炎癥微環(huán)境下miR-140-5p靶向作用Smad3影響軟骨細(xì)胞生物學(xué)特性的機(jī)制研究
發(fā)布時(shí)間:2021-12-10 11:02
顳下頜關(guān)節(jié)(Temporomandibular joint,TMJ)是頜面部唯一同時(shí)具備轉(zhuǎn)動(dòng)和滑動(dòng)運(yùn)動(dòng)的左右聯(lián)動(dòng)關(guān)節(jié),參與了咀嚼、吞咽、言語(yǔ)、表情和呼吸等精細(xì)且復(fù)雜的功能活動(dòng)。骨關(guān)節(jié)炎(Osteoarthritis,OA)是關(guān)節(jié)疾病中的典型代表,其發(fā)病率高,對(duì)人體危害大,嚴(yán)重影響了患者的生活質(zhì)量。雖有不少學(xué)者對(duì)這類關(guān)節(jié)疾病進(jìn)行了相關(guān)研究報(bào)道,但目前對(duì)骨關(guān)節(jié)炎,尤其是顳下頜關(guān)節(jié)骨關(guān)節(jié)炎(Temporomandibular joint osteoarthritis,TMJ-OA)復(fù)雜的病理機(jī)制尚缺乏完整認(rèn)識(shí)。以往的研究表明,miR-140-5p在軟骨細(xì)胞分化和軟骨內(nèi)穩(wěn)態(tài)過(guò)程中發(fā)揮著重要的作用,miR-140-5p基因敲除鼠會(huì)出現(xiàn)增齡性骨關(guān)節(jié)炎的癥狀。然而,臨床研究卻發(fā)現(xiàn)骨關(guān)節(jié)炎患者的關(guān)節(jié)軟骨中miR-140-5p表達(dá)顯著增加。雖然越來(lái)越多的研究認(rèn)為miR-140-5p與OA關(guān)系密切,但其功能作用至今仍存在爭(zhēng)議。有關(guān)miR-140-5p在OA,尤其在TMJ-OA中的作用機(jī)制亟需被闡明。本研究第一部分通過(guò)建立炎癥微環(huán)境體外實(shí)驗(yàn)?zāi)P?模擬了 TMJ-OA樣病損改變。第二部分探索了 miR-140...
【文章來(lái)源】:昆明醫(yī)科大學(xué)云南省
【文章頁(yè)數(shù)】:120 頁(yè)
【學(xué)位級(jí)別】:博士
【部分圖文】:
圖1.下頜髁突軟骨細(xì)胞及脛骨生長(zhǎng)板軟骨細(xì)胞染色鑒定??
定波長(zhǎng)450nm,測(cè)定各組軟骨細(xì)胞在Id、3d、5?d、7??d、9?d時(shí)的光密度(表6),其光密度值(Optical?density,?0D)可間接反映活細(xì)??胞數(shù)量。結(jié)果可見(jiàn)下頜髁突軟骨細(xì)胞與脛骨生長(zhǎng)板軟骨細(xì)胞在培養(yǎng)至3?d時(shí)兩組??間的OD值無(wú)明顯差別。當(dāng)培養(yǎng)至5?d后兩種軟骨細(xì)胞開(kāi)始迅速分裂增殖,進(jìn)入對(duì)??數(shù)生長(zhǎng)期,且脛骨生長(zhǎng)板軟骨細(xì)胞的OD值較下頜髁突軟骨細(xì)胞高。第9?d時(shí)0D??值無(wú)明顯增長(zhǎng),細(xì)胞進(jìn)入增殖的平臺(tái)期。兩種軟骨細(xì)胞增殖趨勢(shì)大體一致,均呈??“S”形的生長(zhǎng)曲線(圖2)。經(jīng)兩因素重復(fù)測(cè)量方差分析(整體比較)發(fā)現(xiàn):脛骨??生長(zhǎng)板軟骨細(xì)胞的增殖水平整體上升幅度大于髁突軟骨細(xì)胞。再對(duì)兩組在各個(gè)時(shí)??間點(diǎn)的0D值進(jìn)行單因素方差分析以及各時(shí)間點(diǎn)數(shù)據(jù)差值t檢驗(yàn),結(jié)果提示5?d時(shí)脛??骨生長(zhǎng)板軟骨細(xì)胞組和下頜髁突軟骨細(xì)胞組的OD值差異顯著?傊,統(tǒng)計(jì)結(jié)果??提示脛骨生長(zhǎng)板軟骨細(xì)胞的細(xì)胞增殖能力強(qiáng)于髁突軟骨細(xì)胞。??表6.?CCK-8檢測(cè)的0D值表達(dá)變化([土s)??^MCCs?(n=3)?TGPCs?(n=3)??Td?0.312±0.017?0.290±0.017??3d?0.327±0.0I9?0.359±0.014??5d?0.406±0.0I6*?0.510±0.025??7d?0.497±0.014?0.549±0_031??9d?0.489±0.046?0.574±0.058??注:a’=0.05,正態(tài)分布數(shù)據(jù)。??0.8-1??-?-MCCs??T?-m-?TGPCs??。海??O?r??0.2-??〇?〇?I?i?i?i?i?i?i?i?i?
