夏膝颗粒降压作用及对SHR心肌细胞外基质影响的实验研究
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摘要
目的:
观察夏膝颗粒对自发性高血压大鼠血压及心肌细胞外基质的影响。
方法:
12周龄雄、雌性各25只SHR随机分为SHR空白组、缬沙坦组、夏膝颗粒高剂量组、夏膝中剂量组、夏膝低剂量组,每组10只,另取8只WKY大鼠作为正常对照组,给药8周观察各项指标。
结果:
1夏膝颗粒对SHR血压和心肌结构的影响
1.1对SHR行为和一般状态的影响
研究表明夏膝颗粒能改善SHR行为和一般状态,表明夏膝颗粒对SHR的一般生存状态有一定的调理作用。
1.2对SHR血压的影响
夏膝颗粒能抑制SHR血压随年龄增长而升高的趋势,给药4周到6周后与空白组比较即有显著性差异(P<0.05),且降压效果平稳持久而有效。
1.3对SHR左心室肥厚的影响
夏膝颗粒能降低SHR左心室重量指数(LVWI),与空白组比较有显著性差异(P<0.01),表明夏膝颗粒能在一定程度上逆转左心室肥厚(LVH)。对心脏的形态学观察:给药8周后,夏膝颗粒能明显减轻SHR心肌细胞肥大,减少细胞外基质的增生提示夏膝颗粒能减轻心脏损害。
2夏膝颗粒对SHR心肌细胞外基质的影响
2.1对血清Ⅰ型前胶原羧基端肽(PⅠCP)、血清III型前胶原氨基端肽(PⅢNP)浓度和血清血管紧张素Ⅱ(AngⅡ)的影响
采用酶联免疫吸附法(ELISA)测定。结果表明,SHR血清PICP、PⅢNP、AngⅡ浓度显著高于WKY对照组(P <0.01);给予不同剂量夏膝颗粒后调控PICP、PⅢNP、AngⅡ的水平亦有所不同,即阳性药组和夏膝颗粒高剂量组与WKY组比较无明显差异(P>0.05),但夏膝颗粒中、低剂量组与WKY对照组比较有显著性差异(P<0.05);阳性药组、夏膝高、中剂量组与SHR模型组比较有显著差异(P<0.01),在PICP检查中夏膝低剂量组与SHR比较未见明显差异,而高、中两组有差异。揭示I型胶原、III型胶原升高可能是SHR心室肥厚的物质基础之一,夏膝颗粒能通过降低I型胶原、III型胶原和AngⅡ浓度而发挥对SHR细胞外基质的调理作用,起到改善基质增生的作用。
2.2对血清MMP-1的影响
采用酶联免疫吸附法(ELISA)测定SHR血清MMP-1浓度。SHR血清MMP-1浓度显著低于WKY对照组(P<0.01);给予不同剂量夏膝颗粒后调控MMP-1的水平也存在不同变化,即阳性药组和夏膝颗粒高剂量组与WKY组比较无明显差异(P>0.05),但夏膝颗粒中、低剂量组与WKY对照组比较有显著性差异(P<0.05);阳性药组、夏膝高、中、低剂量组与SHR模型组比较都有显著差异(P<0.05)。
2.3对心肌TGF-β1表达的影响
用免疫组化法(ICH)检测心肌TGF-β1表达,用逆转录聚合酶链反应(RT-PCR)检测心肌TGF-β1基因表达的含量。结果表明夏膝颗粒对SHR具有降低心肌TGF-β1含量的作用,并降低心肌TGF-β1蛋白及基因表达。揭示夏膝颗粒可通过抑制转化生长因子,发挥调节功能而降低SHR的细胞外基质过度表达,恢复细胞外基质合成和降解的平衡,具有类似血管紧张素受体拮抗剂(ARB)的调控机制。
结论:
一、研究显示,调肝肾、解毒通络治法及其方药(夏膝颗粒)在高血压发病不同时段给药,有确切的延缓血压上升的作用,而且作用效果平稳、持久、缓和,以全程给药组作用最佳,从而部分揭示了这一临床组方思路的可行性与合理性。
二、夏膝颗粒能有效改善心肌细胞肥大和细胞外胶原组织的过度合成,提示该药能够逆转高血压左室肥厚现象,具有一定的心脏保护作用。
三、夏膝颗粒能够有效调控血清PICP、PⅢNP、MMP-1的动态平衡,间接反映了对心肌细胞外基质主要成分I、III型胶原蛋白的合成和降解的调控作用。并表明,血清PICP、PⅢNP很有可能成为以后临床作为判断心肌细胞外基质增生程度的判断指标。
四、夏膝颗粒能抑制AngII,TGF-β1蛋白的过度表达,提示其对细胞外基质增生的始动因子有抑制作用。
Purpose:
Effects of Xia xi Particles on blood pressure and extra cellular matrix of myocardial in spontaneously hypertensive rats.
