蒙药森登-4汤抑制胶原诱导性关节炎大鼠骨破坏作用机制及对滑膜成纤维细胞体外增殖的影响
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摘要
目的研究蒙药森登-4汤抑制胶原诱导性关节炎(CIA)大鼠骨破坏作用机制及对滑膜成纤维细胞(FLS)体外增殖的影响。方法本研究首先利用牛Ⅱ型胶原建立SD大鼠CIA模型,造模成功后随机分为模型组、雷公藤多苷组和蒙药森登-4汤高、中、低剂量组,各10只;正常组10只大鼠不造模。各组大鼠给予相应药物,观察各组大鼠踝关节组织病理学改变,检测各组大鼠血清白细胞介素1(IL-1)、IL-6、IL-17、肿瘤坏死因子α(TNF-α)等细胞因子含量,检测Wnt信号通路关键因子DKK1蛋白在爪关节中的表达。随机取1只CIA大鼠分离FLS行体外培养,实验分为空白对照组、雷公藤多苷组及高、中、低剂量蒙药森登-4汤含药血清组,干预24、48、72 h后检测CIA-FLS增殖情况。结果各用药组大鼠滑膜增生、细胞浸润及骨破坏评分较模型组均有不同程度下调,其中雷公藤多苷组和蒙药森登-4汤高剂量组下调最为明显[(1. 10±0. 57)、(1. 20±0. 42)分比(2. 80±0. 42)分;(1. 10±0. 88)、(1. 10±0. 32)分比(2. 90±0. 32)分;(0. 90±0. 57)、(1. 30±0. 48)分比(2. 70±0. 48)分](均P <0. 01)。各用药组大鼠血清IL-1、IL-6、IL-17、TNF-α水平和爪关节DKK1蛋白表达水平与模型组比较均有所下降。高、中剂量蒙药森登-4汤含药血清组和雷公藤多苷组干预24、48、72 h后,以及低剂量蒙药森登-4汤含药血清组干预48、72 h后,均可明显抑制CIA-FLS增殖,与空白对照组比较差异均有统计学意义(均P <0. 05)。结论蒙药森登-4汤对CIA大鼠具有明显的骨破坏抑制作用,对CIA-FLS体外增殖有明显抑制作用,其作用存在量效关系,蒙药森登-4汤高剂量作用优于中剂量和低剂量。
Objective To explore the mechanism of Mongolian medicine Sendeng-4 decoction in inhibiting bone destruction in collagen-induced arthritis( CIA) rats and its effect on fibroblast-like synoviocyte( FLS) culture in vitro. Methods CIA model of SD rats was established using bovine collagen type Ⅱ. SD rats were randomly divided into normal group,model control group,tripterygium wilfordii polyglycoside group and high,medium,low dose of Mongolian medicine Sendeng-4 decoction groups,with 10 rats in each group. Histopathological changes of ankle joint were observed. Levels of serum interleukin-1( IL-1),IL-6,IL-17 and tumor necrosis factor-α( TNF-α)were detected. Expression of DKK1 protein,the key factor of Wnt signaling pathway,in claw joint was detected.CIA-FLS was isolated from 1 rat,cultured in vitro and divided into blank control group,tripterygium wilfordii polyglycoside group and high,medium,low dose of Mongolian medicine Sendeng-4 decoction groups. Proliferation of CIA-FLS was detected 24,48,72 h after drug intervention. Results Scores of synovial hyperplasia,cell infiltration and bone damage were down-regulated in drug intervention groups compared to those in model control group,especially in tripterygium wilfordii group and high dose of Sendeng-4 decoction group [( 1. 10 ± 0. 57),( 1. 20 ± 0. 42) vs( 2. 80 ± 0. 42);( 1. 10 ± 0. 88),( 1. 10 ± 0. 32) vs( 2. 90 ± 0. 32);( 0. 90 ± 0. 57),( 1. 30 ±0. 48) vs( 2. 70 ± 0. 48) ]( all P < 0. 01). Levels of IL-1,IL-6,IL-17,TNF-α and expression of DKK1 protein in drug intervention groups were lower than those in model control group. Compared to blank control group,CIA-FLS proliferation was significantly inhibited in high and medium dose Sendeng-4 decoction groups at 24,48,72 h after intervention and in low dose Sendeng-4 decoction group at 48 and 72 h after intervention( all P < 0. 01).Conclusion Mongolian medicine Sendeng-4 decoction shows dose-dependent inhibition on bone destruction in CIA rats and vitro culture of CIA-FLS.