p-CREB?CREB??-151?.邊?151??re'l'??c?tj?*?—?g?ns??P:illi?1JIUI??S????1???1???£????I?j?I?4?1^1^??o?o??o?o??MCCs?TGPCs?MCCs?TGPCs??圖7.炎癥微環(huán)境下軟骨細(xì)胞表型相關(guān)蛋白表達(dá)降低??Fig.?7?Reduction?of?chondrocyte?phenotype-related?proteins?in?inflammatory??microenvironment.?MCCs?and?TGPCs?were?treated?with?IL-ip?for?24?h,?then?the??chondrocyte?phenotype-related?proteins,?including?Sox9??CREB,?p-CREB,?TGF-P3,??were?determined?by?Western?blotting.?All?values?are?presented?as?means?士?SD?(n=3).??*P<0.05?compared?with?control?group?in?MCCs.?**尸<0.01?compared?with?control??group?in?MCCs.?#尸<0.05?compared?with?control?group?in?TGPCs.?##P<0.01??compared?with?control?group?in?TGPCs.?&P<0.05?comparison?between?MCCs?and??TGPCs?in?the?infla
【參考文獻(xiàn)】:
期刊論文
[1]Bone–cartilage crosstalk:a conversation for understanding osteoarthritis[J]. David M Findlay,Julia S Kuliwaba. Bone Research. 2016(03)
[2]Articulated dental cast analysis of asymptomatic and symptomatic populations[J]. Frank E Cordray. International Journal of Oral Science. 2016(02)
[3]Current research on pharmacologic and regenerative therapies for osteoarthritis[J]. Wei Zhang,Hongwei Ouyang,Crispin R Dass,Jiake Xu. Bone Research. 2015(04)
[4]TGF- signaling and the development of osteoarthritis[J]. Jie Shen,Shan Li,Di Chen. Bone Research. 2014(02)
[5]Effects of sleep bruxism on functional and occlusal parameters:a prospective controlled investigation[J]. Michelle Alicia Ommerborn,Maria Giraki,Christine Schneider,Lars Michael Fuck,Jrg Handschel,Matthias Franz,Wolfgang Hans-Michael Raab,Ralf Schfer. International Journal of Oral Science. 2012(03)
本文編號(hào):3532494
【文章來(lái)源】:昆明醫(yī)科大學(xué)云南省
【文章頁(yè)數(shù)】:120 頁(yè)
【學(xué)位級(jí)別】:博士
【部分圖文】:
圖1.下頜髁突軟骨細(xì)胞及脛骨生長(zhǎng)板軟骨細(xì)胞染色鑒定??
