黄芪桂枝五物汤对骨关节炎大鼠血管新生的作用
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摘要
目的:研究黄芪桂枝五物汤对阳虚寒凝骨关节炎(osteoarthritis,OA)模型大鼠血管内皮生长因子(VEGF)及相关因子的影响。方法:60只SPF级SD大鼠随机分为正常组,模型组,塞来昔布组(20.82 mg·kg~(-1)),黄芪桂枝五物汤低、中、高剂量组(3.24,6.48,12.96 g·kg~(-1))。除正常组外,其余各组通过冷固法复合寒冷环境刺激42 d,复制OA模型。造模后,给药组分别灌胃相应药物,正常组和模型组给同等剂量蒸馏水,每天1次,连续28 d。末次给药24 h后,酶联免疫吸附测定(ELISA)检测血清中VEGF,前列腺素E_2(PGE_2),转化生长因子β_1(TGF-β_1)水平;免疫组化染色检测左膝关节滑膜和软骨内VEGF表达;实时荧光定量聚合酶链式反应(Real-time PCR)检测右膝关节滑膜白细胞介素-17(IL~(-1)7)和VEGF mRNA表达。结果:与正常组比较,模型组血清、软骨和滑膜组织中VEGF表达均显著性升高(P<0.01),同时,血清中PGE_2和TGF-β_1,滑膜组织IL~(-1)7水平均显著升高(P<0.01);与模型组比较,各用药组血清、软骨和滑膜组织中VEGF表达均显著性降低(P<0.05),同时,血清中PGE_2和TGF-β_1水平,滑膜组织中IL~(-1)7水平亦显著降低(P<0.05);各用药组间比较,黄芪桂枝五物汤高剂量组膝关节滑膜组织IL~(-1)7较塞来昔布组升高(P<0.05),黄芪桂枝五物汤中、高剂量组VEGF较塞来昔布组显著升高(P<0.01),其余各组间比较均无显著性差异。结论:黄芪桂枝五物汤可能通过减轻骨关节炎大鼠血管新生相关细胞因子PGE_2和TGF-β_1等的表达而作用于VEGF,从而抑制膝骨关节处血管新生,减轻软骨损伤。
Objective:To study the effect of Huangqi Guizhi Wuwu Tang on angiogenesis of osteoarthritis with Yang deficiency and cold coagulation.Method:Totally 60 female SD rats were randomly divided into control group,model group,celecoxib group(20.82 mg·kg~(-1))and low,medium,high-dose Huangqi Guizhi Wuwu Tang groups(3.24,6.48,12.96 g·kg~(-1)).All groups,except for control group,were involved in duplicating the osteoarthritis(OA)model through frozen and knee fixation,as well as 42-day cold environmental stimulation.After modeling,all drug-group rats were respectively administrated with corresponding drugs for 28 days,once a day.Meanwhile,control group and model group were given equivalent distilled water by gavage.24 hours after the last gavage,vascular endothelial growth factor(VEGF),prostaglandin E_2(PGE_2)and transforming growth factor-β_1(TGF-β_1)in serum were detected with enzyme linked immunosorbent assay(ELISA)method,VEGF expressions in cartilage and synovial with immunohistochemical method,and interleukin-17(IL~(-1)7)and VEGF levels in synovial with Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).Result:Compared with normal group,the expression of VEGF in serum,cartilage and synovial were significantly increased(P<0.01),and PGE_2and TGF-β_1expressions,IL~(-1)7 level were increased significantly(P<0.01)as well.Compared with model group,VEGF in serum,cartilage and synovial were significantly decreased in the 4 drug groups.Meanwhile,PGE_2,TGF-β_1and IL~(-1)7 level were decreased significantly(P<0.05).Among drug groups,IL~(-1)7 of high-dose group in synovial was higher than that of celecoxib group(P<0.05),and VEGF of medium and high-dose Huangqi Guizhi Wuwu Tang groups in synovial was higher than that of celecoxib group(P<0.05).There was no significant difference among other drug groups.Conclusion:Huangqi Guizhi Wuwu Tang has an effect in suppressing angiogenesis of knee and alleviate cartilage lesion by regulating VEGF and its upstream cytokines PGE_2and TGF-β_1.
