夏膝口服液对SHR儿茶酚胺、粘附分子及RAS影响的实验研究
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摘要
高血压病是是以体循环动脉压升高为主要表现的一种疾病。可以造成心、脑、肾等重要脏器的严重损害。本文从文献综述、理论研究、实验研究方面对夏膝口服液治疗高血压病进行较系统的研究。该方以调整肝肾,通络解毒为原则而设立,提出“毒伤血络”是高血压病的病机关键,在治疗高血压病方面具有创新性。在实验研究中探讨中药具有ACEI及ARB样降压作用,并通过中药对粘附分子的影响揭示毒伤血络的病理基础,尚未见报导,有较强的创新性和实用性。
1文献综述
对中医药治疗高血压病从病因病机、辨证分型、单味中药、中成药及辨证论治方面进行了系统的总结,并对目前辨证分型、疗效评定标准和科研等方面存在的问题进行了评述和展望。
从现代医学的角度对高血压病的诊断、发病机理,治疗现状及近年来中医中药防治高血压病机理的实验研究成果、研究动态进行了综述。
2理论研究运用中医理论引经据典,对高血压病的病名、病位、病因病机、识毒要点、毒与络关系等进行了探讨,提出肝肾阴阳失调,毒伤血络的病机理论,论述了调整肝肾,通络解毒是治疗高血压病的有效途径。
3实验研究
3.1夏膝口服液对SHR降压及心血管保护作用
采用SHR为高血压模型,设WKY对照组、空白对照组、卡托普利组、厄贝沙坦组和中药实验组(夏膝口服液),实验观察各项指标,结果表明,夏膝口服液能明显改善SHR的行为和一般状态,降低SHR血压,与空白组比较在给药4周后有显著性统计学意义(P<0.05),给药8周后有极显著性差异(P<0.01);能够使LVW/BW平均降低了24. 05% (P <0. 0l);能减轻心肌细胞的肥大,保护血管内皮,减轻中膜平滑肌细胞的增殖。提示夏膝口服液对SHR具有良好的降压及保护心血管的作用。
3.2夏膝口服液对SHR血清儿茶酚胺影响的实验研究本实验用酶联免疫法(ELISA)测定SHR血清儿茶酚胺水平,结果表明,SHR血清儿茶酚胺水平显著高于WKY对照组(P<0.01);给予夏膝口服液灌胃20天即可明显降低大鼠血清儿茶酚胺水平(P<0.05),至第40天降至正常水平,与SHR空白组比较有显著性差异(P<0.01),并稳定维持至第60天。说明夏膝口服液能降低SHR儿茶酚胺水平。揭示儿茶酚胺水平增高可能是SHR兴奋、肝阳上亢及血压升高的内在基础。夏膝口服液通过降低SHR儿茶酚胺水平从而降低其交感神经活性,起到降低血压及改善大鼠行为状态的作用。
3.3夏膝口服液对SHR血浆sICAM-1浓度及肾小球ICAM-1表达的影响本实验采用ELISA法及免疫组化法分别检测各组动物血浆sICAM-1浓度及肾小球ICAM-1蛋白的表达。结果显示,夏膝口服液能降低血浆sICAM-1浓度。及小球内膜ICAM-1蛋白表达。揭示①粘附分子的高表达所引起的内皮损伤可能是毒伤血络的内在病理基础之一。②夏膝口服液的通络解毒作用可能是通过降低粘附分子的表达,保护血管内皮而实现。
3.4夏膝口服液对SHR循环及局部RAS的影响
采用放免法、免疫组化法及逆转录聚合酶链反应(RT-PCR)方法检测各组动物血浆肾素、血管紧张素Ⅱ、血清ACE活性及主动脉AT1受体蛋白及mRNA表达。结果表明,夏膝口服液能降低血浆肾素、AngⅡ含量及ACE活性,同时降低SHR主动脉AT1R蛋白含量及mRNA表达。具有类似ACEI及ARB样的降压作用。
Essential hypertension is a disease which represented mainly by the rising blood pressure of systemic circulation, it can cause grave damage to heart, brain, kidney and other important viscera. This essay systemic study the treatment of essential hypertension with xiaxi oral liquid from reference review, theory study and experimental study. This prescription is established by the principle of modulating liver and kidney, and putting forward that superficial vessel’s damnification by toxin is the crux of essential hypertension. It has innovation in Hypertension Therapy. Through the experimental study of Chinese medicine’s effect of reducing pressure as ACEI and ARB, superficial vessel’s damnification by toxin basis of pathology is explained by study of the impact of the Chinese medicine to cell adhesion molecules. this has never been reported before, so this essay is practical and innovative.
