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[1]趙泉,吳劍.富血小板血漿對硝普鈉誘導的軟骨細胞Wnt/β-catenin信號通路的影響[J].中醫(yī)正骨,2018,30(04):4-7,12.
　ZHAO Quan,WU Jian.Effect of platelet-rich plasma on Wnt/β-catenin signaling pathway in chondrocytes induced by nitroprusside[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2018,30(04):4-7,12.
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富血小板血漿對硝普鈉誘導的軟骨細胞Wnt/β-catenin信號通路的影響()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第30卷
期數(shù):
2018年04期
頁碼:
4-7,12
欄目:
基礎研究
出版日期:
2018-04-20

文章信息/Info

Title:
Effect of platelet-rich plasma on Wnt/β-catenin signaling pathway in chondrocytes induced by nitroprusside
作者:
趙泉; 吳劍
湖北省咸寧市中心醫(yī)院湖北科技學院附屬第一醫(yī)院,湖北 咸寧 437100
Author(s):
ZHAO Quan; WU Jian
Xianning Central Hospital,Xianning 437100,Hubei,China
關鍵詞:
富血小板血漿;  骨關節(jié)炎;  軟骨細胞;  硝普鈉;  Wnt信號通路;  β連環(huán)素;  糖原合成酶激酶3
Keywords:
Keywords platelet-rich plasma;  osteoarthritis;  chondrocytes;  nitroprusside;  wnt signaling pathway;  beta catenin;  glycogen synthase kinase 3
摘要:
目的:觀察富血小板血漿(platelet-rich plasma,PRP)對硝普鈉誘導的軟骨細胞Wnt/β-catenin信號通路的影響。方法:取5月齡新西蘭大白兔耳靜脈血制成富血小板血漿。取兔雙膝關節(jié)軟骨,分離、培養(yǎng)軟骨細胞。取培養(yǎng)的第2代軟骨細胞,分為空白對照組、硝普鈉對照組、PRP組和Wnt蛋白生成抑制劑2(the inhibitor of Wnt production-2,IWP-2)組,空白對照組與硝普鈉對照組加入生理鹽水、PRP組加入PRP、IWP-2組加入IWP-2干預1 h后,除空白對照組外,其余3組再加入硝普鈉干預24 h。采用實時熒光定量PCR法檢測各組軟骨細胞Wnt1、β-catenin和糖原合成酶激酶-3β(glycogen synthase kinase-3β,GSK-3β)mRNA的表達情況,采用Western-blot法檢測各組軟骨細胞Wnt1、β-catenin和GSK-3β蛋白表達情況。結(jié)果:經(jīng)甲苯胺藍染色及Ⅱ型膠原免疫組化染色,鑒定所培養(yǎng)細胞為軟骨細胞。干預后,4組軟骨細胞Wnt1、β-catenin和GSK-3β mRNA相對表達量組間總體比較,差異均有統(tǒng)計學意義(0.429±0.015,0.968±0.058,0.693±0.019,0.228±0.012,F=4.673,P=0.013; 0.434±0.021,0.948±0.067,0.702±0.024,0.191±0.011,F=3.918,P=0.008; 0.816±0.039,0.225±0.017,0.459±0.018,0.929±0.118,F=3.015,P=0.005)。PRP組軟骨細胞Wnt1、β-catenin mRNA相對表達量均較空白對照組和IWP-2組高(P=0.036,P=0.000; P=0.026,P=0.000),但均較硝普鈉對照組低(P=0.017; P=0.013)。PRP組軟骨細胞GSK-3β mRNA相對表達量較空白對照組和IWP-2組低(P=0.001,P=0.000),但較硝普鈉對照組高(P=0.016)。干預后,4組軟骨細胞Wnt1、β-catenin和GSK-3β蛋白相對表達量組間總體比較,差異均有統(tǒng)計學意義(0.565±0.017,0.941±0.134,0.725±0.031,0.251±0.013,F=5.062,P=0.017; 0.526±0.024,0.872±0.016,0.653±0.016,0.262±0.018,F=3.592,P=0.006; 0.753±0.014,0.262±0.015,0.579±0.024,0.823±0.025,F=2.384,P=0.002)。PRP組軟骨細胞Wnt1和β-catenin蛋白相對表達量較空白對照組和IWP-2組高(P=0.031,P=0.000; P=0.041,P=0.002),但低于硝普鈉對照組(P=0.032,P=0.025); PRP組軟骨細胞中GSK-3β蛋白相對表達量較空白對照組和IWP2組低(P=0.035,P=0.011),但高于硝普鈉對照組(P=0.010)。結(jié)論:PRP可抑制兔膝關節(jié)軟骨細胞中Wnt1、β-catenin的表達,促進GSK-3β的表達。PRP可抑制硝普鈉誘導的軟骨細胞Wnt/β-catenin通路活性; 但與Wnt蛋白生成抑制劑–2相比,PRP對軟骨細胞Wnt/β-catenin通路活性的抑制作用較弱。
Abstract:
ABSTRACT Objective:To observe the effect of platelet-rich plasma(PRP)on Wnt/β-catenin signaling pathway in chondrocytes induced by nitroprusside.Methods:Five-month-old New Zealand rabbits were selected and their blood was drawn from auricular veins for making PRP.Then the rabbits were executed and their articular cartilages were fetched out from both knees for separating and culturing chondrocytes.The second-generation chondrocytes of New Zealand rabbits cultured in vitro were divided into blank control group,nitroprusside control group,PRP group and the inhibitor of Wnt