Effect Of Zhenganxifeng Decoction On Vascular Remodeling In Spontaneously Hypertensive Rats And Its Mechanism

Purpose:By using experimental techniques such as behavioral science, histomorphology andmolecular biology, the study observes Zhen-gan Xi-feng Soup Decoction’s influence onprotein and mRNA expression of PPAR, MAPK-P38, PCNA Raf-1and Bcl-2inspontaneously hypertensive rats vessel. The thesis aims to investigate Zhen-gan Xi-feng SoupDecoction’s influence on vascular remodeling in spontaneously hypertensive rats and itsmechanism, clarifying Zhen-gan Xi-feng Soup Decoction’s pharmacological mechanisms inspontaneously hypertension, as well as providing experimental evidence for the classicprescription.Materials and Methods:Fifty15-week-old male SHR rats were randomly divided into five groups: model group(saline), Traditional Chinese Medicine (TCM) low-dose group (Zhen-gan Xi-feng SoupDecoction), TCM middle-dose group (Zhen-gan Xi-feng Soup Decoction), TCM high-dosegroup (Zhen-gan Xi-feng Soup Decoction), and Compound Apocynum group. Ten15-week-old homologous male WKY rats were used as the normal control group (saline). Those ratswere given adaptive feeding for five weeks, and then to measure their tail arterial pressurebefore the experiment in order to determine whether SHR hereditary hypertensive rats fall ill.At the beginning of the experiment, TCM group was given Zhen-gan Xi-feng Soup Decoctionby gavage, model group and normal control group were given normal saline by gavage, andcompound Apocynum group was administered compound Apocynum by gavage. Gastricperfusion lasted for10weeks.1. After ten weeks of administration, we measured the rats’ tail arterial pressure in themorning for continuous five days using the method of non-invasive blood pressure byBP-98A machine.2. To sample blood from the rats’ abdominal aorta, put them into procoagulant separatortube to coagulate and centrifuge, and then to measure cholesterol (CHOL), triglycerides (TG),low density lipoprotein (LDL) in serum.3. To take the aortic arch and4%paraformaldehyde fixed, and perform histomorphometry using hematoxylin-eosin (HE) color; to determine the aortic arch apoptoticchanges with TUNEL method.4. To take the aortic arch, and measure PPAR GAMMA, MAPK-P38, PCNA, Raf-1,Bcl-2protein expression changes by Western-blotting, and measure PPAR GAMMA mRNA,MAPK-P38mRNA, PCNA mRNA, Raf-1mRNA, Bcl-2mRNA expression with Real TimeRT-PCR.Results:1. Compared with model group, each group’s mean arterial blood pressure (MABP)reduced significantly(P <0.01); compared with normal group, MABP of TCM middle-dosegroup, TCM high-dose group and Compound Apocynum group showed no significant change(P>0.01); compared with normal group, MABP of TCM low-dose group rised significantly(P<0.01).2. Compared with model group, each group’s cholesterol (CHOL), triglycerides (TG),low density lipoprotein (LDL) decreased significantly (P<0.01); compared with normal group,cholesterol, triglycerides, and low density lipoprotein of TCM middle-dose group, TCMhigh-dose group and Compound Apocynum group decreased significantly (P<0.05);compared with normal group, cholesterol, triglycerides, and low density lipoprotein of TCMlow-dose group showed no significant change (P>0.05).3. Aortic arch HE staining showed that compared with model group, the TCM group’svascular wall thickness, muscle fibers and vascular smooth muscle cell proliferationdecreased.4. Compared with model group, the number of vascular smooth muscle cell of aorticarch apoptosis decreased significantly in each group (P<0.01); compared with TCMhigh-dose group and Compound Apocynum group, the number of vascular smooth musclecell of aortic arch apoptosis in TCM middle-dose group increased significantly(P <0.01);compared with Compound Apocynum group, the number of vascular smooth muscle cell ofaortic arch apoptosis in TCM high-dose group showed no significant change (P>0.05).5. The results of Raf-1mRNA expression showed that: compared with the model group,Raf-1mRNA expression in each group increased significantly(P <0.01); compared with the normal group, Raf-1mRNA expression showed no significant change in TCM middle-dosegroup, TCM high-dose group and Compound Apocynum group (P>0.05).6. The results of MAPK-P38mRNA expression showed that: compared with the modelgroup, MAPK-P38mRNA expression in each group increased significantly(P<0.01);compared