1 Reference review
This essay systematically summarizes the treatment of essential hypertension with TCM as follows: the etiology and pathogenesis, syndrome differentiation with differentiation of symptoms and signs, Single Chinese Medical Herb, Traditional Chinese Patent Medicines , different treatment with differentiation of symptoms and signs. Some problems such as syndrome differentiation, criterion for the evaluation of therapeutic effect and scientific research were reviewed and prospected.This essay also summarized the experimental study results and advances on the principle of preventing and treating in EH according to the dialectic, pathologenesis, treatment, experimental study and advances.
2 Theory study
By discussing the name, localization,pathogenicfactors and pathogenesis, the important point of knowing poison and the relationship between poison and superficial venules and lymph vessels of EH with Chinese medicine theories quoting from classics works and combining modern research results,we put forward the theory that the main mechanism of EH is the maladjustment of liver and kidney and damnification of superficial venules and lymph vessels,and the valid path of treat EH is the way of adjustment of liver and kidney and removing toxicity and obstruction.
3 Experimental study
3.1 The antihypertension and cardiovascular protective effect of Xiaxi oral liquid in SHR
40 12-week-old male SHR were divided randomly into blank control group ,captopril control group,Irbesartanl control group ,and Xiaxi oral liquid control group and 10 wister-kyoto rats were set as normal control(WKY group).The various markets were observed.The results showed that Xiaxi oral liquid obviously improve the SHR’s behavior and general condition and had a certain effect on lowered blood pressure and extremely significant statistic sense after dministration 8 weeks(P<0.01). Xiaxi oral liquid can reduce the ratio of LVW to BW by 24.05%(P<0.01). It can also alleviate the hypertrophy of cadiocyte, protecte blood vessel endothelium and reduce the proliferation of media vascular smooth muscle cells.The result showed that Xiaxi oral liquid has the function of lowering blood pressure and protecting to cardiovascular system in SHR.
3.2 The experimental study of effect of Xiaxi oral liquid on SHR serum catecholamine (CA)
Studying the density of CA in serum of SHR with Enzyme-Linked Immunosorbnent Assay (ELISA) .The results showed that the density of serum CA in SHR is obviously higher than that in WKY rats(P<0.01). Xiaxi oral liquid can reduce the level of serum CA in SHR ,which had a significant statistic sense in comparing with blank control group after administration 20 days (P<0.05),and can reduce the level to normal standard after 40 days (P<0.01) , the tendency lasting for 60 days. The result showed that Xiaxi oral liquid has the function of reducing the density of CA of serum of SHR,and indicate that the increasing catecholamine standard may be the inner basis for excitement,hyperactive liver-yang ascending and hypertension in SHR. Xiaxi oral liquid can depress the activity of sympathetic nerve, lower blood pressure and improve behavior and general condition through lowering the density of serum CA in SHR.
3.3 The experimental study of the influence of Xiaxi oral liquid on solubol intercellular adhesion molecule-1 (sICAM-1) density in plasma and intercellular adhesion molecule-1 (ICAM-1) expression in kidney arteriole in SHR
Studying the density of sICAM-1 in plasma with ELISA and ICAM-1 expression in kidney arteriole with immunohistochemistery(IMH) . The results showed that Xiaxi oral liquid can reduce sICAM-1 density in plasma and ICAM-1 expression in kidney arteriole in SHR. The results revealed that the injury of endothelial cell induced by higher expression of ICAM-1may be one of the pathology basis about damnification of superficial venules and lymph vessels by poison.And Xiaxi oral liquid can alleviate the injury of endothelial cell through reduce the expression of ICAM-1 in SHR so that educe the function of removing toxicity and obstruction.
3.4 The study of effects of Xiaxi oral liquid on recirculating and local ennin-angiotensin system(RAS) in SHR
Detecting the plasma renin activity(PRA) and plasma angiotensinⅡ(AngⅡ) content with radio-immunity, serum Angiotensin-Converting enzyme (ACE)activity with spectrophotometer, angiotensinⅡreceptor type 1(AT1R) expression in abdominal aorta with immunohistochemistery(ICH) and the gene expressive content of AT1R in thoracic aorta with Reverse Transcription -polymerasechain reaction(RT-PCR) in rats. The results showed that Xiaxi oral liquid can reduce PRA , AngⅡcontent, ACE activity, AT1R expression in abdominal aorta and the gene expressive content of AT1R in thoracic aorta.It revealed that Xiaxi oral liquid has the function of lower blood pressure as Angiotensin-Converting enzyme inhibitor(ACEI) and angiotensin receptor block(ARB).
