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[1]葉錦霞,付長(zhǎng)龍,林潔,等.透骨消痛膠囊對(duì)毒胡蘿卜素誘導(dǎo)的內(nèi)質(zhì)網(wǎng)應(yīng)激PEKR信號(hào)通路介導(dǎo)的大鼠體外培養(yǎng)關(guān)節(jié)軟骨細(xì)胞凋亡的影響[J].中醫(yī)正骨,2017,29(06):1-7.
　YE Jinxia,FU Changlong,LIN Jie,et al.Effect of Tougu Xiaotong Jiaonang(透骨消痛膠囊)on apoptosis mediated by endoplasmic reticulum stress(PEKR signaling pathway)and induced by thapsigargin in rat's articular chondrocytes cultured in vitro[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2017,29(06):1-7.
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透骨消痛膠囊對(duì)毒胡蘿卜素誘導(dǎo)的內(nèi)質(zhì)網(wǎng)應(yīng)激PEKR信號(hào)通路介導(dǎo)的大鼠體外培養(yǎng)關(guān)節(jié)軟骨細(xì)胞凋亡的影響()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第29卷
期數(shù):
2017年06期
頁(yè)碼:
1-7
欄目:
基礎(chǔ)研究
出版日期:
2017-06-20

文章信息/Info

Title:
Effect of Tougu Xiaotong Jiaonang(透骨消痛膠囊)on apoptosis mediated by endoplasmic reticulum stress(PEKR signaling pathway)and induced by thapsigargin in rat's articular chondrocytes cultured in vitro
作者:
葉錦霞; 付長(zhǎng)龍; 林潔; 吳廣文; 鄭春松; 陳俊; 葉蕻芝
福建中醫(yī)藥大學(xué),福建 福州 350122
Author(s):
YE Jinxia; FU Changlong; LIN Jie; WU Guangwen; ZHENG Chunsong; CHEN Jun; YE Hongzhi
Fujian University of Traditional Chinese Medicine,Fuzhou 350112,Fujian,China
關(guān)鍵詞:
骨關(guān)節(jié)炎;  透骨消痛膠囊;  軟骨細(xì)胞;  軟骨; 關(guān)節(jié);  凋亡;  內(nèi)質(zhì)網(wǎng)應(yīng)激;  毒胡蘿卜素;  大鼠;  動(dòng)物實(shí)驗(yàn)
Keywords:
Key words osteoarthritis;  Tougu Xiaotong Jiaonang;  chondrocytes;  cartilage; articular;  apoptosis;  endoplasmic reticulum stress;  thapsigargin;  rats;  animal experimentation
摘要:
目的:觀察透骨消痛膠囊對(duì)毒胡蘿卜素(thapsigargin,TG)誘導(dǎo)的內(nèi)質(zhì)網(wǎng)應(yīng)激(PEKR信號(hào)通路)介導(dǎo)的大鼠體外培養(yǎng)關(guān)節(jié)軟骨細(xì)胞凋亡的影響。方法:采用4周齡SD大鼠膝關(guān)節(jié)軟骨建立軟骨細(xì)胞體外培養(yǎng)體系,將培養(yǎng)的第3代軟骨細(xì)胞分為空白組、模型組及透骨消痛膠囊高、中、低劑量組。空白組不進(jìn)行干預(yù); 模型組及透骨消痛膠囊高、中、低劑量組先以2 μmol·L-1 TG干預(yù)4 h,然后模型組更換為正常培養(yǎng)基,透骨消痛膠囊高、中、低劑量組分別更換為濃度為100 μg·mL-1、50 μg·mL-1、25 μg·mL-1的含透骨消痛膠囊培養(yǎng)基干預(yù)24 h。干預(yù)后以MTT法檢測(cè)各組軟骨細(xì)胞活性、以AnnexinⅤ-FITC/PI雙染法流式細(xì)胞術(shù)檢測(cè)各組軟骨細(xì)胞凋亡率、以Western Blot法檢測(cè)各組軟骨細(xì)胞中激活轉(zhuǎn)錄因子4(activating transcription factor 4,ATF4)、結(jié)合免疫球蛋白(binding immunoglobulin protein,BIP)、天冬氨酸特異性半胱氨酸蛋白酶12(cysteine aspartate specific protease 12,Caspase12)和Caspase3的表達(dá)量。結(jié)果:①軟骨細(xì)胞活性。5組軟骨細(xì)胞吸光度比較,差異有統(tǒng)計(jì)學(xué)意義(0.428±0.009,0.226±0.028,0.345±0.025,0.301±0.035,0.269±0.033,F=39.462,P=0.000)。模型組吸光度低于空白組(P=0.000); 透骨消痛膠囊高、中、低劑量組吸光度均高于模型組(P=0.000,P=0.000,P=0.022); 透骨消痛膠囊中、低劑量組的吸光度均低于高劑量組(P=0.019,P=0.000); 透骨消痛膠囊低劑量組與中劑量組比較,差異無統(tǒng)計(jì)學(xué)意義(P=0.085)。②軟骨細(xì)胞凋亡率。5組軟骨細(xì)胞凋亡率比較,差異有統(tǒng)計(jì)學(xué)意義[(3.270±0.231)%,(30.003±4.128)%,(12.383±0.933)%,(15.387±2.961)%,(20.143±3.472)%,F=37.560,P=0.000]。模型組軟骨細(xì)胞凋亡率高于空白組(P=0.000); 透骨消痛膠囊高、中、低劑量組軟骨細(xì)胞凋亡率均低于模型組(P=0.000,P=0.000,P=0.001); 透骨消痛膠囊低劑量組軟骨細(xì)胞凋亡率低于高劑量組(P=0.007),透骨消痛膠囊高、低劑量組與中劑量組比較,差異均無統(tǒng)計(jì)學(xué)意義(P=0.216,P=0.063)。