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[1]林木南,林艷紅,劉獻(xiàn)祥,等.溫?zé)崾杳懿▽οス顷P(guān)節(jié)炎模型大鼠軟骨細(xì)胞 RAS/絲裂原活化蛋白激酶信號(hào)通路的影響[J].中醫(yī)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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溫?zé)崾杳懿▽οス顷P(guān)節(jié)炎模型大鼠軟骨細(xì)胞 RAS/絲裂原活化蛋白激酶信號(hào)通路的影響()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第28卷
期數(shù):
2016年02期
頁碼:
1-6
欄目:
基礎(chǔ)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; 林艷紅2; 劉獻(xiàn)祥2; 張朝春1; 曾西明1; 李西海2; 秦茵1; 郭健紅1; 高暉1; 陳立典2
1.中國人民解放軍南京軍區(qū)福州總醫(yī)院,福建 福州 350025;
2.福建中醫(yī)藥大學(xué),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
關(guān)鍵詞:
骨關(guān)節(jié)炎; 膝;  絲裂原激活蛋白激酶類;  軟骨細(xì)胞;  信號(hào)傳導(dǎo);  大鼠;  Sprague-Dawley;  動(dòng)物實(shí)驗(yàn)
Keywords:
osteoarthritis; knee;  mitogen-activated protein kinases;  chondrocytes;  signal transduction;  warm sparse-dense wave;  rats; Sprague-Dawley;  animal experimentation
摘要:
目的:觀察溫?zé)崾杳懿▽οス顷P(guān)節(jié)炎(knee osteoarthritis,KOA)模型大鼠軟骨細(xì)胞RAS/絲裂原活化蛋白激酶(mitogen- activated protein kinase,MAPK)信號(hào)通路的影響。方法:將120只2月齡SPF級(jí)健康SD大鼠隨機(jī)分為空白組、模型組、對照組、 實(shí)驗(yàn)1組、實(shí)驗(yàn)2組、實(shí)驗(yàn)3組,每組20只。除空白組外,其余各組大鼠均通過在右后膝關(guān)節(jié)腔內(nèi)注射4%木瓜蛋白酶建立KOA模型。 造模結(jié)束后,空白組和模型組不進(jìn)行干預(yù); 對照組以KJ-6200C微波治療儀照射右后膝關(guān)節(jié),每日1次,每次30 min; 實(shí)驗(yàn)1組、實(shí) 驗(yàn)2組和實(shí)驗(yàn)3組大鼠均以膝關(guān)節(jié)炎治療儀進(jìn)行溫?zé)崾杳懿ㄖ委?分別選擇疏密波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。分別于實(shí)驗(yàn)干預(yù)開始4、8周后,從各組隨機(jī)選取10只大鼠,切取右后肢股骨內(nèi)髁負(fù)重面和脛骨平臺(tái)軟骨,分 別采用Western Blot法和Real-time PCR法測定軟骨中ERK、P38、P53、RAS蛋白和mRNA含量。結(jié)果:干預(yù)4周后,模型組、對照組 及實(shí)驗(yàn)組軟骨中ERK mRNA、P53 mRNA、RAS mRNA含量比較,組間差異均無統(tǒng)計(jì)學(xué)意義; 對照組、實(shí)驗(yàn)2組、實(shí)驗(yàn)3組軟骨中P38 mRNA含量均低于模型組(P=0.024,P=0.024,P=0.006),其余各組間兩兩比較,組間差異均無統(tǒng)計(jì)學(xué)意義。對照組、實(shí)驗(yàn)3組軟骨中 ERK蛋白含量均低于模型組(P=0.047,P=0.020); 實(shí)驗(yàn)3組軟骨中P38蛋白含量低于模型組、實(shí)驗(yàn)1組和實(shí)驗(yàn)2組 (P=0.018,P=0.035,P=0.026); 對照組、實(shí)驗(yàn)1組、實(shí)驗(yàn)2組、實(shí)驗(yàn)3組軟骨中P53蛋白含量均低于模型組 (P=0.029,P=0.012,P=0.003,P=0.002); 對照組、實(shí)驗(yàn)1組、實(shí)驗(yàn)2組、實(shí)驗(yàn)3組軟骨中RAS蛋白含量均低于模型組 (P=0.002,P=0.000,P=0.001,P=0.000)。干預(yù)8周后,各組軟骨中ERK mRNA含量比較,差異無統(tǒng)計(jì)學(xué)意義; 實(shí)驗(yàn)2組、實(shí)驗(yàn)3組軟骨 中P38 mRNA含量均低于模型組(P=0.020,P=0.024); 對照組、實(shí)驗(yàn)1組、實(shí)驗(yàn)2組、實(shí)驗(yàn)3組軟骨中P53 mRNA含量均低于模型組 (P=0.009,P=0.001,P=0.004,P=0.001); 對照組、實(shí)驗(yàn)1組、實(shí)驗(yàn)2組、實(shí)驗(yàn)3組軟骨中RAS mRNA含量均低于模型組 (P=0.002,P=0.000,P=0.000,P=0.000),實(shí)驗(yàn)1組、實(shí)驗(yàn)3組軟骨中RAS mRNA含量均低于對照組(P=0.043,P=0.031)。實(shí)驗(yàn)3組軟骨 中ERK蛋白含量低于模型組和實(shí)驗(yàn)1組(P=0.033,P=0.009),實(shí)驗(yàn)2組軟骨中ERK蛋白含量低于實(shí)驗(yàn)1組(P=0.022); 對照組、實(shí)驗(yàn)1 組、實(shí)驗(yàn)2組、實(shí)驗(yàn)3組軟骨中P38蛋白含量均低于模型組(P=0.008,P=0.008,P=0.005,P=0.000),實(shí)驗(yàn)3組軟骨中P38蛋白含量低 于對照組、實(shí)驗(yàn)1組和實(shí)驗(yàn)2組(P=0.014,P=0.015,P=0.022); 實(shí)驗(yàn)1組、實(shí)驗(yàn)2組、實(shí)驗(yàn)3組軟骨中P53蛋白含量均低于模型組和 對照組(P=0.003,P=0.005; P=0.000,P=0.001; P=0.001,P=0.012); 實(shí)驗(yàn)1組、實(shí)驗(yàn)2組、實(shí)驗(yàn)3組RAS蛋白含量均低于模型組 (P=0.000,P=0.030,P=0.000),實(shí)驗(yàn)1組和實(shí)驗(yàn)3組RAS蛋白含量均低于對照組(P=0.000,P=0.000),實(shí)驗(yàn)3組RAS蛋白含量低于實(shí)驗(yàn)1 組和實(shí)驗(yàn)2組(P=0.000,P=0.000)。結(jié)論:溫?zé)崾杳懿ǹ赏ㄟ^抑制RAS和ERK表達(dá),調(diào)節(jié)RAS/MAPK信號(hào)轉(zhuǎn)導(dǎo)通路,抑制炎癥反應(yīng)引起 的軟骨細(xì)胞凋亡,從而延緩OA軟骨退變,其中疏密波1:2模式效果較好,優(yōu)于微波治療。
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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備注/Memo

備注/Memo:
基金項(xiàng)目:福建省自然科學(xué)基金項(xiàng)目(2013J01349); 南京軍區(qū)福州總醫(yī)院院內(nèi)課題(201003)
通訊作者:陳立典 E-mail:[email protected]
更新日期/Last Update: 2016-04-30