黄连解毒汤对幼龄自发性高血压大鼠血管内皮NO通路影响的研究

Effects of Huanglianjiedu Decoction on vascular endothelial NO pathway in young spontaneously hypertensive rats

目的:研究黄连解毒汤(HLJD)对自发性高血压大鼠(SHR)血管内皮保护作用及该药物的作用机制。方法:40只6周龄雄性SHR大鼠随机分为模型组(SHR组)、阳性组(卡托普利组)、黄连解毒汤高剂量组、黄连解毒汤低剂量组每组10只,空白组10只(正常WKY大鼠),灌胃6周并用无创尾套法测量其尾动脉收缩压及舒张压;酶联免疫吸附(ELISA)法测定大鼠血清中一氧化氮(NO)、超氧物歧化酶(SOD)、血管紧张素II(AngII)、钙调素(CaM)含量变化;实时荧光定量PCR(qRT-PCR)、免疫印迹技术(Western blot)测定胸主动脉中Ras同源基因家族成员A(RhoA)、环鸟苷酸依赖性蛋白激酶(cGK)、肌球蛋白结合亚单位(MYPT1)mRNA与蛋白表达情况。结果:给药6周后,与模型组相比,黄连解毒汤各剂量组和阳性组均能显著地降低大鼠的血压。与空白组比较,模型组动物血清中AngII与SOD水平显著升高(P<0.01),NO与CaM水平显著降低(P<0.01);与模型组比较,阳性组和黄连解毒汤各组中的AngII与SOD水平降低(P<0.01或P<0.05),NO与CaM水平提高(P<0.01或P<0.05)。黄连解毒汤能够降低高血压大鼠胸主动脉RhoA、MYPT1的表达(P<0.05),并能提高高血压大鼠胸主动脉cGK的表达(P<0.05)。结论:黄连解毒汤能降低高血压大鼠的血压,调节血管内皮舒缩因子的分泌,并可影响cGK,RhoA,MYPT1等因子的表达,而这些因子的表达与NO的活性密切相关。

AIM:To study the protective effect of Huanglianjiedu Decoction(HLJD)on vascular endothelial endothelium in spontaneously hypertensive rats(SHR)and the mechanism of action of the drug.METHODS:A total of 40 SHR rats were randomly divided into model group(SHR group),positive group(Captopril group),HLJD high dose group,and HLJD low dose group with 10 rats in each group.Simultaneously,there was 10 Wistar rats in control group.All groups were treated by gavage for 6 weeks.Systolic and diastolic blood pressure of the caudal artery was measured by noninvasive tail sleeve method.The changes of nitric oxide(NO),superoxide dismutase(SOD),angiotensin(AngII)and calmodulin(CaM)in rat serum were tested by enzyme linked immunosorbent assay(ELISA)method;the expression of RhoA,cGK,MYPT1 protein and mRNA in thoracic aorta were determined by Western blot and real-time fluorescence quantitative(PCR).RESULTS:After 6 weeks,compared with the model group,HLJD and positive group can significantly reduce the blood pressure of rats.Compare with blank group,the levels of AngII and SOD in serum of the model group were significantly increased(P<0.01),and the levels of NO and CaM were significantly lower(P<0.01).Conversely,the levels of AngII and SOD in the positive and HLJD groups were significantly decreased(P<0.01 or P<0.05),and the levels of NO and CaM were significantly increased(P<0.01 or P<0.05).HLJD could reduce the expression of RhoA and MYPT1 in the thoracic aorta of hypertension rats(P<0.05),and increase the expression of cGK in the thoracic aorta of hypertensive rats(P<0.05).CONCLUSION:HLJD can reduce the blood pressure of hypertensive rats,regulate the secretion of vascular endothelial vasopressin and affect the expression of factors such as cGK,RhoA,MYPT1.The expression of these factors is closely related to the activity of NO.