胡椒碱对槟榔碱促进家兔离体小肠平滑肌运动的影响
|
摘要
目的观察胡椒碱对槟榔碱促进家兔离体小肠平滑肌运动的影响及其作用机制.方法采用离体平滑肌恒温灌流的方法,取家兔离体小肠,通过BL-420生物机能实验系统测定其张力的变化,观察不同浓度(0.06 g/L、0.6 g/L、6 g/L)胡椒碱溶液对正常状态下家兔离体小肠平滑肌自发性收缩的影响;先观察胡椒碱对家兔离体小肠平滑肌自发性收缩的影响,随后观察加入槟榔碱后对家兔离体小肠平滑肌自发性收缩的影响;为研究胡椒碱抑制家兔离体小肠平滑肌收缩的作用机制,应用IP3受体(inositol 1, 4, 5-trisphosphate, IP3)阻断剂肝素(Heparin,HP)、肌浆网ryanodine受体阻断剂钌红(ruthenium red,RR)和一氧化氮合酶抑制剂左旋硝基精氨酸甲酯(N'-nitro-L-arginine-methylesterhydrochloride,L-NAME),共同阐明胡椒碱对家兔离体小肠平滑肌作用的机制.结果胡椒碱抑制家兔离体小肠平滑肌自发性收缩,药物浓度在6 g/L时可显著抑制家兔离体小肠平滑肌收缩的振幅(P<0.01);在此基础上加入0.006 g/L槟榔碱溶液促进家兔离体小肠平滑肌收缩的振幅(P<0.05)且升高小肠收缩幅度小于单个槟榔碱促进家兔离体小肠平滑肌的收缩振幅(P<0.05); IP3受体HP能增强胡椒碱舒张家兔离体小肠平滑肌收缩的作用(P<0.05),而肌浆网ryanodine受体阻断剂钌红对胡椒碱舒张家兔小肠平滑肌的作用无明显影响(P>0.05); L-NAME能够部分阻断胡椒碱舒张家兔离体小肠平滑肌收缩的作用(P<0.05).结论胡椒碱可显著抑制家兔离体小肠平滑肌收缩的振幅;抑制单个槟榔碱促进家兔离体小肠平滑肌的收缩振幅;其机制可能与增加一氧化氮浓度,抑制IP3受体介导的内钙释放有关,但对肌浆网ryanodine受体途径引起的内钙释放无关.
AIM To investigate the effect of piperine on arecoline induced contraction of isolated small intestinal smooth muscle from rabbits and explore the possible mechanism involved. METHODS The method of ex vitro smooth muscle perfusion at a constant temperature was used to collect the rabbit small intestine in vitro. The effect of piperine solution at concentrations of 0.06 g/L, 0.6 g/L, and 6 g/L on the spontaneous contraction of isolated rabbit small intestinal smooth muscle was observed using the BL-420 biofunctional experiment system. Then, the effect of piperine and arecoline, alone or in combination, on spontaneous contraction of isolated small intestinal smooth muscle from rabbits was observed. To explore the mechanism by which piperine affected the contraction of isolated rabbit small intestinal smooth muscle, an inositol 1,4,5-trisphosphate(IP3) receptor antagonist(heparin, HP), a ryanodine receptor antagonist(ruthenium red, RR), and a nitric oxide synthase(NOS) inhibitor(nitro-Larginine methyl ester, L-NAME) were used. RESULTS Piperine inhibited the spontaneous contraction of isolated small intestinal smooth muscle from rabbits. At a concentration of 6 g/L, piperine showed a significant inhibitory effect on the amplitude of spontaneous contractions(P < 0.01). On this basis, arecoline solution at 0.006 g/L significantly increased the amplitude of contraction of isolated rabbit small intestinal smooth muscle(P < 0.05), but the amplitude of small intestinal smooth muscle contraction was smaller than that treated with arecoline alone(P < 0.05). IP3 receptor antagonist heparin could strengthen the relaxation effect of piperine on intestinal smooth muscle(P < 0.05),but ryanodine receptor antagonist ruthenium red had no effect on the relaxation effect of piperine(P > 0.05). L-NAME inhibited the relaxation effect of piperine(P < 0.05). CONCLUSION Piperine can inhibit the amplitude of spontaneous and arecoline induced contraction of rabbit intestinal smooth muscle. The mechanism may be related to the increase of NO concentration in intestinal smooth muscle and the inhibition of intracellular Ca~(2+)release via IP3 of sarcoplasmic reticulum.
