[1]林木南,林艳红,刘献祥,等.温热疏密波对膝骨关节炎模型大鼠软骨细胞 RAS/丝裂原活化蛋白激酶信号通路的影响[J].中医正骨,2016,28(02):1-6.
　LIN Munan,LIN Yanhong,LIU Xianxiang,et al.Effect of warm sparse-dense wave on RAS/mitogen-activated protein kinase signaling pathway in chondrocytes of knee osteoarthritis rat models[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2016,28(02):1-6.
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温热疏密波对膝骨关节炎模型大鼠软骨细胞 RAS/丝裂原活化蛋白激酶信号通路的影响()
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《中医正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第28卷
期数:
2016年02期
页码:
1-6
栏目:
基础研究
出版日期:
2016-02-20

文章信息/Info

Title:
Effect of warm sparse-dense wave on RAS/mitogen-activated protein kinase signaling pathway in chondrocytes of knee osteoarthritis rat models
作者:
林木南1; 林艳红2; 刘献祥2; 张朝春1; 曾西明1; 李西海2; 秦茵1; 郭健红1; 高晖1; 陈立典2
1.中国人民解放军南京军区福州总医院,福建 福州 350025;
2.福建中医药大学,福建 福州 350122
Author(s):
LIN Munan1; LIN Yanhong2; LIU Xianxiang2; ZHANG Zhaochun1; ZENG Ximing1; LI Xihai2; QIN Yin1; GUO Jianhong1; GAO Hui1; CHEN Lidian2
1.Fuzhou General Hospital of Nanjing Military Command of PLA,Fuzhou 350025,Fujian,China, 2.Fujian University of Traditional Chinese Medicine,Fuzhou 350108,Fujian,China
关键词:
骨关节炎; 膝;  丝裂原激活蛋白激酶类;  软骨细胞;  信号传导;  温热疏密波;  大鼠;  Sprague-Dawley;  动物实验
Keywords:
osteoarthritis; knee;  mitogen-activated protein kinases;  chondrocytes;  signal transduction;  warm sparse-dense wave;  rats; Sprague-Dawley;  animal experimentation
摘要:
目的:观察温热疏密波对膝骨关节炎(knee osteoarthritis,KOA)模型大鼠软骨细胞RAS/丝裂原活化蛋白激酶(mitogen- activated protein kinase,MAPK)信号通路的影响。方法:将120只2月龄SPF级健康SD大鼠随机分为空白组、模型组、对照组、 实验1组、实验2组、实验3组,每组20只。除空白组外,其余各组大鼠均通过在右后膝关节腔内注射4%木瓜蛋白酶建立KOA模型。 造模结束后,空白组和模型组不进行干预; 对照组以KJ-6200C微波治疗仪照射右后膝关节,每日1次,每次30 min; 实验1组、实 验2组和实验3组大鼠均以膝关节炎治疗仪进行温热疏密波治疗,分别选择疏密波1:1模式、疏密波2:1模式、疏密波1:2模式,每 日1次,每次30 min。分别于实验干预开始4、8周后,从各组随机选取10只大鼠,切取右后肢股骨内髁负重面和胫骨平台软骨,分 别采用Western Blot法和Real-time PCR法测定软骨中ERK、P38、P53、RAS蛋白和mRNA含量。结果:干预4周后,模型组、对照组 及实验组软骨中ERK mRNA、P53 mRNA、RAS mRNA含量比较,组间差异均无统计学意义; 对照组、实验2组、实验3组软骨中P38 mRNA含量均低于模型组(P=0.024,P=0.024,P=0.006),其余各组间两两比较,组间差异均无统计学意义。对照组、实验3组软骨中 ERK蛋白含量均低于模型组(P=0.047,P=0.020); 实验3组软骨中P38蛋白含量低于模型组、实验1组和实验2组 (P=0.018,P=0.035,P=0.026); 对照组、实验1组、实验2组、实验3组软骨中P53蛋白含量均低于模型组 (P=0.029,P=0.012,P=0.003,P=0.002); 对照组、实验1组、实验2组、实验3组软骨中RAS蛋白含量均低于模型组 (P=0.002,P=0.000,P=0.001,P=0.000)。干预8周后,各组软骨中ERK mRNA含量比较,差异无统计学意义; 实验2组、实验3组软骨 中P38 mRNA含量均低于模型组(P=0.020,P=0.024); 对照组、实验1组、实验2组、实验3组软骨中P53 mRNA含量均低于模型组 (P=0.009,P=0.001,P=0.004,P=0.001); 对照组、实验1组、实验2组、实验3组软骨中RAS mRNA含量均低于模型组 (P=0.002,P=0.000,P=0.000,P=0.000),实验1组、实验3组软骨中RAS mRNA含量均低于对照组(P=0.043,P=0.031)。实验3组软骨 中ERK蛋白含量低于模型组和实验1组(P=0.033,P=0.009),实验2组软骨中ERK蛋白含量低于实验1组(P=0.022); 对照组、实验1 组、实验2组、实验3组软骨中P38蛋白含量均低于模型组(P=0.008,P=0.008,P=0.005,P=0.000),实验3组软骨中P38蛋白含量低 于对照组、实验1组和实验2组(P=0.014,P=0.015,P=0.022); 实验1组、实验2组、实验3组软骨中P53蛋白含量均低于模型组和 对照组(P=0.003,P=0.005; P=0.000,P=0.001; P=0.001,P=0.012); 实验1组、实验2组、实验3组RAS蛋白含量均低于模型组 (P=0.000,P=0.030,P=0.000),实验1组和实验3组RAS蛋白含量均低于对照组(P=0.000,P=0.000),实验3组RAS蛋白含量低于实验1 组和实验2组(P=0.000,P=0.000)。结论:温热疏密波可通过抑制RAS和ERK表达,调节RAS/MAPK信号转导通路,抑制炎症反应引起 的软骨细胞凋亡,从而延缓OA软骨退变,其中疏密波1:2模式效果较好,优于微波治疗。
Abstract:
Objective:To observe the effect of warm sparse-dense wave on RAS/mitogen-activated protein kinase (MAPK)signaling pathway in chondrocytes of knee osteoarthritis(KOA)rat models.Methods:One hundred and twenty healthy 2-month- old SPF-grade SD rats were randomly divided into blank group,model group,control group,experimental group 1,experimental group 2 and experimental group 3,20 rats in each group.The KOA models were built by intraarticular injecting 4% caroid into the right posterior knee of the rats except those in blank group.The rats in blank group and model group were not be treated after the modeling.The rats in control group were treated with irradiation at right posterior knee by using KJ-6200C microwave therapy apparatus,once a day for 30 minutes at a time; while the rats in experimental group 1,2 and 3 were treated with warm sparse-dense wave