with the normal group, MAPK-P38mRNA expression showed no significantchange in TCM middle-dose group, TCM high-dose group and Compound Apocynum group(P>0.05).7. The results of PCNA mRNA expression showed that: compared with the model group,PCNA mRNA expression in each group increased significantly(P <0.01); compared with thenormal group, PCNA mRNA expression showed no significant change in TCM middle-dosegroup, TCM high-dose group and Compound Apocynum group (P>0.05).8. The results of Bcl-2mRNA expression showed that: compared with the model group,Bcl-2mRNA expression in each group increased significantly(P<0.01); compared with thenormal group, Bcl-2mRNA expression showed no significant change in TCM high-dosegroup and Compound Apocynum group (P>0.05).9. The results of PPAR GAMMA mRNA expression showed that: compared with themodel group, PPAR GAMMA mRNA expression in each group increased significantly(P<0.01); compared with the normal group, PPARGAMMAmRNA expression showed nosignificant change in TCM low-dose group, TCM middle-dose group, TCM high-dose groupand Compound Apocynum group (P>0.05).10. The results of Raf-1protein expression showed that: compared with the model group,Raf-1protein expression in each group increased significantly (P <0.01); compared with thenormal group, Raf-1protein expression showed no significant change in TCM middle-dosegroup, TCM high-dose group and Compound Apocynum group (P>0.05).11. The results of MAPK-P38protein expression showed that: compared with the modelgroup, MAPK-P38protein expression in each group increased significantly(P <0.01);compared with the normal group, MAPK-P38protein expression showed no significantchange in TCM middle-dose group, TCM high-dose group and Compound Apocynum group(P>0.05).12. The results of PCNA protein expression showed that: compared with the model group, PCNA protein expression in each group increased significantly(P <0.01); comparedwith the normal group, PCNA protein expression showed no significant change in TCMmiddle-dose group, TCM high-dose group and Compound Apocynum group (P>0.05).13. The results of Bcl-2protein expression showed that: compared with the model group,Bcl-2protein expression in each group increased significantly(P <0.01); compared with thenormal group, Bcl-2protein expression showed no significant change in TCM high-dosegroup and Compound Apocynum group (P>0.05).14. The results of PPAR GAMMA protein expression showed that: compared with themodel group, PPAR GAMMA protein expression in each group increased significantly(P<0.01); compared with the normal group, PPARGAMMAprotein expression showed nosignificant change in TCM low-dose group, TCM middle-dose group, TCM high-dose groupand Compound Apocynum group (P>0.05). Zhen-gan Xi-feng Soup Decoction can activatethe Raf-1factor and the activation of Raf-1factor promote the MAPK P38activation, theactivation of MAPK P38factor is able to activate PCNA factor, PPAR GAMMA factor andthe Bcl-2factor.Conclusions:1. Zhen-gan Xi-feng Soup Decoction can affect blood pressure in spontaneously hyperten-sive rats. It has a good effect on lowering blood pressure in spontaneously hypertensive rats.2. Zhen-gan Xi-feng Soup Decoction can affect blood lipids in spontaneously hypertensi-ve rats, It can reduce the level of blood cholesterol (CHOL), triglycerides (TG) and lowdensity lipoprotein (LDL), and can lower spontaneously hypertensive rats’ lipids, which playsa role in protecting blood vessels and lower blood pressure and reduce blood lipids damage tothe blood vessels.3. Zhen-gan Xi-feng Soup Decoction can effectively improve the structure of spontaneoushypertensive rats vascular pathology, reducing vascular wall injury caused by high bloodpressure, and thus play a role in protecting blood vessels，and reduce the aorta vascularremodeling4. Zhen-gan Xi-feng Soup Decoction can inhibit apoptosis of vascular smooth muscle cellin spontaneously hypertensive rats, reducing the impact of hypertension on apoptosis of vascular smooth muscle cell, which play a role in inhibition of vascular remodeling andreduce the aorta vascular remodeling.5. Zhen-gan Xi-feng Soup Decoction can up-regulate the Raf-1、MAPK-P38、PCNA、PPAR GAMMA、Bcl-2mRNA and protein expression of vascular smooth muscle cell inspontaneously hypertensive rats, so that inhibition of aorta vascular smooth muscle cellapoptosis and reduce the aorta vascular remodeling.