[1]李文超 林一峰 梁祖建 沈国喜 原超 朱辉 池利业 张文财 刘洪江.鹿茸多肽对软骨终板细胞基质蛋白及其降解酶基因表达的影响[J].中国中医骨伤科杂志,2019,27(01):6-10.
　LI Wenchao LIN Yifeng LIANG Zujian SHEN Guoxi YUAN Chao ZHU Hui CHI Liye ZHANG Wencai LIU Hongjiang.Effects of Pilose Antler Polypeptides on Matrix Protein and Matrix Degradation Enzyme mRNA Expression of Rat Cartilage Endplate Cell[J].Chinese Journal of Traditional Medical Traumatology & Orthopedics,2019,27(01):6-10.
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鹿茸多肽对软骨终板细胞基质蛋白及其降解酶基因表达的影响()
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《中国中医骨伤科杂志》[ISSN:1005-0205/CN:42-1340/R]

卷:
第27卷
期数:
2019年01期
页码:
6-10
栏目:
实验研究
出版日期:
2019-01-15

文章信息/Info

Title:
Effects of Pilose Antler Polypeptides on Matrix Protein and Matrix Degradation Enzyme mRNA Expression of Rat Cartilage Endplate Cell
文章编号:
1005-0205(2019)01-0006-05
作者:
李文超1 林一峰1 梁祖建1 沈国喜1 原超1 朱辉1 池利业1 张文财1 刘洪江1
广州中医药大学第三附属医院脊柱外科(广州,510375)
Author(s):
LI Wenchao1 LIN Yifeng1 LIANG Zujian1 SHEN Guoxi1 YUAN Chao1 ZHU Hui1 CHI Liye1 ZHANG Wencai1 LIU Hongjiang1
The Third Hospital, Affiliated to Guangzhou University of Traditional Chinese Medicine, Guangzhou 510375, China.
关键词:
鹿茸多肽;  软骨终板;  椎间盘退变;  白介素-1β;  基质金属蛋白酶-13
Keywords:
pilose antler polypeptides;  cartilage endplate;  intervertebral disc degeneration;  IL-1β;  matrix metalloproteinase-13
分类号:
R-33
文献标志码:
A
摘要:
观察鹿茸多肽对 IL-1β 诱导的大鼠椎间盘软骨终板细胞的增殖及基质蛋白(Ⅱ, Ⅹ型胶原)、基质降解酶(金属蛋白酶-13)基因表达的影响。方法:选取1月龄的健康SD大鼠,对其椎间盘软骨终板细胞进行分离与培养,取第3代软骨终板细胞实验,空白对照组不做任何处置,诱导组单加入10 μg/L 的IL-1β,药物处理组分三组,分别在培养基中加入10 μg/L 的IL-1β和10 μg/mL,30 μg/mL,50 μg/mL的鹿茸多肽。MTT比色法检测三个时间节点(24 h,48 h,72 h)各组椎间盘软骨终板细胞的增殖情况; qPCR检测法对软骨终板细胞中基质Ⅱ,Ⅹ型胶原蛋白、基质金属蛋白酶-13基因的表达情况。结果:在24 h,48 h,72 h时间点,MTT 法检测10 μg/mL鹿茸多肽组在各时间点软骨终板细胞增殖情况与IL-1β组比较,差异无统计学意义(P>0.05); 而30 μg/mL组和50 μg/mL组均能减轻IL-1β对软骨终板细胞増殖的抑制作用,差异有统计学意义(P<0.05,P<0.01)。qPCR检测结果显示:IL-1β诱导组的Ⅱ型胶原蛋白基因的表达减弱,Ⅹ型胶原蛋白、基质金属蛋白酶-13基因的表达增强,与空白组对比差异有统计学意义(P<0.05); 鹿茸多肽各浓度组均能上升Ⅱ型胶原蛋白基因的表达,下调Ⅹ型胶原蛋白、基质金属蛋白酶-13基因的表达水平,差异有统计学意义(P<0.05,P<0.01),以50 μg/mL组效果显著,差异有统计学意义(P<0.01)。结论:一定浓度的鹿茸多肽(50 μg/mL组)能有效地拮抗IL-1β对软骨终板细胞增殖的抑制作用,上升Ⅱ型胶原蛋白基因的表达,下调Ⅹ型胶原蛋白、基质金属蛋白酶-13基因的表达,改善了基质的代谢,起到延缓椎间盘软骨终板退变的作用。
Abstract:
To observe effects of pilose antler polypeptides(PAP)at different concentrations on matrix protein(collagen type Ⅱ and type Ⅹ)and Matrix degradation enzyme(MMP-13)mRNA expression of rat cartilage endplate cell induced by the IL-1β.Methods: Chondrocytes from endplate of 4-weeks-old SD rat's intervertebral discs were obtained and cultured. Chondrocytes at passage 3 were divided into five groups: normal group without any treatment, induced group with IL-1β(10 μg/L)and PAP groups with IL-1β(10 μg/L)divided into three groups adding 10 μg/mL, 30 μg/mL and 50 μg/mL PAP respectively. MTT method was used to measure the proliferation at three time points(24 h, 48 h, and 72 h), while qPCR method was used to detect the expression of collagen type Ⅱ, type Ⅹ and MMP-13 mRNA. Results:At 24 h, 48 h, and 72 h time point, the proliferation of 10 μg/mL PAP group and induced group were measured by MTT and no significant differences were found(P>0.05); while the proliferation were increased in 30ug/ml and 50 μg/mL PAP groups with significant differences(P<0.05,P<0.01).By qPCR, collagen type Ⅱ mRNA expression was decreased, while collagen type Ⅹ mRNA and MMP-13 mRNA expression were increased in induced group. Compared to the normal group, there was significant difference(P<0.05). Compared to the induced group, Collagen type Ⅱ mRNA expression was increased, while collagen type Ⅹ mRNA and MMP-13 mRNA expression were decreased in all the PAP groups with significant differences(P<0.05,P<0.01), especially the 30 μg/mL PAP group and 50 μg/mL PAP group(P<0.01). Conclusion: This study demonstrated PAP at certain concentrations can decrease the inhibition of endplate chondrocytes induced by IL-1β, increase the expression of collagen type Ⅱ mRNA and inhibit the expression of collagen type Ⅹ mRNA and MMP-13 mRNA, improving matrix metabolism, resulting in relief the cartilage endplate of intervertebral disc degeneration.

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备注/Memo

备注/Memo:
1广州中医药大学第三附属医院脊柱外科(广州,510375)
更新日期/Last Update: 2019-01-15