The Experimental, Clinical Study And Effect On Microvascular Rarefaction In Hypertension With Modified ZhenXuanFang

In China, with the improvement of living standards and lifestyle changes, the incidence of hypertension increased steadily, the 2002 National Nutrition and Health Survey data show that adult prevalence rate was 18.8%, there are about 160 million patients with hypertension. As the most common chronic disease, hypertension is one of most important basis disease led to stroke, coronary heart disease and renal failure and other diseases.Although Western medicine could lower blood pressure quickly, efficacy and the advantages of good compliance, but the side effects of antihypertensive drugs was more frequent, many patients had to give up treatment. Chinese medicine treatment of hypertension has unique advantages, such as blood pressure stable and durable, while alleviating symptoms, less side effects. Professor Chen Baotian according to the "dizzy all the Feng, all belonging to the liver," "do not dizzy without Tan " and related theories of Chinese medicine, combined with decades of clinical experience, create Modified ZhenXuanFang for the treatment of hypertension, and had a good effect. If rational development and wide clinical application, it will help complement the disadvantage of western medicine in decreasing blood pressure stablely and alleviating symptoms in patients with hypertension, reducing blood pressure medication side effects, improve health quality of nearly 200 million people hypertensive patients in chinathe pathophysiology of microcirculation has paly a more important role in hypertension. Reduction in microvessel density, not only led to decreased blood parallel pathway, and lead to increased peripheral vascular resistance, thereby promoting the continued increase in arterial blood pressure, reduced microvessel density, but also enables organizations to the level of physical energy metabolism, storage capacity and material exchange capacity decreased, leading to target organ damage at the same time. The main purpose of this study:first, to explore effect of Modified ZhenXuanFang on blood pressure in experimental and clinical research,senong, to explore effects of Modified ZhenXuanFang on microvascular system and other mechanism.ChapterⅠThe study of the effect of Modified ZhenXuanFang on blood pressure of spontaneously hypertensive ratsObjective To explore the effect of Modified ZhenXuanFang on blood pressure of spontaneously hypertensive ratsMethods 50 male SHRs were randomly assigned to 5 groups namely the low-dose,the middle-dose and high-dose Modified ZhenXuanFang group,the captopril group and SHR control group, Blood pressure and heart rate were recorded from the tail at the beginning and every 2 weeks during 12 weeks treatment. At the end of the experiment, animals were sacrificed.the plasma AngⅡconcentration were measured.Rusults Test of between-subjects effect in decreasing blood pressure indicate: significant differences were seen in the differences groups of SHR (F=54.557,P= 0.000),When compared with control group, the low-dose groups (P=0.001),the middle-dose (P=0.000) and high-dose (P=0.000) Modified ZhenXuanFang group,the captopril group (P=0.000) all show significant differences in decreasing blood pressure of spontaneously hypertensive rats.When compared with high-dose Modified ZhenXuanFang group, the low-dose groups show significantly higher blood pressure (P=0.029), the captopril group show significantly lower blood pressure (P =0.001),and no significant differences were seen in middle-dose group (P= 0.249).Significant differences were seen in differences time (F= 209.221,P=0.000).There were no significant differences in blood pressure at the beginning (F=0.850,P=0.501) and significant differences were seen in blood pressure in differences group on second week (F=59.646,P=0.000),fourth week (F=56.970,P=0.000),sixth week (F=45.251,P=0.000),eighth week (F= 45.546,P=0.000),tenth week (F=57.358,P=0.000),twelfth week (F= 75.150,P=0.000).When compared with control group, the low-dose (second week P =0.089,fifth week P=0.002,sixth week P=0.000,eighth week P=0.000,ten week P=0.000,twelfth week P=0.000),the middle-dose (second week P=0.001,fifth week P=0.000,sixth week P=0.000,eighth week P=0.000,ten week P=0.000,twelfth week P=0.000),high-dose (second week P=0.000,fifth week P=0.000,sixth week P=0.000,eighth week P=0.000,ten week P=0.000,twelfth week P=0.000) Modified ZhenXuanFang group and the captopril group (second week P=0.000,fifth week P=0.000,sixth week P=0.000,eighth week P=0.000,ten week P=0.000,twelfth week P=0.000) all show significant in lowering blood pressure.The high-dose group show the best effect in reduction in blood pressure and show significant differences compared with the low-dose (P=0.000) and middle-dose groups (P=0.001) and no significant differences compared with captoril group (P=0.020)in decreasing blood pressure in 12 weeks. the captopril group were more significant in reduction in blood pressure at other times (second week P= 0.000,fifth week P=0.000,sixth week P=0.000,eighth week P=0.000,ten week P=0.000).Ttraditional Chinese medicine group could steady decrease blood pressure in the experiment,and captoril group could quickly decrease blood pressure. Time and group show significant crossover effect (F=25.646,P=0.000)Test of between-subjects effect in decreasing blood pressure indicate:significant differences were seen in the differences groups of SHR(F=11.594,P=0.000); When compared with control group, the low-dose groups (P=0.004),the middle-dose (P =0.000) and high-dose (P=0.000) Modified ZhenXuanFang group all show significant differences in decreasing heart rate of spontaneously hypertensive rats. no significant differences was seen in captoril group(P=0.297). When compared with high-dose Modified ZhenXuanFang group, the low-dose group (P=0.209) and middle-dose group (P=0.596) show no significant differences in heart rate of spontaneously hypertensive rats.Significant differences were seen in differenent time (F=40.907,P=0.000).There were no significant differences in heart rate at the beginning (F= 0.981,P=0.427) and significant differences were seen in heart rate in differences group on second week (F=3.911,P=0.008).fourth week (F=7.220,P=0.000),sixth week (F=8.949,P=0.000),eighth week (F=9.179,P=0.000),tenth week (F= 12.621,P=0.000),twelfth week (F=30.622,P=0.000).When compared with control group, the low-dose (second week P=0.045,fifth week P=0.006,sixth week P=0.005,eighth week P=0.000,ten week P=0.000,twelfth week P=0.000),the middle-dose (second week P=0.002,fifth week P=0.001,sixth week P=0.000,eighth week P=0.000,ten week P=0.000,twelfth week P=0.000),high-dose (second week P =0.002,fifth week P=0.000,sixth week P=0.000,eighth week P=0.000,ten week P=0.000,twelfth week P=0.000) Modified ZhenXuanFang group all show significant in decreasing heart rate. the captopril group show no significant differences (second week P=0.068,fifth week P=0.343,sixth week P=0.448,eighth week P=0.210,ten week P=0.416,twelfth week P=0.358) at any time in the experiment.Time and group show significant crossover effect (F=7.449,P=0.000) Test of between-subjects effect in plasma AngⅡconcentration indicate: significant differences were seen in differenent in different group (F= 7.047,P=0.000), When compared with control group, the low-dose (P=0.015), middle-dose (P=0.006), high-dose (P=0.000) Modified ZhenXuanFang group and captopril group (P=0.000) all show significant differences in plasma AngⅡconcentration of spontaneously hypertensive rats. When compared with high-dose Modified ZhenXuanFang group, the low-dose group (P=0.142),the middle-dose (P=0.257) and the captopril group (P=0.359) show no significant differences in decreasing plasma AngⅡconcentration of spontaneously hypertensive ratsConclusion Modified zhenxuan granules could decrease blood pressure of spontaneously hypertensive rats, perhaps by decreasing heart rate and plasma AngⅡconcentration.ChapterⅡThe study of the effect of Modified ZhenXuanFang on endothelial function in spontaneously hypertensive ratsObjective To explore the effect of Modified ZhenXuanFang on blood pressure of spontaneously hypertensive rats By measuring noninvasively endothelial function in the auricular skin microcirculation by monitoring with a laser-Doppler system the blood flow changes induced by warming of the skin and biosynthesis and bioavailability of NO.Methods 50 male SHRs were randomly assigned to 5 groups namely the low-dose,the middle-dose and high-dose groups Modified zhenxuan fang group,the captopril group and SHR control group.At the end of the experiment, animals were anesthetized and measured auricular skin blood flow changes by Laser Doppler system before and after local warming.Then animals were sacrificed. The plasma NO concentration,arteriae aorta nitric oxide synthase (NOS) activity and arteriae aorta cyclic guanylic acid (cGMP) concentration were measured. Rusults The baseline auricular skin blood flow perfusion units show no Significant differences in the differences groups of SHR (F=0.633,P=0.641).The basis of blood flow as a covariate, using analysis of covariance, we find auricular skin blood flow perfusion units were significantly different in each group (F=55.40,P=0.000). Compared with the control group, the low-dose (P=0.000),the middle-dose (P=0.000), high-dose group (P=0.000) Modified zhenxuan fang group and the captopril group (P=0.000) could improve auricular skin blood flow perfusion units of SHR. high-dose group could improve auricular skin blood flow perfusion units compared with other groups (P=0.000).captopril and low dose group show no significant difference in auricular skin blood perfusion units (P= 0.450).the rate of change of the auricular microvascular perfusion units differences (χ2=39.692, P=0.000) before and after local warming, the control group increased blood flow perfusion units at least, followed by captopril group, low dose group, middle dose group and high dose group.The plasma NO (F=3.400, P=0.016), aortic NOS activity (F=2.892, P= 0.033) and aortic cGMP content (F=8.195, P=0.000) were significantly different.Compared with the control group, Modified ZhenXuanFang low dose group SHR plasma NO (P=0.058), aortic NOS activity (P=0.070) had no significant difference, Modified ZhenXuanFang middle dose group (P=0.007), high dose group (P=0.006) and captopril group (P=0.002) were able to significantly increase the plasma NO concentration in SHR,modified ZhenXuanFang middle dose (P=0.022), high dose group (P=0.009) and captopril group (P=0.004) were able to significantly increase aortic NOS activity. Compared with the high dose group, modified Zhenxuan low-dose (P=0.254), middle dose group (P=0.958) and captopril group (P=0.731) showed no significant difference in plasma NO, modified Zhenxuan low-dose group (P=0.393), middle dose group (P=0.734), captopril group (P= 0.737) show no significant difference in aortic NOS activity.Compared with the control group, modified Zhenxuan low-dose group (P= 0.000), middle dose group(P=0.000), high dose group (P=0.000) and captopril group (P=0.009) could increase aortic cGMP concentration; Compared with Modified ZhenXuanFang high dose group, low dose group (P=0.306), middle dose group (P=0.905) show no significant differences in aortic cGMP, the captopril group was lower(P=0.035) in Aortic cGMP concentration,Conclusion Modified ZhenXuanFang could affecte NO/NOS/cGMP biosynthesis and bioavailability of SHR, increase the SHR rat auricular microvascular perfusion to improve endothelial function.ChapterⅢThe study of modified ZhenXuanFang on SHR myocardial matrix metalloproteinase-2 and left ventricular remodelingObjective:To explore the effect of Modified ZhenXuanFang on left ventricular remodeling and possible mechanisms by observing spontaneously hypertensive rats left ventricular mass, left ventricular mass index, left ventricular collagen concentration, myocardial expression of MMP-2 after treatment.Methods 50 male SHRs were randomly assigned to 5 groups namely the low-dose,the middle-dose and high-dose Modified ZhenXuanFang group,the captopril group and SHR control group, At the end of the experiment, animals were sacrificed. electronic balance to take the quality of left ventricular and ventricular septal left ventricular mass, left ventricular mass index (LVMD=left ventricular mass (g)/ body weight (kg). Weighed 50mg of left ventricular tissue to detect hydroxyproline, other left ventricular tissue was washed in 4℃saline,dried, weighed, and 4% paraformaldehyde solution fixed and paraffin embedded and detected myocardial MMP-2 protein by immunohistochemistry.Results:Weight:animals'weight differ among groups (F=3.548, P=0.013), compared with captopril group, Modified ZhenXuanFang low dose group (P= 0.003),middle dose group (P=0.034), high dose group (P=0.004) and control group (P=0.004) weight heavier, compared with Modified ZhenXuanFang high dose group, low dose group (P=0.904), middle dose group (P=0.401), control group (P=1.000) weight show no significant difference.Left ventricular mass:Left ventricular mass differ among the groups (F= 10.868, P=0.000); compared with the control group, modified ZhenXuanFang low dose group (P=0.001), middle dose group (P=0.000), high dose group (P=0.000), captopril group (P=0.000) decreased left ventricular mass significantly; Compared with modified ZhenXuanFang high dose group, low dose group (P=0.013) left ventricular mass increased, the middle dose group (P=0.237), captopril group (P= 0.082) left ventricular mass show no signifcant differences.Left ventricular mass index:the group left ventricular mass index differ among the groups (F=9.550, P=0.000); compared with the control group, modified ZhenXuanFang low dose group (P=0.001), middle dose group (P=0.000), high dose group (P=0.000), captopril group (P=0.001) significantly reduced left ventricular mass index; compared with modified ZhenXuanFang high dose group, low dose group (P=0.019), captopril group (P=0.017) left ventricular mass index increase,middle dose group (P=0.198) show no significant difference in left ventricular mass index.Myocardial expression of MMP-2 protein were significantly different among the groups (F=170.854, P=0.000), compared with the control group, modified ZhenXuanFang low dose group (P=0.037), middle group (P=0.000), high dose group (P=0.000), captopril group (P=0.000) were able to reduce the myocardial expression of MMP-2 protein; and compared with the high dose group, low dose group (P=0.000), middle dose group (P=0.000), captopril group (P=0.000) myocardial MMP-2 protein expression were up regulation.Left ventricular myocardial collagen concentration were significantly different (F=25.535, P=0.000), compared with the control group, modified ZhenXuanFang low dose group (P=0.000), middle dose group(P=0.000), high dose group (P= 0.000), captopril group (P=0.000) left ventricular collagen concentration decreased. Compared with modified ZhenXuanFang low dose group, low dose group (P= 0.012), captopril group (P=0.004) increase in the concentration of collagen, the middle dose group show no significant difference (P=0.285)Left ventricular mass was significantly correlated with end blood pressure (r= 0.645, P=0.000),left ventricular mass index was significantly correlated with end blood pressure (r=0.580, P=0.000).Conclusion:Modified ZhenXuanFang could reduce the expression of matrix metalloproteinase-2, lower left ventricular mass, reduced left ventricular mass index and left ventricular collagen concentrationChapterⅣThe study of modified ZhenXuanFang on microvascular rarefaction in heart,brain and kidney of SHRObjective To explore Modified ZhenXuanFang on microvessel density in hypertension and its possible mechanism by observing the Modified ZhenXuanFang group, captopril group, spontaneously hypertensive rats control group myocardial, cerebral and renal microvessel density.Methods 50 male SHRs were randomly assigned to 5 groups namely the low-dose,the middle-dose and high-dose Modified ZhenXuanFang group,the captopril group and SHR control group, At the end of the experiment, animals were sacrificed, left ventricular tissue was washed in 4℃saline,dried, weighed, and 4% paraformaldehyde solution fixed and paraffin embedded, At the same time separating the brain, kidney, respectively, after weighing, cut along the coronal plane, and immediately put into 4% paraformaldehyde solution fixed and paraffin-embedded sections,CD34 immunohistochemical staining to detect myocardial, cerebral, renal MVDResults Myocardial microvessel density in each group were significantly different (F=5.212, P=0.002). Compared with the control group, the low dose fang group (P=0.029), middle dose group (P=0.024), high dose Modified zhenxuan group (P=0.000), captopril group (P=0.032) were able to improve the SHR cerebral microvessel density. Compared with high-dose Modified ZhenXuanFang group,low dose group (P=0.026), middle dose group (P=0.031), captopril group (P=0.023) SHR cerebral microvessel density show no significant differenceCerebral microvessel density in each group were significantly different (F=22.793,P=0.000). Compared with the control group, the low dose group (P= 0.000), middle dose group(P=0.000), high dose Modified ZhenXuanFang group (P= 0.000), captopril group (P=0.007) were able to improve the cerebral microvessel density. Compared with high-dose Modified ZhenXuanFang group, low dose group (P=0.030), captopril group (P=0.000) cerebral microvessel density decreased,the middle dose group show no significant difference (P=0.080).Peritubular microvessel density in each group were significantly different (F= 26.820, P=0.000). Compared with the control group, the low dose group (P=0.000), middle dose group (P=0.000),high dose group (P=0.000), captopril group (P= 0.000) were able to improve the SHR Peritubular microvessel density. Compared with high-dose Modified ZhenXuanFang group, low dose group (P=0.003), captopril group (P=0.001) Peritubular microvessel density decreased, the middle dose group show no significant difference (P=0.093).Glomerular microvessel density in each group were significantly different (F =29.766, P=0.000). Compared with the control group, the low dose group (P= 0.000), middle dose group (P=0.000),high dose group (P=0.000), captopril group (P=0.000) were able to improve the SHR glomerular microvessel density. Compared with high-dose Modified ZhenXuanFang group, low dose group (P=0.003), captopril group (P=0.001) SHR glomerular microvessel density decreased, the middle dose group show no significant difference (P=0.076).Peritubular and glomerular microvessel density all correlated with end blood pressure, respectively (r=-0.427, P=0.003) and (r=-0.500, P=0.000), myocardial (r=-0.172, P=0.247), cerebral tissue (r=-0.255, P=0.084) microvessel density was not related with end blood pressure.Conclusion All dose Modified ZhenXuanFang group can improve the main target organs cardiac, cerebral, renal microvessel density, high-dose group improve cerebral microvascular best, middle and high dose group improve myocardial, peritubular, glomerular microvessel density best. Modified ZhenXuanFang was superior to captopril in improving microvessel density. glomerular and peritubular microvessel density correlated with end blood pressure.Clinical part The study of the effect of Modified ZhenXuanFang on essential hypertensionObjective To explore the clinical effectiveness of lowering blood pressure and safety of Modified ZhenXuanFang by study the effect of Modified ZhenXuanFang on essential hypertension and TCM Symptom scores.Methods All patients from the Henan hypertension hospital (Henan Academy of Traditional Chinese Medicine Hospital) out-patient department and in-patient department, either sex, aged 18-65 years old, a total of 90 cases. Test line with clinical trials of medical ethics, all patients enter clinical trials after informed consent.All the hypertensive patients should be consistent with low-risk or middle-risk group(1-2) diagnosis and ganhuokangsheng or tanshiyongshen of Chinese medicine diagnosis.90 cases were randomly divided into treatment group and captopril group,45 cases of Chinese medicine treatment group and 45 cases of captopril group, treatment for 4 weeks. Medicines for the Treatment of Modified ZhenXuanFang, the control group for captopril tablets (12.5mg,.b.i.d) for 4 weeks. The blood pressure, the symptom score and the safety index were measured before and after treatment. the gender (χ2=3.455,P=0.063), age (t=0.429, P=0.669)and Course of disease (t=-1.157, P=0.251), systolic blood pressure (Z=-0.543, P= 0.587), diastolic blood pressure (Z=-1.091, P=0.275),TCM syndrome score (Z=-0.734, P=0.463)show no significant difference in two groups before treatment.Results Systolic blood pressure were significantly different at different times (F=369.678, P=0.000), systolic blood pressure change between the two groups was not significantly different (F=0.132, P=0.719), Time and group show no significant crossover effect (F=1.120, P=0.297). Diastolic blood pressure were significantly different at different times (F=50.230, P=0.000), diastolic blood pressure change between the two groups was not significantly different (F=4.010, P=0.053). Time and group show no significant crossover effect (F=0.005, P=0.944).TCM syndrome score in different time points were significantly different (F= 368.841, P=0.000), TCM syndrome score between the two groups were significantly different (F=10.462,P=0.003). Time and group show significant crossover effect (F =59.588, P=0.000). Antihypertensive effect in two groups show no significant difference (Z=-0.759, P=-0.448). Treatment group was superior to the control group in relieving syndromes (Z=-5.147, P=0.000). Safety evaluation:two patients laboratory indexes were safe before and after trail, the captopril group removed 1 case because of erythra,6 cases of loss due to cough,7 cases were removed because of stopping to take chinese medicine in Modified ZhenXuanFang.Conclusion Modified ZhenXuanFang and captopril (12.5mg/d) could lower blood pressure, the two groups showed no significant difference in lower systolic and diastolic blood pressure. Two medicine could relieving syndromes both, modified ZhenXuanFang was superior to captopril in relieving syndromes...