③軟骨細(xì)胞內(nèi)質(zhì)網(wǎng)應(yīng)激(PEKR信號(hào)通路)關(guān)鍵蛋白含量。5組軟骨細(xì)胞中ATF4含量比較,差異有統(tǒng)計(jì)學(xué)意義(0.257±0.028,0.435±0.033,0.315±0.023,0.276±0.038,0.351±0.043,F=13.057,P=0.001)。空白組及透骨消痛膠囊高、中、低劑量組的ATF4含量均低于模型組(P=0.000,P=0.001,P=0.000,P=0.012); 透骨消痛膠囊低劑量組的ATF4含量高于中劑量組(P=0.022); 透骨消痛膠囊高劑量組的ATF4含量與透骨消痛膠囊中、低劑量組比較,差異均無統(tǒng)計(jì)學(xué)意義(P=0.188,P=0.229)。5組軟骨細(xì)胞中BIP含量比較,差異有統(tǒng)計(jì)學(xué)意義(0.227±0.026,0.432±0.022,0.294±0.035,0.263±0.020,0.339±0.032,F=25.416,P=0.000)。空白組及透骨消痛膠囊高、中、低劑量組的BIP含量均低于模型組(P=0.000,P=0.000,P=0.000,P=0.002); 透骨消痛膠囊低劑量組的BIP含量高于中劑量組(P=0.006); 透骨消痛膠囊高劑量組的BIP含量與透骨消痛膠囊中、低劑量組比較,差異均無統(tǒng)計(jì)學(xué)意義(P=0.185,P=0.072)。5組軟骨細(xì)胞中Caspase12含量比較,差異有統(tǒng)計(jì)學(xué)意義(0.252±0.027,0.346±0.028,0.294±0.011,0.315±0.024,0.325±0.019,F=7.345,P=0.005)。模型組的Caspase12含量高于空白組(P=0.001); 透骨消痛膠囊高劑量組的Caspase12含量低于模型組(P=0.018); 透骨消痛膠囊中、低劑量組的Caspase12含量與模型組比較,差異均無統(tǒng)計(jì)學(xué)意義(P=0.126,P=0.277); 透骨消痛膠囊中、低劑量組的Caspase12含量與高劑量組比較,差異均無統(tǒng)計(jì)學(xué)意義(P=0.277,P=0.126); 透骨消痛膠囊中、低劑量組的Caspase12含量比較,差異無統(tǒng)計(jì)學(xué)意義(P=0.614)。5組軟骨細(xì)胞中Caspase3含量比較,差異有統(tǒng)計(jì)學(xué)意義(0.265±0.028,0.380±0.017,0.317±0.026,0.304±0.019,0.333±0.022,F=10.507,P=0.001)。模型組的Caspase3含量高于空白組(P=0.000); 透骨消痛膠囊高、中、低劑量組的Caspase3含量均低于模型組(P=0.006,P=0.002,P=0.029); 透骨消痛膠囊中、低劑量組的Caspase3含量與高劑量組比較,差異均無統(tǒng)計(jì)學(xué)意義(P=0.496,P=0.387),透骨消痛膠囊中、低劑量組的Caspase3含量比較,差異無統(tǒng)計(jì)學(xué)意義(P=0.138)。結(jié)論:透骨消痛膠囊能抑制TG誘導(dǎo)的大鼠體外培養(yǎng)關(guān)節(jié)軟骨細(xì)胞發(fā)生由內(nèi)質(zhì)網(wǎng)應(yīng)激(PEKR信號(hào)通路)介導(dǎo)的細(xì)胞凋亡,其作用效果與藥物劑量無明顯關(guān)系。
Abstract:
ABSTRACT Objective:To observe the effect of Tougu Xiaotong Jiaonang(透骨消痛膠囊,TGXTJN)on apoptosis mediated by endoplasmic reticulum stress(PEKR signaling pathway)and induced by thapsigargin(TG)in rat's articular chondrocytes cultured in vitro.Methods:The knee cartilage of 4-week-old SD rats were used to build the in-vitro cultivation system of chondrocytes,and the third-generation chondrocytes of SD rats cultured in vitro were divided into blank group,model group,TGXTJN high-dose group,TGXTJN middle-dose group and TGXTJN low-dose group.The cells in blank group were not be treated and the cells in other groups were treated with TG(2 μmol/L)for 4 hours.Then the cells in model group were cultured in normal nutritive medium,and the cells in TGXTJN high-,middle- and low-dose group were cultured in nutritive medium supplemented with TGXTJN with concentration of 100,50 and 25 μg/mL respectively for 24 hours.After the end of intervention,the chondrocyte activities were detected by using MTT method,the apoptosis rates of chondrocytes were detected by using AnnexinⅤ-FITC/PI double-staining flow cytometry(FCM),and the expressions of activating transcription factor 4(ATF4),binding immunoglobulin protein(BIP),cysteine aspartate specific protease 