Methods:
25 12-week-old male or female SHR were randomly divided into blank SHR group, valsartan group, the Xia Xi Particle high-dose group, middle dose group, low-dose groups of 10, and the other from 8 WKY rats as a normal control group, administration of 8 weeks to observe the following indicators.
Results:
1 Effects of Xia xi Particles on blood pressure and SHR cardiac function and structure
1.1 behavior and general condition effect:Xia xi Particles can improve the behavior and general state of SHR, That indicates that Xia Xi Particles has regulation function.
1.2 Decreasing blood effect: Xia xi Particles can inhibit the SHR blood pressure increased with age trends,4～6 weeks later compared with the blank group that there is significant difference (P <0.05), and antihypertensive effect of a smooth durable and effective.
1.3 Reversing left ventricular hypertrophy(LVH) effect: Xia xi Particles reduced SHR left ventricular mass index (LVWI), compared with blank control group were significantly different (P <0.01), can reverse LVH to a degree. Morphological observation of the heart: 8 weeks after delivery, Xia xi particles can significantly reduce the SHR myocardial hypertrophy, reduction of extracellular matrix hyperplasia prompted and can reduce heart damage.
2 Effects of Xia xi Particles on myocardial extracellular matrix
2.1 PICP、PⅢNP、AngII effect: Studying the density of PICP、PⅢNP、AngII in serum of SHR with Enzyme-Linked Immunosorbnent Assay (ELISA) . The results show that, SHR serum PICP, PⅢNP, AngⅡconcentration was significantly higher than WKY control group (P <0.01); Given different doses of Xia xi Particles control PICP, PⅢNP, AngⅡlevels are also different,positive medicine group and the Xia xi Particles high dose group compared with the WKY group was no significant difference (P > 0.05),but the medium dose , low-dose group compared with the WKY control group, there is significant difference (P <0.05), in PICP low dose group compared with no significant difference in SHR, while the other two groups have differences. Reveals the I-type collagen, III collagen may be increased SHR ventricular hypertrophy, one of the material basis. Xia xi Particles can reduce the I-type collagen, III collagen and AngⅡconcentration exert on the extracellular matrix SHR opsonization played the role of improving the stromal hyperplasia.
2.2 MMP-1 effect: Studying the density of MMP-1 in serum of SHR with Enzyme-Linked Immunosorbnent Assay (ELISA) . SHR serum MMP-1 concentration was significantly lower than WKY control group (P <0.01); Positive drug group and the Xia xi Particles high dose group compared with the WKY group was no significant difference (P > 0.05), but medium and low-dose group compared with the WKY control group, there is significant difference (P <0.05);
2.3 The influence of Xia Xi Particles to myocardial TGF-β1 expression:
By immunohistochemical method (ICH) Detect Myocardial myocardial expression of TGF-β1, by reversing transcription polymerase chain reaction (RT-PCR) detection of myocardial TGF-β1 gene expression levels. The results of Xia Xi Particles of SHR with reduced myocardial TGF-β1 content in the role, and reduce myocardial TGF-β1 protein and gene expression. Xia Xi Particles can be revealed by inhibiting transforming growth factor, exert systemic regulation of the function of the SHR to lower over-expression of extracellular matrix, with similar angiotensin receptor antagonist (ARB) of the lower blood pressure mechanism.
Conclusion:
一、The studies show that transfer of liver and kidney, Tongluo detoxification treatment and prescriptions (XiaXi particles) at different times of administration of hypertension, there is the exact role of slow rise in blood pressure and effect a smooth, durable, ease to the whole administration the role of the best groups in order to reveal some of the clinical group thought to the feasibility and reasonableness.
二、XiaXi particles Can effectively improve myocardial hypertrophy and extracellular excessive collagen synthesis, suggesting that the drug can reverse the situation left ventricular hypertrophy, the heart must have a protective effect.
三、XiaXi particles be able to effectively control serum PICP, PⅢNP, MMP-1 of the dynamic balance, and indirectly reflects the myocardial cells of the major components of extracellular matrix I, III collagen protein synthesis and degradation of the regulation. And show that serum PICP, PⅢNP is likely to be judged after clinical myocardial extracellular matrix to determine the extent of target hyperplasia.
