中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2013年
11期
1917-1924
,共8页
张业勇%程文丹%陈哲峰%范卫民%刘锋
張業勇%程文丹%陳哲峰%範衛民%劉鋒
장업용%정문단%진철봉%범위민%류봉
组织构建%软骨组织构建%人参皂苷 Rg1%Ⅱ型胶原%环氧合酶 2%软骨细胞%白细胞介素 1β%反转录聚合酶链反应%骨关节炎%CCK-8%组织构建图片文章
組織構建%軟骨組織構建%人參皂苷 Rg1%Ⅱ型膠原%環氧閤酶 2%軟骨細胞%白細胞介素 1β%反轉錄聚閤酶鏈反應%骨關節炎%CCK-8%組織構建圖片文章
조직구건%연골조직구건%인삼조감 Rg1%Ⅱ형효원%배양합매 2%연골세포%백세포개소 1β%반전록취합매련반응%골관절염%CCK-8%조직구건도편문장
tissue construction%cartilage tissue construction%ginsenoside Rg1%type Ⅱ col agen%cyclooxygenase-2%chondrocytes%interleukin-1 beta%reverse transcription-PCR%osteoarthritis%Cel Counting Kit-8%tissue construction photographs-containing paper
背景:人参是具有抗炎、抗应激、调节免疫等广泛药理学活性的中草药,其主要药理活性成分是人参皂苷,而 Rg1是含量较多的活性成分.目的:探讨人参皂苷 Rg1对白细胞介素1β诱导的人骨关节炎模型中软骨细胞Ⅱ型胶原及环氧合酶2 mRNA 表达的影响.方法:取因骨关节炎接受全膝关节置换患者的膝关节软骨进行体外培养,取第2代体外培养的软骨细胞, CCK-8检测0.001,0.01,0.1,1,10,100 mg/L 人参皂苷 Rg1对软骨细胞增殖率的影响.再将第2代体外培养的关节软骨细胞,随机分为空白组、对照组和实验组,分别加入 DMEM 培养液、10μg/L 白细胞介素1β,10μg/L 白细胞介素1β+0.1,1,10,100 mg/L 人参皂苷 Rg1,培养24 h 后反转录 PCR检测各组细胞中Ⅱ型胶原和环氧合酶2 mRNA 的表达.结果与结论:与对照组相比,人参皂苷 Rg1质量浓度为0.001,0.01,0.1,1 mg/L 时促进软骨细胞的增殖作用不明显,差异无显著性意义(P >0.05);当人参皂苷 Rg1质量浓度为10,100 mg/L 时,促进软骨细胞的增殖作用明显(P <0.05).与空白组相比,对照组的软骨细胞Ⅱ型胶原的 mRNA 表达明显下降,而环氧合酶2 mRNA 表达明显升高(P <0.05);与对照组相比,联合加入人参皂苷 Rg1质量浓度为0.1和1 mg/L 时,人软骨细胞中Ⅱ型胶原和环氧合酶2 mRNA 表达没有明显变化(P >0.05);而联合加入人参皂苷 Rg1质量浓度为10和100 mg/L 时,人软骨细胞中Ⅱ型胶原 mRNA 表达增加,而环氧合酶2 mRNA 表达降低(P <0.05).说明一定浓度的人参皂苷 Rg1可以拮抗白细胞介素1β引起的人软骨细胞中Ⅱ型胶原的 mRNA 表达的降低和环氧合酶2 mRNA 表达的升高.
揹景:人參是具有抗炎、抗應激、調節免疫等廣汎藥理學活性的中草藥,其主要藥理活性成分是人參皂苷,而 Rg1是含量較多的活性成分.目的:探討人參皂苷 Rg1對白細胞介素1β誘導的人骨關節炎模型中軟骨細胞Ⅱ型膠原及環氧閤酶2 mRNA 錶達的影響.方法:取因骨關節炎接受全膝關節置換患者的膝關節軟骨進行體外培養,取第2代體外培養的軟骨細胞, CCK-8檢測0.001,0.01,0.1,1,10,100 mg/L 人參皂苷 Rg1對軟骨細胞增殖率的影響.再將第2代體外培養的關節軟骨細胞,隨機分為空白組、對照組和實驗組,分彆加入 DMEM 培養液、10μg/L 白細胞介素1β,10μg/L 白細胞介素1β+0.1,1,10,100 mg/L 人參皂苷 Rg1,培養24 h 後反轉錄 PCR檢測各組細胞中Ⅱ型膠原和環氧閤酶2 mRNA 的錶達.結果與結論:與對照組相比,人參皂苷 Rg1質量濃度為0.001,0.01,0.1,1 mg/L 時促進軟骨細胞的增殖作用不明顯,差異無顯著性意義(P >0.05);噹人參皂苷 Rg1質量濃度為10,100 mg/L 時,促進軟骨細胞的增殖作用明顯(P <0.05).與空白組相比,對照組的軟骨細胞Ⅱ型膠原的 mRNA 錶達明顯下降,而環氧閤酶2 mRNA 錶達明顯升高(P <0.05);與對照組相比,聯閤加入人參皂苷 Rg1質量濃度為0.1和1 mg/L 時,人軟骨細胞中Ⅱ型膠原和環氧閤酶2 mRNA 錶達沒有明顯變化(P >0.05);而聯閤加入人參皂苷 Rg1質量濃度為10和100 mg/L 時,人軟骨細胞中Ⅱ型膠原 mRNA 錶達增加,而環氧閤酶2 mRNA 錶達降低(P <0.05).說明一定濃度的人參皂苷 Rg1可以拮抗白細胞介素1β引起的人軟骨細胞中Ⅱ型膠原的 mRNA 錶達的降低和環氧閤酶2 mRNA 錶達的升高.
