化痰除湿祛瘀剂对人骨关节炎软骨细胞凋亡的影响

发布时间：2018-02-05 20:14

本文关键词： 化痰除湿祛瘀剂人骨关节炎细胞凋亡　出处：《中华中医药学刊》2017年06期 　论文类型：期刊论文

【摘要】：目的:研究化痰除湿祛瘀剂对人骨关节炎(osteoarthritis,OA)软骨细胞凋亡的影响。方法:(1)在苏州中医院骨伤科协助下,取因外伤急性截肢患者的软骨组织以及全膝关节置换手术OA患者的软骨组织,采用酶消化培养法培养人软骨细胞。采用甲苯胺蓝染色鉴定细胞来源,观察显微镜观察其形态变化。(2)将软骨细胞分成OA软骨细胞组、OA软骨细胞组+化痰除湿祛瘀剂(0.016、0.08、0.4、2、10 mg/mL)组,根据分组情况分别培养24 h或48 h后,应用MTT法比较各组细胞的增殖情况。(3)根据增殖结果将软骨细胞分成OA软骨细胞组、OA软骨细胞组+化痰除湿祛瘀剂(0.05、0.1、2 mg/mL)组,用原位末端标记法(TUNEL)检测各组细胞凋亡情况。结果:(1)成功建立了人骨关节炎软骨细胞有限细胞系,细胞生长状态良好;甲苯胺蓝证明所培养的细胞为来自中胚层的软骨细胞。(2)与对照组比较(24 h+),不同浓度的化痰除湿祛瘀剂对OA软骨细胞活力无影响。与对照组(OD值:0.393±0.010,λ=450 nm)比较(24 h+),化痰除湿祛瘀剂(1 mg/mL,OD值:0.441±0.009)和(2mg/mL,OD值:0.430±0.012)组可明显促进软骨细胞增殖(均P0.001)。(3)OA软骨细胞经TUNEL标记后的结果显示,对照组的凋亡指数为(42.21±2.67)%,化痰除湿祛瘀剂(0.1 mg/mL,37.14%±4.01%;2 mg/mL,34.21%±5.00%)可明显抑制软骨细胞早期凋亡与对照组比较(P0.05或P0.01)。结论:化痰除湿祛瘀剂促进细胞增殖,抑制软骨细胞过度凋亡,这可能是抑制软骨破坏的重要机制之一。
[Abstract]:Objective: to study the effect of phlegm removing dampness and removing blood stasis on the apoptosis of chondrocytes in osteoarthritis osteoarthritis OAA. Methods: 1) with the assistance of Department of Orthopedics and Trauma, Suzhou traditional Chinese Medicine Hospital. The cartilage tissue of acute amputation due to trauma and the cartilage tissue of OA patients undergoing total knee arthroplasty were collected and cultured by enzyme digestion. The origin of the cells was identified by toluidine blue staining. The chondrocytes were divided into OA chondrocyte group and OA chondrocyte group. The 10 mg / mL group was cultured for 24 h or 48 h according to the group condition. According to the results of proliferation, chondrocytes were divided into OA chondrocyte group and OA chondrocyte group. In 2 mg / mL group, Tunel was used to detect the apoptosis of human osteoarthritis chondrocytes. Results: 1) Human osteoarthritis chondrocyte limited cell line was successfully established. Cell growth was good; Toluidine blue showed that the cultured cells were chondrocytes from mesoderm. The activity of OA chondrocytes was not affected by different concentrations of phlegm removing dampness and removing blood stasis, and the OD value of OA chondrocytes was 0.393 卤0.010, 位 450nm compared with the control group for 24 h). The OD value of phlegm removing dampness and removing blood stasis was 0. 441 卤0. 009 and the OD value of 1 mg / mLX was 0. 441 卤0. 009) and 2 mg / mL. OD: 0.430 卤0.012) group could significantly promote the proliferation of chondrocytes (all P0.001 ~ (3) OA chondrocytes were labeled with TUNEL. The apoptotic index of the control group was 42.21 卤2.67, and that of the phlegm removing dampness and removing blood stasis was 37.14% 卤4.01%, and that of removing phlegm and removing dampness and removing blood stasis was 37.14% 卤4.01; 2 mg/mL. (34.21% 卤5.00) can significantly inhibit the early apoptosis of chondrocytes compared with the control group, P0.05 or P0.01.Conclusion: phlegm removing dampness and removing stasis can promote the proliferation of chondrocytes. Inhibition of excessive apoptosis of chondrocytes may be one of the important mechanisms to inhibit cartilage damage.
【作者单位】：苏州吴门医派研究院;上海中医药大学附属曙光医院;苏州市中医医院骨伤科;
【基金】：苏州市科技局计划项目(SYS201420)
【分类号】：R274.9

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