Objective To explore the mechanism of Mongolian medicine Sendeng-4 decoction in inhibiting bone destruction in collagen-induced arthritis( CIA) rats and its effect on fibroblast-like synoviocyte( FLS) culture in vitro. Methods CIA model of SD rats was established using bovine collagen type Ⅱ. SD rats were randomly divided into normal group,model control group,tripterygium wilfordii polyglycoside group and high,medium,low dose of Mongolian medicine Sendeng-4 decoction groups,with 10 rats in each group. Histopathological changes of ankle joint were observed. Levels of serum interleukin-1( IL-1),IL-6,IL-17 and tumor necrosis factor-α( TNF-α)were detected. Expression of DKK1 protein,the key factor of Wnt signaling pathway,in claw joint was detected.CIA-FLS was isolated from 1 rat,cultured in vitro and divided into blank control group,tripterygium wilfordii polyglycoside group and high,medium,low dose of Mongolian medicine Sendeng-4 decoction groups. Proliferation of CIA-FLS was detected 24,48,72 h after drug intervention. Results Scores of synovial hyperplasia,cell infiltration and bone damage were down-regulated in drug intervention groups compared to those in model control group,especially in tripterygium wilfordii group and high dose of Sendeng-4 decoction group [( 1. 10 ± 0. 57),( 1. 20 ± 0. 42) vs( 2. 80 ± 0. 42);( 1. 10 ± 0. 88),( 1. 10 ± 0. 32) vs( 2. 90 ± 0. 32);( 0. 90 ± 0. 57),( 1. 30 ±0. 48) vs( 2. 70 ± 0. 48) ]( all P < 0. 01). Levels of IL-1,IL-6,IL-17,TNF-α and expression of DKK1 protein in drug intervention groups were lower than those in model control group. Compared to blank control group,CIA-FLS proliferation was significantly inhibited in high and medium dose Sendeng-4 decoction groups at 24,48,72 h after intervention and in low dose Sendeng-4 decoction group at 48 and 72 h after intervention( all P < 0. 01).Conclusion Mongolian medicine Sendeng-4 decoction shows dose-dependent inhibition on bone destruction in CIA rats and vitro culture of CIA-FLS.
引文
[1]施桂英.关节炎概要[M].北京:中国医药科技出版社,2000:675.Shi GY. Synopsis of arthritis[M]. Beijing:Medicine Science and Technology Press of China,2000:675.
[2]宇妥·元旦贡布.四部医典[M].呼和浩特:内蒙古人民出版社,1959:394.Yutuo YDGB. The Four Medical Tantras[M]. Hohhot:Inner Mongolia Peoples Publishing House,1959:394.
[3]刘玉林,张琰,胡玉珍.基础医学动物实验技术[M].西安:第四军医大学出版社,2008:106-115.Liu YL,Zhang Y,Hu YZ. Animal experimental techniques in Basic Medicine[M]. Xi'an:Fourth Military Medical University Press,2008:106-115.
[4] Trentham DE,Townes AS,Kang AH. Autoimmunity to typeⅡcollagen an experimental model of arthritis[J]. J Exp Med,1977,146(3):857-868.
[5]王燕,赵宏艳,刘梅洁,等.类风湿关节炎肾虚证大鼠模型的建立[J].中西医结合学报,2011,9(9):973-982. DOI:10. 3736/jcim20110908.Wang Y,Zhao HY,Liu MJ,et al. Establishment of a rat model of rheumatoid arthritis with kidney deficiency syndrome[J]. Journal of Chinese Integrative Medicine,2011,9(9):973-982. DOI:10.3736/jcim20110908.
[6] Rosloniec EF,Cremer M,Kang AH,et al. Collagen-induced arthritis[J]. Curr Protoc Immunol,2010,89:15. 5. 1-15. 5. 25.DOI:10. 1002/0471142735. im1505s89.
[7]陈奇.中药药理研究方法学[M]. 2版.北京:人民卫生出版社,2006:31-32.Chen Q. Methodology of Pharmacological Research of Traditional Chinese Medicine[M]. 2nd ed. Beijing:People's Medical Publishing House,2006:31-32.
[8]高皖皎,邓秋狄,白殊同,等.佐剂型关节炎大鼠滑膜成纤维细胞模型建立及特征分析[J].中国药理学通报,2015,31(12):1693-1698. DOI:10. 3969/j. issn. 1001-1978. 2015. 12. 013.Gao WJ,Deng QD,Bai ST,et al. Establishment and characteristic analysis of fibroblast-like synoviocytes in rats with adjuvant arthritis[J].Chinese Pharmacological Bulletin,2015,31(12):1693-1698.DOI:10. 3969/j. issn. 1001-1978. 2015. 12. 013.