with a ratio of 1:1,2:1 and 1:2 respectively by using knee arthritis therapy apparatus,once a day for 30 minutes at a time.At 4 and 8 weeks after the beginning of the treatment,10 rats were randomly selected from each group,and the cartilages were fetched out from the weight-bearing surface of the medial femoral condyle and tibial plateau in the right posterior limbs,then the protein and mRNA contents of ERK,P38,P53 and RAS in cartilages were measured by using Western Blot assays and Real-time PCR assays respectively.Results:(1)After 4-week treatment,there was no statistical difference in the contents of ERK mRNA,P53 mRNA and RAS mRNA in the cartilages between model group,control group and experimental groups; and the contents of P38 mRNA in cartilages were lower in control group,experimental group 2 and 3 compared to model group (P=0.024,P=0.024,P=0.006),while there was no statistical difference between the rest paired groups.The contents of ERK protein in cartilages were lower in control group and experimental group 3 compared to model group(P=0.047,P=0.020).The contents of P38 protein in cartilages were lower in experimental group 3 compared to model group,experimental group 1 and 2(P=0.018,P=0.035,P=0.026).The contents of P53 protein in cartilages were lower in control group,experimental group 1,2 and 3 compared to model group (P=0.029,P=0.012,P=0.003,P=0.002).The contents of RAS protein in cartilages were lower in control group,experimental group 1,2 and 3 compared to model group(P=0.002,P=0.000,P=0.001,P=0.000).(2)After 8-week treatment,there was no statistical difference in the contents of ERK mRNA in cartilages between different groups.The contents of P38 mRNA in cartilages were lower in experimental group 2 and 3 compared to model group(P=0.020,P=0.024).The contents of P53 mRNA in cartilages were lower in control group,experimental group 1,2 and 3 compared to model group(P=0.009,P=0.001,P=0.004,P=0.001).The contents of RAS mRNA in cartilages were lower in control group,experimental group 1,2 and 3 compared to model group (P=0.002,P=0.000,P=0.000,P=0.000).The contents of RAS mRNA in cartilages were lower in experimental group 1 and 3 compared to control group(P=0.043,P=0.031).The contents of ERK protein in cartilages were lower in experimental group 3 compared to model group and experimental group 1(P=0.033,P=0.009).The contents of ERK protein in cartilages were lower in experimental group 2 compared to experimental group 1(P=0.022).The contents of P38 protein in cartilages were lower in control group,experimental group 1,2 and 3 compared to model group(P=0.008,P=0.008,P=0.005,P=0.000).The contents of P38 protein in cartilages were lower in experimental group 3 compared to control group,experimental group 1 and 2(P=0.014,P=0.015,P=0.022).The contents of P53 protein in cartilages were lower in experimental group 1,2 and 3 compared to model group and control group(P=0.003,P=0.005; P=0.000,P=0.001; P=0.001,P=0.012).The contents of RAS protein in cartilages were lower in experimental group 1,2 and 3 compared to model group(P=0.000,P=0.030,P=0.000).The contents of RAS protein in cartilages were lower in experimental group 1 and 3 compared to control group (P=0.000,P=0.000).The contents of RAS protein in cartilages were lower in experimental group 3 compared to experimental group 1 and 2(P=0.000,P=0.000).Conclusion:Warm sparse-dense wave can regulate the RAS/MAPK signaling pathway and inhibit the chondrocyte apoptosis induced by inflammatory reaction through inhibiting the expression of RAS and ERK,thereby it can delay cartilage degeneration in the process of OA.Better effect can be obtained by using warm sparse-dense wave with a ratio of 1:2,which surpasses microwave treatment in curative effect.

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备注/Memo

备注/Memo:
基金项目:福建省自然科学基金项目(2013J01349); 南京军区福州总医院院内课题(201003)
通讯作者:陈立典 E-mail:lmnan_fzzyy@126.com
更新日期/Last Update: 2016-04-30