白芍总苷对人骨关节炎软骨细胞增殖及p38MAPK蛋白表达的影响
|
摘要
目的:研究白芍总苷对人骨关节炎软骨细胞增殖和p38MAPK表达的影响。方法:分离膝骨关节炎患者的关节软骨细胞,分为4组,对照组只加培养基,白芍总苷组加入白芍总苷终浓度为8μmol/L的培养基,激动剂组加入p38MAPK信号通路激动剂anisomycin终浓度为2μg/L的培养基,联合组加入含白芍总苷和anisomycin的培养基(终浓度同前)。分组培养24、48、72 h后,MTT法检测细胞增殖能力;分组培养48 h后,碘化丙啶单染法检测细胞周期,qRT-PCR法检测细胞中Ki67、PCNA mRNA表达水平,Western blot法检测细胞中p38MAPK和磷酸化p38MAPK(p-p38MAPK)蛋白表达水平。结果:Anisomycin单独作用可抑制骨关节炎软骨细胞的增殖,阻滞细胞周期进展,促进p38MAPK和p-p38MAPK蛋白的表达(P <0. 05);白芍总苷单独作用则促进细胞增殖和细胞周期进展,抑制p38MAPK和p-p38MAPK蛋白的表达(P <0. 05);两者联用,白芍总苷可以拮抗anisomycin诱导的骨关节炎软骨细胞p38MAPK和p-p38MAPK的表达增加和增殖抑制作用(P <0. 05)。结论:白芍总苷可能通过抑制p38MAPK信号通路,促进人骨关节炎软骨细胞的增殖。
Aim: To study the effects of total glucosides of paeonia(TGP) on the proliferation and p38 MAPK protein expression of human osteoarthritic chondrocytes. Methods: Human osteoarthritic chondrocytes were isolated from the patients with knee osteoarthritis,and were allocated into 4 groups,TGP group(cultured with 8 μmol/L TGP),agonist group(cultured with 2 μg/L anisomycin which was p38 MAPK signal pathway agonist),combination group(cultured with 8μmol/L TGP and 2 μg/L anisomycin),blank control group(cultured without TGP or anisomycin). After 24,48,72 hour culture,cell proliferation was detected by MTT assay. After 48 hour culture,the cell cycle was detected by PI monochrome staining,the expressions of Ki67 and PCNA mRNA was detected by qRT-PCR,and the p38 MAPK and phosphorylatedp38 MAPK(p-p38 MAPK) proteins were detected by Western blot. Results: When treated with anisomycin,the proliferation ability of human osteoarthritic chondrocytes was decreased,the cell cycle was blocked at G0/G1 phase,and the expression levels of p38 MAPK and p-p38 MAPK were increased(P < 0. 05). When treated with TGP,the proliferation ability of human osteoarthritic chondrocytes was increased,the cell cycle was progressing,and the expression levels of p38 MAPK and pp38 MAPK were decreased(P < 0. 05). The combination of the two drugs antagonized each other(P < 0. 05). Conclusion:TGP may promote the proliferation of human osteoarthritic chondrocytes by inhibiting p38 MAPK signaling pathway.
Aim: To study the effects of total glucosides of paeonia(TGP) on the proliferation and p38 MAPK protein expression of human osteoarthritic chondrocytes. Methods: Human osteoarthritic chondrocytes were isolated from the patients with knee osteoarthritis,and were allocated into 4 groups,TGP group(cultured with 8 μmol/L TGP),agonist group(cultured with 2 μg/L anisomycin which was p38 MAPK signal pathway agonist),combination group(cultured with 8μmol/L TGP and 2 μg/L anisomycin),blank control group(cultured without TGP or anisomycin). After 24,48,72 hour culture,cell proliferation was detected by MTT assay. After 48 hour culture,the cell cycle was detected by PI monochrome staining,the expressions of Ki67 and PCNA mRNA was detected by qRT-PCR,and the p38 MAPK and phosphorylatedp38 MAPK(p-p38 MAPK) proteins were detected by Western blot. Results: When treated with anisomycin,the proliferation ability of human osteoarthritic chondrocytes was decreased,the cell cycle was blocked at G0/G1 phase,and the expression levels of p38 MAPK and p-p38 MAPK were increased(P < 0. 05). When treated with TGP,the proliferation ability of human osteoarthritic chondrocytes was increased,the cell cycle was progressing,and the expression levels of p38 MAPK and pp38 MAPK were decreased(P < 0. 05). The combination of the two drugs antagonized each other(P < 0. 05). Conclusion:TGP may promote the proliferation of human osteoarthritic chondrocytes by inhibiting p38 MAPK signaling pathway.