[9]石淙.细胞增殖的检测方法[J].实验与检验医学,2012,30(2):153-155,168. DOI:10. 3969/j. issn. 1674-1129. 2012. 02. 016.Shi C. Detection of cell proliferation[J]. Experimental and Laboratory Medicine,2012,30(2):153-155,168. DOI:10. 3969/j.issn. 1674-1129. 2012. 02. 016.
[10] Kong JS,Yoo SA,Kim JW,et al. Anti-neuropilin-1 peptide inhibition of synoviocyte survival,angiogenesis,and experimental arthritis[J].Arthritis Rheum,2010,62(1):179-190. DOI:10. 1002/art. 27243.
[11] Gupta SC,Tyagi AK,Deshmukh-Taskar P,et al. Downregulation of tumor necrosis factor and other proinflammatory biomarkers by polyphenols[J]. Arch Biochem Biophys,2014,559:91-99. DOI:10. 1016/j. abb. 2014. 06. 006.
[12] Jules J,Zhang P,Ashley JW,et al. Molecular basis of requirement of receptor activator of nuclear factorκB signaling for interleukin1-mediated osteoclastogenesis[J]. J Biol Chem,2012,287(19):15728-15738. DOI:10. 1074/jbc. M111. 296228.
[2]宇妥·元旦贡布.四部医典[M].呼和浩特:内蒙古人民出版社,1959:394.Yutuo YDGB. The Four Medical Tantras[M]. Hohhot:Inner Mongolia Peoples Publishing House,1959:394.
[3]刘玉林,张琰,胡玉珍.基础医学动物实验技术[M].西安:第四军医大学出版社,2008:106-115.Liu YL,Zhang Y,Hu YZ. Animal experimental techniques in Basic Medicine[M]. Xi'an:Fourth Military Medical University Press,2008:106-115.
[4] Trentham DE,Townes AS,Kang AH. Autoimmunity to typeⅡcollagen an experimental model of arthritis[J]. J Exp Med,1977,146(3):857-868.
[5]王燕,赵宏艳,刘梅洁,等.类风湿关节炎肾虚证大鼠模型的建立[J].中西医结合学报,2011,9(9):973-982. DOI:10. 3736/jcim20110908.Wang Y,Zhao HY,Liu MJ,et al. Establishment of a rat model of rheumatoid arthritis with kidney deficiency syndrome[J]. Journal of Chinese Integrative Medicine,2011,9(9):973-982. DOI:10.3736/jcim20110908.
[6] Rosloniec EF,Cremer M,Kang AH,et al. Collagen-induced arthritis[J]. Curr Protoc Immunol,2010,89:15. 5. 1-15. 5. 25.DOI:10. 1002/0471142735. im1505s89.
[7]陈奇.中药药理研究方法学[M]. 2版.北京:人民卫生出版社,2006:31-32.Chen Q. Methodology of Pharmacological Research of Traditional Chinese Medicine[M]. 2nd ed. Beijing:People's Medical Publishing House,2006:31-32.
[8]高皖皎,邓秋狄,白殊同,等.佐剂型关节炎大鼠滑膜成纤维细胞模型建立及特征分析[J].中国药理学通报,2015,31(12):1693-1698. DOI:10. 3969/j. issn. 1001-1978. 2015. 12. 013.Gao WJ,Deng QD,Bai ST,et al. Establishment and characteristic analysis of fibroblast-like synoviocytes in rats with adjuvant arthritis[J].Chinese Pharmacological Bulletin,2015,31(12):1693-1698.DOI:10. 3969/j. issn. 1001-1978. 2015. 12. 013.
[9]石淙.细胞增殖的检测方法[J].实验与检验医学,2012,30(2):153-155,168. DOI:10. 3969/j. issn. 1674-1129. 2012. 02. 016.Shi C. Detection of cell proliferation[J]. Experimental and Laboratory Medicine,2012,30(2):153-155,168. DOI:10. 3969/j.issn. 1674-1129. 2012. 02. 016.
[10] Kong JS,Yoo SA,Kim JW,et al. Anti-neuropilin-1 peptide inhibition of synoviocyte survival,angiogenesis,and experimental arthritis[J].Arthritis Rheum,2010,62(1):179-190. DOI:10. 1002/art. 27243.
[11] Gupta SC,Tyagi AK,Deshmukh-Taskar P,et al. Downregulation of tumor necrosis factor and other proinflammatory biomarkers by polyphenols[J]. Arch Biochem Biophys,2014,559:91-99. DOI:10. 1016/j. abb. 2014. 06. 006.
[12] Jules J,Zhang P,Ashley JW,et al. Molecular basis of requirement of receptor activator of nuclear factorκB signaling for interleukin1-mediated osteoclastogenesis[J]. J Biol Chem,2012,287(19):15728-15738. DOI:10. 1074/jbc. M111. 296228.