[1]陈后煌,邵翔,叶蕻芝,等.独活寄生汤对脂多糖诱导的大鼠软骨细胞炎症模型 IL-1β、TNF-α表达的影响[J].中华老年骨科与康复电子杂志,2017,(02):77-84.[doi:10.3877/cma.j.issn.2096-0263.2017.02.003]
　Chen Houhuang,Shao Xiang,Ye Hongzhi,et al.Effects of DuhuoJisheng decoction on the expression of IL-1β and TNF-α of inflammatory Chondrocytes[J].Chin J Geriatr Orthop Rehabil(Electronic Edition),2017,(02):77-84.[doi:10.3877/cma.j.issn.2096-0263.2017.02.003]
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独活寄生汤对脂多糖诱导的大鼠软骨细胞炎症模型 IL-1β、TNF-α表达的影响()
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中华老年骨科与康复电子杂志[ISSN:1674-3911/CN:11-9292/R]

卷:
期数:
2017年02期
页码:
77-84
栏目:
论著
出版日期:
2017-03-25

文章信息/Info

Title:
Effects of DuhuoJisheng decoction on the expression of IL-1β and TNF-α of inflammatory Chondrocytes
作者:
陈后煌 1; 邵翔 1; 叶蕻芝 1; 2; 李西海 1
350122 福建中医药大学中西医结合研究院 1；福建省中西医结合老年性疾病重点实验室 2
Author(s):
Chen Houhuang1;  Shao Xiang1;  Ye Hongzhi2;  Li Xihai1
1Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, Fuzhou 350122, China; 2Fujian Province Key Laboratory of Integrative age-related diseases, Fuzhou 350122, China
关键词:
骨关节炎;  软骨细胞;  脂多糖;  白细胞介素 1β;  肿瘤坏死因子α
Keywords:
Osteoarthritis;  Chondrocytes;  Lipopolysaccharides;  Interleukin- 1beta;  Tumor necrosis factor-alpha
DOI:
10.3877/cma.j.issn.2096-0263.2017.02.003
文献标志码:
A
摘要:
目的 观察独活寄生汤水提物对体外培养的炎症模型大鼠软骨细胞 IL-1β和 TNF-α的影响。方法 采用 4周龄 SD大鼠分离培养体外软骨细胞，Ⅱ型胶原免疫组织化学染色法对第二代软骨细胞进行鉴定。脂多糖诱导软骨细胞，建立软骨细胞的炎症模型，确定最佳干预浓度和时间；模型复制成功后，随机分为空白对照组（脂多糖浓度为 0 ng/ml）、模型组（脂多糖浓度为 10 ng/ml）和独活寄生汤组，分别干预软骨细胞，elisa法检测各组细胞上清液中白细胞介素 1β（IL-1β）和肿瘤坏死因子α（TNF-α）的表达变化。结果 造模实验中，各组干预 8 h 后，脂多糖浓度为 10 ng/ml、100 ng/ml、1 000 ng/ml、 2 000 ng/ml 时 ，细胞上清液中 IL-1β和 TNF- α表达高于空白对照组。脂多糖浓度为 10 ng/ml 时，IL-1β和 TNF- α表达高于 100 ng/ml、1 000 ng/ml、2 000 ng/ml，差异具有统计学意义（P＜0.01）； 10 ng/ml 脂多糖分别干预软骨细胞 4 h，8 h，12 h 和 24 h，细胞上清液中 IL-1β（t=-30.869，t=-8.422， t=-13.999，t=-19.532，P＜0.05）和 TNF-α（t=-9.441，t=-14.053，t=-4.238，t=-4.124，P＜0.05）的表达均高于空白对照组，且干预 8 h 时细胞上清液中 IL-1β（F=21.48，P＜0.01）和 TNF-α（F=104.15，P＜0.01）的表达高于 4 h，12 h和 24 h。独活寄生汤组应用独活寄生汤干预 8 h造模细胞后，独活寄生汤浓度为 100 ?g/ml、200 ?g/ml、300 ?g/ml 时，细胞上清液中 IL-1β和 TNF- α的表达均低于空白对照组，差异 具 有 统 计 学 意 义（F=323.95，F=2789.80，P＜0.01），浓度 300 ?g/ml 时 IL-1β和 TNF- α表达低于 100 ?g/ml 和 200 ?g/ml 时，差异具有统计学意义（F=34.46，F=4.373，P＜0.01）。结论 脂多糖浓度为 10 ng/ml时，可成功建立体外软骨细胞炎症模型，独活寄生汤浓度为300 ?g/ml时可有效抑制软骨细胞炎症模型IL-1β、TNF-α的表达。
Abstract:
Objective To observe the efficacy of Duhuo Jisheng decoction (DHJSD) on the expression of IL- 1β and TNF- α in lipopolysaccharide (LPS)- treated Chondrocytes in vitro. Methods Chondrocytes was isolated from 4-week Sprague-Dawley rats and cells were cultured in vitro. The second generation of chondrocytes were identified with Type II collagen immunohistochemical staining. Chondrocytes were incubated with different concentrations of LPS in different time intervals to establish the optimal inflammatory model. Once it was duplicated successfully, three series groups were randomly divided into blank control group (0 ng/ml LPS), model group (10 ng/ml LPS) and DHJSD group to compare the expression levels of IL-1β and TNF-α using enzyme-linked immunosorbent assay (ELISA). Results In the modeling experiment, each group were initially intervened with different concentrations of LPS (100, 200, 300 ?g/ml) for 8 h. Expression of IL-1 beta and TNF- alpha were detected and higher expression level were observed compare to the blank control group and chondrocytes intervened with 10 ng/ml LPS for 4 h, 8 h, 12 h and 24 h, the expression of supernatant IL-1 beta (P<0.01）and TNF- alpha (P<0.01) were all higher than the blank control group, the highest expression level occurred at 8 h after intervention (P<0.01). In the DHJSD group, 100 ?g/ml, 200 ?g/ml and 300 ?g/ml DHJSD were used to intervene chondrocytes for 8 h, the expression of supernatant IL-1 beta (t=-30.869, t=-8.422, t=-13.999, t=-19.532, P<0.01) and TNFalpha (t=- 9.441, t=- 14.053, t=- 4.238, t=- 4.124, P<0.05) were lower than the blank control group, difference was statistically significant. The expression of supernatant IL-1 beta (F=21.48, P<0.01) and TNFalpha (F=104.15, P<0.01) were higher than 4 h, 12 h and 24 h compared to 8 h. And the expression of IL-1 beta (F=34.46, P<0.01) and TNF- alpha (F=4.373, P<0.05) at 300 ?g/ml DHJSD was lower than 100 ?g/ml and 200 ?g/ml (P<0.01). Conclusion Chondrocyte inflammatory model can establish successfully with 10 ng/mL LPS, and DHJSD can effevtively reduce the expression of IL-1β and TNF-α at 300 ?g/ml .

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备注/Memo

备注/Memo:
基金项目：国家自然科学基金（81573998）；福建省卫生系统中青年骨干人才培养项目 (2014-ZQN-JC-29)；福建中医药大学校管课题-科研创新平台专项（X2015021-平台）
更新日期/Last Update: 2017-04-01