| Purposes:This study is meant to observe the influence of qishenjianpifang on the SHR about the caudal artery’s systolic pressure and the content of NO, PGI2, EDHF, AngⅡ, ET-1by which we can value its antihypertensive effect and also investigate the vessals endodermis vasomotor releasing factors’anti-influence on the prescription. Meanwhile, we can even inspect this prescription’s repairing situation towards the vessal endothelium and pursue the amelioration system of the endothelial dysfunction during the process of hypertension.At the same time, we will also discuss how.the qishenjianpifang regulates the expression of kidney Kallikrein and influences the content of signaling molecule which are in the SHR ACE2-Ang(1-7)-Mas-AKT thoroughfare, and the mutual action on the adjustment of vessals endodermis vasomotor releasing process will also be included. Besides, we will further do research about how the qishenjianpifang influences SHR thoracic aorta collagen Ⅰ, Ⅲ, and the expression of TGF-β1, meanwhile, we will also explore the reconstruction system of the vessals during the process of hypertension. All these experiments have offered the scientific basis and theories for the use of qishenjianpifang of the clinical treatment and also partially explain its molecule process of protecting the vessals.Materials and methods:Using the randomized controlled method to divide the6024-week-old SHR into the model group (giving water, SHR group), perindopril group [perindopril,0.4mg/(kg·d)], perindopril associated with herb group [perindopril0.4mg/(kg·d)+medium dose of herb], and another3groups individually high level, middle level and low level dose of herb (the dose is decided according to the100g weight which are4g,2g, and1g) and the normal group is10normal arterial pressure WKY at the same age and the same kind (giving pure water, WKY). The rats are fed once a day. We will detect the systolic pressure’s change of their caudal artery with BP-98A intelligent sphygmomanometer before and4hours,2weeks, and6weeks after giving them the medicine, and the measuring time is4hours after the medicine giving. It will last6weeks.The rats will be fed only water and after24hours, these groups of rats will be weighed after given10%chloral hydrate’s anesthetic. We will fix the rats on the desk after anesthetic and sterilize their belly skin with0.5%Iodophor, then we will cut off their belly skin to make the abdominal aorta seeable, we draw off the blood and drop gelation inhibitor into it, and let the blood stand2hours with the temperature of4℃, and then off-centre it with3000rpm. After all these steps, we will draw off the upper class jasmine plasma and keep it under the experiment of-20℃temperature. We will detect the dose of plasma endodermis vasomotor and releasing factors and Ang(1-7). We will widen the thoracic cavity along the abdominal cavity to make the aorta thoracica seeable, and then ligate it about1cm at the both ends of the aorta theoracica to make sure that there is blood in it. Next we will cut off the vessals at the ligature and put it into4%paraformal-dehyde to carry on the observation of pathology images and at the same time detect the expression of SHR thoracic aorta collagen Ⅰ, Ⅲ and TGF-β1,We will take other vascular tissue about50mg to put it in the EP canal in the refrigerator under the temperature of-70℃, and the Western method will be used to detect the expression of the vessal TGF-β1, To cut off the abdominal cavity to make the kidney seeable, and take about50mg tissue of the left kidney and also put it into the EP canal combined with1mL Trizol in the refrigerator under the temperature of-70℃gene of the kidney; We will also take another50mg tissue which will be kept and dealt with in the same way to detect the expression of AKT with the Western way. The data will be checked with one factor analysis of variance using the SPSS16.0software, and LSD method will be used to make comparison in the way of (x±s), if P<0.05, it will have significant difference. Results:1. The influence of qishenjianpifang on the SHR about the caudal artery’s systolic pressureThe statistical results explanation:before giving the medicine, the systolic pressure of these groups of rats are all higher than the group WKY (P<0.01);4hours after giving them the medicine, there is no obvious change about the systolic pressure; After2weeks, the pressure is obvious getting down for the perindopril group and the perindopril associated with herb group, and comparing with the model group, it has the statistical meaning (P<0.01); after4weeks and6weeks, the pressire is continuously getting down and it has the statistical meaning comparing with the model group (P<0.01). While for the3level dose of herb groups, the pressure is also getting down at some extent after2weeks of using the medicine, however, after4and6weeks, the pressure is not getting down clearly, and comparing with the model group, it has no statistical meaning.2. The influence of qishenjianpifang on the dose of SHR Vessals Endodermis Vasomotor and Releasing FactorsThe statistical results explanation:The content of NO in SHR group、PGI2、 EDHF is prominently lower than the WKY group; the dose of NO of the perindopril group, the perindopril associated with herb group and the high and middle level of the herb groups is clearly higher than the SHR group (P<0.05); For the PGI2the perindopril group, the perindopril associated with herb group and the high level of the herb groups are higher than the SHR group (P<0.05); For the EDHF, the perindopril associated with herb group and the high and middle level of the herb groups is clearly higher than the SHR group (P<0.05); while for the dose of releasing factors, the the perindopril associated with herb group and the perindopril group has no statistical meaning. The dose of AngⅡand