12(Caspase12)and Caspase3 in chondrocytes were detected by using Western Blot assays.Results:There was statistical difference in the absorbance of chondrocytes between the 5 groups(0.428+/-0.009,0.226+/-0.028,0.345+/-0.025,0.301+/-0.035,0.269+/-0.033,F=39.462,P=0.000).The absorbance of chondrocytes were lower in model group compared to blank group(P=0.000)and were higher in TGXTJN high-,middle- and low-dose group compared to model group(P=0.000,P=0.000,P=0.022)and were lower in TGXTJN middle- and low-dose group compared to TGXTJN high-dose group(P=0.019,P=0.000).There was no statistical difference in absorbance of chondrocytes between TGXTJN low-dose group and TGXTJN middle-dose group(P=0.085).There was statistical difference in apoptosis rate of chondrocytes between the 5 groups(3.270+/-0.231,30.003+/-4.128,12.383+/-0.933,15.387+/-2.961,20.143+/-3.472%,F=37.560,P=0.000).The apoptosis rate of chondrocytes was higher in model group compared to blank group(P=0.000)and were lower in TGXTJN high-,middle- and low-dose group compared to model group(P=0.000,P=0.000,P=0.001)and was lower in TGXTJN low-dose group compared to TGXTJN high-dose group(P=0.007).There was no statistical difference in apoptosis rate of chondrocytes between TGXTJN high-dose group and TGXTJN middle-dose group and between TGXTJN low-dose group and TGXTJN middle-dose group(P=0.216,P=0.063).There was statistical difference in contents of ATF4 in chondrocytes between the 5 groups(0.257+/-0.028,0.435+/-0.033,0.315+/-0.023,0.276+/-0.038,0.351+/-0.043,F=13.057,P=0.001).The ATF4 contents were lower in blank group and TGXTJN high-, middle- and low-dose group compared to model group(P=0.000,P=0.001,P=0.000,P=0.012)and were higher in TGXTJN low-dose group compared to TGXTJN middle-dose group(P=0.022).There was no statistical difference in ATF4 contents between TGXTJN high-dose group and TGXTJN middle-dose group and between TGXTJN high-dose group and TGXTJN low-dose group(P=0.188,P=0.229).There was statistical difference in contents of BIP in chondrocytes between the 5 groups(0.227+/-0.026,0.432+/-0.022,0.294+/-0.035,0.263+/-0.020,0.339+/-0.032,F=25.416,P=0.000).The BIP contents were lower in blank group,TGXTJN high-,middle- and low-dose group compared to model group(P=0.000,P=0.000,P=0.000,P=0.002).The BIP contents were higher in TGXTJN low-dose group compared to TGXTJN middle-dose group(P=0.006).There was no statistical difference in BIP contents between TGXTJN high-dose group and TGXTJN middle-dose group and between TGXTJN high-dose group and TGXTJN low-dose group(P=0.185,P=0.072).There was statistical difference in contents of Caspase 12 in chondrocytes between the 5 groups(0.252+/-0.027,0.346+/-0.028,0.294+/-0.011,0.315+/-0.024,0.325+/-0.019,F=7.345,P=0.005).The Caspase 12 contents were higher in model group compared to blank group(P=0.001)and TGXTJN high-dose