The Study About Defatigation Caused By Forced Swimming Induced The Activation Of TAS And The Damage Of Vascular Endothelium And The Effect Of Tongluo Intervention

Objectives: Directed by the theroy of TCM and the collateral disease, established the model of vascular endothlial dysfunction (VED)by feeding rats homomethionin and taken it as teference, established the model of defatigation by forced swimming, and then combined homomethionin and forced swimming to establish complex model. Through observing the changes of the structure and function of vascular endothelium, the renin–angiotensin-system(RAS), and correlated index of oxidative stress, we explored the mechanism of defatigation caused by forced swimming on VED and the effect of Tongxinluo. All of the study will provide the experimental foundation of the effect about social psychic factor on vasculopathy and the prevention of it .Methods:1.The effect of forced swimming on the stucture and function of vascular endothelial cell (VEC)Male Wistar rats were randomly divided into the following four groups :①control group;②hyperhomocysteinemia group (Hcy group);③forced swimming group;④complex model group (hyperhomocysteinemia +forced swimming);to evaluate objectively the defatigation condition acorrding with genenral condition observation, the score of biological superficial sighs, and the climbing time of rats; to observe the morphologic change of aorta by light microscope and transmission electron microscope(TEM);to analysis the VED through detecting the expression of endothelin receptor (ET1) endothelial nitric oxide synthase (eNOS)by the technology of immunohistochemistry, and the level of endothelin (ET)and nitro oxygen (NO)in blood.2. The effect of forced swimming on RASThe method of model establishment and grouping were the same as above. Gathered the blood of rats and separated plasma and serum, detected plasma rennin activity (PRA), angiotensinⅡ( AngⅡ) by radio-immunity (RI); serum superoxide dismutase (SOD) and malondialdehyde (MDA) by hydroxylamine and TBA, and the expression of AngⅡ, AngiotensinⅡreceptor (AT1R), angiotensin–convertion enxymeⅠ(ACEⅠ)and p22phox by immunohistochemistry.3. The activation of RAS and the damage of VEC caused by forced swimming and the effect of Tongluo intervention.Divided randomly into four groups :①control group;②complex model group;③Tongxinluo group;④Benazepril group. Index detected and the method were the same as above. In this part, we explored that defatigation caused by forced swimming induced the activation of RAS and the damage of vascular endothelium and effect of Tongxinluo. Results:1. The effect of forced swimming on the structure and function of VEC1.1 The result of the morphology of aorta endothelium1.1.1 Results analysised by light microscopeHcy group, EC were edema and amotic, inflammatory cell infiltration could be seen, minor hyperplasia could be found in the subendothelial layer, edema could also be observed in the smooth muscle cell(SMC);Forced swimming group, EC were edema and maldistribution, lymph cell infiltration could be seen, hyperplasia could be found in the subendothelial layer, the SMC ranked disordery;Complex model group, EC were edema, maldistribution and partly disappeared, the intima were partly thickening, inflammatory cell infiltration and fragmentation could be seen in the intima, the SMC were edema and ranked disorderly.1.1.2 Results analysised by TEMHcy group, bulk crests of mitochondria of VEC fused with membrane, and even disapeared; dilataion and degranulation could be found in the rough surfaced endoplasmic reticulum; outer the layer of caryotheca disappeared, the counts of pinocytotic cells could be found decreased; perinuclear cisterna could be seen partly dilated;Forced swimming group, bulk creats of mitochondria of VEC fused with membrane, and even disapeared; dilatation and degranulation could be found in the rough surfaced endoplasmic reticulum; outer the layer of caryotheca disappeared, the counts of pinoctytoic cells could be found decreased; perinuclear cisterna could be seen partly dilated;Complex model group, bulk cresrs of mitochindria of VEC fused with membrane, and even disapeared; obvious dilatation and degranulation could be found in the tough surfaced endoplasmic reticulum, oedema could be seen in the cell-substance and cell nucleus, membranolysis and even disapperaed, the count of them were outside.1.2 The changes of correlated index of VED.1.2.1 The changes of NO and ET: Compared with control group, the content of NO in setum decreased significantly(P<0.001) and that of ET in blood plasma heightened significantly (P<0.05,P<0.001) in Hcy group, forced swimming group and complex model group. Compared with Hcy group, the changes of NO and ET had no statistic difference (P>0.05), the content of NO decreased singnificantly and that of ET heightened significantly (P<0.05) in complex model group.1.2.2 The expression of ET1 and eNOS in aortaThe stained yellow signals of ET1 enhanced and that of eNOS decreased in the Hcy group;The stained yellow signals of ET1 enhanced and that of eNOS decreased in the forced swimming group;The stained yellow signals of ET1 enhanced more than the Hcy group, no stained yellow signals of eNOS could be seen.2. The effect of forced swimming on RAS2.1 The effect on cycloid type RASCompared with control