배경:인삼시구유항염、항응격、조절면역등엄범약이학활성적중초약,기주요약리활성성분시인삼조감,이 Rg1시함량교다적활성성분.목적:탐토인삼조감 Rg1대백세포개소1β유도적인골관절염모형중연골세포Ⅱ형효원급배양합매2 mRNA 표체적영향.방법:취인골관절염접수전슬관절치환환자적슬관절연골진행체외배양,취제2대체외배양적연골세포, CCK-8검측0.001,0.01,0.1,1,10,100 mg/L 인삼조감 Rg1대연골세포증식솔적영향.재장제2대체외배양적관절연골세포,수궤분위공백조、대조조화실험조,분별가입 DMEM 배양액、10μg/L 백세포개소1β,10μg/L 백세포개소1β+0.1,1,10,100 mg/L 인삼조감 Rg1,배양24 h 후반전록 PCR검측각조세포중Ⅱ형효원화배양합매2 mRNA 적표체.결과여결론:여대조조상비,인삼조감 Rg1질량농도위0.001,0.01,0.1,1 mg/L 시촉진연골세포적증식작용불명현,차이무현저성의의(P >0.05);당인삼조감 Rg1질량농도위10,100 mg/L 시,촉진연골세포적증식작용명현(P <0.05).여공백조상비,대조조적연골세포Ⅱ형효원적 mRNA 표체명현하강,이배양합매2 mRNA 표체명현승고(P <0.05);여대조조상비,연합가입인삼조감 Rg1질량농도위0.1화1 mg/L 시,인연골세포중Ⅱ형효원화배양합매2 mRNA 표체몰유명현변화(P >0.05);이연합가입인삼조감 Rg1질량농도위10화100 mg/L 시,인연골세포중Ⅱ형효원 mRNA 표체증가,이배양합매2 mRNA 표체강저(P <0.05).설명일정농도적인삼조감 Rg1가이길항백세포개소1β인기적인연골세포중Ⅱ형효원적 mRNA 표체적강저화배양합매2 mRNA 표체적승고.
BACKGROUND: Ginseng has a wide range of pharmacological activities, such as anti-inflammatory, anti-stress, and immunomodulatory roles. Its major pharmacological active ingredient is the ginsenoside, and Rg1 is an active ingredient with a higher content. OBJECTIVE: To investigate the effect of ginsenoside Rg1 on type Ⅱ col agen and cyclooxygenase-2 mRNA expression in chondrocytes of an interleukin-1β-induced osteoarthritis model. METHODS: Undamaged cartilage from osteoarthritis patients undergoing total knee arthroplasty was harvested and cultured. The effect of ginsenoside Rg1 (0.001, 0.01, 0.1, 1, 10, 100 mg/L) on proliferation rate of passage 2 chondrocytes was analyzed by Cel Counting Kit-8. Then the passage 2 chondrocytes were divided into blank group, control group and experimental group randomly. Dulbecco’s modified Eagle’s medium was added into the blank group alone. Interleukin-1β at a dose of 10 μg/L was added into the control group to establish an osteoarthritis model. While 10 μg/L interleukin-1β and ginsenoside Rg1 with different concentrations (0.1, 1, 10, 100 mg/L) were added into the experimental group concomitantly. After 24-hour in vitro culture, the expressions of type Ⅱ col agen and cyclooxygenase-2 gene in human articular chondrocytes were analyzed by reverse transcription-PCR. RESULTS AND CONCLUSION: Promotive effect of the certain concentration of ginsenoside Rg1 on chondrocyte proliferation was observed by the result of cel counting kit-8 analysis. However, compared with the blank control group, the lower concentration of ginsenoside Rg1 (0.001, 0.01, 0.1, 1 mg/L) could not stimulate the proliferation rate of chondrocytes significantly (P > 0.05); only the higher concentration of ginsenoside Rg1 (10 and 100 mg/L) could stimulate the proliferation rate of chondrocytes significantly (P < 0.05). From the results of cel counting kit-8 analysis, compared with the blank group, the type Ⅱ col agen mRNA expression significantly decreased and cyclooxygenase-2 mRNA expression significantly increased in chondrocytes of the control group (P < 0.05). When 10 μg/L interleukin-1β and ginsenoside Rg1 of different concentrations (0.1 and 1 mg/L) were added concomitantly in the experimental group, the type Ⅱ col agen and cyclooxygenase-2 mRNA expression had no obvious changes compared with the control group (P > 0.05). Meanwhile, when 10 μg/L interleukin-1βand ginsenoside Rg1 of different concentrations (10, 100 mg/L) were added concomitantly in the experimental group, the type Ⅱ col agen mRNA expression significantly increased and cyclooxygenase-2 mRNA expression significantly decreased compared with the control group (P < 0.05). These findings showed that interleukin-1βinduced decreased mRNA expression of type Ⅱ col agen and increased mRNA expression of cyclooxygenase-2 could be antagonized by somewhat concentration of ginsenoside Rg1.