Objective:To study the effect of Huangqi Guizhi Wuwu Tang on angiogenesis of osteoarthritis with Yang deficiency and cold coagulation.Method:Totally 60 female SD rats were randomly divided into control group,model group,celecoxib group(20.82 mg·kg~(-1))and low,medium,high-dose Huangqi Guizhi Wuwu Tang groups(3.24,6.48,12.96 g·kg~(-1)).All groups,except for control group,were involved in duplicating the osteoarthritis(OA)model through frozen and knee fixation,as well as 42-day cold environmental stimulation.After modeling,all drug-group rats were respectively administrated with corresponding drugs for 28 days,once a day.Meanwhile,control group and model group were given equivalent distilled water by gavage.24 hours after the last gavage,vascular endothelial growth factor(VEGF),prostaglandin E_2(PGE_2)and transforming growth factor-β_1(TGF-β_1)in serum were detected with enzyme linked immunosorbent assay(ELISA)method,VEGF expressions in cartilage and synovial with immunohistochemical method,and interleukin-17(IL~(-1)7)and VEGF levels in synovial with Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).Result:Compared with normal group,the expression of VEGF in serum,cartilage and synovial were significantly increased(P<0.01),and PGE_2and TGF-β_1expressions,IL~(-1)7 level were increased significantly(P<0.01)as well.Compared with model group,VEGF in serum,cartilage and synovial were significantly decreased in the 4 drug groups.Meanwhile,PGE_2,TGF-β_1and IL~(-1)7 level were decreased significantly(P<0.05).Among drug groups,IL~(-1)7 of high-dose group in synovial was higher than that of celecoxib group(P<0.05),and VEGF of medium and high-dose Huangqi Guizhi Wuwu Tang groups in synovial was higher than that of celecoxib group(P<0.05).There was no significant difference among other drug groups.Conclusion:Huangqi Guizhi Wuwu Tang has an effect in suppressing angiogenesis of knee and alleviate cartilage lesion by regulating VEGF and its upstream cytokines PGE_2and TGF-β_1.
引文
[1] Daniel P A,Judge A,Javaid M K,et al. Incidence and risk factors for clinically diagnosed knee,hip and hand osteoarthritis:influences of age, gender and osteoarthritis affecting other joints[J]. Ann Rheum Dis,2014,73(9):1659-1664.
[2]黄志,郭卫兵,王小虎,等.早期骨关节炎慢性滑膜炎的发病机制研究进展[J].实用骨科杂志,2016,22(5):428-431.
[3] Landry J P,FEI Y Y,ZHU X D,et al. Discovering small molecule ligands of vascular endothelial growth factor that block VEGF-KDR binding using label-free microarray-based assays[J]. Assay Drug Dev Technol,2013,11(5):326-332.
[4]刘秀丽,滕蔚然,金立伦.血管内皮生长因子在骨关节炎中的研究进展[J].医学综述,2014,20(17):3092-3094.
[5]郑洁,王瑞辉,寇久社.血管生成及神经生长在骨关节炎中的作用[J].中国老年学杂志,2015,35(18):5339-5342.
[6]徐慧,陆进明. VEGF在骨关节炎发病机制中的研究进展[J].安徽医学,2016,37(7):923-925.
[7]李冀,孙新雨,毕珺辉.黄芪桂枝五物汤的临证应用及实验研究进展[J].中医药学报,2014,24(5):108-111.
[8]赵统秀,王煜,王自立.王自立主任医师运用黄芪桂枝五物汤化裁验案举隅[J].新中医,2014,46(8):247-248.
[9]陈文超.骨灵膏及其拆方制剂对骨关节炎软骨细胞凋亡和胞外基质降解的影响[D].武汉:湖北中医药大学,2014.
[10] Trounson A,Mc Donald C. Stem cell therapies in clinical trials:progress and chanllenges[J]. Cell Stem Cell,2015,17(1):11-22.
[11]陈世杰,劳文艳,周艳丽,等.骨关节炎动物模型建立在功能食品研究中的研究[J].中国组织工程研究,2017,21(28):4536-4542.
[12]李艳彦,赵乐,王永辉,等.黄芪桂枝五物汤对阳虚寒凝型骨关节炎大鼠免疫及瘀血状态的影响[J].中国实验方剂学杂志,2016,22(23):149-154.
[13]赵乐,李艳彦,王永辉,等.黄芪桂枝五物汤对阳虚寒凝型骨关节炎大鼠免疫相关细胞因子的影响[J].中国实验方剂学杂志,2017,23(7):160-166.
[14]袁芳,何晓瑾,石俊,等.骨痹方治疗膝骨关节炎肾虚络痹证临床观察[J].中国实验方剂学杂志,2018,24(7):207-211.
[15]彭全利.加味黄芪桂枝五物汤治疗膝骨性关节炎80例临床观察[J].临床医学研究与实践,2017,2(23):116-117.
[16]蔡辉,常文静,商玮.血管新生在类风湿关节炎中的作用[J].安徽医药,2015,19(3):417-421.
[17]郑双融.痹祺胶囊对CIA大鼠滑膜炎症及血管新生相关因子的影响[D].天津:天津医科大学,2016.
[18]周建林,方洪松,彭昊,等.血管内皮生长因子和缺氧诱导因子对软骨细胞凋亡的作用[J].中国组织工程研究,2015,19(29):4705-4709.
[19] Meini S,Cucchi P,Tinti L,et al. Fasitibant prevents the bradykinin and interleukin 1βsynergism on prostaglandin E2release and cyclooxygenase 2 expression in human fibroblalst-like synociocytes[J]. Springer,2012,358(8):777-786.
[20]梁江,马武开,刘正奇,等.黑骨藤乙醇提取物影响人类风湿关节炎滑膜成纤维细胞增殖及COX-2、PGE2表达的研究[J].时珍国医国药,2015,26(1):24-27.
[21]郭亚春,高亚贤,宋鸿儒,等.薯蓣皂苷元体外对大鼠滑膜RSC-364细胞株STAT3及VEGF影响的研究[J].现代预防医学,2016,43(1):128-131.
[22]陆斌,李建武,杨艳,等.膝骨关节炎患者血浆中IL-17、IL-22的表达水平及其临床意义[J].山西医科大学学报,2014,45(3):183-185.
[23]谈冰,刘健,章平衡.新风胶囊通过抑制NF-κB信号通路改善膝骨关节炎患者高凝状态[J].免疫学杂志,2016,32(9):781-789.
[2]黄志,郭卫兵,王小虎,等.早期骨关节炎慢性滑膜炎的发病机制研究进展[J].实用骨科杂志,2016,22(5):428-431.
[3] Landry J P,FEI Y Y,ZHU X D,et al. Discovering small molecule ligands of vascular endothelial growth factor that block VEGF-KDR binding using label-free microarray-based assays[J]. Assay Drug Dev Technol,2013,11(5):326-332.
[4]刘秀丽,滕蔚然,金立伦.血管内皮生长因子在骨关节炎中的研究进展[J].医学综述,2014,20(17):3092-3094.
[5]郑洁,王瑞辉,寇久社.血管生成及神经生长在骨关节炎中的作用[J].中国老年学杂志,2015,35(18):5339-5342.
[6]徐慧,陆进明. VEGF在骨关节炎发病机制中的研究进展[J].安徽医学,2016,37(7):923-925.
[7]李冀,孙新雨,毕珺辉.黄芪桂枝五物汤的临证应用及实验研究进展[J].中医药学报,2014,24(5):108-111.
[8]赵统秀,王煜,王自立.王自立主任医师运用黄芪桂枝五物汤化裁验案举隅[J].新中医,2014,46(8):247-248.
[9]陈文超.骨灵膏及其拆方制剂对骨关节炎软骨细胞凋亡和胞外基质降解的影响[D].武汉:湖北中医药大学,2014.
[10] Trounson A,Mc Donald C. Stem cell therapies in clinical trials:progress and chanllenges[J]. Cell Stem Cell,2015,17(1):11-22.
[11]陈世杰,劳文艳,周艳丽,等.骨关节炎动物模型建立在功能食品研究中的研究[J].中国组织工程研究,2017,21(28):4536-4542.
[12]李艳彦,赵乐,王永辉,等.黄芪桂枝五物汤对阳虚寒凝型骨关节炎大鼠免疫及瘀血状态的影响[J].中国实验方剂学杂志,2016,22(23):149-154.
[13]赵乐,李艳彦,王永辉,等.黄芪桂枝五物汤对阳虚寒凝型骨关节炎大鼠免疫相关细胞因子的影响[J].中国实验方剂学杂志,2017,23(7):160-166.
[14]袁芳,何晓瑾,石俊,等.骨痹方治疗膝骨关节炎肾虚络痹证临床观察[J].中国实验方剂学杂志,2018,24(7):207-211.
[15]彭全利.加味黄芪桂枝五物汤治疗膝骨性关节炎80例临床观察[J].临床医学研究与实践,2017,2(23):116-117.
[16]蔡辉,常文静,商玮.血管新生在类风湿关节炎中的作用[J].安徽医药,2015,19(3):417-421.
[17]郑双融.痹祺胶囊对CIA大鼠滑膜炎症及血管新生相关因子的影响[D].天津:天津医科大学,2016.
[18]周建林,方洪松,彭昊,等.血管内皮生长因子和缺氧诱导因子对软骨细胞凋亡的作用[J].中国组织工程研究,2015,19(29):4705-4709.
[19] Meini S,Cucchi P,Tinti L,et al. Fasitibant prevents the bradykinin and interleukin 1βsynergism on prostaglandin E2release and cyclooxygenase 2 expression in human fibroblalst-like synociocytes[J]. Springer,2012,358(8):777-786.
[20]梁江,马武开,刘正奇,等.黑骨藤乙醇提取物影响人类风湿关节炎滑膜成纤维细胞增殖及COX-2、PGE2表达的研究[J].时珍国医国药,2015,26(1):24-27.
[21]郭亚春,高亚贤,宋鸿儒,等.薯蓣皂苷元体外对大鼠滑膜RSC-364细胞株STAT3及VEGF影响的研究[J].现代预防医学,2016,43(1):128-131.
[22]陆斌,李建武,杨艳,等.膝骨关节炎患者血浆中IL-17、IL-22的表达水平及其临床意义[J].山西医科大学学报,2014,45(3):183-185.
[23]谈冰,刘健,章平衡.新风胶囊通过抑制NF-κB信号通路改善膝骨关节炎患者高凝状态[J].免疫学杂志,2016,32(9):781-789.
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