[1]郑春松,叶蕻芝,李西海,等.独活寄生汤含药血清对白细胞介素1β诱导的 退变关节软骨细胞中基质金属蛋白酶 和环氧化酶2表达的影响[J].中医正骨,2015,27(12):1-6.
　ZHENG Chunsong,YE Hongzhi,LI Xihai,et al.Impact of Duhuo Jisheng Tang(独活寄生汤)medicated serum on expression of matrix metalloproteinase and cyclooxygenase 2 in degenerative articular chondrocytes induced by interleukin-1 beta[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(12):1-6.
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独活寄生汤含药血清对白细胞介素1β诱导的 退变关节软骨细胞中基质金属蛋白酶 和环氧化酶2表达的影响()
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《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第27卷
期数:
2015年12期
页码:
1-6
栏目:
基础研究
出版日期:
2015-12-30

文章信息/Info

Title:
Impact of Duhuo Jisheng Tang(独活寄生汤)medicated serum on expression of matrix metalloproteinase and cyclooxygenase 2 in degenerative articular chondrocytes induced by interleukin-1 beta
作者:
郑春松1; 叶蕻芝1; 李西海2; 付长龙1; 吴广文2; 陈星强2; 刘献祥1
1.福建中医药大学中西医结合研究院,福建 福州 350122;
2.福建省中西医结合老年性疾病重点实验室,福建 福州 350122
Author(s):
ZHENG Chunsong1; YE Hongzhi1; LI Xihai2; FU Changlong1; WU Guangwen2; CHEN Xingqiang2; LIU Xianxiang1
1.Academy of Integrated Medicine affiliated to Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian,China
2.Fujian Key Laboratory of Integrated Medicine on Geriatrics,Fuzhou 350122,Fujian,China
关键词:
骨关节炎;  独活寄生汤;  软骨细胞;  基质金属蛋白酶类;  环氧化酶2;  大鼠; Sprague-Dawley;  动物实验
Keywords:
osteoarthritis;  Duhuo Jisheng Tang;  chondrocytes;  matrix metalloproteinases;  cyclooxygenase 2;  rats; Sprague-Dawley;  animal experimentation
摘要:
目的:观察独活寄生汤含药血清对白细胞介素1β(interleukin-1 beta,IL-1β)诱导的退变关节软骨细胞中基质金属蛋白酶(matrix metalloproteinase,MMP)和环氧化酶2(cyclooxygenase 2,COX-2)表达的影响。方法:将36只2月龄SD大鼠随机分为独活寄生汤血清组、壮骨关节丸血清组和空白血清组,每组12只。分别以独活寄生汤、壮骨关节丸溶液和生理盐水灌胃,每次0.6 mL,每天2次,连续7 d。末次给药后采血,分离血清备用。将体外培养的SD大鼠第3代关节软骨细胞分为空白组、模型组、独活寄生汤组和壮骨关节丸组。模型组、独活寄生汤组和壮骨关节丸组软骨细胞加入IL-1β干预24 h后,分别采用含10%空白血清组血清、独活寄生汤血清组血清和壮骨关节丸血清组血清的DMEM培养基培养; 空白组软骨细胞不加IL-1β,直接以含10%空白血清组血清的DMEM培养基培养。培养72 h后采用Western Blot法测定各组细胞中MMP-1、MMP-3、MMP-9、MMP-13和COX-2的含量。结果:4组软骨细胞的MMP-1、MMP-3、MMP-9、MMP-13和COX-2含量比较,组间差异均有统计学意义(0.135±0.007,0.324±0.006,0.174±0.007,0.234±0.007,F=266.333,P=0.000; 0.150±0.028,0.346±0.027,0.250±0.028,0.293±0.028,F=26.855,P=0.000; 0.131±0.014,0.283±0.009,0.148±0.014,0.158±0.015,F=50.442,P=0.000; 0.173±0.035,0.691±0.051,0.318±0.038,0.286±0.039,F=91.860,P=0.000; 0.201±0.033,0.796±0.031,0.370±0.033,0.327±0.034,F=125.270,P=0.000)。空白组的MMP-1、MMP-3、MMP-9、MMP-13和COX-2含量均低于模型组(P=0.000,P=0.000,P=0.000,P=0.000,P=0.000)。模型组的MMP-1、MMP-3、MMP-9、MMP-13和COX-2含量均高于独活寄生汤组和壮骨关节丸组(P=0.000,P=0.003,P=0.000,P=0.000,P=0.000; P=0.000,P=0.047,P=0.000,P=0.000,P=0.000)。独活寄生汤组的MMP-1含量低于壮骨关节丸组(P=0.000); 2组的MMP-3、MMP-9、MMP-13和COX-2含量比较,组间差异均无统计学意义(P=0.094,P=0.497,P=0.380,P=0.220)。结论:独活寄生汤含药血清可抑制IL-1β诱导的退变关节软骨细胞中MMP-1、MMP-3、MMP-9、MMP-13和COX-2的表达,其抑制MMP-1表达的作用优于壮骨关节丸含药血清。
Abstract:
Objective:To observe the impact of Duhuo Jisheng Tang(独活寄生汤,DHJST)medicated serum on expression of matrix metalloproteinase(MMP)and cyclooxygenase 2(COX-2)in degenerative articular chondrocytes induced by interleukin-1 beta(IL-1β).Methods:Thirty-six 2-month-old SD rats were randomly divided into DHJST serum group,Zhuanggu Guanjie Wan(壮骨关节丸, ZGGJW)serum group and blank serum group,12 cases in each group.The rats were intragastric administrated with DHJST,ZGGJW solution and normal saline respectively(0.6 mL at a time,twice a day)for 7 consecutive day.After the last dose,the blood was collected and the serum was separated for further use.The third-generation articular chondrocytes of SD rats cultured in vitro were divided into blank group,model group,DHJST group and ZGGJW group.The chondrocytes in the latter 3 groups were cultured in Dulbecco's Modified Eagle Medium(DMEM)supplemented with 10% serum of blank serum group,DHJST serum group and ZGGJW serum group after 24-hour intervention with IL-1β.The chondrocytes in blank group were cultured in DMEM supplemented with 10% serum of blank serum group and didn't intervented by IL-1β.The contents of MMP-1,MMP-3,MMP-9,MMP-13 and COX-2 in the chondrocytes of each group were measured by using Western Blot after 72-hour culture.Results:There was statistical difference in the contents of MMP-1,MMP-3,MMP-9,MMP-13 and COX-2 in the chondrocytes between the 4 groups(0.135+/-0.007,0.324+/-0.006,0.174+/-0.007,0.234+/-0.007,F=266.333,P=0.000; 0.150+/-0.028,0.346+/-0.027,0.250+/-0.028,0.293+/-0.028,F=26.855,P=0.000; 0.131+/-0.014,0.283+/-0.009,0.148+/-0.014,0.158+/-0.015,F=50.442,P=0.000; 0.173+/-0.035,0.691+/-0.051,0.318+/-0.038,0.286+/-0.039,F=91.860,P=0.000; 0.201+/-0.033,0.796+/-0.031,0.370+/-0.033,0.327+/-0.034,F=125.270,P=0.000).The contents of MMP-1,MMP-3,MMP-9,MMP-13 and COX-2 were lower in blank group compared to model group(P=0.000,P=0.000,P=0.000,P=0.000,P=0.000).The contents of MMP-1,MMP-3,MMP-9,MMP-13 and COX-2 were higher in model group compared to DHJST group and ZGGJW group(P=0.000,P=0.003,P=0.000,P=0.000,P=0.000; P=0.000,P=0.047,P=0.000,P=0.000,P=0.000).The content of MMP-1 was lower in DHJST group compared to ZGGJW group(P=0.000)and there was no statistical difference in the contents of MMP-3,MMP-9,MMP-13 and COX-2 between the 2 groups(P=0.094,P=0.497,P=0.380,P=0.220).Conclusion:DHJST medicated serum can inhibit the expression of MMP-1,MMP-3,MMP-9,MMP-13 and COX-2 in degenerative articular chondrocytes induced by IL-1β,and it surpasses ZGGJW medicated serum in inhibiting the expression of MMP-1.

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备注/Memo

备注/Memo:
2015-09-23收稿 2015-11-19修回
基金项目:福建省自然科学基金项目(2015J01338,2014J01357); 福建省中青年教师教育科研项目(JA14152); 陈可冀中西医结合发展基金(CKJ2015010)
通讯作者:叶蕻芝 E-mail:yelin0930@163.com
更新日期/Last Update: 2015-12-30