1文献综述
对中医药治疗高血压病从病因病机、辨证分型、单味中药、中成药及辨证论治方面进行了系统的总结,并对目前辨证分型、疗效评定标准和科研等方面存在的问题进行了评述和展望。
从现代医学的角度对高血压病的诊断、发病机理,治疗现状及近年来中医中药防治高血压病机理的实验研究成果、研究动态进行了综述。
2理论研究运用中医理论引经据典,对高血压病的病名、病位、病因病机、识毒要点、毒与络关系等进行了探讨,提出肝肾阴阳失调,毒伤血络的病机理论,论述了调整肝肾,通络解毒是治疗高血压病的有效途径。
3实验研究
3.1夏膝口服液对SHR降压及心血管保护作用
采用SHR为高血压模型,设WKY对照组、空白对照组、卡托普利组、厄贝沙坦组和中药实验组(夏膝口服液),实验观察各项指标,结果表明,夏膝口服液能明显改善SHR的行为和一般状态,降低SHR血压,与空白组比较在给药4周后有显著性统计学意义(P<0.05),给药8周后有极显著性差异(P<0.01);能够使LVW/BW平均降低了24. 05% (P <0. 0l);能减轻心肌细胞的肥大,保护血管内皮,减轻中膜平滑肌细胞的增殖。提示夏膝口服液对SHR具有良好的降压及保护心血管的作用。
3.2夏膝口服液对SHR血清儿茶酚胺影响的实验研究本实验用酶联免疫法(ELISA)测定SHR血清儿茶酚胺水平,结果表明,SHR血清儿茶酚胺水平显著高于WKY对照组(P<0.01);给予夏膝口服液灌胃20天即可明显降低大鼠血清儿茶酚胺水平(P<0.05),至第40天降至正常水平,与SHR空白组比较有显著性差异(P<0.01),并稳定维持至第60天。说明夏膝口服液能降低SHR儿茶酚胺水平。揭示儿茶酚胺水平增高可能是SHR兴奋、肝阳上亢及血压升高的内在基础。夏膝口服液通过降低SHR儿茶酚胺水平从而降低其交感神经活性,起到降低血压及改善大鼠行为状态的作用。
3.3夏膝口服液对SHR血浆sICAM-1浓度及肾小球ICAM-1表达的影响本实验采用ELISA法及免疫组化法分别检测各组动物血浆sICAM-1浓度及肾小球ICAM-1蛋白的表达。结果显示,夏膝口服液能降低血浆sICAM-1浓度。及小球内膜ICAM-1蛋白表达。揭示①粘附分子的高表达所引起的内皮损伤可能是毒伤血络的内在病理基础之一。②夏膝口服液的通络解毒作用可能是通过降低粘附分子的表达,保护血管内皮而实现。
3.4夏膝口服液对SHR循环及局部RAS的影响
采用放免法、免疫组化法及逆转录聚合酶链反应(RT-PCR)方法检测各组动物血浆肾素、血管紧张素Ⅱ、血清ACE活性及主动脉AT1受体蛋白及mRNA表达。结果表明,夏膝口服液能降低血浆肾素、AngⅡ含量及ACE活性,同时降低SHR主动脉AT1R蛋白含量及mRNA表达。具有类似ACEI及ARB样的降压作用。
Essential hypertension is a disease which represented mainly by the rising blood pressure of systemic circulation, it can cause grave damage to heart, brain, kidney and other important viscera. This essay systemic study the treatment of essential hypertension with xiaxi oral liquid from reference review, theory study and experimental study. This prescription is established by the principle of modulating liver and kidney, and putting forward that superficial vessel’s damnification by toxin is the crux of essential hypertension. It has innovation in Hypertension Therapy. Through the experimental study of Chinese medicine’s effect of reducing pressure as ACEI and ARB, superficial vessel’s damnification by toxin basis of pathology is explained by study of the impact of the Chinese medicine to cell adhesion molecules. this has never been reported before, so this essay is practical and innovative.
