黄连解毒汤对自发性高血压大鼠血管内皮因子及RhoA蛋白表达的影响（英文）
|
摘要
本文观察黄连解毒汤(HLJD)对自发性高血压大鼠(SHR)血管内皮的保护作用及胸主动脉中RhoA蛋白表达的影响,探讨黄连解毒汤对血管内皮的作用机制。将40只SHR大鼠随机分为模型组(SHR组)、阳性组(卡托普利组)、HLJD高剂量组、HLJD低剂量组,每组10只,同时空白组使用10只Wistar Kyoto大鼠,各组均给予灌胃6周。采用酶联免疫吸附法(ELISA)检测大鼠血清一氧化氮合酶(NOS)、假性血友病因子(vWF)、内皮素(ET-1)和钙调素(CAM)的变化,蛋白印记(WB)和real-time荧光定量(PCR)检测RhoA及其mR NA在胸主动脉中的表达。6周后,与空白组比较模型组动物血清中ET-l与VWF水平显著升高(P<0.01),NOS与CAM水平显著降低(P<0.01)。与模型组比较阳性组和HLJD各组中的ET-1与vW F水平降低(P<0.01或P<0.05),NOS与CAM水平提高(P<0.01或P<0.05)。HLJD高剂量组大鼠胸主动脉中RhoA的mR NA和蛋白表达均明显下降(P<0.05)。由此可得, HLJD提高大鼠血管内皮舒张因子(CAM和NOS),降低收缩因子(ET-1和vW F),能调节血管内皮(BVE)因子的分泌,改善血管内皮功能,揭示了早期高血压血管内皮功能障碍的预防作用,其机制可能与RhoA蛋白因子活性的抑制有关。
To investigate the mechanism of effect of Huanglian Jiedu Decoction(HLJD) on vascular endothelium, we assessed the protective effect of HLJD on vascular endothelium in spontaneously hypertensive rats(SHR) and the expression of RhoA in thoracic aorta. A total of 40 SHR rats were randomly and evenly divided into model group(SHR group), positive group(captopril group), HLJD high-dose group, and HLJD low-dose group. Simultaneously, 10 Wistar Kyoto rats were used in the blank group. All groups were treated by gavage for 6 weeks. The changes of nitric oxide synthase(NOS), von Willebrand factor(vWF), endothelin(ET-1) and calmodulin(CAM) in rat serum were tested by enzyme linked immunosorbent assay(ELISA) method. The expression of RhoA at the protein and mRNA levels in thoracic aorta was determined by Western blotting(WB) and quantitative real-time PCR, respectively. Compare with the blank group after 6 weeks, the levels of ET-l and VWF in serum of the model group were significantly increased(P<0.01), and the levels of NOS and CAM were significantly decreased(P<0.01). Conversely, the levels of ET-1 and vWF in the positive and HLJD groups were significantly decreased(P<0.01 or P<0.05), and the levels of NOS and CAM were significantly increased(P<0.01 or P<0.05) compared with the model group. The expression of Rho A at the mRNA and protein levels was decreased obviously(P<0.05) in HLJD high-dose group. The results shown that HLJD increased diastolic factor(CAM and NOS) in the vascular endothelial of rats, leading to reduced contraction factor(ET-1 and vWF). HLJD revealed the preventive function in the vascular endothelial dysfunction of the early stage hypertension through adjusting secretion of blood vessel endothelium(BVE) relaxing factor and improving vascular endothelial function. The mechanism might be associated with the inhibition of the activity of RhoA protein factor.
To investigate the mechanism of effect of Huanglian Jiedu Decoction(HLJD) on vascular endothelium, we assessed the protective effect of HLJD on vascular endothelium in spontaneously hypertensive rats(SHR) and the expression of RhoA in thoracic aorta. A total of 40 SHR rats were randomly and evenly divided into model group(SHR group), positive group(captopril group), HLJD high-dose group, and HLJD low-dose group. Simultaneously, 10 Wistar Kyoto rats were used in the blank group. All groups were treated by gavage for 6 weeks. The changes of nitric oxide synthase(NOS), von Willebrand factor(vWF), endothelin(ET-1) and calmodulin(CAM) in rat serum were tested by enzyme linked immunosorbent assay(ELISA) method. The expression of RhoA at the protein and mRNA levels in thoracic aorta was determined by Western blotting(WB) and quantitative real-time PCR, respectively. Compare with the blank group after 6 weeks, the levels of ET-l and VWF in serum of the model group were significantly increased(P<0.01), and the levels of NOS and CAM were significantly decreased(P<0.01). Conversely, the levels of ET-1 and vWF in the positive and HLJD groups were significantly decreased(P<0.01 or P<0.05), and the levels of NOS and CAM were significantly increased(P<0.01 or P<0.05) compared with the model group. The expression of Rho A at the mRNA and protein levels was decreased obviously(P<0.05) in HLJD high-dose group. The results shown that HLJD increased diastolic factor(CAM and NOS) in the vascular endothelial of rats, leading to reduced contraction factor(ET-1 and vWF). HLJD revealed the preventive function in the vascular endothelial dysfunction of the early stage hypertension through adjusting secretion of blood vessel endothelium(BVE) relaxing factor and improving vascular endothelial function. The mechanism might be associated with the inhibition of the activity of RhoA protein factor.