production-2(IWP-2)group.The chondrocytes in blank control group,nitroprusside control group,PRP group and IWP-2 group were intervened with normal salin(NS),NS,PRP and IWP-2 respectively for 1 hour,moreover,the chondrocytes in nitroprusside control group,PRP group and IWP-2 group were intervened with nitroprusside for 24 hours.After intervention,the mRNA and protein expression levels of Wnt1,β-catenin and glycogen synthase kinase-3β(GSK-3β)in chondrocytes of each group were detected by using Real-time fluorescent quantitative PCR and Western-blot assays respectively.Results:The cultured cells were identified as chondrocytes through toluidine blue staining and typeⅡcollagen immunohistochemical staining.After intervention,there was statistical difference in relative expression levels of Wnt1 mRNA,β-catenin mRNA and GSK-3β mRNA in chondrocytes between the 4 groups in general(0.429+/-0.015,0.968+/-0.058,0.693+/-0.019,0.228+/-0.012,F=4.673,----------------------------------------------- 基金項目:湖北省咸寧市中心醫(yī)院科研重點資助項目(2016XYA008) 通訊作者:吳劍 E-mail:[email protected] P=0.013; 0.434+/-0.021,0.948+/-0.067,0.702+/-0.024,0.191+/-0.011,F=3.918,P=0.008; 0.816+/-0.039,0.225+/-0.017,0.459+/-0.018,0.929+/-0.118,F=3.015,P=0.005).The relative expression levels of Wnt1 mRNA and β-catenin mRNA in chondrocytes were higher in PRP group compared to blank control group and IWP-2 group(P=0.036,P=0.000; P=0.026,P=0.000)and were lower in PRP group compared to nitroprusside control group(P=0.017; P=0.013).The relative expression level of GSK-3β mRNA in chondrocytes was lower in PRP group compared to blank control group and IWP-2 group(P=0.001,P=0.000)and was higher in PRP group compared to nitroprusside control group(P=0.016).After intervention,there was statistical difference in the relative expression levels of Wnt1 protein,β-catenin protein and GSK-3β protein in chondrocytes between the 4 groups in general(0.565+/-0.017,0.941+/-0.134,0.725+/-0.031,0.251+/-0.013,F=5.062,P=0.017; 0.526+/-0.024,0.872+/-0.016,0.653+/-0.016,0.262+/-0.018,F=3.592,P=0.006; 0.753+/-0.014,0.262+/-0.015,0.579+/-0.024,0.823+/-0.025,F=2.384,P=0.002).The relative expression levels of Wnt1 protein and β-catenin protein in chondrocytes were higher in PRP group compared to blank control group and IWP-2 group(P=0.031,P=0.000; P=0.041,P=0.002)and were lower in PRP group compared to nitroprusside control group(P=0.032,P=0.025).The relative expression level of GSK-3β protein in chondrocytes was lower in PRP group compared to blank control group and IWP-2 group(P=0.035,P=0.011)and was higher in PRP group compared to nitroprusside control group(P=0.010).Conclusion:PRP can inhibit the expression of Wnt1 and β-catenin and promote the expression of GSK-3β in chondrocytes of rabbit knees.Meanwhile,PRP can inhibit the activity of Wnt/β-catenin pathway in chondrocytes induced by nitroprusside.However,PRP is inferior to IWP-2 in inhibiting the activity of Wnt/β-catenin pathway in chondrocytes.

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備注/Memo

備注/Memo:
基金項目:湖北省咸寧市中心醫(yī)院科研重點資助項目(2016XYA008) 通訊作者:吳劍 E-mail:[email protected]
更新日期/Last Update: 2018-08-24