引文
[1]MALFAIT AM.Osteoarthritis year in review 2015:biology[J].Osteoarthritis Cartilage,2016,24(1):21
[2]HERRERO-BEAUMONT G,ROMAN-BLAS JA,BRUYEREO,et al.Clinical settings in knee osteoarthritis:pathophysiology guides treatment[J].Maturitas,2017,96(13):54
[3]闫虎.骨关节炎相关信号通路的研究[J].中华临床医师杂志:电子版,2016,10(12):1823
[4]ZHANG ZM,SHEN C,LI H,et al.Leptin induces the apoptosis of chondrocytes in an in vitro model of osteoarthritis via the JAK2STAT3 signaling pathway[J].Mol Med Rep,2016,13(4):3684
[5]王涛,殷红,廖江龙,等.骨关节炎与MAPK信号通路关系的研究概况[J].中国民族民间医药,2017,26(19):27
[6]SUN HY,HU KZ,YIN ZS.Inhibition of the p38-MAPKsignaling pathway suppresses the apoptosis and expression of proinflammatory cytokines in human osteoarthritis chondrocytes[J].Cytokine,2017,90(11):135
[7]LUO J,JIN DE,YANG GY,et al.Total glucosides of paeony for rheumatoid arthritis:a systematic review of randomized controlled trials[J].Complement Ther Med,2017,34(10):46
[8]张洪峰,肖卫国,侯平.白芍总苷治疗系统性红斑狼疮的临床研究[J].中国中西医结合杂志,2011,31(4):476
[9]胥方元,何成松,干锦华,等.白芍总苷治疗膝关节骨关节炎[J].中国康复,2004,19(3):166
[10]PIAO T,MA Z,LI X,et al.Taraxasterol inhibits IL-1β-induced inflammatory response in human osteoarthritic chondrocytes[J].Eur J Pharmacol,2015,756(12):38
[11]游必凯,王平,孙克民.中药防治骨性关节炎实验研究进展[J].医学综述,2010,16(2):295
[12]HOU PW,FU PK,HSU HC,et al.Traditional Chinese medicine in patients with osteoarthritis of the knee[J].J Tradit Complement Med,2015,5(4):182
[13]LI LF,LIU HQ,SHI WM,et al.Insights into the action mechanisms of traditional Chinese medicine in osteoarthritis[J].Evid Based Complement Alternat Med,2017.doi:10.1155/2017/5190986
[14]杜旭召,杨豪,邓素玲,等.白芍总苷对骨关节炎软骨细胞增殖及分泌表达的影响[J].中国骨质疏松杂志,2016,22(11):1375
[15]杨海波.白芍总苷对兔膝骨性关节炎滑膜基质金属蛋白酶变化的实验研究[D].沈阳:辽宁中医药大学,2015.
