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[1]邱慈鑫,余添賜,楊樹華,等.龜鹿二仙膠不同提取部位對豚鼠軟骨細胞活性的影響[J].中醫(yī)正骨,2014,26(10):7-11.
　Qiu Cixin*,Yu Tianci,Yang Shuhua,et al.Effect of different extract fraction of GUILU ERXIAN gelatin on the chondrocytes activity in Guinea pigs[J].The Journal of Traditional Chinese Orthopedics and Traumatology,2014,26(10):7-11.
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龜鹿二仙膠不同提取部位對豚鼠軟骨細胞活性的影響()
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《中醫(yī)正骨》[ISSN:1001-6015/CN:41-1162/R]

卷:
第26卷
期數(shù):
2014年10期
頁碼:
7-11
欄目:
基礎(chǔ)研究
出版日期:
2014-10-30

文章信息/Info

Title:
Effect of different extract fraction of GUILU ERXIAN gelatin on the chondrocytes activity in Guinea pigs
作者:
邱慈鑫; 余添賜; 楊樹華; 魏迎辰; 陳澤華; 林海英; 王和鳴; 李楠
福建中醫(yī)藥大學,福建 福州 350122
Author(s):
Qiu Cixin*; Yu Tianci; Yang Shuhua; Wei Yingchen; Chen Zehua; Lin Haiying; Wang Heming; Li Nan.*
Fujian University of Traditional Chinese Medicine,Fuzhou 350122,Fujian,China
關(guān)鍵詞:
龜鹿二仙膠 提取部位 軟骨細胞 膠原Ⅱ型 Bcl-2 Bax 基質(zhì)金屬蛋白酶類 動物實驗
Keywords:
GUILU ERXIAN gelatin;  Extract fraction;  Chondrocytes;  Collagen TypeⅡ;  Bcl-2;  Bax;  Matrix Metalloproteinases;  Animal experimentation
摘要:
目的:探討龜鹿二仙膠不同提取部位對豚鼠軟骨細胞活性的影響。方法:采用溶劑極性依次遞增分離法分別制備龜鹿二仙 膠的三氯甲烷提取物、正丁醇提取物和水提取物。然后分別以三氯甲烷提取物(烷提組)、正丁醇提取物(醇提組)和水提取物(水 提組)及生理鹽水(對照組)對4組Wistar大鼠進行灌胃,制備含藥血清,再分別以這些含藥血清對體外培養(yǎng)的豚鼠軟骨細胞進行干 預(yù)。干預(yù)結(jié)束后分別采用免疫組織化學技術(shù)、實時熒光定量聚合酶鏈式反應(yīng)法及酶聯(lián)免疫吸附法測定經(jīng)4種含藥血清干預(yù)后的豚 鼠軟骨細胞中Ⅱ型膠原蛋白、Bcl-2和Bax基因及MMP-3水平。結(jié)果:①Ⅱ型膠原蛋白水平。4組細胞Ⅱ型膠原蛋白水平比較,差異 有統(tǒng)計學意義[(0.178±0.002),(0.188±0.003),(0.177±0.003),(0.173±0.001),F=32.905,P=0.000]; 對 照組Ⅱ型膠原蛋白水平低于其余3組(P=0.003,P=0.000,P=0.023); 醇提組高于烷提組和水提組(P=0.000,P=0.000); 烷提組和水 提組比較,差異無統(tǒng)計學意義(P=0.367)。②Bcl-2基因和Bax基因水平。4組細胞Bcl-2基因水平比較,差異有統(tǒng)計學意義 [(0.569±0.052),(1.000±0.000),(0.636±0.072),(0.387±0.061),F=136.767,P=0.000]; 對照組低于其余 3組(P=0.000,P=0.000,P=0.000); 醇提組高于烷提組和水提組(P=0.000,P=0.000); 烷提組低于水提組(P=0.045)。4組細胞Bax 基因水平比較,差異有統(tǒng)計學意義[(1.789±0.240),(1.000±0.000),(1.374±0.226), (2.251±0.196),F=47.445,P=0.000]; 對照組高于其余3組(P=0.000,P=0.000,P=0.000); 醇提組低于烷提組和水提組 (P=0.000,P=0.003); 烷提組高于水提組(P=0.000)。③基質(zhì)金屬蛋白酶3水平。4組細胞基質(zhì)金屬蛋白酶3水平比較,差異有統(tǒng)計 學意義[(11.828±0.363)ng·mL-1,(10.685±0.317)ng·mL-1,(11.397±0.314) ng·mL-1,(13.136±0.379)ng·mL-1,F=71.401,P=0.000]; 對照組高于其余3組 (P=0.000,P=0.000,P=0.000); 醇提組低于烷提組和水提組(P=0.000,P=0.000); 烷提組高于水提組(P=0.019)。結(jié)論:龜鹿二仙 膠的三氯甲烷、正丁醇和水3種提取部位均能提高軟骨細胞中Ⅱ型膠原和Bcl-2基因水平,降低Bax基因和基質(zhì)金屬蛋白酶3水平, 其中以正丁醇提取部位的作用最強。
Abstract:
Objective:To study the effect of different extract fraction of GUILU ERXIAN gelatin on the chondrocytes activity in Guinea pig.Methods:The trichloromethane extractive,n-butanol extractive and water extractive were made from GUILU ERXIAN gelatin respectively by the separation method of increasing solvent polarity progressively.The Wistar rats in 4 groups were intragastric administrated with trichloromethane extractive,n-butanol extractive,water extractive and normal saline respectively.Then the chondrocytes of Guinea pigs cultured in vitro were intervened by the medicated serums extracted from the Wistar rats respectively.After intervention,the levels of Collagen TypeⅡ,Bcl-2 gene,Bax gene and MMP-3 in the chondrocytes of Guinea pigs were determined by using immunohistochemical technique,real-time fluorescent quantitative polymerase chain reaction and enzyme linked immunosorbent assay respectively.Results:There was statistical difference in the level of Collagen TypeⅡbetween the 4 groups(0.178+/-0.002,0.188+/-0.003,0.177+/- 0.003,0.173+/-0.001,F=32.905,P=0.000).The level of Collagen TypeⅡwas lower in the control group compared with the other 3 groups(P=0.003,P=0.000,P=0.023)and was higher in the n-butanol extractive group compared with trichloromethane extractive group and water extractive group(P=0.000,P=0.000)and there was no statistical difference between n-butanol extractive group and water extractive group(P=0.367).There was statistical difference in the level of Bcl-2 gene between the 4 groups(0.569+/-0.052,1.000+/-0.000,0.636+/-0.072,0.387+/- 0.061,F=136.767,P=0.000).The level of Bcl-2 gene was lower in the control group comk pared with the other 3 groups(P=0.000,P=0.000,P=0.000),and was higher in the n-butanol extractive group compared with trichloromethane extractive group and water extractive group(P=0.000,P=0.000)and was lower in the trichloromethane extractive group compared with water extractive group(P=0.045).There was statistical difference in the level of Bax gene between the 4 groups(1.789+/-0.240,1.000+/-0.000,1.374+/-0.226,2.251+/- 0.196,F=47.445,P=0.000).The level of Bax gene was higher in the control group compared with the other 3 groups (P=0.000,P=0.000,P=0.000)and was lower,in the n-butanol extractive group compared with trichloromethane extractive group and water extractive group(P=0.000,P=0.003)and was higher in the trichloromethane extractive group compared with the water extractive group(P=0.000).There was statistical difference in the level of MMP-3 between the 4 groups(11.828+/-0.363,10.685+/-0.317,11.397+/-0.314,13.136+/-0.379 ng/mL,F=71.401,P=0.000).The level of MMP-3 was higher in the control group compared with the other 3 groups(P=0.000,P=0.000,P=0.000)and was lower in the n-butanol extractive group compared with trichloromethane extractive group and water extract group(P=0.000,P=0.000)and was higher in the trichloromethane extractive group compared with water extractive group(P=0.019).Conclusion:The extract fraction of GUILU ERXIAN gelatin including trichloromethane,n-butanol and water can up-regulate the levels of Collagen TypeⅡand Bcl-2 gene and down-regulate the levels of Bax gene and MMP-3,and the effect of n-butanol is the strongest.

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備注/Memo

備注/Memo:
基金項目:國家自然科學基金項目(30772814),福建省教育廳重點項目(JA13153)
通訊作者:李楠 E-mail:mr.linan@126.com
更新日期/Last Update: 2014-10-30