大黄酸抑制气道平滑肌收缩机制研究
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摘要
目的研究大黄酸是否抑制气道平滑肌收缩及其机制。方法取SPF级BALB/c♂小鼠气管环,将气管环置于恒温灌流系统中,通过JH-2型张力换能器记录气管环张力值。使用80 mmol/L钾(K+)收缩气管环,待气管环张力达到稳定后,分别加入终浓度为1. 00、3. 16、10.00、31. 60、100.00、177. 80、200. 00μmol/L的大黄酸,加入等量的溶剂作为对照。用膜片钳记录L-型电压依赖性钙通道(L-type voltage-dependent calcium channel,LVDCC)电流,大黄酸可以抑制LVDCC电流。用TILL钙成像系统测量平滑肌细胞内钙浓度变化。结果当加入177. 80μmol/L大黄酸时,气管环舒张比率达到最大,为(92. 7±8. 0)%,抑制LVDCC介导的电流、LVDCC介导的钙内流,以及钙内流引起的收缩。结论大黄酸可通过抑制LVDCC抑制气道平滑肌收缩。
Objective To investigate the relaxant effect of rhein on mouse airway smooth muscle and the underlying mechanism. Methods SPF BALB/c male mice were sacrificed to isolate the tracheal rings(TRs). After balancing in the perfusion system,the contraction tension of the TRs was recorded with JH-2 tension recording system. The TRs were precontracted using 80 mmol/L K+. When the muscle tension of the TRs reached a steady state, the rhein was accumulatively added at concentrations of 1. 00,3. 16,10. 00,31. 60,100. 00,177. 80,200. 00 μmol/L,respectively,and the tension changes were observed. The control experiments were also performed. The effects of rhein on the currents of the L-type voltage-dependent Ca~(2+)channels(LVDCC)were recorded using patchclamp technique. Similarly,the effects of rhein at different concentrations on the levels of intracellular Ca~(2+)([Ca~(2+)]i)in airway smooth muscle cells were recorded using TILL calcium imaging system.Results The relaxant effects induced by rhein of 177. 80 μmol/L reached the maximal value(92. 7±8. 0)%.Rhein inhibited LVDCC-mediated currents and LVDCC-mediated increases of[Ca~(2+)]i,as well as contraction caused by calcium influx. Conclusion Rhein inhibits contraction of airway smooth muscle via inhibiting LVDCC.
Objective To investigate the relaxant effect of rhein on mouse airway smooth muscle and the underlying mechanism. Methods SPF BALB/c male mice were sacrificed to isolate the tracheal rings(TRs). After balancing in the perfusion system,the contraction tension of the TRs was recorded with JH-2 tension recording system. The TRs were precontracted using 80 mmol/L K+. When the muscle tension of the TRs reached a steady state, the rhein was accumulatively added at concentrations of 1. 00,3. 16,10. 00,31. 60,100. 00,177. 80,200. 00 μmol/L,respectively,and the tension changes were observed. The control experiments were also performed. The effects of rhein on the currents of the L-type voltage-dependent Ca~(2+)channels(LVDCC)were recorded using patchclamp technique. Similarly,the effects of rhein at different concentrations on the levels of intracellular Ca~(2+)([Ca~(2+)]i)in airway smooth muscle cells were recorded using TILL calcium imaging system.Results The relaxant effects induced by rhein of 177. 80 μmol/L reached the maximal value(92. 7±8. 0)%.Rhein inhibited LVDCC-mediated currents and LVDCC-mediated increases of[Ca~(2+)]i,as well as contraction caused by calcium influx. Conclusion Rhein inhibits contraction of airway smooth muscle via inhibiting LVDCC.
引文
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[2] MUNGAN D,AYDIN O,MAHBOUB B,et al. Burden of disease associated with asthma among the adult general population of five Middle eastern countries:results of the snapshot program[J]. Respir Med,2018,139:55-64.
[3] CHAPMAN D G,IRVIN C G. Mechanisms of airway hyper-responsiveness in asthma:the past,present and yet to come[J]. Clin Exp Allergy,2015,45(4):706-719.
[4] ABRAMSON M J,WALTERS J,WALTERS E H. Adverse effects of beta-agonists:are they clinically relevant?[J]. Am J Respir Med,2003,2(4):287-297.
[5] CAZZOLA M,MATERA M G,DONNER C F. Inhaled beta2-adrenoceptor agonists:cardiovascular safety in patients with obstructive lung disease[J]. Drugs,2005,65(12):1595-1610.
[6] NINO G,HU A,GRUNSTEIN J S,et al. Mechanism regulating proasthmatic effects of prolonged homologous beta2-adrenergic receptor desensitization in airway smooth muscle[J]. Am J Physiol Lung Cell Mol Physiol,2009,297(4):L746-L757.
[7] SAI W B,YU M F,WEI M Y,et al. Bitter tastants induce relaxation of rat thoracic aorta precontracted with high K+[J].Clin Exp Pharmacol Physiol,2014,41(4):301-308.
[8] HE D,LEE L,YANG J,et al. Preventive effects and mechanisms of rhein on renal interstitial fibrosis in obstructive nephropathy[J]. Biol Pharm Bull,2011,34(8):1219-1226.
[9]何烨,马力扬,李志强,等.大黄酸对大鼠离体子宫平滑肌运动的影响[J].沈阳药科大学学报,2007,24(4):242-244.
[10] KARAKI H,OZAKI H,HORI M,et al. Calcium movements,distribution,and functions in smooth muscle[J]. Pharmacol Rev,1997,49(2):157-230.
[11]李晓红,李蒙,陶艳蓉.大黄酸及其衍生物药理作用研究新进展[J].现代药物与临床,2010,25(6):417-421.