四、XiaXi particles can inhibit AngII, TGF-β1 protein over-expression, suggesting that the proliferation of extracellular matrix has the only fixed factor inhibition
观察夏膝颗粒对自发性高血压大鼠血压及心肌细胞外基质的影响。
方法:
12周龄雄、雌性各25只SHR随机分为SHR空白组、缬沙坦组、夏膝颗粒高剂量组、夏膝中剂量组、夏膝低剂量组,每组10只,另取8只WKY大鼠作为正常对照组,给药8周观察各项指标。
结果:
1夏膝颗粒对SHR血压和心肌结构的影响
1.1对SHR行为和一般状态的影响
研究表明夏膝颗粒能改善SHR行为和一般状态,表明夏膝颗粒对SHR的一般生存状态有一定的调理作用。
1.2对SHR血压的影响
夏膝颗粒能抑制SHR血压随年龄增长而升高的趋势,给药4周到6周后与空白组比较即有显著性差异(P<0.05),且降压效果平稳持久而有效。
1.3对SHR左心室肥厚的影响
夏膝颗粒能降低SHR左心室重量指数(LVWI),与空白组比较有显著性差异(P<0.01),表明夏膝颗粒能在一定程度上逆转左心室肥厚(LVH)。对心脏的形态学观察:给药8周后,夏膝颗粒能明显减轻SHR心肌细胞肥大,减少细胞外基质的增生提示夏膝颗粒能减轻心脏损害。
2夏膝颗粒对SHR心肌细胞外基质的影响
2.1对血清Ⅰ型前胶原羧基端肽(PⅠCP)、血清III型前胶原氨基端肽(PⅢNP)浓度和血清血管紧张素Ⅱ(AngⅡ)的影响
采用酶联免疫吸附法(ELISA)测定。结果表明,SHR血清PICP、PⅢNP、AngⅡ浓度显著高于WKY对照组(P <0.01);给予不同剂量夏膝颗粒后调控PICP、PⅢNP、AngⅡ的水平亦有所不同,即阳性药组和夏膝颗粒高剂量组与WKY组比较无明显差异(P>0.05),但夏膝颗粒中、低剂量组与WKY对照组比较有显著性差异(P<0.05);阳性药组、夏膝高、中剂量组与SHR模型组比较有显著差异(P<0.01),在PICP检查中夏膝低剂量组与SHR比较未见明显差异,而高、中两组有差异。揭示I型胶原、III型胶原升高可能是SHR心室肥厚的物质基础之一,夏膝颗粒能通过降低I型胶原、III型胶原和AngⅡ浓度而发挥对SHR细胞外基质的调理作用,起到改善基质增生的作用。
2.2对血清MMP-1的影响
采用酶联免疫吸附法(ELISA)测定SHR血清MMP-1浓度。SHR血清MMP-1浓度显著低于WKY对照组(P<0.01);给予不同剂量夏膝颗粒后调控MMP-1的水平也存在不同变化,即阳性药组和夏膝颗粒高剂量组与WKY组比较无明显差异(P>0.05),但夏膝颗粒中、低剂量组与WKY对照组比较有显著性差异(P<0.05);阳性药组、夏膝高、中、低剂量组与SHR模型组比较都有显著差异(P<0.05)。
2.3对心肌TGF-β1表达的影响
用免疫组化法(ICH)检测心肌TGF-β1表达,用逆转录聚合酶链反应(RT-PCR)检测心肌TGF-β1基因表达的含量。结果表明夏膝颗粒对SHR具有降低心肌TGF-β1含量的作用,并降低心肌TGF-β1蛋白及基因表达。揭示夏膝颗粒可通过抑制转化生长因子,发挥调节功能而降低SHR的细胞外基质过度表达,恢复细胞外基质合成和降解的平衡,具有类似血管紧张素受体拮抗剂(ARB)的调控机制。
结论:
一、研究显示,调肝肾、解毒通络治法及其方药(夏膝颗粒)在高血压发病不同时段给药,有确切的延缓血压上升的作用,而且作用效果平稳、持久、缓和,以全程给药组作用最佳,从而部分揭示了这一临床组方思路的可行性与合理性。
二、夏膝颗粒能有效改善心肌细胞肥大和细胞外胶原组织的过度合成,提示该药能够逆转高血压左室肥厚现象,具有一定的心脏保护作用。
三、夏膝颗粒能够有效调控血清PICP、PⅢNP、MMP-1的动态平衡,间接反映了对心肌细胞外基质主要成分I、III型胶原蛋白的合成和降解的调控作用。并表明,血清PICP、PⅢNP很有可能成为以后临床作为判断心肌细胞外基质增生程度的判断指标。
四、夏膝颗粒能抑制AngII,TGF-β1蛋白的过度表达,提示其对细胞外基质增生的始动因子有抑制作用。
Purpose:
Effects of Xia xi Particles on blood pressure and extra cellular matrix of myocardial in spontaneously hypertensive rats.
Methods:
25 12-week-old male or female SHR were randomly divided into blank SHR group, valsartan group, the Xia Xi Particle high-dose group, middle dose group, low-dose groups of 10, and the other from 8 WKY rats as a normal control group, administration of 8 weeks to observe the following indicators.
Results:
1 Effects of Xia xi Particles on blood pressure and SHR cardiac function and structure
1.1 behavior and general condition effect:Xia xi Particles can improve the behavior and general state of SHR, That indicates that Xia Xi Particles has regulation function.
1.2 Decreasing blood effect: Xia xi Particles can inhibit the SHR blood pressure increased with age trends,4～6 weeks later compared with the blank group that there is significant difference (P <0.05), and antihypertensive effect of a smooth durable and effective.
1.3 Reversing left ventricular hypertrophy(LVH) effect: Xia xi Particles reduced SHR left ventricular mass index (LVWI), compared with blank control group were significantly different (P <0.01), can reverse LVH to a degree. Morphological observation of the heart: 8 weeks after delivery, Xia xi particles can significantly reduce the SHR myocardial hypertrophy, reduction of extracellular matrix hyperplasia prompted and can reduce heart damage.
2 Effects of Xia xi Particles on myocardial extracellular matrix
2.1 PICP、PⅢNP、AngII effect: Studying the density of PICP、PⅢNP、AngII in serum of SHR with Enzyme-Linked Immunosorbnent Assay (ELISA) . The results show that, SHR serum PICP, PⅢNP, AngⅡconcentration was significantly higher than WKY control group (P <0.01); Given different doses of Xia xi Particles control PICP, PⅢNP, AngⅡlevels are also different,positive medicine group and the Xia xi Particles high dose group compared with the WKY group was no significant difference (P > 0.05),but the medium dose , low-dose group compared with the WKY control group, there is significant difference (P <0.05), in PICP low dose group compared with no significant difference in SHR, while the other two groups have differences. Reveals the I-type collagen, III collagen may be increased SHR ventricular hypertrophy, one of the material basis. Xia xi Particles can reduce the I-type collagen, III collagen and AngⅡconcentration exert on the extracellular matrix SHR opsonization played the role of improving the stromal hyperplasia.
2.2 MMP-1 effect: Studying the density of MMP-1 in serum of SHR with Enzyme-Linked Immunosorbnent Assay (ELISA) . SHR serum MMP-1 concentration was significantly lower than WKY control group (P <0.01); Positive drug group and the Xia xi Particles high dose group compared with the WKY group was no significant difference (P > 0.05), but medium and low-dose group compared with the WKY control group, there is significant difference (P <0.05);
2.3 The influence of Xia Xi Particles to myocardial TGF-β1 expression:
By immunohistochemical method (ICH) Detect Myocardial myocardial expression of TGF-β1, by reversing transcription polymerase chain reaction (RT-PCR) detection of myocardial TGF-β1 gene expression levels. The results of Xia Xi Particles of SHR with reduced myocardial TGF-β1 content in the role, and reduce myocardial TGF-β1 protein and gene expression. Xia Xi Particles can be revealed by inhibiting transforming growth factor, exert systemic regulation of the function of the SHR to lower over-expression of extracellular matrix, with similar angiotensin receptor antagonist (ARB) of the lower blood pressure mechanism.
Conclusion:
一、The studies show that transfer of liver and kidney, Tongluo detoxification treatment and prescriptions (XiaXi particles) at different times of administration of hypertension, there is the exact role of slow rise in blood pressure and effect a smooth, durable, ease to the whole administration the role of the best groups in order to reveal some of the clinical group thought to the feasibility and reasonableness.
二、XiaXi particles Can effectively improve myocardial hypertrophy and extracellular excessive collagen synthesis, suggesting that the drug can reverse the situation left ventricular hypertrophy, the heart must have a protective effect.
三、XiaXi particles be able to effectively control serum PICP, PⅢNP, MMP-1 of the dynamic balance, and indirectly reflects the myocardial cells of the major components of extracellular matrix I, III collagen protein synthesis and degradation of the regulation. And show that serum PICP, PⅢNP is likely to be judged after clinical myocardial extracellular matrix to determine the extent of target hyperplasia.
四、XiaXi particles can inhibit AngII, TGF-β1 protein over-expression, suggesting that the proliferation of extracellular matrix has the only fixed factor inhibition
引文
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