苗药验方“四大血”对胶原诱导关节炎大鼠IL-17和VEGF表达的影响
|
摘要
目的:探讨苗药验方"四大血"(SX)对牛Ⅱ型胶原蛋白诱导关节炎(CIA)大鼠白细胞介素17(IL-17)及血管内皮生长因子(VEGF)表达水平的影响。方法:42只SD大鼠分为空白对照组(NG组,n=7)及造模组(n=35),除NG组外,所有大鼠用牛Ⅱ型胶原与等量不完全弗氏佐剂混合乳化剂于尾根部皮下注射致炎、复制CIA动物模型,造模第14天将造模组大鼠均分为模型对照组(Mod组),SX高、中、低药物治疗组(分别给予SX 40、20及10 g/kg灌胃,21 d,n=7)及阳性药物对照组(GTW组,给予雷公藤多甙片40 g/kg灌胃,21 d,n=7);于造模第14天、给药治疗第7、14及21天时检测足肿胀度(ER);给药第21天时,观察大鼠膝关节滑膜组织病理改变,采用实时荧光定量PCR(qRT-PCR)法测定大鼠膝关节滑膜组织VEGF mRNA的表达,ELISA法检测测大鼠血清IL-17及VEGF水平。结果:与Mod组相比,SX 40、20及10 g/kg组、GTW组大鼠足关节ER减轻,关节滑膜组织炎性细胞浸润明显下降,纤维组织增生及水肿程度减轻;治疗第21天时,与Mod组比较,SX 40、20及10 g/kg组大鼠膝关节滑膜组织VEGF mRNA水平、血清VEGF含量均显著降低(P<0.01),SX 40、20 g/kg组血清IL-17含量显著降低(P<0.01)。结论:SX能有效缓解CIA病情,其机制可能与降低IL-17和VEGF含量有关。
Objective: To study the effect of "Sidaxue" of Miao medicine prescription on the expression levels of IL-17 and VEGF in CIA rats. Methods: 42 SD rats were divided into blank control group(NG group, n=7) and module group(n=35). In addition to the NG group, all the rats were subcutaneously injected with bovine type II collagen and equivalent Freund's adjuvant to induce inflammation and replicate CIA animal model. On the 14 th day, the rats in the model group were divided into model control group(Mod group), SX high, medium and low dose treatment group(respectively give SX 40, 20 and 10 g/kg to fill the stomach, 21 d, n=7) and positive drug control(GTW group, give GTW 40 g/kg to fill the stomach, 21 d, n=7). The degree of swelling was detected on the 14 th day of modeling and the 7 th, 14 th and 21 day of administration. After 21 th day of administration, synovial tissue of the knee joint was taken to prepare pathological tissue sections. The expression of VEGF mRNA in rat synovial tissue was detected by qRT-PCR and the expression levels of IL-17 and VEGF in rat serum were detected by ELISA. Results: Compared with the Mod group, the swelling of the rat foot joints in the SX 40, 20 and 10 g/kg groups and the GTW group was alleviated; the infiltration of inflammatory cells was significantly decreased; the fibrous tissue hyperplasia and edema extent were alleviated. Compared with the Mod group on 21 th day, mRNA levels of VEGF in SX 40, 20 and 10 g/kg serum were significantly down-regulated(P<0.01), and IL-17 levels in SX 40 g/kg and 20 g/kg groups were decreased(P<0.01). Conclusions: SX can effectively alleviate the condition of CIA by reducing the expression levels of IL-17 and VEGF.
Objective: To study the effect of "Sidaxue" of Miao medicine prescription on the expression levels of IL-17 and VEGF in CIA rats. Methods: 42 SD rats were divided into blank control group(NG group, n=7) and module group(n=35). In addition to the NG group, all the rats were subcutaneously injected with bovine type II collagen and equivalent Freund's adjuvant to induce inflammation and replicate CIA animal model. On the 14 th day, the rats in the model group were divided into model control group(Mod group), SX high, medium and low dose treatment group(respectively give SX 40, 20 and 10 g/kg to fill the stomach, 21 d, n=7) and positive drug control(GTW group, give GTW 40 g/kg to fill the stomach, 21 d, n=7). The degree of swelling was detected on the 14 th day of modeling and the 7 th, 14 th and 21 day of administration. After 21 th day of administration, synovial tissue of the knee joint was taken to prepare pathological tissue sections. The expression of VEGF mRNA in rat synovial tissue was detected by qRT-PCR and the expression levels of IL-17 and VEGF in rat serum were detected by ELISA. Results: Compared with the Mod group, the swelling of the rat foot joints in the SX 40, 20 and 10 g/kg groups and the GTW group was alleviated; the infiltration of inflammatory cells was significantly decreased; the fibrous tissue hyperplasia and edema extent were alleviated. Compared with the Mod group on 21 th day, mRNA levels of VEGF in SX 40, 20 and 10 g/kg serum were significantly down-regulated(P<0.01), and IL-17 levels in SX 40 g/kg and 20 g/kg groups were decreased(P<0.01). Conclusions: SX can effectively alleviate the condition of CIA by reducing the expression levels of IL-17 and VEGF.
引文
[1] ANDERSSON K M E,WASéN C,JUZOKAITE L,et al.Inflammation in the hippocampus affects IGF1 receptor signaling and contributes to neurological sequelae in rheumatoid arthritis[J].Proc Natl Acad Sci,2018,115(51):12063-12072.
[2] NAKAMURA H,SHIMAMURA S,YASUDA S,et al.Ectopic RASGRP2 (CalDAG-GEFI) expression in rheumatoid synovium contributes to the development of destructive arthritis[J].Ann Rheum Dis,2018,77(12):1765-1772.
[3] FEARON U,CANAVAN M,BINIECKA M,et al.Hypoxia,mitochondrial dysfunction and synovial invasiveness in rheumatoid arthritis[J].Nat Rev Rheumatol,2016,12(7):385-397.
[4] KAN H J,DYAGILEV K,SCHULAM P,et al.Factors associated with physicians' prescriptions for rheumatoid arthritis drugs not filled by patients[J].Arthritis Research & Therapy,2018,20(1):1-12.
[5] FIKRY E M,GAD A M,EID A H,et al.Caffeic acid and ellagic acid ameliorate adjuvant-induced arthritis in rats via targeting inflammatory signals,chitinase-3-like protein-1 and angiogenesis[J].Biomed Pharmacother,2019,110:878-886.
[6] 莫晓川,冯光霞,曾佳,等.苗药四大血对佐剂性关节炎大鼠抗炎作用研究[J].湖北中医药大学学报,2015,17(1):15-18.
[7] 吴昌学,吴宁,冯光霞,等.苗药方四大血对佐剂性类风湿关节炎大鼠TNF-α及IL-1β表达的影响[J].贵州医科大学学报,2014,39(1):27-29.
[8] 吴宁,曾佳,王时敏,等.苗药四大血对大鼠佐剂性关节炎作用的研究[J].中国实验方剂学杂志,2014,20(2):120-123.
[9] 罗薇,任继刚,周海燕,等.中医药研究中类风湿性关节炎动物模型建立的思考[J].中华中医药学刊,2016,34(2):292-294.
[10]FEUERHERM A J,DENNIS E A,JOHANSEN B.Cytosolic group IVA phospholipase A2 inhibitors,AVX001 and AVX002,ameliorate collagen-induced arthritis[J] .Arthritis Res Ther,2019,21(1):29-42.
[11]HUNG L H,WU C H,LIN B F,et al.Hyperimmune colostrum alleviates rheumatoid arthritis in a collagen-induced arthritis murine model[J].Journal of Dairy Science,2018,101(5):3778-3787.
[12]LEE Y H,BAE S C.Correlation between circulating VEGF levels and disease activity in rheumatoid arthritis:a meta-analysis[J].Z Rheumatol,2018,77(3):240-248.
[13]赵智明,蔡辉.类风湿关节炎表观遗传学研究进展[J].中国免疫学杂志,2017(4):634-637.
[14]VAN MULLIGEN E,DE JONG PHP,KUIJPER T M,et al.Gradual tapering TNF inhibitors versus conventional synthetic DMARDs after achieving controlled disease in patients with rheumatoid arthritis:first-year results of the randomised controlled TARA study[J] .Ann.Rheum Dis,2019,78(6):746-753.
[15]郑美思,江启煜,赵琦瑶,等.基于生物信息学的桂枝芍药知母汤治疗RA的作用机制[J].中国实验方剂学杂志,2017(10):195-200.
[16]MARWA O S,KALTHOUM T,WAJIH K,et al.Association of IL17A and IL17F genes with Rheumatoid Arthritis disease and the impact of genetic polymorphisms on response to treatment[J].Immunology Letters,2017,183:24-36.
[17]SVENSSON M N,DOODY K M,SCHMIEDEL B J,et al.Reduced expression of phosphatase PTPN2 promotes pathogenic conversion of Tregs in autoimmunity[J].J Clin Invest,2019,129(3):1193-1210.
[18]DOSS H M,GANESAN R.An herbal compound ameliorates rheumatoid arthritis by the suppression of inflammatory immune responses in rats with adjuvant-induced arthritis and on cultured fibroblast like synoviocytes via the inhibition of the NF-κB signaling pathway[J].Chem Biol Interact,2016,258:175-186.
[19]BOUSSEAU S,VERGORI L,SOLETI R,et al.Glycosylation as new pharmacological strategies for diseases associated with excessive angiogenesis[J] .Pharmacol Ther,2018,191:92-122.
