84年鼠女哪年财运最旺,857comvvv色九欧美激情|85PO_87国产精品欲av国产av资源

[1]陳澤華,王毅,申震,等.共培養(yǎng)條件下肌衛(wèi)星細(xì)胞C2C12對(duì)軟骨細(xì)胞ATDC5活性的影響[J].中醫(yī)正骨,2024,36(02):17-22,31.
　CHEN Zehua,WANG Yi,SHEN Zhen,et al.Effect of skeletal muscle satellite cells C2C12 on the activity of chondrocytes ATDC5 in the co-cultured condition[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2024,36(02):17-22,31.
點(diǎn)擊復(fù)制

共培養(yǎng)條件下肌衛(wèi)星細(xì)胞C2C12對(duì)軟骨細(xì)胞ATDC5活性的影響()
分享到：

《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第36卷
期數(shù):
2024年02期
頁(yè)碼:
17-22,31
欄目:
基礎(chǔ)研究
出版日期:
2024-02-20

文章信息/Info

Title:
Effect of skeletal muscle satellite cells C2C12 on the activity of chondrocytes ATDC5 in the co-cultured condition
作者:
陳澤華1; 王毅2; 申震3; 李俊毅2; 歐梁4
(1.株洲市中醫(yī)傷科醫(yī)院,湖南 株洲 412007; 2.廣州中醫(yī)藥大學(xué)第五臨床醫(yī)學(xué)院,廣東 廣州 510095; 3.昆明市中醫(yī)醫(yī)院,云南 昆明 650599; 4.湖南省中醫(yī)藥研究院,湖南 長(zhǎng)沙 410006)
Author(s):
CHEN Zehua1; WANG Yi2; SHEN Zhen3; LI Junyi2; OU Liang4
1.The Orthopedics Hospital of Traditional Chinese Medicine Zhuzhou city,Zhuzhou 412007,Hunan,China 2.The Fifth Clinical Medical College of Guangzhou University of Chinese Medicine,Guangzhou 510095,Guangdong,China 3.Kunming Municipal Hospital of Traditional Chinese Medicine,Kunming 650599,Yunnan,China 4.Hunan Academy of Chinese Medicine,Changsha 410006,Hunan,China
關(guān)鍵詞:
衛(wèi)星細(xì)胞; 骨骼肌;  軟骨細(xì)胞;  骨關(guān)節(jié)炎;  肌萎縮;  共同培養(yǎng)技術(shù)
Keywords:
satellite cells; skeletal muscle;  chondrocytes;  osteoarthritis;  muscular atrophy;  coculture techniques
摘要:
目的:觀察共培養(yǎng)條件下肌衛(wèi)星細(xì)胞C2C12對(duì)軟骨細(xì)胞ATDC5活性的影響。方法:①將肌衛(wèi)星細(xì)胞C2C12分為對(duì)照組和地塞米松組,對(duì)照組常規(guī)培養(yǎng),地塞米松組采用地塞米松干預(yù),通過(guò)CCK8法和BCA法測(cè)定地塞米松對(duì)C2C12細(xì)胞增殖和蛋白合成的影響,以劃痕實(shí)驗(yàn)和Transwell小室觀察地塞米松對(duì)C2C12細(xì)胞修復(fù)和遷移能力的影響。②采用Transwell小室將正常肌衛(wèi)星細(xì)胞C2C12(共培養(yǎng)組)和經(jīng)地塞米松預(yù)處理的肌衛(wèi)星細(xì)胞C2C12(預(yù)處理組)分別與軟骨細(xì)胞ATDC5共培養(yǎng),對(duì)照組Transwell下層小室內(nèi)接種ATDC5細(xì)胞、上層小室內(nèi)為無(wú)細(xì)胞的常規(guī)培養(yǎng)基,采用CCK8法和二氯二氫熒光素-乙酰乙酸酯熒光探針檢測(cè)ATDC5細(xì)胞增殖率和細(xì)胞內(nèi)活性氧含量。結(jié)果:①C2C12細(xì)胞增殖率和蛋白含量測(cè)定結(jié)果。地塞米松組C2C12細(xì)胞增殖率和蛋白含量均低于對(duì)照組[(78.402±5.401)%,(100.000±3.096)%,t=8.498,P=0.000;(5 080.367±296.657)μg·mL-1,(5 775.577±150.476)μg·mL-1,t=3.620,P=0.022]。②C2C12細(xì)胞傷口修復(fù)率測(cè)定結(jié)果。地塞米松組C2C12細(xì)胞傷口修復(fù)率低于對(duì)照組[(53.173±1.800)%,(79.979±10.176)%,t=4.493,P=0.011]。③C2C12細(xì)胞遷移數(shù)量測(cè)定結(jié)果。培養(yǎng)24 h和48 h時(shí),地塞米松組C2C12細(xì)胞遷移數(shù)量均少于對(duì)照組[24 h:(24.200±5.630)個(gè),(57.000±2.449)個(gè),t=11.945,P=0.000; 48 h:(57.600±8.820)個(gè),(91.000±4.743)個(gè),t=7.457,P=0.000]。④ATDC5細(xì)胞增殖率測(cè)定結(jié)果。3組ATDC5細(xì)胞增殖率比較,差異有統(tǒng)計(jì)學(xué)意義[(100.000±1.663)%,(116.894±7.917)%,(89.130±2.980)%,F=23.700,P=0.001]。對(duì)照組和共培養(yǎng)組ATDC5細(xì)胞增殖率均高于預(yù)處理組(P=0.037,P=0.006),共培養(yǎng)組ATDC5細(xì)胞增殖率高于對(duì)照組(P=0.001)。⑤ATDC5細(xì)胞內(nèi)活性氧含量測(cè)定結(jié)果。3組ATDC5細(xì)胞內(nèi)活性氧含量比較,差異有統(tǒng)計(jì)學(xué)意義(20.148±5.636,13.959±4.110,40.691±3.146,F=30.096,P=0.001)。預(yù)處理組ATDC5細(xì)胞內(nèi)活性氧含量高于對(duì)照組和共培養(yǎng)組(P=0.001,P=0.000),對(duì)照組和共培養(yǎng)組ATDC5細(xì)胞內(nèi)活性氧含量的差異無(wú)統(tǒng)計(jì)學(xué)意義(P=0.137)。結(jié)論:地塞米松可抑制肌衛(wèi)星細(xì)胞C2C12增殖,應(yīng)用地塞米松干預(yù)肌衛(wèi)星細(xì)胞C2C12可體外模擬肌肉萎縮條件下肌衛(wèi)星細(xì)胞的生長(zhǎng)狀態(tài)。正常狀態(tài)下,肌衛(wèi)星細(xì)胞C2C12的代謝產(chǎn)物可促進(jìn)軟骨細(xì)胞ATDC5增殖; 肌衛(wèi)星細(xì)胞C2C12活力降低,可抑制軟骨細(xì)胞ATDC5增殖,同時(shí)會(huì)增加軟骨細(xì)胞ATDC5氧化性損傷。
Abstract:
Objective:To observe the effect of skeletal muscle satellite cells(MuSCs)C2C12 on the activity of chondrocytes ATDC5 in the case of co-cultured condition.Methods:①The C2C12 cells were cultured,and the third-generation cells were collected and divided into control group and dexamethasone(DEX)group.The C2C12 cells in the control group were cultured in the conventional Dulbecco's Modified Eagle's Medium(DMEM),while the ones in DEX group were intervened with DEX.After that,the effects of DEX on proliferation and protein synthesis in C2C12 cells were determined by using the cell counting kit-8(CCK8)assay and bicinchoninic acid(BCA)assay,respectively,meanwhile,the wound scratch assay and Transwell chamber were employed for observing the effects of DEX on repairing and migration ability of C2C12 cells.②The normal C2C12 cells(co-cultured group)and DEX-pretreated C2C12 cells(pre-treatment group)were co-cultured with ATDC5 chondrocytes,respectively,in the Transwell chambers; In the Transwell chamber of control group,the upper chamber was the cells-free conventional culture medium,and the lower was inoculated with ATDC5 cells.After that,the proliferation rate and intracellular reactive oxygen species(ROS)level of ATDC5 cells were detected by using CCK8 assay and 2',7'-dichlorofluorescin diacetate(DCFH-DA)fluorescent probe,respectively.Results:①The proliferation rate and protein level of C2C12 cells were lower in DEX group compared to control group(78.402±5.401 vs 100.000±3.096%,t=8.498,P=0.000; 5 080.367±296.657 vs 5 775.577±150.476 μg/mL,t=3.620,P=0.022).②The wound repairing rate of C2C12 cells was lower in DEX group in contrast to control group(53.173±1.800 vs 79.979±10.176%,t=4.493,P=0.011).③On hour 24 and 48,the number of migrated C2C12 cells was less in DEX group compared with that of control group(on hour 24:24.200±5.630 vs 57.000±2.449 cells,t=11.945,P=0.000; on hour 48:57.600±8.820 vs 91.000±4.743 cells,t=7.457,P=0.000).④There was statistical difference in proliferation rate of ATDC5 cells among the 3 groups(100.000±1.663,116.894±7.917,89.130±2.980%,F=23.700,P=0.001).The proliferation rate of ATDC5 cells was higher in control group and co-cultured group compared to pre-treatment group(P=0.037,P=0.006),and was highest in co-cultured group(P=0.001).⑤There was statistical difference in intracellular ROS level of ATDC5 cells among the 3 groups(20.148±5.636,13.959±4.110,40.691±3.146,F=30.096,P=0.001).The intracellular ROS level of ATDC5 cells was higher in pre-treatment group compared to control group and co-cultured group(P=0.001,P=0.000),while,the difference between control group and co-cultured group was not statistically significant(P=0.137).Conclusion:DEX can inhibit the proliferation of C2C12 cells,and the growth state of MuSCs under the condition of muscle atrophy in vitro can be simulated by intervening C2C12 cells with DEX.In the case of normal conditions,the metabolic products of C2C12 cells can promote the proliferation of chondrocytes ATDC5,and the reduction in viability of C2C12 cells can inhibit the proliferation of chondrocytes ATDC5,as well as increase the oxidative damage to chondrocytes ATDC5.

參考文獻(xiàn)/References:

[1] COLLAO N,AKOHENE-MENSAH P,NALLABELLI J,et al.The role of L-type amino acid transporter 1(Slc7a5)during in vitro myogenesis[J].Am J Physiol Cell Physiol,2022,323(2):C595-C605.
[2] 陳曉萍,范明.肌衛(wèi)星細(xì)胞研究進(jìn)展[J].生理科學(xué)進(jìn)展,2003,34(2):136-139.
[3] 李景龍,劉紅菊,王飛,等.肌衛(wèi)星細(xì)胞在失重肌萎縮中的可塑性變化及機(jī)制[J].生物化學(xué)與生物物理進(jìn)展,2016,43(6):607-615.
[4] 王震,藺海旗,何霏,等.運(yùn)動(dòng)激活骨骼肌衛(wèi)星細(xì)胞:增齡性肌衰減癥及肌肉損傷修復(fù)的運(yùn)動(dòng)預(yù)防和治療[J].中國(guó)組織工程研究,2021,25(23):3752-3759.
[5] IKEDA S,TSUMURA H,TORISU T.Age-related quadriceps-dominant muscle atrophy and incident radiographic knee osteoarthritis[J].J Orthop Sci,2005,10(2):121-126.
[6] KRISHNASAMY P,HALL M,ROBBINS S R.The role of skeletal muscle in the pathophysiology and management of knee osteoarthritis[J].Rheumatology(Oxford),2018,57(suppl 4):iv22-iv33.
[7] SHORTER E,SANNICANDRO A J,POULET B,et al.Skeletal muscle wasting and its relationship with osteoarthritis:a mini-review of mechanisms and current interventions[J].Curr Rheumatol Rep,2019,21(8):40.
[8] NOEHREN B,KOSMAC K,WALTON R G,et al.Alterations in quadriceps muscle cellular and molecular properties in adults with moderate knee osteoarthritis[J].Osteoarthritis Cartilage,2018,26(10):1359-1368.
[9] LAURENT M R,DUBOIS V,CLAESSENS F,et al.Muscle-bone interactions:from experimental models to the clinic?A critical update[J].Mol Cell Endocrinol,2016,432:14-36.
[10] 黃蓓,劉隆福,蔣鐵建.糖皮質(zhì)激素與肌少癥[J].中華骨質(zhì)疏松和骨礦鹽疾病雜志,2020,13(5):486-492.
[11] 王叢叢,李軍,魯飛翔,等.尿石素B對(duì)地塞米松誘導(dǎo)小鼠肌萎縮的改善作用及機(jī)制[J].中國(guó)老年學(xué)雜志,2018,38(8):1963-1966.
[12] SHEN S,LIAO Q,LIU J,et al.Myricanol rescues dexa-methasone-induced muscle dysfunction via a sirtuin 1-dependent mechanism[J].J Cachexia Sarcopenia Muscle,2019,10(2):429-444.
[13] CHANG J S,KONG I D.Irisin prevents dexamethasone-induced atrophy in C2C12 myotubes[J].Pflugers Arch,2020,472(4):495-502.
[14] LARSON A A,SYVERUD B C,FLORIDA S E,et al.Effects of dexamethasone dose and timing on tissue-engineered skeletal muscle units[J].Cells Tissues Organs,2018,205(4):197-207.
[15] CAIRNS D M,UCHIMURA T,KWON H,et al.Muscle cells enhance resistance to pro-inflammatory cytokine-induced cartilage destruction[J].Biochem Biophys Res Commun,2010,392(1):22-28.
[16] CAIRNS D M,LEE P G,UCHIMURA T,et al.The role of muscle cells in regulating cartilage matrix production[J].J Orthop Res,2010,28(4):529-536.
[17] RAINBOW R S,KWON H,FOOTE A T,et al.Muscle cell-derived factors inhibit inflammatory stimuli-induced damage in hMSC-derived chondrocytes[J].Osteoarthritis Cartilage,2013,21(7):990-998.
[18] PEAKE J M,DELLA GATTA P,SUZUKI K,et al.Cytokine expression and secretion by skeletal muscle cells:regulatory mechanisms and exercise effects[J].Exerc Immunol Rev,2015,21:8-25.

備注/Memo

備注/Memo:
基金項(xiàng)目:湖南省中醫(yī)藥科研課題(B2023153); 貴州省中醫(yī)藥管理局中醫(yī)藥、民族醫(yī)藥科學(xué)技術(shù)研究課題(QZYY-2019-031)
通訊作者:陳澤華 E-mail:[email protected]
更新日期/Last Update: 1900-01-01