group, the content of PRA,AngⅡin blood plasma had the tendency to heighten but had no statistic difference(P>0.05)in the Hcy group; the content of PRA,AngⅡall increased significantly in forced swimming group and complex model group(P<0.01, P<0.05); the content of PRA,AngⅡall increased significantly in complex model group when compared with Hcy group(P<0.01).2.2 The effect on tissue type RASThe expression of AngⅡ,ACE,AT1R enhanced, and stained yellow signals could be seen in the kytoplasm of the aorta tissue in the Hcy group;Stained yellow signals could be seen in the aorta tissue of the forced swimming group;The stained yellow signals of AngⅡ,ACE,AT1R enhanced in the complex model group.2.3 The changes of SOD, MDA and p22phoxThe content of MDA in serum all increased significantly(P<0.01, P<0.05), the content of SOD in serum decreased(P<0.01, P<0.05)in the Hcy group, the expression of p22phox all enhanced indifferent degree and enhanced significantly in the complex model group.3. The activation of RAS and the damage of VEC caused by forced swimming and the effect of Tongxinluo intervention 3.1 The improvement of Tongxinluo to the damage of endothelium caused by forced swimming3.1.1 The effect of Tongxinluo on the morphologic change of endothelium caused by forced swimming①R esults analysised by light microscopeThe two drugs could relieve the edema, ablation of EC and the hyperplasy of philo-eosin material in intima in different degree,the effect of philo-eosin material in intima in different degree, the effect of Tongxinluo was the same as Benazepril.②R esults analysised by TEMThe two drugs could relieve the above ultrastructural changes in different degree and the effect of Tongxinluo was the same as Benazepril.3.1.2 The effect of Tongxinluo to the changes of endothelial function caused by forced swimming①T he effect to NO and ETThe content of NO increased significantly (P<0.001) in Tongxinluo group when compared with complex model group, but that was no satiatic difference in Benazepril group; The content of ET decreased significantly(P<0.001, P<0.01) in the two drugs group, and there were no significant statistic difference between the two drugs group(P>0.05).②T he expression of ET 1 and eNOS in the aorta tissueET1: The expression of ET1 in the aorta weakened in the two drugs group when compared with complex model group.eNOS: The two drugs could enhance the expression of eNOS when compared with complex model group.3.2 The effect of Tongxinluo to RAS3.2.1 The effect of Tongxinluo to cycloid type RASWhen compared with complex model group, the content of PRA and AngⅡdecresed significantly(P<0.05, P<0.001) in Tongxinluo group and the content of AngⅡdecresed significantly(P<0.01) in Benazepril group.3.2.2 The effect of Tongxinluo to tissue type RASWhen compared with complex model group, Tongxinluo and Benazepril could relieve the expression of AngⅡ,ACE,AT1R in different degree.3.2.3 The effect of Tongxinluo to the change of SOD,MDA and p22phox①The changes of SOD and MDAThe two drugs could all decrese the content of MDA and increase that of SOD in serum of rats(P<0.05) when compared with control group, and the effect of Tongxinluo was better than Benazepril (P<0.05).②T he changes of p22phoxThe stained yellow signals of p22phox in the aorta of rats decreased in two groups and the change was significant in Tongxinluo group.Conclusion:1. This study was introduced on the background of the effect of social psychic factor such as defatigation in the development of vasculopathy and in accordance with the damage of vascular endothelium by feeding rats homomenthionin, we expored the effect of defatigation caused by forced swimming to the structure and function of vascular endothelium, our experimental results indicated that forced swimming could injure the structure and function of VEC obviously, this indicated that defatigation caused damage to the vascular endothelium, when combined forced swimming with feeding rats homomenthionin, the damage of the vascular endothelium could aggravate.2. We also explore that the effect of force swimming to cycloid and tissue type RAS, and then we discovered that forced swimming could activate cycoid type RAS but had no significant effecton tissue type RAS of rats; and the effect of feeding rats homomenthionin was just opposite; however, the two RAS could be seen all activated in the complex model group. All above indicated that forced swimming could induce the excess activation of cycloid type RAS, and then enhanced the activation of tissue type RAS which caused by feeding rats homomenthionin.3. As forced swimming caused the activation of RAS, the oxidation-antioxygen was disequilibrium, all of those aggravated in the complex model group. Which indicated that the activation of RAS caused by forced swimming was correlated with the oxidative stress which induced by the way of the activation of p22phox.4. Tongxinluo could decrease the content of PRA,AngⅡin plasma of rats in the complex model group, and then restrained the excess activation of cycloid type RAS. However, the effect of Benazepril to cycloid type RAS was not significant, meanwhile, Tongxinluo could also restrain the excess activation of tissue type RAS, and then retrained the autoxidation caused by the activation of RAS, thus protected the structure and function of vascular endothelium.