AIM To investigate the effect of piperine on arecoline induced contraction of isolated small intestinal smooth muscle from rabbits and explore the possible mechanism involved. METHODS The method of ex vitro smooth muscle perfusion at a constant temperature was used to collect the rabbit small intestine in vitro. The effect of piperine solution at concentrations of 0.06 g/L, 0.6 g/L, and 6 g/L on the spontaneous contraction of isolated rabbit small intestinal smooth muscle was observed using the BL-420 biofunctional experiment system. Then, the effect of piperine and arecoline, alone or in combination, on spontaneous contraction of isolated small intestinal smooth muscle from rabbits was observed. To explore the mechanism by which piperine affected the contraction of isolated rabbit small intestinal smooth muscle, an inositol 1,4,5-trisphosphate(IP3) receptor antagonist(heparin, HP), a ryanodine receptor antagonist(ruthenium red, RR), and a nitric oxide synthase(NOS) inhibitor(nitro-Larginine methyl ester, L-NAME) were used. RESULTS Piperine inhibited the spontaneous contraction of isolated small intestinal smooth muscle from rabbits. At a concentration of 6 g/L, piperine showed a significant inhibitory effect on the amplitude of spontaneous contractions(P < 0.01). On this basis, arecoline solution at 0.006 g/L significantly increased the amplitude of contraction of isolated rabbit small intestinal smooth muscle(P < 0.05), but the amplitude of small intestinal smooth muscle contraction was smaller than that treated with arecoline alone(P < 0.05). IP3 receptor antagonist heparin could strengthen the relaxation effect of piperine on intestinal smooth muscle(P < 0.05),but ryanodine receptor antagonist ruthenium red had no effect on the relaxation effect of piperine(P > 0.05). L-NAME inhibited the relaxation effect of piperine(P < 0.05). CONCLUSION Piperine can inhibit the amplitude of spontaneous and arecoline induced contraction of rabbit intestinal smooth muscle. The mechanism may be related to the increase of NO concentration in intestinal smooth muscle and the inhibition of intracellular Ca~(2+)release via IP3 of sarcoplasmic reticulum.
引文
1 Mittal R,Gupta RL.In vitro antioxidant activity of piperine.Methods Find Exp Clin Pharmacol 2000;22:271-274[PMID:11031726]
2莫峥嵘,张岐.胡椒碱的抗氧化活性及稳定性研究.海南师范学院学报(自然科学版)2006;19:52-54[DOI:10.3969/j.issn.1674-4942.2006.01.016]
3裴印权,张仲一,李军,崔广智.胡椒碱对中枢神经系统功能的影响.中国药学杂志2003;38:268-270[DOI:10.3321/j.issn:1001-2494.2003.04.011]
4包兰兰,金桩,博·格日勒图.胡椒碱降血脂作用的实验研究.中国民族医药杂志2004;10:22-23[DOI:10.3969/j.issn.1006-6810.2004.01.016]
5刘屏,索婧侠,于腾飞.胡椒碱药理作用的研究进展.中国药物应用与监测2007;4:7-9[DOI:10.3969/j.issn.1672-8157.2007.03.003]
6杨阳,姚晓慧,周玉东,宋疆霞,买尼沙古丽·巴吾东,张晓红,余佳琳,赵红琼.辣椒、花椒和胡椒联合用药对小鼠离体小肠运动影响的研究.黑龙江畜牧兽医2016;13[DOI:10.13881/j.cnki.hljxmsy.2016.1211]
7陈潮燕.槟榔碱的提取分离及其对胃肠道平滑肌收缩作用的影响.广东药学2000;10:48-50[DOI:10.3969/j.issn.1674-229X.2000.02.026.]
8杜志敏,万新祥,伍爱婵,陈一岳.槟榔碱对离体肠自发性蠕动的影响.解放军广州医高专学报1998;2:98-99
9朱大年,王庭槐.生理学.8版.北京:人民卫生出版社2013:187-209
10王玢,左明雪.人体及动物生理学.2版.北京:高等教育出版2001:277-313
11吕锦芳,陈燕.不同处理因素对离体兔小肠平滑肌运动的影响.安徽科技学院学报2002;16:17-22[DOI:10.3969/j.issn.1673-8772.2002.01.007]
12张三印,董宇,杨鹏.胃肠运动障碍发病机制与证治.中医药学刊2003;21:209-211[DOI:10.3969/j.issn.1673-7717.2003.02.022]
13付立波,邵凯,景丽娜.白酒对家兔离体小肠平滑肌生理特性影响的研究.长春师范大学学报(自然科学版)2008;27:72-74[DOI:10.3969/j.issn.1008-178X-B.2008.04.022]
14付立波,付立山,张静菊,曹婧.丹参对家兔离体小肠平滑肌生理特性的影响.长春师范大学学报(自然科学版)2009;28:35-39[DOI:10.3969/j.issn.1008-178X-B.2009.03.011]