[20]BAE S C,LEE Y H.Association between the functional pTpN22 G788A (R263Q) polymorphism and susceptibility to autoimmune diseases:A meta-analysis[J].Cellular and Molecular Biology,2018,64(5):46-51.
[2] NAKAMURA H,SHIMAMURA S,YASUDA S,et al.Ectopic RASGRP2 (CalDAG-GEFI) expression in rheumatoid synovium contributes to the development of destructive arthritis[J].Ann Rheum Dis,2018,77(12):1765-1772.
[3] FEARON U,CANAVAN M,BINIECKA M,et al.Hypoxia,mitochondrial dysfunction and synovial invasiveness in rheumatoid arthritis[J].Nat Rev Rheumatol,2016,12(7):385-397.
[4] KAN H J,DYAGILEV K,SCHULAM P,et al.Factors associated with physicians' prescriptions for rheumatoid arthritis drugs not filled by patients[J].Arthritis Research & Therapy,2018,20(1):1-12.
[5] FIKRY E M,GAD A M,EID A H,et al.Caffeic acid and ellagic acid ameliorate adjuvant-induced arthritis in rats via targeting inflammatory signals,chitinase-3-like protein-1 and angiogenesis[J].Biomed Pharmacother,2019,110:878-886.
[6] 莫晓川,冯光霞,曾佳,等.苗药四大血对佐剂性关节炎大鼠抗炎作用研究[J].湖北中医药大学学报,2015,17(1):15-18.
[7] 吴昌学,吴宁,冯光霞,等.苗药方四大血对佐剂性类风湿关节炎大鼠TNF-α及IL-1β表达的影响[J].贵州医科大学学报,2014,39(1):27-29.
[8] 吴宁,曾佳,王时敏,等.苗药四大血对大鼠佐剂性关节炎作用的研究[J].中国实验方剂学杂志,2014,20(2):120-123.
[9] 罗薇,任继刚,周海燕,等.中医药研究中类风湿性关节炎动物模型建立的思考[J].中华中医药学刊,2016,34(2):292-294.
[10]FEUERHERM A J,DENNIS E A,JOHANSEN B.Cytosolic group IVA phospholipase A2 inhibitors,AVX001 and AVX002,ameliorate collagen-induced arthritis[J] .Arthritis Res Ther,2019,21(1):29-42.
[11]HUNG L H,WU C H,LIN B F,et al.Hyperimmune colostrum alleviates rheumatoid arthritis in a collagen-induced arthritis murine model[J].Journal of Dairy Science,2018,101(5):3778-3787.
[12]LEE Y H,BAE S C.Correlation between circulating VEGF levels and disease activity in rheumatoid arthritis:a meta-analysis[J].Z Rheumatol,2018,77(3):240-248.
[13]赵智明,蔡辉.类风湿关节炎表观遗传学研究进展[J].中国免疫学杂志,2017(4):634-637.
[14]VAN MULLIGEN E,DE JONG PHP,KUIJPER T M,et al.Gradual tapering TNF inhibitors versus conventional synthetic DMARDs after achieving controlled disease in patients with rheumatoid arthritis:first-year results of the randomised controlled TARA study[J] .Ann.Rheum Dis,2019,78(6):746-753.
[15]郑美思,江启煜,赵琦瑶,等.基于生物信息学的桂枝芍药知母汤治疗RA的作用机制[J].中国实验方剂学杂志,2017(10):195-200.
[16]MARWA O S,KALTHOUM T,WAJIH K,et al.Association of IL17A and IL17F genes with Rheumatoid Arthritis disease and the impact of genetic polymorphisms on response to treatment[J].Immunology Letters,2017,183:24-36.
[17]SVENSSON M N,DOODY K M,SCHMIEDEL B J,et al.Reduced expression of phosphatase PTPN2 promotes pathogenic conversion of Tregs in autoimmunity[J].J Clin Invest,2019,129(3):1193-1210.
[18]DOSS H M,GANESAN R.An herbal compound ameliorates rheumatoid arthritis by the suppression of inflammatory immune responses in rats with adjuvant-induced arthritis and on cultured fibroblast like synoviocytes via the inhibition of the NF-κB signaling pathway[J].Chem Biol Interact,2016,258:175-186.
[19]BOUSSEAU S,VERGORI L,SOLETI R,et al.Glycosylation as new pharmacological strategies for diseases associated with excessive angiogenesis[J] .Pharmacol Ther,2018,191:92-122.
[20]BAE S C,LEE Y H.Association between the functional pTpN22 G788A (R263Q) polymorphism and susceptibility to autoimmune diseases:A meta-analysis[J].Cellular and Molecular Biology,2018,64(5):46-51.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |