治疗颈性眩晕的细胞生物力学效应的研究
|
摘要
通过培养人血管内皮细胞株(CRL-1730)来建立颈性眩晕血瘀证患者血清诱导的细胞损伤模型,对细胞进行形态学鉴定后,实施体外压力刺激,建立对细胞的力学加载模型,同时用丹参酮ⅡA对细胞模型干预进行对比研究,检测细胞外液NO、ET,游离钙浓度、内皮损伤标志物(血栓调节蛋白)含量变化。实验表明:与颈性眩晕血瘀对照组相比,该组压力组与加药组细胞形态变化不甚明显,且细胞活性、细胞内钙离子浓度、NO、ET含量比值及内皮损伤标志物含量差异具有统计学意义(p<0.01);颈性眩晕血瘀低压组与颈性眩晕血瘀高压组相比,低压力刺激干预在减少细胞损伤标志物的含量方面效果更佳,证实对血管内皮细胞的损伤起一定的保护作用,效果甚至优于药物组。
The model of serum induced cell injury in patients with cervical vertigo and blood stasis syndrome was established by culturing human vascular endothelial cell line(CRL-1730). After the morphological identification of the cells, in vitro pressure stimulation was carried out to establish the mechanical loading model. The levels of NO,ET, free calcium concentration and endothelial injury markers(v WF) were measured by Tan Ⅱ A intervention cell model. The results showed that compared with the cervical vertigo and blood stasis control group, the cell morphology of the pressure group and the drug group was not obvious, and the cell activity, intracellular calcium concentration and endothelial injury marker content difference(p<0.01); compared with the cervical vertigo and blood stasis high pressure group, low pressure stimulation intervention had better effect in reducing the content of cell injury markers, and confirmed the protective effect on the damage of vascular endothelial cells, and the effect is even better than that of the medication group.
The model of serum induced cell injury in patients with cervical vertigo and blood stasis syndrome was established by culturing human vascular endothelial cell line(CRL-1730). After the morphological identification of the cells, in vitro pressure stimulation was carried out to establish the mechanical loading model. The levels of NO,ET, free calcium concentration and endothelial injury markers(v WF) were measured by Tan Ⅱ A intervention cell model. The results showed that compared with the cervical vertigo and blood stasis control group, the cell morphology of the pressure group and the drug group was not obvious, and the cell activity, intracellular calcium concentration and endothelial injury marker content difference(p<0.01); compared with the cervical vertigo and blood stasis high pressure group, low pressure stimulation intervention had better effect in reducing the content of cell injury markers, and confirmed the protective effect on the damage of vascular endothelial cells, and the effect is even better than that of the medication group.
引文
Aju bi N.E.,Klein-Nulend J.,Alblas M.J.,Burger E.H.,and Nijweide P.J.,1999,Signal transduction pathways involved in fluid flow-induced PGE2 production by cultured osteocytes,Am.J.Physiol.,276(1):E171-E178
Ali M.H.,Pearlstein D.P.,Mathieu C.E.,and Schumacker P.T.,2004,Mitochondrial requirement for endothelial responses to cyclic strain:implications for mechanotransduction,Am.J.Physiol.Lung Cell.Mol.Physiol.,287(3):486-496
Brandt T.,and Bronstein A.M.,2001,Cervical Vertigo,J.Neurol Neurosurg Psychiatry.,71(1):8-12
Chapman K.E.,Sinclair S.E.,Zhuang D.,Hassid A.,Desai L.P.,and Waters C.M.,2005,Cyclic mechanical strain increases reactive oxygen species production in pulmonary epithelial cells,Am.J.Physiol.Lung Cell.Mol.Physiol.,289(5):834-841
Chen B.,Xie X.M.,Jia Y.,and Lv M.Z.,2008,Study on the effects of extracorporeal pressure stimulation on the release of NO and IL-1βin rat fascial tissue cel ls,Zhonghua Zhongyiyao Zazhi(Chinese Journal of Traditional Chinese Medicine),23(7):622-624(陈波,谢西梅,贾莹,吕明庄,2008,体外压力刺激对大鼠筋膜组织细胞合成释放NO和IL-1β影响的研究,中华中医药杂志,23(7):622-624)
Fan Z.Y.,Zha H.P.,Chen L.G.,and Peng X.M.,2011,Research on the construction and mechanism of cell massage model,Shanghai Zhongyiyao Daxue Xuebao(Journal of Shanghai University of Traditional Chinese Medicine),25(5):100-103(范志勇,查和萍,陈利国,彭旭明,2011,细胞推拿模型的构建及作用机制研究进展,上海中医药大学学报,25(5):100-103)
Fu P.L.,Fang Z.Y.,Zhang J.Z.,Li L.,Peng Z.Y.,Tian N.,Su B.Y.,and Wu S.,2016,Effect of stress stimulation of manipulation on endothelial cells with serum induced injury in patients with blood stasis syndrome of cervical vertigo,Zhongguo Zhongyi Jizheng(Journal of Emergency in Traditional Chinese Medicine),25(9):1656-1659(傅品来,范志勇,张竞之,李黎,彭志允,田宁,苏碧莹,吴栓,2016,模拟手法压力刺激对颈性眩晕血瘀证患者血清损伤后内皮细胞的影响,中国中医急症,25(9):1656-1659)
Hu X.Q.,Chen L.G.,and Zeng X.W.,2009,Hypertension patients with blood stasis on vascular endothelial cell cytoplasm Ca2+concentration in serum and the influence of TanⅡA intervention effect,Liaoning Zhongyi Zazhi(Liao Ning Journal of Traditional Chinese Medicine),36(5):836-838(胡小勤,陈利国,曾学文,2009,高血压病血瘀证患者血清对血管内皮细胞胞浆内游离钙浓度的影响及TanⅡA的干预作用,辽宁中医杂志,36(5):836-838)
Hu X.Q.,Chen L.G.,Jia H.X.,Peng Z.Y.,and Zhang J.Z.,2008,Effects of tanshinoneⅡA on activity of serum ECV-304in patients with blood stasis syndrome of hypertension,Liaoning Zhongyi Zazhi(Liao Ning Journal of Traditional Chinese Medicine),35(7):964-966(胡小勤,陈利国,贾会欣,彭志允,张竞之,2008,丹参酮ⅡA对高血压病血瘀证患者血清致伤ECV-304细胞活性的影响,辽宁中医杂志,35(7):964-966)
Kessler L.,Wiesel M.L.,Attali P.,Mossard J.M.,Cazenave J.P.,and Pinget M.,1998,Von Willebrand factor in diabetic angiopathy,Diabetes Metab.,24(4):327-336
Li T.,Chen L.G.,Li Q.,Zhou Y.H.,Tang H.L.,and Chen S.B.,2011,The protective effects of Tetramethylpyrazine(TMP)combined with Astragalus Polysaccharides(APS)on vascular endothelial cells(VECs),Zhonghua Zhongyiyao Zahi(Chinese Journal of Traditional Chinese Medicine),26(11):2672-2675(李眺,陈利国,李权,周永红,唐海兰,程少冰,2011,川芎嗪配伍黄芪多糖对血管内皮细胞的保护作用,中华中医药杂志,26(11):2672-2675)
Li Y.C.,and Peng B.J.,2015,Pathogenesis,diagnosis,and treatment of cervical vertigo,Zhongguo Jiaoxing Waike Zazhi(Orthopedic Journal of China),23(3):250-253(李永超,彭宝淦,2015,颈性眩晕的发病机制及诊治新进展,中国矫形外科杂志,23(3):250-253)
Ma H.S.,Zhang H.,Yan J.T.,and Shen G.Q.,2010,Design and construction of loadi ng equipment suitable to manipulation for cell mechanics,Zh ongguo Zuzhi Gongcheng Yanjiu(Journal of Clinical Rehabilitative Tissue Engineering Research),14(26):4751-4754(马惠昇,张宏,严隽陶,沈国权,2010,适于推拿手法细胞力学加载装置模型的设计与构建,中国组织工程研究,14(26):4751-4754)
Peng Z.Y.,Chen L.G.,Jia H.X.,Hu X.Q.,and Chen S.B.,2012,Effect of tanshinoneⅡA on endothelial cell injured by serum of patients with hypertension,Zhongchengyao(Chinese Traditional Patent Medicine),34(2):199-203(彭志允,陈利国,贾会欣,胡小勤,程少冰,2012,丹参酮Ⅱ_A对高血压病患者血清损伤后的内皮细胞的影响,中成药,34(2):199-203)
Yacovino D.A.,and Hain T.C.,2013,Clinical characteristics of cervicogenic-related dizziness and vertigo,Semin.Neu rol.,33(3):244-255
You Y.,and Yang H.J.,2008,Research progress of the transient receptor potential cationic channel in vascular endothelial cells,Zhongguo Yaowu Yu Linchuang(Chinese Remedies and Clinics),8(10):760-763(游云,杨洪军,2008,血管内皮细胞瞬时感受器电位阳离子通道研究进展,中国药物与临床,8(10):760-763)
Zhou Y.H.,Chen L.G.,Qu Y.,Yan L.,Tang H.L.,and Cheng S.B.,2009,Vascular endothelial cell lesion model of blood-stasis syndrome of typle 2 diabetes mellitus,Beijing Zhongyiyao Daxue Xuebao(Journal of Beijing University of Traditional Chinese Medicine),32(11):742-746(周永红,陈利国,屈援,颜亮,唐海兰,程少冰,2009,2型糖尿病血瘀证血管内皮细胞损伤模型的研究,北京中医药大学学报,32(11):742-746)
Ali M.H.,Pearlstein D.P.,Mathieu C.E.,and Schumacker P.T.,2004,Mitochondrial requirement for endothelial responses to cyclic strain:implications for mechanotransduction,Am.J.Physiol.Lung Cell.Mol.Physiol.,287(3):486-496
Brandt T.,and Bronstein A.M.,2001,Cervical Vertigo,J.Neurol Neurosurg Psychiatry.,71(1):8-12
Chapman K.E.,Sinclair S.E.,Zhuang D.,Hassid A.,Desai L.P.,and Waters C.M.,2005,Cyclic mechanical strain increases reactive oxygen species production in pulmonary epithelial cells,Am.J.Physiol.Lung Cell.Mol.Physiol.,289(5):834-841
Chen B.,Xie X.M.,Jia Y.,and Lv M.Z.,2008,Study on the effects of extracorporeal pressure stimulation on the release of NO and IL-1βin rat fascial tissue cel ls,Zhonghua Zhongyiyao Zazhi(Chinese Journal of Traditional Chinese Medicine),23(7):622-624(陈波,谢西梅,贾莹,吕明庄,2008,体外压力刺激对大鼠筋膜组织细胞合成释放NO和IL-1β影响的研究,中华中医药杂志,23(7):622-624)
Fan Z.Y.,Zha H.P.,Chen L.G.,and Peng X.M.,2011,Research on the construction and mechanism of cell massage model,Shanghai Zhongyiyao Daxue Xuebao(Journal of Shanghai University of Traditional Chinese Medicine),25(5):100-103(范志勇,查和萍,陈利国,彭旭明,2011,细胞推拿模型的构建及作用机制研究进展,上海中医药大学学报,25(5):100-103)
Fu P.L.,Fang Z.Y.,Zhang J.Z.,Li L.,Peng Z.Y.,Tian N.,Su B.Y.,and Wu S.,2016,Effect of stress stimulation of manipulation on endothelial cells with serum induced injury in patients with blood stasis syndrome of cervical vertigo,Zhongguo Zhongyi Jizheng(Journal of Emergency in Traditional Chinese Medicine),25(9):1656-1659(傅品来,范志勇,张竞之,李黎,彭志允,田宁,苏碧莹,吴栓,2016,模拟手法压力刺激对颈性眩晕血瘀证患者血清损伤后内皮细胞的影响,中国中医急症,25(9):1656-1659)
Hu X.Q.,Chen L.G.,and Zeng X.W.,2009,Hypertension patients with blood stasis on vascular endothelial cell cytoplasm Ca2+concentration in serum and the influence of TanⅡA intervention effect,Liaoning Zhongyi Zazhi(Liao Ning Journal of Traditional Chinese Medicine),36(5):836-838(胡小勤,陈利国,曾学文,2009,高血压病血瘀证患者血清对血管内皮细胞胞浆内游离钙浓度的影响及TanⅡA的干预作用,辽宁中医杂志,36(5):836-838)
Hu X.Q.,Chen L.G.,Jia H.X.,Peng Z.Y.,and Zhang J.Z.,2008,Effects of tanshinoneⅡA on activity of serum ECV-304in patients with blood stasis syndrome of hypertension,Liaoning Zhongyi Zazhi(Liao Ning Journal of Traditional Chinese Medicine),35(7):964-966(胡小勤,陈利国,贾会欣,彭志允,张竞之,2008,丹参酮ⅡA对高血压病血瘀证患者血清致伤ECV-304细胞活性的影响,辽宁中医杂志,35(7):964-966)
Kessler L.,Wiesel M.L.,Attali P.,Mossard J.M.,Cazenave J.P.,and Pinget M.,1998,Von Willebrand factor in diabetic angiopathy,Diabetes Metab.,24(4):327-336
Li T.,Chen L.G.,Li Q.,Zhou Y.H.,Tang H.L.,and Chen S.B.,2011,The protective effects of Tetramethylpyrazine(TMP)combined with Astragalus Polysaccharides(APS)on vascular endothelial cells(VECs),Zhonghua Zhongyiyao Zahi(Chinese Journal of Traditional Chinese Medicine),26(11):2672-2675(李眺,陈利国,李权,周永红,唐海兰,程少冰,2011,川芎嗪配伍黄芪多糖对血管内皮细胞的保护作用,中华中医药杂志,26(11):2672-2675)
Li Y.C.,and Peng B.J.,2015,Pathogenesis,diagnosis,and treatment of cervical vertigo,Zhongguo Jiaoxing Waike Zazhi(Orthopedic Journal of China),23(3):250-253(李永超,彭宝淦,2015,颈性眩晕的发病机制及诊治新进展,中国矫形外科杂志,23(3):250-253)
Ma H.S.,Zhang H.,Yan J.T.,and Shen G.Q.,2010,Design and construction of loadi ng equipment suitable to manipulation for cell mechanics,Zh ongguo Zuzhi Gongcheng Yanjiu(Journal of Clinical Rehabilitative Tissue Engineering Research),14(26):4751-4754(马惠昇,张宏,严隽陶,沈国权,2010,适于推拿手法细胞力学加载装置模型的设计与构建,中国组织工程研究,14(26):4751-4754)
Peng Z.Y.,Chen L.G.,Jia H.X.,Hu X.Q.,and Chen S.B.,2012,Effect of tanshinoneⅡA on endothelial cell injured by serum of patients with hypertension,Zhongchengyao(Chinese Traditional Patent Medicine),34(2):199-203(彭志允,陈利国,贾会欣,胡小勤,程少冰,2012,丹参酮Ⅱ_A对高血压病患者血清损伤后的内皮细胞的影响,中成药,34(2):199-203)
Yacovino D.A.,and Hain T.C.,2013,Clinical characteristics of cervicogenic-related dizziness and vertigo,Semin.Neu rol.,33(3):244-255
You Y.,and Yang H.J.,2008,Research progress of the transient receptor potential cationic channel in vascular endothelial cells,Zhongguo Yaowu Yu Linchuang(Chinese Remedies and Clinics),8(10):760-763(游云,杨洪军,2008,血管内皮细胞瞬时感受器电位阳离子通道研究进展,中国药物与临床,8(10):760-763)
Zhou Y.H.,Chen L.G.,Qu Y.,Yan L.,Tang H.L.,and Cheng S.B.,2009,Vascular endothelial cell lesion model of blood-stasis syndrome of typle 2 diabetes mellitus,Beijing Zhongyiyao Daxue Xuebao(Journal of Beijing University of Traditional Chinese Medicine),32(11):742-746(周永红,陈利国,屈援,颜亮,唐海兰,程少冰,2009,2型糖尿病血瘀证血管内皮细胞损伤模型的研究,北京中医药大学学报,32(11):742-746)