天津医药

天津医药 ›› 2019, Vol. 47 ›› Issue (12): 1210-1214.doi: 10.11958/20191918

• 细胞与分子生物学 • 上一篇    下一篇

葫芦素B对IL-1β诱导的软骨细胞损伤及 胞外基质代谢紊乱的影响

王西彬1，左瑞庭2   

  1. 1河南省中医院（河南中医药大学第二附属医院）脊柱科（邮编450002），2风湿科
  • 收稿日期:2019-06-25 修回日期:2019-08-19 出版日期:2019-12-15 发布日期:2019-12-15
  • 通讯作者: 左瑞庭 E-mail:zuort_hn@163.com
  • 作者简介:王西彬（1976），男，硕士，副主任医师，主要从事中医骨伤脊柱相关疾病方面研究
  • 基金资助:
     

Effects of cucurbitacin on IL-1β-induced cell injury and metabolism dysfunction in chondrocytes

WANG Xi-bin1, ZUO Rui-ting2   

  1. 1 Department of Spine, 2 Department of Rheumatology, Henan Province Hospital of TCM, Zhengzhou 450002, China
  • Received:2019-06-25 Revised:2019-08-19 Published:2019-12-15 Online:2019-12-15
  • Supported by:
     

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摘要： 目的 明确葫芦素B对白细胞介素（IL）-1β诱导的软骨细胞损伤及胞外基质代谢紊乱的影响。方法 用 不同浓度（0.5、1、5、10、20 µmol/L）的葫芦素B处理小鼠软骨细胞，随后用IL-1β（10 µg/L）处理。采用CCK-8法检测 细胞活性，试剂盒检测乳酸脱氢酶（LDH）及Caspase-3的活性，确定葫芦素B最佳处理浓度。采用流式细胞术检测细 胞凋亡情况；qRT-PCR检测细胞胞外基质代谢相关的Ⅱ型胶原（CollagenⅡ）及蛋白多糖aggrecan、基质金属蛋白酶 （MMP）-3及MMP-13的mRNA水平；酶联免疫吸附测定法（ELISA）检测MMP-3、MMP-13、前列腺素E2（PGE2）的水 平，同时检测一氧化氮（NO）的产生；采用Western blot分析葫芦素B对IL-1β介导的软骨细胞中核因子（NF）-κB信号 通路活化的影响。结果 葫芦素B剂量依赖性地减弱IL-1β对软骨细胞存活的抑制效应（P＜0.05），同时抑制IL-1β 诱导的LDH释放、细胞凋亡及Caspase-3活性（P＜0.05），10 µmol/L是其最佳实验剂量。与IL-1β处理组相比，葫芦 素B处理可减缓IL-1β对CollagenⅡ、aggrecan的mRNA表达抑制效应（P＜0.05）。此外，葫芦素B还可抑制IL-1β诱 导的MMP-3及MMP-13的mRNA表达及释放（P＜0.05），降低炎性介质NO、PGE2的产生（P＜0.05）。同时，IL-1β可 明显上调软骨细胞中p-IκB、p-p65 NF-κB的表达，而葫芦素B则可抑制上述分子的表达（P＜0.05）。结论 葫芦素 B可能通过阻断NF-κB信号通路的激活抑制IL-1β诱导的软骨细胞损伤及代谢紊乱，提示其具有潜在的抗骨关节炎 的功效。

关键词: 软骨细胞, 骨关节炎, 白细胞介素1β, NF-κB, 炎性介质, 葫芦素B

Abstract:

Objective To explore the function of cucurbitacin B (CuB) in cell injury and metabolism dysfunction in
chondrocytes upon interleukin (IL)-1β conditions. Methods Mouse chondrocytes were pre-treated with various doses of
CuB (0.5,1,5,10 and 20 µmol/L), prior to expose to IL-1β (10 µg/L). Cell viability was then determined by CCK-8 assay.
The commercial kits were used to detect lactate dehydrogenase (LDH) and Caspase-3 activity. The cell apoptosis was
evaluated by flow cytometer. The mRNA levels of metabolism dysfunction-related Collagen Ⅱ , aggrecan, matrix
metalloproteinase (MMP)-3 and MMP-13 were analyzed by qRT-PCR. ELISA assay was performed to measure the contents
of MMP-3, MMP-13 and PGE-2. The levels of nitrous oxide (NO) were determined by a commercial kit. Western blot assay
was conducted to investigate the effects of CuB on the activation of NF -
κB signaling in IL-
1β - treated chondrocytes
.
Results CuB dose-dependently ameliorated the suppressive roles of IL-
1β on chondrocyte viability
(P＜0.05).
Simultaneously, CuB suppressed IL-1β-induced LDH release, cell apoptosis and Caspase-3 activity (
P＜0.05). In contrast
to IL-1β - treated groups, administration with CuB antagonized the inhibitory effects of IL-
1β on the mRNA levels of
Collagen Ⅱ and aggrecan
(
P＜0.05). Furthermore, CuB treatment attenuated IL-1β- induced transcripts and releases of
MMP-
3 and MMP-
13 (
P＜
0.05), concomitant with reductions in the production of inflammatory mediator NO and PGE
2
(
P＜0.05
). Additionally
, IL-1β incubation enhanced the expression of p-I
κB and p-p
65 NF - κB (
P＜0.05), which were
reversed following CuB treatment (
P＜0.05). Conclusion CuB may suppress IL-
1β-induced cell injury and metabolism
dysfunction in chondrocytes by blocking the NF-
κB signaling
, implying a potential function in osteoarthritis therapy
.

Key words: chondrocytes, osteoarthritis, interleukin-1beta, NF-kappa B, inflammatory mediator, cucurbitacin B

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