膝眼穴针灸对大鼠膝关节软骨细胞表型以及PGC-1α信号通路的影响
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摘要
目的研究膝眼穴针灸疗法对膝关节软骨细胞表型改变及其参与线粒体生成氧自由基(ROS)的过氧化物酶体增殖物活化受体γ协同刺激因子1α(PGC-1α)信号通路变化的影响。方法通过免疫组化技术检测膝眼穴针灸后大鼠膝关节软骨细胞表型的主要特征分子指标(col 1、col 2、aggrecan)以及PGC-1α、线粒体转录因子A(TFAM)、解耦联蛋白2(UCP 2)和NADPH氧化酶1/4(NOX 1/4)因子的表达变化。进一步检测向大鼠膝关节腔内注射ZN005L(PGC-1α激活剂)激活PGC-1α表达后大鼠软骨细胞表型特征分子指标的表达变化,以及软骨细胞内ROS生成量的改变。结果对大鼠进行膝眼穴针灸后,PGC-1α表达增加;UCP 2以及软骨细胞表型主要指标aggrecan表达下降。激活PGC-1α表达后,软骨细胞内ROS的生成量降低,并且软骨细胞表型丢失现象被抑制。结论膝眼穴针灸疗法不能改善大鼠膝关节软骨细胞的功能,相反会促进软骨细胞表型特征分子的丢失,并损害软骨细胞功能。
Objective To study the effects of acupuncture for the knee point on the change of chondrocyte phenotype and the peroxisome proliferators activated receptor gamma co-activator 1 alpha( PGC-1α) signaling pathway involved in mitochondrial oxygen free radical( ROS). Methods The main indicators( col 1,col 2,aggrecan) of chondrocyte phenotype of the rat knee cartilage,PGC-1α,mitochondrial transcriptional factor A( TFAM),uncoupling protein 2(UCP 2) and NADPH oxidase1/4(NOX 1/4) were detected by immunohistochemical technique afer the knee joints of the rats were acupuncture for the knee point. Moreover,after ZN005 L was injected into the knee joint of the rat to activate the expression of PGC-1α,the changes of chondrocyte phenotype and ROS were detected. Results The upregulation of PGC-1α could inhibit the loss of chondrocyte phenotype and reduced the production of ROS. Although acupuncture for the knee point could increase the expression of PGC-1α,it reduced the mitochondrial function and the effect of the coupling,and leaded to the loss of chondrocyte phenotype. Conclusions Acupunture therapy can not improve the function of chondrocytes,on the contrary,it can promote the loss of chondrocyte phenotype and destroy the function of the chondrocytes.
Objective To study the effects of acupuncture for the knee point on the change of chondrocyte phenotype and the peroxisome proliferators activated receptor gamma co-activator 1 alpha( PGC-1α) signaling pathway involved in mitochondrial oxygen free radical( ROS). Methods The main indicators( col 1,col 2,aggrecan) of chondrocyte phenotype of the rat knee cartilage,PGC-1α,mitochondrial transcriptional factor A( TFAM),uncoupling protein 2(UCP 2) and NADPH oxidase1/4(NOX 1/4) were detected by immunohistochemical technique afer the knee joints of the rats were acupuncture for the knee point. Moreover,after ZN005 L was injected into the knee joint of the rat to activate the expression of PGC-1α,the changes of chondrocyte phenotype and ROS were detected. Results The upregulation of PGC-1α could inhibit the loss of chondrocyte phenotype and reduced the production of ROS. Although acupuncture for the knee point could increase the expression of PGC-1α,it reduced the mitochondrial function and the effect of the coupling,and leaded to the loss of chondrocyte phenotype. Conclusions Acupunture therapy can not improve the function of chondrocytes,on the contrary,it can promote the loss of chondrocyte phenotype and destroy the function of the chondrocytes.
引文
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[14]Mailloux R J,Harper M E.Uncoupling proteins and the control of mitochondrial reactive oxygen species production[J].Free Radic Biol Med,2011,51(6):1106-1115.
[15]Robson-Doucette C A,Sultan S,Allister E M,et al.Beta-cell uncoupling protein 2 regulates reactive oxygen species production,which influences both insulin and glucagon secretion[J].Diabetes,2011,60(11):2710-2719.
[2]Shuler F D,Conjeski J,Kendall D,et al.Understanding the burden of osteoporosis and use of the World Health Organization FRAX[J].Orthopedics,2012,35(9):798-805.
[3]Dy P,Wang W,Bhattaram P,et al.Sox9 directs hypertrophic maturation and blocks osteoblast differentiation of growth plate chondrocytes[J].Dev Cell,2012,22(3):597-609.
[4]Han Y,Lefebvre V.L-Sox5 and Sox6 drive expression of the aggrecan gene in cartilage by securing binding of Sox9 to a far-upstream enhancer[J].Mol Cell Biol,2008,28(16):4999-5013.
[5]Schulze-Tanzil G.Activation and dedifferentiation of chondrocytes:implications in cartilage injury and repair[J].Ann Anat,2009,191(4):325-338.
[6]彭之莲,黄剑.针灸治疗膝关节骨性关节炎经穴研究[J].河南中医,2008,28(7):104-105.
[7]St9r W,Irnich D.Acupuncture:Basics,practice,and evidence[J].Anaesthesist,2009,58(3):311-323.
[8]田力欣,王卫,郭义,等.国外针灸治疗膝骨关节炎的随机对照研究概览[J].针灸临床杂志,2010,26(6):76-79.
[9]Wang Y,Zhao X,Lotz M,et al.Mitochondrial biogenesis is impaired in osteoarthritis chondrocytes but reversible via peroxisome proliferator-activated receptorγcoactivator 1α[J].Arthritis Rheumatol,2015,67(8):2141-2153.
[10]Rudofsky G Jr,Schr9dter A,Voron'ko O E,et al.Promoter polymorphisms of UCP l,UCP 2,and UCP 3 are not associated with diabetic microvascular complications in type 2 diabetes[J].Horm Metab Res,2007,39(4):306-309.
[11]Mailloux R J,Harper M E.Uncoupling proteins and the control of mitochondrial reactive oxygen species production[J].Free Radic Biol Med,2011,51(6):1106-1115.
[12]Olmos Y,Valle I,Borniquel S,et al.Mutual dependence of Foxo3a and PGC-1alpha in the induction of oxidative stress genes[J].J Biol Chem,2009,284(21):14476-14484.
[13]Kang C,Li J L.Role of PGC-1αsignaling in skeletal muscle health and disease[J].Ann N Y Acad Sci,2012,1271(1):110-117.
[14]Mailloux R J,Harper M E.Uncoupling proteins and the control of mitochondrial reactive oxygen species production[J].Free Radic Biol Med,2011,51(6):1106-1115.
[15]Robson-Doucette C A,Sultan S,Allister E M,et al.Beta-cell uncoupling protein 2 regulates reactive oxygen species production,which influences both insulin and glucagon secretion[J].Diabetes,2011,60(11):2710-2719.