[16]王欢,王庆甫,唐学章,等.白芍总苷对滑膜成纤维细胞增殖、凋亡及周期影响的实验研究[J].天津中医药,2017,34(12):841
[17]ZHU L,WEI W,ZHENG YQ,et al.Effects and mechanisms of total glucosides of paeony on joint damage in rat collageninduced arthritis[J].Inflamm Res,2005,54(5):211
[18]LI Z,MENG D,LI G,et al.Celecoxib combined with diacerein effectively alleviates osteoarthritis in rats via regulating JNK and p38MAPK signaling pathways[J].Inflammation,2015,38(4):1563
[19]赵晓,黄飞麒,姚乃婕,等.S100A11-RAGE通过P38MAPK信号转导通路调控小鼠骨关节炎软骨细胞肥大和细胞外基质代谢[J].中国现代医学杂志,2016,26(16):6
[20]潘建科,谢辉,刘军,等.龙鳖胶囊对骨关节炎滑膜细胞p38MAPK信号通路及NF-κB p65的调控研究[J].中华中医药学刊,2017,35(1):65
[2]HERRERO-BEAUMONT G,ROMAN-BLAS JA,BRUYEREO,et al.Clinical settings in knee osteoarthritis:pathophysiology guides treatment[J].Maturitas,2017,96(13):54
[3]闫虎.骨关节炎相关信号通路的研究[J].中华临床医师杂志:电子版,2016,10(12):1823
[4]ZHANG ZM,SHEN C,LI H,et al.Leptin induces the apoptosis of chondrocytes in an in vitro model of osteoarthritis via the JAK2STAT3 signaling pathway[J].Mol Med Rep,2016,13(4):3684
[5]王涛,殷红,廖江龙,等.骨关节炎与MAPK信号通路关系的研究概况[J].中国民族民间医药,2017,26(19):27
[6]SUN HY,HU KZ,YIN ZS.Inhibition of the p38-MAPKsignaling pathway suppresses the apoptosis and expression of proinflammatory cytokines in human osteoarthritis chondrocytes[J].Cytokine,2017,90(11):135
[7]LUO J,JIN DE,YANG GY,et al.Total glucosides of paeony for rheumatoid arthritis:a systematic review of randomized controlled trials[J].Complement Ther Med,2017,34(10):46
[8]张洪峰,肖卫国,侯平.白芍总苷治疗系统性红斑狼疮的临床研究[J].中国中西医结合杂志,2011,31(4):476
[9]胥方元,何成松,干锦华,等.白芍总苷治疗膝关节骨关节炎[J].中国康复,2004,19(3):166
[10]PIAO T,MA Z,LI X,et al.Taraxasterol inhibits IL-1β-induced inflammatory response in human osteoarthritic chondrocytes[J].Eur J Pharmacol,2015,756(12):38
[11]游必凯,王平,孙克民.中药防治骨性关节炎实验研究进展[J].医学综述,2010,16(2):295
[12]HOU PW,FU PK,HSU HC,et al.Traditional Chinese medicine in patients with osteoarthritis of the knee[J].J Tradit Complement Med,2015,5(4):182
[13]LI LF,LIU HQ,SHI WM,et al.Insights into the action mechanisms of traditional Chinese medicine in osteoarthritis[J].Evid Based Complement Alternat Med,2017.doi:10.1155/2017/5190986
[14]杜旭召,杨豪,邓素玲,等.白芍总苷对骨关节炎软骨细胞增殖及分泌表达的影响[J].中国骨质疏松杂志,2016,22(11):1375
[15]杨海波.白芍总苷对兔膝骨性关节炎滑膜基质金属蛋白酶变化的实验研究[D].沈阳:辽宁中医药大学,2015.
[16]王欢,王庆甫,唐学章,等.白芍总苷对滑膜成纤维细胞增殖、凋亡及周期影响的实验研究[J].天津中医药,2017,34(12):841
[17]ZHU L,WEI W,ZHENG YQ,et al.Effects and mechanisms of total glucosides of paeony on joint damage in rat collageninduced arthritis[J].Inflamm Res,2005,54(5):211
[18]LI Z,MENG D,LI G,et al.Celecoxib combined with diacerein effectively alleviates osteoarthritis in rats via regulating JNK and p38MAPK signaling pathways[J].Inflammation,2015,38(4):1563
[19]赵晓,黄飞麒,姚乃婕,等.S100A11-RAGE通过P38MAPK信号转导通路调控小鼠骨关节炎软骨细胞肥大和细胞外基质代谢[J].中国现代医学杂志,2016,26(16):6
[20]潘建科,谢辉,刘军,等.龙鳖胶囊对骨关节炎滑膜细胞p38MAPK信号通路及NF-κB p65的调控研究[J].中华中医药学刊,2017,35(1):65