定波長(zhǎng)450nm,測(cè)定各組軟骨細(xì)胞在Id、3d、5?d、7??d、9?d時(shí)的光密度(表6),其光密度值(Optical?density,?0D)可間接反映活細(xì)??胞數(shù)量。結(jié)果可見(jiàn)下頜髁突軟骨細(xì)胞與脛骨生長(zhǎng)板軟骨細(xì)胞在培養(yǎng)至3?d時(shí)兩組??間的OD值無(wú)明顯差別。當(dāng)培養(yǎng)至5?d后兩種軟骨細(xì)胞開(kāi)始迅速分裂增殖,進(jìn)入對(duì)??數(shù)生長(zhǎng)期,且脛骨生長(zhǎng)板軟骨細(xì)胞的OD值較下頜髁突軟骨細(xì)胞高。第9?d時(shí)0D??值無(wú)明顯增長(zhǎng),細(xì)胞進(jìn)入增殖的平臺(tái)期。兩種軟骨細(xì)胞增殖趨勢(shì)大體一致,均呈??“S”形的生長(zhǎng)曲線(圖2)。經(jīng)兩因素重復(fù)測(cè)量方差分析(整體比較)發(fā)現(xiàn):脛骨??生長(zhǎng)板軟骨細(xì)胞的增殖水平整體上升幅度大于髁突軟骨細(xì)胞。再對(duì)兩組在各個(gè)時(shí)??間點(diǎn)的0D值進(jìn)行單因素方差分析以及各時(shí)間點(diǎn)數(shù)據(jù)差值t檢驗(yàn),結(jié)果提示5?d時(shí)脛??骨生長(zhǎng)板軟骨細(xì)胞組和下頜髁突軟骨細(xì)胞組的OD值差異顯著?傊,統(tǒng)計(jì)結(jié)果??提示脛骨生長(zhǎng)板軟骨細(xì)胞的細(xì)胞增殖能力強(qiáng)于髁突軟骨細(xì)胞。??表6.?CCK-8檢測(cè)的0D值表達(dá)變化([土s)??^MCCs?(n=3)?TGPCs?(n=3)??Td?0.312±0.017?0.290±0.017??3d?0.327±0.0I9?0.359±0.014??5d?0.406±0.0I6*?0.510±0.025??7d?0.497±0.014?0.549±0_031??9d?0.489±0.046?0.574±0.058??注:a’=0.05,正態(tài)分布數(shù)據(jù)。??0.8-1??-?-MCCs??T?-m-?TGPCs??。海??O?r??0.2-??〇?〇?I?i?i?i?i?i?i?i?i?
p-CREB?CREB??-151?.邊?151??re'l'??c?tj?*?—?g?ns??P:illi?1JIUI??S????1???1???£????I?j?I?4?1^1^??o?o??o?o??MCCs?TGPCs?MCCs?TGPCs??圖7.炎癥微環(huán)境下軟骨細(xì)胞表型相關(guān)蛋白表達(dá)降低??Fig.?7?Reduction?of?chondrocyte?phenotype-related?proteins?in?inflammatory??microenvironment.?MCCs?and?TGPCs?were?treated?with?IL-ip?for?24?h,?then?the??chondrocyte?phenotype-related?proteins,?including?Sox9??CREB,?p-CREB,?TGF-P3,??were?determined?by?Western?blotting.?All?values?are?presented?as?means?士?SD?(n=3).??*P<0.05?compared?with?control?group?in?MCCs.?**尸<0.01?compared?with?control??group?in?MCCs.?#尸<0.05?compared?with?control?group?in?TGPCs.?##P<0.01??compared?with?control?group?in?TGPCs.?&P<0.05?comparison?between?MCCs?and??TGPCs?in?the?infla
【參考文獻(xiàn)】:
期刊論文
[1]Bone–cartilage crosstalk:a conversation for understanding osteoarthritis[J]. David M Findlay,Julia S Kuliwaba. Bone Research. 2016(03)
[2]Articulated dental cast analysis of asymptomatic and symptomatic populations[J]. Frank E Cordray. International Journal of Oral Science. 2016(02)
[3]Current research on pharmacologic and regenerative therapies for osteoarthritis[J]. Wei Zhang,Hongwei Ouyang,Crispin R Dass,Jiake Xu. Bone Research. 2015(04)
[4]TGF- signaling and the development of osteoarthritis[J]. Jie Shen,Shan Li,Di Chen. Bone Research. 2014(02)
[5]Effects of sleep bruxism on functional and occlusal parameters:a prospective controlled investigation[J]. Michelle Alicia Ommerborn,Maria Giraki,Christine Schneider,Lars Michael Fuck,Jrg Handschel,Matthias Franz,Wolfgang Hans-Michael Raab,Ralf Schfer. International Journal of Oral Science. 2012(03)
本文編號(hào):3532494
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