引文
[1]Taddei,S.;Ghiadoni,L.;Virdis,A.;Versari,D.;Salvetti,A.Clinical significance of the assessment of endothelial function.Ital.Heart J.Suppl.2004,5,357.
[2]Yue,G.H.;Zhuo,S.Y.;Xia,M.;Zhang,Z.;Gao,Y.W.Effect of Huanglian Jiedu Decoction on Thoracic Aorta Gene Expression in Spontaneous Hypertensive Rats.Res.Article.2014,11,1-9.
[3]Ridley,A.J.Rho family proteins:coordinating cell responses.Trends Cell Biol.2011,11,471-477.
[4]Rolfe,B.E.;Worth,N.F.;World,C.J.;Campbell,J.H.;Campbell,G.R.Rhoa and vascular disease.Atherosclerosis.2005,183,1-16.
[5]Zhu,H.X.;Pan,L.M.;Li,H.Chin.Tradit.Pat.Med.2010,32,1815-1818.
[6]Cheng,J.;Wan,L.H.;Fan,A.L.;Xiong,W.B.Study on the protective effect of Pearl Capsule on vascular endothelial cells in SHR.Pharmacol.Clin.Chin.Mater.Med.2009,25,68.
[7]Taddei,S.;Ghiadoni,L.;Virdis,A.;Versari,D.;Salvetti,A.Clinical significance of the assessment of endothelial function.Ital.Heart J.Suppl.2004,5,357.
[8]Ma,Q.;Liao,H.L.Advances in research of vascular remodeling in hypertension.Asia Pac.Tradit.Med.2016,12,58-59.
[9]Yue,G.H.;Mo,L.;Luo,Y.Effects of Yiqi Huoxue Jiedu Recipe on inflammatory factors and vascular von Willebrand factor levels in spontaneously hypertensive rats.J.Emerg.Tradit.Chin.Med.2010,19,1365-1367.
[10]Chu,Y.F.;Gao,P.J.Pathway and hypertension.Chin.J.Hypertension.2007,15,714-716.
[11]Berridge,M.J.Smooth muscle cell calcium activation mechanisms.J.Physiol.2008,586,047-5061.
[12]Lu,W.;Hu,K.;Yu,X.;Lan,X.Research progress of RhoA/ROCK pathway in pathogenesis of hypertension.China Med.Herald.2015,12,28-32.
[13]Friel,A.M.;Sexton,D.J.;O’reilly,N.W.RhoA/Rho kinase:human umbilical artery m RNA expression in normal and preeclamptic pregnancies and functional role in isoprost ane-induced vasoconstriction.Reproduction.2006,132,169-176.
[14]Yi,S.L.;Kant ores,C.;Belcast ro,R.;Cabacungan,J.;Tanswell,A.K.Jankov,R.P.8-Is op rost ane-induced endothelin-1 production by infant rat pulmonary artery smooth muscle cells is mediated by Rho-kinase.Free Radic.Biol.Med.2006,41,942-949.
[15]Watanabe,Y.;Faraci,F.M.;Heistad,D.D.Activation of Rho-associated kinase during augment edcont raction of the basilar artery to serotonin after subarachnoid hemorrhage.Am.J.Physiol.Heart Circ.Physiol.2005,288,2653-2658.
[16]Zhang,N.;Gao,Y.W.;Yue,G.H.;Zhang,Z.Effect of Huanglian Jiedu Decoction on ultrastructure of vascular structure in young spontaneously hypertensive rats.Lishizhen Med.J.Med.2013,24,2333-2335.