1 Reference review
This essay systematically summarizes the treatment of essential hypertension with TCM as follows: the etiology and pathogenesis, syndrome differentiation with differentiation of symptoms and signs, Single Chinese Medical Herb, Traditional Chinese Patent Medicines , different treatment with differentiation of symptoms and signs. Some problems such as syndrome differentiation, criterion for the evaluation of therapeutic effect and scientific research were reviewed and prospected.This essay also summarized the experimental study results and advances on the principle of preventing and treating in EH according to the dialectic, pathologenesis, treatment, experimental study and advances.
2 Theory study
By discussing the name, localization,pathogenicfactors and pathogenesis, the important point of knowing poison and the relationship between poison and superficial venules and lymph vessels of EH with Chinese medicine theories quoting from classics works and combining modern research results,we put forward the theory that the main mechanism of EH is the maladjustment of liver and kidney and damnification of superficial venules and lymph vessels,and the valid path of treat EH is the way of adjustment of liver and kidney and removing toxicity and obstruction.
3 Experimental study
3.1 The antihypertension and cardiovascular protective effect of Xiaxi oral liquid in SHR
40 12-week-old male SHR were divided randomly into blank control group ,captopril control group,Irbesartanl control group ,and Xiaxi oral liquid control group and 10 wister-kyoto rats were set as normal control(WKY group).The various markets were observed.The results showed that Xiaxi oral liquid obviously improve the SHR’s behavior and general condition and had a certain effect on lowered blood pressure and extremely significant statistic sense after dministration 8 weeks(P<0.01). Xiaxi oral liquid can reduce the ratio of LVW to BW by 24.05%(P<0.01). It can also alleviate the hypertrophy of cadiocyte, protecte blood vessel endothelium and reduce the proliferation of media vascular smooth muscle cells.The result showed that Xiaxi oral liquid has the function of lowering blood pressure and protecting to cardiovascular system in SHR.
3.2 The experimental study of effect of Xiaxi oral liquid on SHR serum catecholamine (CA)
Studying the density of CA in serum of SHR with Enzyme-Linked Immunosorbnent Assay (ELISA) .The results showed that the density of serum CA in SHR is obviously higher than that in WKY rats(P<0.01). Xiaxi oral liquid can reduce the level of serum CA in SHR ,which had a significant statistic sense in comparing with blank control group after administration 20 days (P<0.05),and can reduce the level to normal standard after 40 days (P<0.01) , the tendency lasting for 60 days. The result showed that Xiaxi oral liquid has the function of reducing the density of CA of serum of SHR,and indicate that the increasing catecholamine standard may be the inner basis for excitement,hyperactive liver-yang ascending and hypertension in SHR. Xiaxi oral liquid can depress the activity of sympathetic nerve, lower blood pressure and improve behavior and general condition through lowering the density of serum CA in SHR.
3.3 The experimental study of the influence of Xiaxi oral liquid on solubol intercellular adhesion molecule-1 (sICAM-1) density in plasma and intercellular adhesion molecule-1 (ICAM-1) expression in kidney arteriole in SHR
Studying the density of sICAM-1 in plasma with ELISA and ICAM-1 expression in kidney arteriole with immunohistochemistery(IMH) . The results showed that Xiaxi oral liquid can reduce sICAM-1 density in plasma and ICAM-1 expression in kidney arteriole in SHR. The results revealed that the injury of endothelial cell induced by higher expression of ICAM-1may be one of the pathology basis about damnification of superficial venules and lymph vessels by poison.And Xiaxi oral liquid can alleviate the injury of endothelial cell through reduce the expression of ICAM-1 in SHR so that educe the function of removing toxicity and obstruction.
3.4 The study of effects of Xiaxi oral liquid on recirculating and local ennin-angiotensin system(RAS) in SHR
Detecting the plasma renin activity(PRA) and plasma angiotensinⅡ(AngⅡ) content with radio-immunity, serum Angiotensin-Converting enzyme (ACE)activity with spectrophotometer, angiotensinⅡreceptor type 1(AT1R) expression in abdominal aorta with immunohistochemistery(ICH) and the gene expressive content of AT1R in thoracic aorta with Reverse Transcription -polymerasechain reaction(RT-PCR) in rats. The results showed that Xiaxi oral liquid can reduce PRA , AngⅡcontent, ACE activity, AT1R expression in abdominal aorta and the gene expressive content of AT1R in thoracic aorta.It revealed that Xiaxi oral liquid has the function of lower blood pressure as Angiotensin-Converting enzyme inhibitor(ACEI) and angiotensin receptor block(ARB).
引文
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