| 大鼠血浆血清对软骨细胞纤维化转变的影响 |
| 投稿时间：2018-01-09 修订日期：2018-02-05 点此下载全文 |
| 引用本文：贺自克,丁道芳,王上增,薛艳,郑昱新,王学宗.大鼠血浆血清对软骨细胞纤维化转变的影响[J].医学研究杂志,2018,47(12):31-36 |
| DOI：
10.11969/j.issn.1673-548X.2018.12.009 |
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| 基金项目:国家自然科学基金资助项目（81503598） |
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| 中文摘要:目的 观察大鼠的血清和血浆对大鼠软骨细胞的增殖，形态改变，基质蛋白及基质调控基因的表达变化。方法 取出生24h SD大鼠关节处软骨，Ⅱ型胶原酶多次消化后获得软骨细胞，取P1代细胞进行实验。软骨细胞分别培养于含10%大鼠血浆（rat plasma，RP）和10%大鼠血清（rat serum，RS）的DMEM低糖培养基中，分别为血浆组和血清组。分别培养24h，镜下观察细胞形态变化，CCK8方法检测24h、48h和72h软骨细胞增殖（A450）。细胞免疫荧光、Western blot和定量PCR检测软骨指标Sox9、Ⅱ型胶原、MMP13、Col2a1、A-can、ADAMTS5、MMP9、MMP3的表达。结果 培养于大鼠血清和血浆的培养基中，24h后血清组的软骨细胞由鹅卵石样排列的多角形变成长梭形。血清组24h（0.67±0.01），48h（1.29±0.05）和72h（1.92±0.03）A450值，明显高于血浆组24h（0.43±0.01），48h（0.72±0.01）和72h（1.27±0.04）A450值，差异均有统计学意义（P<0.01）。血清组软骨细胞MMP13和Col1a1表达显著上调，Ⅱ型胶原和Sox9的表达显著下调。PCR结果表明，血清组A-can（0.24±0.01）和Sox9（0.56±0.04）基因表达相对于大鼠血浆组Sox9（1.00±0.05）和A-can（1.00±0.08）表达明显降低，而血清组ADAMTS5（34.51±2.25）和MMP3（40.33±2.43）表达相对于大鼠血浆组ADAMTS5（1.02±0.21）和MMP3（1.00±0.11）表达明显增强，两组间基因表达差异均有统计学意义（P<0.01）。结论 血清促进软骨细胞转变成纤维样软骨细胞。 |
| 中文关键词:血清和血浆 软骨细胞 细胞增殖 形态改变 成纤维样的软骨细胞 |
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| Effects of Rat RP and RS on the Fibroblastic Transformation from Rat Chondrocytes |
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| Abstract:Objective To investigate the effects of rat plasma and rat serum on the proliferation, morphology change, the expression of both extracellular matrix and associated proteins.Methods Chondrocytes were isolated from articular cartilages of newly born (within 24 hours) SD rats, and digested with type Ⅱ collagenase. Chondrocytes at passage one were used for the experiments. Chondrocytes were cultured with10%RP and 10% RS in low glucose DMEM for 24 hours. Cell morphology was investigated under inverted microscope. Chondrocytes proliferation was detected by CCK8 method(A450). The protein expression levels of Sox9 and type Ⅱ collagen were analyzed by immunofluorescence staining. Protein and RNA was extracted from the rat chondrocytes treated with RP or RS.The protein expression levels of MMP13 and typeⅠ collagen were examined by Western blot. mRNA levels of A-can, ADAMTS5, MMP3 and Sox9 were detected by Real-Time PCR.Results The cell morphology was changed from polygon to spindle after rat chondrocytes were cultured with medium containing RP or RS.The A450 values at 24h(0.67±0.01),48h(1.29±0.05)and 72h(1.92±0.03)in RS group were much higher than those A450 values 24h(0.43±0.01),48h(0.72±0.01)and 72h(1.27±0.04)in RP group, the differences were statistically significant between RS group and RP group(P<0.01). The results of Western blot showed that the expression levels of MMP13 and type I collagen were increased significantly in RS group. The mRNA levels of A-can(0.24±0.01)and Sox9(0.56±0.04)in RS group were down-regulated, the expression levels of ADMATS5(34.51±2.25)and MMP3(40.33±2.43) were up-regulated in RS group compared with the expression levels of Sox9(1.00±0.05),A-can(1.00±0.08), ADAMTS5(1.02±0.21)and MMP3(1.00±0.11)in RP group, the differences were statistically significant between RS group and RP group(P<0.01).Conclusion RS promotes the transformation of rat chondrocytes into fibroblastic chondrocytes. |
| keywords:RS and RP Chondrocytes Cell proliferation Morphology change Fibroblastic chondrocytes |
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