龟鹿二仙胶不同提取部位对豚鼠软骨细胞活性的影响

目的：探讨龟鹿二仙胶不同提取部位对豚鼠软骨细胞活性的影响.方法：采用溶剂极性依次递增分离法分别制备龟鹿二仙胶的三氯甲烷提取物、正丁醇提取物和水提取物.然后分别以三氯甲烷提取物（烷提组）、正丁醇提取物（醇提组）和水提取物（水提组）及生理盐水（对照组）对4组Wistar大鼠进行灌胃,制备含药血清,再分别以这些含药血清对体外培养的豚鼠软骨细胞进行干预.干预结束后分别采用免疫组织化学技术、实时荧光定量聚合酶链式反应法及酶联免疫吸附法测定经4种含药血清干预后的豚鼠软骨细胞中Ⅱ型胶原蛋白、Bcl-2和Bax基因及MMP-3水平.结果：①Ⅱ型胶原蛋白水平.4组细胞Ⅱ型胶原蛋白水平比较,差异有统计学意义[（0.178±0.002）,（0.188±0.003）,（0.177±0.003）,（0.173 ±0.001）,F=32.905,P=0.000];对照组Ⅱ型胶原蛋白水平低于其余3组（P=0.003,P=0.000,P=0.023）;醇提组高于烷提组和水提组（P=0.000,P=0.000）;烷提组和水提组比较,差异无统计学意义（P =0.367）.②Bcl-2基因和Bax基因水平.4组细胞Bcl-2基因水平比较,差异有统计学意义[（0.569±0.052）,（l.000±0.000）,（0.636±0.072）,（0.387±0.061）,F=136.767,P=0.000];对照组低于其余3组（P =0.000,P=0.000,P=0.000）;醇提组高于烷提组和水提组（P=0.000,P=0.000）;烷提组低于水提组（P=0.045）.4组细胞Bax基因水平比较,差异有统计学意义[（1.789±0.240）,（1.000 ±0.000）,（1.374±0.226）,（2.251±0.196）,F=47.445,P=0.000];对照组高于其余3组（P=0.000,P=0.000,P=0.000）;醇提组低于烷提组和水提组（P=0.000,P=0.003）;烷提组高于水提组（P=0.000）.③基质金属蛋白酶3水平.4组细胞基质金属蛋白酶3水平比较,差异有统计学意义[（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];对照组高于其余3组（P =0.000

Effect of different extract fraction of GUILU ERXIAN gelatin on the chondrocytes activity in Guinea pigs

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 respec- tively 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 cul- tured in vitro were intervened by the medicated serums extracted from the Wistar rats respectively. After intervention, the levels of Collagen Type II ,Bel- 2 gene, Bax gene and MMP -3 in the chondrocytes of Guinea pigs were determined by using immunohistoehemical tech- nique, real - time fluorescent quantitative polymerase chain reaction and enzyme linked immunosorbent assay respectively. Results ： There was statistical difference in the level of Collagen Type 1I between the 4 groups （0. 178 ＋/-. 002,0. 188 ＋/-. 003,0. 177 ＋/-. 003, 0. 173 ＋/-0. 001, F = 32. 905, P = 0. 000）. The level of Collagen Type lI was 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 comkpared 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 trichlo- romethane extractive group and water extractive group （ P = 0. 000, P = 0. 000 ） and wa