ET-1are prominently higher than the WKY group (P<0.05); The dose of AngⅡand ET-1of the perindopril group, the perindopril associated with herb group and the high, middle and low level of the herb groups are also much lower than the SHR group (P<0.01); For the AngⅡ, the dose of WHK, the perindopril associated with herb group and the high level of the herb groups have no satatistical meaning comparing with perindopril group, and for the dose of ET-1, the dose of WHK, the perindopril associated with herb group and the three levels of the herb groups have no statistical meaning comparing with perindopril group.3. The influence of the qishenjianpifang on the dose of signaling molecule which are in the SHR ACE2-Ang(1-7)-Mas-AKT thoroughfare.The content of Ang(1-7) of SHR group is clearly lower than that of WKY group(P<0.01); The content of the perindopril group, the perindopril associated with herb group and the high, middle and low levels of the herb groups is clearly higher than the SHR group (F<0.05). The content of Ang (1-7) of the3levels groups of the herb is lower than the perindopril group (P<0.05). Comparing with the WKY group, the kidney ACE2, Mas and Kallikrein mRNA’s expression quantity of the SHR group is obviously getting down; the kidney ACE2, Mas and Kallikrein mRNA’s expression quantity of the perindopril associated with herb group and the perindopril group is much higher than those of the SHR group (P<0.01, P<0.05); the Mas mRNA’s expression quantity of the high level and middle level groups of the herb is much higher than those of the SHR group (P<0.01); the ACE2and Kallikrein mRNA’s expression quantity of the high level of the herb is much higher than those of the SHR group (P<0.05); comparing with the SHR group, the kidney AKT protein expression is decreasing at a great extent(P<0.01); while for the perindopril associated with herb group, the high level of herb group and the perindopril group, the expression of the AKT protein is prominently higher than that of the SHR group (P<0.01, P<0.05), while for the middle and low level of the herb groups, the expression has no statistical meaning comparing with the SHR group.4. The influence of qishenjianpi fang on the SHR thoracic aortic wall morphologyFor the aorta thoracica vessal endangium of the WKY group, the inner cells didn’t drop off, and the blood corpuscle also didn’t adhere to the endangium. The cells rank compactly and there is no cells hyperplasia, and the vascular wall isn’t getting thicker. For the SHR group, the endothelial cells of the aorta thoracica are partially dropping off, and the intima isn’t smooth enough, and there is also blood corpuscle compacting. The elastic fiber of the middle membrane becomes straight, declined, and ruptured and the smooth muscle cells are also becoming hypertrophied; the ectoblast cells hypertrophies actively, and the karyon reaches higher density, and the vessal inner wall is getting thicker. While for the Chinese associated western medicine group, the rats aorta thoracica the inner cells didn’t drop off and the blood corpuscle also didn’t adhere to the endangium and the smooth muscle cells aren’t becoming hypertrophied either, and the elastic fiber is much in the order than that of the SHR group. After observing the vessal pathology forms, we get rather similar results between the high level of herb group and the perindopril associated with herb group. For the perindopril group, the middle and low level of herb group, the aorta thoracica endothelial cells are dropping off sometimes, while, comparing with the SHR group, the elastic fiber of the middle membrane can be called in a good order, and the smooth muscle cells’hypertrophy is controlled5. The influence of qishenjianpifang on the SHR aorta thoracica collagen Ⅰ, Ⅲ and the expression of TGF-β,The expression of collagen Ⅰ, Ⅲ and TGF-β1of the SHR group is much more than that of the WKY group (P<0.01); while for the perindopril group, the perindopril associated with herb group and the high, middle and low levels of the herb groups, it becomes less than that of the SHR group (P<0.01). The difference of the collagen Ⅰ, Ⅲ and TGF-0, expression between the perindopril associated with herb group and the WKY group has no statistical meaning. For the SHR group, the expression of the TGF-β1, is much more than that of the WKY group (P<0.01); while for the perindopril group, the perindopril associated with herb group and the high, middle and low levels of the herb groups, it is much less than that of the SHR group (P<0.01) Conclusions:1. The middle level dose of the qishenjianpifang combined with perindopril shows the better lowing pressure effect on the caudal artery systolic pressure, after4weeks’treatment, it will reach the stable level; it has the similar treating effect with the perindopril group.2. By regulating the releasing balance of NO、PGI2、EDHF、AngⅡand ET-1of the vessals endodermis vasomotor and relaxing factors, the qishenjianpifang can modify the vessals’function obstacle which is caused by the high blood pressure.3. The qishenjianpi fang combining with perindopril and high level dose of folk prescription can stimulate the expression of the signal molecule which are in the ACE2-Ang(1-7)-Mas-AKT thoroughfare. At the same time, it also increases the expression quantity of the kallikrein gene so as to regulate the releasing and secreting of the vessals endodermis vasomotor and relaxing factors to exert its function of lowing blood pressure and protecting the vessal endodermis.4. The qishenjianpi fang combining with perindopril and high level dose of folk prescription can improve SHR vessal pathology images conformation. By inhibiting the expression of collagen Ⅰ, Ⅲ and TGF-β1, so as to reverse the reconstruction of the vessal. |
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