[17]Gao,Y.W.;Zhang,N.;Zhang,Z.;Yue,G.H.Effect of Huanglian Jiedu Decoction on the expression of endothelial nitric oxide synthase in aorta of young spontaneously hypertensive rats.Lishizhen Med.J.Med.2012,23,2684-2685.
[18]Yue,G.H.;Zhuo,S.Y.Effects of Yiqi Huoxue Jiedu Recipe on endothelial function in young spontaneously hypertensive rats.Chin.J.Exp.Tradit.Med.Form.2010,16,154-156.
[2]Yue,G.H.;Zhuo,S.Y.;Xia,M.;Zhang,Z.;Gao,Y.W.Effect of Huanglian Jiedu Decoction on Thoracic Aorta Gene Expression in Spontaneous Hypertensive Rats.Res.Article.2014,11,1-9.
[3]Ridley,A.J.Rho family proteins:coordinating cell responses.Trends Cell Biol.2011,11,471-477.
[4]Rolfe,B.E.;Worth,N.F.;World,C.J.;Campbell,J.H.;Campbell,G.R.Rhoa and vascular disease.Atherosclerosis.2005,183,1-16.
[5]Zhu,H.X.;Pan,L.M.;Li,H.Chin.Tradit.Pat.Med.2010,32,1815-1818.
[6]Cheng,J.;Wan,L.H.;Fan,A.L.;Xiong,W.B.Study on the protective effect of Pearl Capsule on vascular endothelial cells in SHR.Pharmacol.Clin.Chin.Mater.Med.2009,25,68.
[7]Taddei,S.;Ghiadoni,L.;Virdis,A.;Versari,D.;Salvetti,A.Clinical significance of the assessment of endothelial function.Ital.Heart J.Suppl.2004,5,357.
[8]Ma,Q.;Liao,H.L.Advances in research of vascular remodeling in hypertension.Asia Pac.Tradit.Med.2016,12,58-59.
[9]Yue,G.H.;Mo,L.;Luo,Y.Effects of Yiqi Huoxue Jiedu Recipe on inflammatory factors and vascular von Willebrand factor levels in spontaneously hypertensive rats.J.Emerg.Tradit.Chin.Med.2010,19,1365-1367.
[10]Chu,Y.F.;Gao,P.J.Pathway and hypertension.Chin.J.Hypertension.2007,15,714-716.
[11]Berridge,M.J.Smooth muscle cell calcium activation mechanisms.J.Physiol.2008,586,047-5061.
[12]Lu,W.;Hu,K.;Yu,X.;Lan,X.Research progress of RhoA/ROCK pathway in pathogenesis of hypertension.China Med.Herald.2015,12,28-32.
[13]Friel,A.M.;Sexton,D.J.;O’reilly,N.W.RhoA/Rho kinase:human umbilical artery m RNA expression in normal and preeclamptic pregnancies and functional role in isoprost ane-induced vasoconstriction.Reproduction.2006,132,169-176.
[14]Yi,S.L.;Kant ores,C.;Belcast ro,R.;Cabacungan,J.;Tanswell,A.K.Jankov,R.P.8-Is op rost ane-induced endothelin-1 production by infant rat pulmonary artery smooth muscle cells is mediated by Rho-kinase.Free Radic.Biol.Med.2006,41,942-949.
[15]Watanabe,Y.;Faraci,F.M.;Heistad,D.D.Activation of Rho-associated kinase during augment edcont raction of the basilar artery to serotonin after subarachnoid hemorrhage.Am.J.Physiol.Heart Circ.Physiol.2005,288,2653-2658.
[16]Zhang,N.;Gao,Y.W.;Yue,G.H.;Zhang,Z.Effect of Huanglian Jiedu Decoction on ultrastructure of vascular structure in young spontaneously hypertensive rats.Lishizhen Med.J.Med.2013,24,2333-2335.
[17]Gao,Y.W.;Zhang,N.;Zhang,Z.;Yue,G.H.Effect of Huanglian Jiedu Decoction on the expression of endothelial nitric oxide synthase in aorta of young spontaneously hypertensive rats.Lishizhen Med.J.Med.2012,23,2684-2685.
[18]Yue,G.H.;Zhuo,S.Y.Effects of Yiqi Huoxue Jiedu Recipe on endothelial function in young spontaneously hypertensive rats.Chin.J.Exp.Tradit.Med.Form.2010,16,154-156.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |