圣愈汤对胶原诱导性关节炎小鼠踝关节组织SDF-1/CXCR4信号通路的影响  

Effect of Shengyu decoction on SDF-1/CXCR4 signaling pathway in ankle joint of mice with collagen-induced arthritis

作　　者：封彦齐[1] 马少骋 姚若愚 邬学群[1] 许理忠[1] 万洪波 FENG Yan-qi;MA Shao-cheng;YAO Ruo-yu;WU Xue-qun;XU Li-zhong;WAN Hong-bo(Longhua Hospital Affiliatedl to Shanghai University of Traditional Chinese Medicine,Shanghai 200032,Chinar)

机构地区：[1]上海中医药大学附属龙华医院,上海200032

出　　处：《解剖学研究》2021年第3期257-263,共7页Anatomy Research

基　　金：上海市卫生健康委员会科研计划项目(201940231)。

摘　　要：目的探讨圣愈汤对胶原诱导性关节炎(CIA)小鼠的治疗作用,及对基质细胞衍生因子1(SDF-1)/趋化因子受体4(CXCR4)信号轴的调控作用。方法取DAB/1小鼠72只,用随机数字表法分为正常对照组、模型组、阳性对照组(甲氨蝶呤,0.1 mg/kg)组和低(9 g/kg)、中(18 g/kg)、高(36 g/kg)剂量圣愈汤组,每组12只。除正常对照组外,其余各组均给予鸡Ⅱ型胶原蛋白(CⅡ)乳化液建立小鼠CIA模型,并于造模成功后开始灌胃给药,各组均连续给药28 d,1次/d。末次给药结束12 h后,观察小鼠一般行为变化,并对关节炎指数进行评分;酶联免疫吸附法检测踝关节液中IL-1、IL-6、TNF-α水平;苏木精-伊红(HE)染色观察踝关节病理变化;免疫组化法染色观察踝关节组织SDF-1及CXCR4表达情况;蛋白免疫印迹法(WB)分别检测踝关节组织SDF-1/CXCR4相关通路蛋白-细胞外信号调节酶(Erk)及其磷酸化Erk(p-Erk)、细胞外信号调节酶相关激酶(p38MAPK)及其磷酸化p38MAPK(pp38MAPK)、基质金属蛋白酶-3(MMP-3)、基质金属蛋白酶-9(MMP-9)表达情况。结果与正常对照组相比,模型组小鼠踝关节肿胀、滑膜增生及软骨退变等病理损伤严重,关节炎评分及滑膜增厚评分、关节腔液IL-1、IL-6及TNF-α水平、踝关节组织SDF-1、CXCR4阳性表达水平、滑膜及软骨组织中p-Erk/Erk、p-p38MAPK/p38MAPK、MMP-3、MMP-9表达水平均升高(P<0.05)。与模型组相比,圣愈汤低、中、高剂量组及阳性对照组小鼠踝关节肿胀、滑膜增生及软骨退变等病理损伤减轻,且SDF-1、CXCR4阳性表达及相关通路蛋白、炎症细胞因子等上述指标均降低(P<0.05),且随着圣愈汤剂量升高上述指标降低越明显(P<0.05);圣愈汤中剂量组与阳性对照组相比差异无统计学意义(P>0.05)。结论圣愈汤可能通过抑制SDF-1/CXCR4激活,减弱Erk/p38MAPK通路活化,减少基质金属酶和炎症细胞因子等释放,改善CIA小鼠踝关节红肿及滑膜增生、软骨�Objective To investigate the therapeutic effect of Shengyu decoction on collagen-induced arthritis(CIA)mice and the regulation on SDF-1/CXCR4 signal axis.Methods Seventy-two DAB/1 mice were randomly divided into normal control group,model group low(9 g/kg),medium(18 g/kg),high(36 g/kg)dose,Shengyu decoction group and positive control group(methotrexate,0.1 mg/kg),with 12 mice in each group;except the normal control group,the other groups were given chicken typeⅡcollagen(CⅡ)emulsion to establish CIA model in mice,after successful modeling,the rats in each group were given drugs by gavage for 28 days,once a day.At 12 hours after the last administration,the general behavior changes of mice were observed,and the arthritis index was scored;the levels of IL-1,IL-6,TNF-αin ankle joint fluid were detected by ELISA;the pathological changes of ankle joint were observed by hematoxylin eosin(HE)staining;the expression of SDF-1 and CXCR4 in ankle joint was observed by immunohistochemistry;the expression of extracellular signal regulated kinase(Erk)and phosphorylated Erk(p-Erk),extracellular signal regulated enzyme related kinase(p38MAPK)and phosphorylated p38MAPK(P-P38MAPK),matrix metalloproteinase-3(MMP-3)and matrix metalloproteinase-9(MMP-9)in ankle joint tissues was detected by Western blot.Results Compared with those in the normal control group,the ankle joint swelling,synovial hyperplasia and cartilage degeneration were serious in the model group,the arthri-tis score and synovial thickening score,levels of IL-1,IL-6 and TNF-αin synovial fluid,positive expression levels of SDF-1 and CXCR4 in ankle joint tissue,expression levels of p-Erk/Erk,p-P38MAPK/p38MAPK,MMP-3 and MMP-9 in synovial and cartilage tissue were all increased(P<0.05).Compared with those in the model group,the ankle swelling,synovial hyperplasia and cartilage degeneration and other pathological damage were reduced in low-,medium-and high-dose Shengyu Decoction groups and positive control group,the positive expression of SDF-1 and CXCR4,related pathway

关 键 词：圣愈汤 胶原诱导性关节炎 基质细胞衍生因子1 趋化因子受体4 

分 类 号：R285.5[医药卫生—中药学]