group(P=0.018).There was no statistical difference in Caspase 12 contents between TGXTJN middle-dose group and model group and between TGXTJN low-dose group and model group(P=0.126,P=0.277).There was no statistical difference in Caspase 12 contents between TGXTJN middle-dose group and TGXTJN high-dose group and between TGXTJN low-dose group and TGXTJN high-dose group(P=0.277,P=0.126).There was no statistical difference in Caspase 12 contents between TGXTJN middle-dose group and TGXTJN low-dose group(P=0.614).There was statistical difference in contents of Caspase 3 in chondrocytes between the 5 groups(0.265+/-0.028,0.380+/-0.017,0.317+/-0.026,0.304+/-0.019,0.333+/-0.022,F=10.507,P=0.001).The Caspase 3 contents were higher in model group compared to blank group(P=0.000)and were lower in TGXTJN high-,middle- and low-dose group compared to model group(P=0.006,P=0.002,P=0.029).There was no statistical difference in Caspase 3 contents between TGXTJN middle-dose group and TGXTJN high-dose group and between TGXTJN low-dose group and TGXTJN high-dose group(P=0.496,P=0.387).There was no statistical difference in Caspase 3 contents between TGXTJN middle-dose group and TGXTJN low-dose group(P=0.138).Conclusion:TGXTJN can inhibit the apoptosis mediated by endoplasmic reticulum stress(PEKR signaling pathway)and induced by TG in rat's articular chondrocytes cultured in vitro,and its effect bears no obvious relation to its dosage.

參考文獻(xiàn)/References:

[1] 劉獻(xiàn)祥.基于陳可冀學(xué)術(shù)思想之骨性關(guān)節(jié)炎研究[J].康復(fù)學(xué)報(bào),2016,26(1):2-5.
[2] 吳廣文,陳俊,潘彩彬,等.膝關(guān)節(jié)炎治療儀治療對(duì)兔膝骨性關(guān)節(jié)炎關(guān)節(jié)軟骨形態(tài)結(jié)構(gòu)及NF-κB炎癥通路的影響[J].康復(fù)學(xué)報(bào),2016,26(2):28-32.
[3] 林木南,劉獻(xiàn)祥.透骨消痛方治療膝骨性關(guān)節(jié)炎30例[J].福建中醫(yī)藥,2005,36(4):15-16.
[4] 李民,陳文列,劉獻(xiàn)祥,等.透骨消痛膠囊干預(yù)膝骨性關(guān)節(jié)炎軟骨下骨重塑分子機(jī)制探討[J].福建中醫(yī)學(xué)院學(xué)報(bào),2009,19(6):35-39.
[5] 李西海.透骨消痛膠囊含藥血清對(duì)軟骨細(xì)胞周期與凋亡影響的研究[D].福州:福建中醫(yī)藥大學(xué),2010.
[6] 吳廣文,褚劍鋒,許惠鳳,等.透骨消痛膠囊含藥血清對(duì)退變關(guān)節(jié)軟骨細(xì)胞表達(dá)TNF-α、IL-1β的影響[J].世界中西醫(yī)結(jié)合雜志,2012,7(4):295-298.
[7] ANDERSON JW,DIWADKAR VA,BRIDGES SR.Selective effects of different antioxidants on oxidation of lipoproteins from rats[J].Proc Soc Exp Biol Med,1998,218(4):376-381.
[8] WAN S,JIANG L.Erratum to:Endoplasmic reticulum(ER)stress and the unfolded protein response(UPR)in plants[J].Protoplasma,2016,253(3):753-764.
[9] GARDNER BM,PINCUS D,GOTTHARDT K,et al.Endoplasmic reticulum stress sensing in the unfolded protein response[J].Cold Spring Harb Perspect Biol,2013,5(3):a013169.
[10] ROZPEDEK W,PYTEL D,MUCHA B,et al.The role of the PERK/eIF2α/ATF4/CHOP signaling pathway in tumor progression during endoplasmic reticulum stress[J].Curr Mol Med,2016,16(6):533-544.
[11] 李載權(quán),周愛儒,唐朝樞.內(nèi)質(zhì)網(wǎng)應(yīng)激反應(yīng)分子機(jī)理研究進(jìn)展[J].中國(guó)生物化學(xué)與分子生物學(xué)報(bào),2004,20(3):283-288.
[12] 方歡,申宗候.內(nèi)質(zhì)網(wǎng)應(yīng)激[J].醫(yī)學(xué)分子生物學(xué)雜志,2004,1(1):36-39.
[13] OYADOMARI S,MORI M.Roles of CHOP/GADD153 in endoplasmic reticulum stress[J].Cell Death Differ,2004,11(4):381-389.
[14] YANG L,CARLSON SG,MCBURNEY D,et al.Multiple signals induce endoplasmic reticulum stress in both primary and immortalized chondrocytes resulting in loss of differentiation,impaired cell growth,and apoptosis[J].J Biol Chem,2005,280(35):31156-31165.
[15] 葉錦霞,吳廣文,賴舜淼,等.透骨消痛膠囊對(duì)膝骨關(guān)節(jié)炎大鼠內(nèi)質(zhì)網(wǎng)應(yīng)激影響的研究[J].風(fēng)濕病與關(guān)節(jié)炎,2015,4(11):5-11.
[16] TAKADA K,HIROSE J,SENBA K,et al.Enhanced apoptotic and reduced protective response in chondrocytes following endoplasmic reticulum stress in osteoarthritic cartilage[J].Int J Exp Pathol,2011,92(4):232-242.

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　LIANG Zhao,CAI Jingyi,YAN Li,et al.Evaluation of the curative effect of needle-knife therapy for relieving knee pain in patients with early knee osteoarthritis[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(06):9.
[5]王建武,黨建軍,李強(qiáng),等.四聯(lián)療法治療膝骨關(guān)節(jié)炎[J].中醫(yī)正骨,2015,27(08):44.
[6]劉紅娟,郭會(huì)利,郭樹農(nóng).云克聯(lián)合中藥治療膝骨關(guān)節(jié)炎的護(hù)理[J].中醫(yī)正骨,2015,27(08):75.
[7]陳衛(wèi)衡.探索建立系統(tǒng)的膝骨關(guān)節(jié)炎中醫(yī)臨床科研范式 和理論體系[J].中醫(yī)正骨,2015,27(07):1.
[8]帥波,沈霖,楊艷萍,等.加味青娥丸治療膝骨關(guān)節(jié)炎的作用機(jī)制研究[J].中醫(yī)正骨,2015,27(07):15.
　SHUAI Bo,SHEN Lin,YANG Yanping,et al.Study on the mechanism of action of Jiawei Qing'e Wan(加味青娥丸)for the treatment of knee osteoarthritis[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(06):15.
[9]梅其杰,袁長(zhǎng)深,段戡,等.壯藥骨痹方燙熨聯(lián)合運(yùn)動(dòng)療法治療膝骨關(guān)節(jié)炎的臨床研究[J].中醫(yī)正骨,2015,27(07):27.
　MEI Qijie,YUAN Changshen,DUAN Kan,et al.Clinical study of the curative effect of hot compressing and rubbing with packet of Gubi Fang(骨痹方)combined with exercise therapy in the treatment of knee osteoarthritis[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(06):27.
[10]王丹輝,張燕,劉麗娟,等.重組人Ⅱ型腫瘤壞死因子受體-抗體融合蛋白 關(guān)節(jié)腔注射聯(lián)合中藥薰洗治療膝骨關(guān)節(jié)炎的臨床研究[J].中醫(yī)正骨,2015,27(07):31.
　WANG Danhui,ZHANG Yan,LIU Lijuan,et al.Clinical study on intra-articular injection of TypeⅡrecombinant human tumor necrosis factor receptor-Fc fusion protein combined with Chinese herbal steaming and washing therapy for treatment of knee osteoarthritis[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(06):31.
[11]鄭春松,葉蕻芝,李西海,等.透骨消痛膠囊中補(bǔ)腎柔肝藥和活血祛風(fēng)藥治療 骨關(guān)節(jié)炎作用方式的計(jì)算機(jī)模擬比較[J].中醫(yī)正骨,2015,27(07):6.
　ZHENG Chunsong,YE Hongzhi,LI Xihai,et al.Comparison of the mode of action of Bushen Rougan(補(bǔ)腎柔肝)drugs versus Huoxue Qufeng(活血祛風(fēng))drugs contained in Tougu Xiaotong Jiaonang(透骨消痛膠囊)for the treatment of osteoarthritis:A computer simulation study[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2015,27(06):6.

備注/Memo

備注/Memo:
基金項(xiàng)目:國(guó)家自然科學(xué)基金青年科學(xué)基金項(xiàng)目(81202836); 福建省自然科學(xué)基金項(xiàng)目(2016J01779) 通訊作者:葉蕻芝 E-mail:[email protected]
更新日期/Last Update: 2017-06-20