摘要
目的:探讨牛膝对人关节软骨细胞体外增殖的影响。方法:膝骨关节炎患者口服不同浓度牛膝醇提物,收集不同浓度牛膝含药血清,取膝骨关节炎行关节置换手术患者的关节软骨,剪碎后,采用Ⅱ型胶原酶溶解消化软骨细胞,甲苯胺蓝染色确定软骨细胞的存在。将第3代软骨细胞分别接种于4组:空白对照组、含药血清高剂量组、含药血清中剂量组、含药血清低剂量组。荧光倒置相差显微镜观察软骨细胞形态和结构,采用CCK-8法检测软骨细胞增殖情况,RT-PCR法分析软骨细胞基因Ⅱ型胶原mRNA表达水平。结果:牛膝含药血清高、中剂量组细胞增殖率高于对照组(P<0.05),含药血清低剂量组与空白对照组比较差异无统计学意义(P>0.05);RT-PCR法检测结果显示牛膝含药血清高、中、低剂量组均高于空白对照组(P<0.05)。结论:牛膝含药血清能促进人软骨细胞体外培养增殖和Ⅱ型胶原的合成。
Objective:To observe the effect of Niuxi(Radix Achyranthes Bidentata)on proliferation of human articular chondrocytes.Methods:Patients with knee osteoarthritis were given orally different concentrations of alcohol extractive of Radix Achyranthis Bidentatae.Different concentrations of Achyranthes containing serum and articular cartilage from performed knee replacement surgery patient was collected.After crushing articular cartilage,typeⅡcollagenase was used to dissolve digestive articular cartilage.Toluidine blue staining confirms the existence of chondrocytes.Third-generation chondrocytes were seeded into 4 groups,blank control group,low dosage of serum group,median dosage of serum group and high dosage of serum group.The morphology and structure of chondrocytes were observed by fluorescence inverted phase contrast microscope.The proliferation rate of chondrocytes was detected by CCK-8.The expression of typeⅡcollagen gene in chondrocytes was analyzed by reverse transcription-PCR.Results:The high and median dosage of Radix Achyranthes Bidentata groups showed higher chondrocytes proliferation rate than the blank control group(P<0.05),and there was no significant difference between the low dosage of Radix Achyranthes Bidentata group and the blank control group(P>0.05);The results of RT-PCR showed that the high,medium,and low dosage of Radix Achyranthes Bidentata groups showed higher expression of typeⅡcollagen gene than the blank control group(P<0.05).Conclusion:Radix Achyranthes Bidentata serum can promote the proliferation of human chondrocytes in vitro and the synthesis of typeⅡcollagen.
作者
胡烈奎
王立新
彭力平
马笃军
廖州伟
李媛
HU Lie-kui;WANG Li-xin;PENG Li-ping;MA Du-jun;LIAO Zhou-wei;LI Yuan(The Fourth Clinical Medical College,Guangzhou University of Chinese Medicine,Shenzhen Guangdong 518033,China;Shenzhen Traditional Chinese Medical Hospital,Shenzhen Guangdong 518033,China)
出处
《中医药导报》
2019年第3期42-45,共4页
Guiding Journal of Traditional Chinese Medicine and Pharmacy
基金
广东省深圳市卫生科研项目(201601050)
关键词
牛膝
人软骨细胞
增殖
Ⅱ型胶原
Niuxi(Radix Achyranthes Bidentata)
human chondrocytes
proliferation
typeⅡcollagen