15 Bolton TB,Prestwich SA,Zholos AV,Gordienko DV.Excitation-contraction coupling in gastrointestinal and other smooth muscles.Annu Rev Physiol 1999;61:85-115[PMID:10099683 DOI:10.1146/annurev.physiol.61.1.85]
16 Bolton TB,Gordienko DV,PucovskyV,Parsons S,Povstyan O.Calcium release events in excitation-contraction coupling in smooth muscle.Novartis Found Symp 2002;246:154-168;discussion 168-173,221-227[PMID:12164307]
17 Mc Carron JG,Chalmers S,Bradley KN,Mac Millan D,Muir TC.Ca2+microdomains in smooth muscle.Cell Calcium 2006;40:461-493[PMID:17069885 DOI:10.1016/j.ceca.2006.08.010]
18 Frei E,Huster M,Smital P,Schlossmann J,Hofmann F,Wegener JW.Calcium-dependent and calcium-independent inhibition of contraction by cGMP/cGKI in intestinal smooth muscle.Am J Physiol Gastrointest Liver Physiol 2009;297:G834-G839[PMID:19628652 DOI:10.1152/ajpgi.00095.2009]
19 Hofmann F.The biology of cyclic GMP-dependent protein kinases.J Biol Chem 2005;280:1-4[PMID:15545263 DOI:10.1074/jbc.R400035200]
20 Murthy KS.Signaling for contraction and relaxation in smooth muscle of the gut.Annu Rev Physiol 2006;68:345-374[PMID:16460276 DOI:10.1146/annurev.physiol.68.040504.094707]
2莫峥嵘,张岐.胡椒碱的抗氧化活性及稳定性研究.海南师范学院学报(自然科学版)2006;19:52-54[DOI:10.3969/j.issn.1674-4942.2006.01.016]
3裴印权,张仲一,李军,崔广智.胡椒碱对中枢神经系统功能的影响.中国药学杂志2003;38:268-270[DOI:10.3321/j.issn:1001-2494.2003.04.011]
4包兰兰,金桩,博·格日勒图.胡椒碱降血脂作用的实验研究.中国民族医药杂志2004;10:22-23[DOI:10.3969/j.issn.1006-6810.2004.01.016]
5刘屏,索婧侠,于腾飞.胡椒碱药理作用的研究进展.中国药物应用与监测2007;4:7-9[DOI:10.3969/j.issn.1672-8157.2007.03.003]
6杨阳,姚晓慧,周玉东,宋疆霞,买尼沙古丽·巴吾东,张晓红,余佳琳,赵红琼.辣椒、花椒和胡椒联合用药对小鼠离体小肠运动影响的研究.黑龙江畜牧兽医2016;13[DOI:10.13881/j.cnki.hljxmsy.2016.1211]
7陈潮燕.槟榔碱的提取分离及其对胃肠道平滑肌收缩作用的影响.广东药学2000;10:48-50[DOI:10.3969/j.issn.1674-229X.2000.02.026.]
8杜志敏,万新祥,伍爱婵,陈一岳.槟榔碱对离体肠自发性蠕动的影响.解放军广州医高专学报1998;2:98-99
9朱大年,王庭槐.生理学.8版.北京:人民卫生出版社2013:187-209
10王玢,左明雪.人体及动物生理学.2版.北京:高等教育出版2001:277-313
11吕锦芳,陈燕.不同处理因素对离体兔小肠平滑肌运动的影响.安徽科技学院学报2002;16:17-22[DOI:10.3969/j.issn.1673-8772.2002.01.007]
12张三印,董宇,杨鹏.胃肠运动障碍发病机制与证治.中医药学刊2003;21:209-211[DOI:10.3969/j.issn.1673-7717.2003.02.022]
13付立波,邵凯,景丽娜.白酒对家兔离体小肠平滑肌生理特性影响的研究.长春师范大学学报(自然科学版)2008;27:72-74[DOI:10.3969/j.issn.1008-178X-B.2008.04.022]
14付立波,付立山,张静菊,曹婧.丹参对家兔离体小肠平滑肌生理特性的影响.长春师范大学学报(自然科学版)2009;28:35-39[DOI:10.3969/j.issn.1008-178X-B.2009.03.011]
15 Bolton TB,Prestwich SA,Zholos AV,Gordienko DV.Excitation-contraction coupling in gastrointestinal and other smooth muscles.Annu Rev Physiol 1999;61:85-115[PMID:10099683 DOI:10.1146/annurev.physiol.61.1.85]
16 Bolton TB,Gordienko DV,PucovskyV,Parsons S,Povstyan O.Calcium release events in excitation-contraction coupling in smooth muscle.Novartis Found Symp 2002;246:154-168;discussion 168-173,221-227[PMID:12164307]
17 Mc Carron JG,Chalmers S,Bradley KN,Mac Millan D,Muir TC.Ca2+microdomains in smooth muscle.Cell Calcium 2006;40:461-493[PMID:17069885 DOI:10.1016/j.ceca.2006.08.010]
18 Frei E,Huster M,Smital P,Schlossmann J,Hofmann F,Wegener JW.Calcium-dependent and calcium-independent inhibition of contraction by cGMP/cGKI in intestinal smooth muscle.Am J Physiol Gastrointest Liver Physiol 2009;297:G834-G839[PMID:19628652 DOI:10.1152/ajpgi.00095.2009]
19 Hofmann F.The biology of cyclic GMP-dependent protein kinases.J Biol Chem 2005;280:1-4[PMID:15545263 DOI:10.1074/jbc.R400035200]
20 Murthy KS.Signaling for contraction and relaxation in smooth muscle of the gut.Annu Rev Physiol 2006;68:345-374[PMID:16460276 DOI:10.1146/annurev.physiol.68.040504.094707]