017,37(4):458-463.
[4] 卜祥伟,张红红,张建萍,等.藤莓汤联合传统改善病情抗风湿药治疗中/高活动度类风湿关节炎患者的疗效观察[J].中国中西医结合杂志,2017,37(11):1320-1324.
[5] 国家药典委员会.中华人民共和国药典[M].北京:中国医药科技出版社,2015:208-509.
[6] 时圣明,潘明佳,王文倩,等.虎杖的化学成分及药理作用研究进展[J].药物评价研究,2016,39(2):317-321.
[7] 王永萍,杨长福,谭芸,等.虎杖大黄素对RA大鼠炎症和新生血管形成的影响[J].中国实验方剂学杂志,2015,21(17):111-115.
[8] 朱丽琴,马露,熊彪,等.虎杖大黄素对RA大鼠Cu-ZnSOD的影响[J].世界最新医学信息文摘,2016,16(54):23-24.
[9] 曾家顺,董晓,刘俊,等.虎杖苷对类风湿关节炎大鼠的治疗机制探究[J].天然产物研究与开发,2018,30(10):1681-1686.
[10] 叶茂高,高祥福.虎杖治疗痛风的作用机制研究进展[J].黑龙江中医药,2015,44(5):71-73.
[11] 刘玮,钱善军,黄平,等.虎杖对大鼠酒精性脂肪肝的作用及机制[J].中成药,2018,40(1):184-186.
[12] 陈瑞林,黄文辉,黄成辉,等.PPAR-γ在类风湿关节炎患者外周血单个核细胞中的表达及与疾病活动度的关系[J].实用临床医药杂志,2015,19(7):65-67,90.
[13] 徐蓓,厉小梅.过氧化物酶体增殖物激活受体与自身免疫性疾病[J].中华风湿病学杂志,2011,15 (3 ) :197-200.
[14] Xiao ZW,Jiang X,Fu J,et al.Polydatin protects myocardiocytes from acute myocardial infarction injury in mice[J].Acta Med Univ Sci Tech Huazhong,2016,45(2):176-180.
[15] Gui TC,Ru YW,Wei LZ.Screening and identification of pancreatic lipase inhibitors in Polygonum cuspidatum with enzyme-immobilized magnetic nanoparticles and LC-MS/M[J].Nat Prod Res Dev,2017,29(2):198-205.
[16] Wu YT,Liu Y,Liu M,et al.Expression and function of PTEN in fibroblast-like synoviocytes of rheumatoid arthritis[J].Chin J Path,2016,32:978-983.
[17] Zhen YY,Dai DX,Pan Z,et al.Inhibitory effect of salidroside on proliferation of HFLS-RA induced by TNF-α and its significance[J].J Jilin Univ Med,2017,43(3):485-490.
[18] 张力力,孙铀.白细胞介素17在类风湿关节炎中的致炎作用及靶向治疗进展[J].医学综述,2017 ,23 (10):1873-1877.
[19] 黄艳,冯辉,刘世敏,等.针灸对CIA大鼠的脾脏单个核细胞IL-17、IL-12表达的影响[J].世界科学技术-中医药现代化,2018,20(2):208-214.
[20] 马红爽.γδT细胞及其细胞功能分子在系统性红斑狼疮和类风湿关节炎中的研究[D].长春:吉林大学,2015.
[2] 陆妍,王亚南,刘慧,等.中药藤莓汤对Ⅱ型胶原诱导性大鼠关节炎性病理损伤的影响[J].中国比较医学杂志,2015,25(4):48-57.
[3] 王洁,王亚南,陆妍,等.藤莓汤对胶原诱导性关节炎大鼠滑膜PPARγ/NF-κB信号途径的影响[J].中国中西医结合杂志,2017,37(4):458-463.
[4] 卜祥伟,张红红,张建萍,等.藤莓汤联合传统改善病情抗风湿药治疗中/高活动度类风湿关节炎患者的疗效观察[J].中国中西医结合杂志,2017,37(11):1320-1324.
[5] 国家药典委员会.中华人民共和国药典[M].北京:中国医药科技出版社,2015:208-509.
[6] 时圣明,潘明佳,王文倩,等.虎杖的化学成分及药理作用研究进展[J].药物评价研究,2016,39(2):317-321.
[7] 王永萍,杨长福,谭芸,等.虎杖大黄素对RA大鼠炎症和新生血管形成的影响[J].中国实验方剂学杂志,2015,21(17):111-115.
[8] 朱丽琴,马露,熊彪,等.虎杖大黄素对RA大鼠Cu-ZnSOD的影响[J].世界最新医学信息文摘,2016,16(54):23-24.
[9] 曾家顺,董晓,刘俊,等.虎杖苷对类风湿关节炎大鼠的治疗机制探究[J].天然产物研究与开发,2018,30(10):1681-1686.
[10] 叶茂高,高祥福.虎杖治疗痛风的作用机制研究进展[J].黑龙江中医药,2015,44(5):71-73.
[11] 刘玮,钱善军,黄平,等.虎杖对大鼠酒精性脂肪肝的作用及机制[J].中成药,2018,40(1):184-186.
[12] 陈瑞林,黄文辉,黄成辉,等.PPAR-γ在类风湿关节炎患者外周血单个核细胞中的表达及与疾病活动度的关系[J].实用临床医药杂志,2015,19(7):65-67,90.
[13] 徐蓓,厉小梅.过氧化物酶体增殖物激活受体与自身免疫性疾病[J].中华风湿病学杂志,2011,15 (3 ) :197-200.
[14] Xiao ZW,Jiang X,Fu J,et al.Polydatin protects myocardiocytes from acute myocardial infarction injury in mice[J].Acta Med Univ Sci Tech Huazhong,2016,45(2):176-180.
[15] Gui TC,Ru YW,Wei LZ.Screening and identification of pancreatic lipase inhibitors in Polygonum cuspidatum with enzyme-immobilized magnetic nanoparticles and LC-MS/M[J].Nat Prod Res Dev,2017,29(2):198-205.
[16] Wu YT,Liu Y,Liu M,et al.Expression and function of PTEN in fibroblast-like synoviocytes of rheumatoid arthritis[J].Chin J Path,2016,32:978-983.
[17] Zhen YY,Dai DX,Pan Z,et al.Inhibitory effect of salidroside on proliferation of HFLS-RA induced by TNF-α and its significance[J].J Jilin Univ Med,2017,43(3):485-490.
[18] 张力力,孙铀.白细胞介素17在类风湿关节炎中的致炎作用及靶向治疗进展[J].医学综述,2017 ,23 (10):1873-1877.
[19] 黄艳,冯辉,刘世敏,等.针灸对CIA大鼠的脾脏单个核细胞IL-17、IL-12表达的影响[J].世界科学技术-中医药现代化,2018,20(2):208-214.
[20] 马红爽.γδT细胞及其细胞功能分子在系统性红斑狼疮和类风湿关节炎中的研究[D].长春:吉林大学,2015.