MTT法检测通络止痛方对人膝骨关节炎滑膜成纤维细胞增殖的影响

摘要

目的:探讨通络止痛方对人膝关节骨性关节炎(Knee Osteoarthritis,KOA)滑膜成纤维细胞增殖的影响.方法:取本院行全膝关节置换术患者术中废弃滑膜组织,经I型胶原酶消化后进行原代培养,取3~4代成纤维细胞进行高、中、低浓度通络止痛方干预.应用四甲基偶氮唑盐法测定(MTT)法检测KOA滑膜细胞的增殖水平,并用倒置显微镜下观察各浓度下的细胞形态.结果:加药组组间与加药组与空白组比较,差异有统计学意义(P<0.05).MTT法检测随通络止痛方浓度减小KOA滑膜细胞在490 nm波长处吸光OD值数越大;高中浓度组抑制率为正值,随浓度增加抑制率增大,低浓度抑制率为负值,随浓度降低抑制率减少.结论:高、中浓度通络止痛方对滑膜细胞增殖有抑制作用,低浓度组的通络止痛方对滑膜细胞有增殖有促进作用.%Objective:To explore the effect of Tongluo Zhitong Decoction on the proliferation of knee osteoarthritis (KOA) synovial fibroblasts.Methods:The abandoned synovial tissues were taken from patients who had knee replacement surgery in the hospital.The tissues were primarily cultured after digestion in I type collagenase.Three to four generation fibroblasts were taken for high-medium-low concentration intervention.The proliferation of KOA synovial cells was detected by 4-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and the cell morphology was observed under the inverted microscope.Results:Comparisons between the drug groups and comparisons between the drug groups and the control group,the difference was statistically significant (P<0.05).MTT assay showed that s the light OD number of KOA synovial cells became greatest in the wavelength of 490 nm when Tongluo Zhitong Decoction concentration decreased.The inhibition rates of high-medium concentration groups were positive.With the increase of the concentration,the inhibition rate increased and the inhibition rate of the low concentration group was negative.With the concentration decreased,the inhibition rate reduced.Conclusion:The medium-high concentrated Tongluo Zhitong Decoction may inhibit the proliferation of synovial cells,while low concentrated Tongluo Zhitong Decoction may promote the proliferation of synovial cells.