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Objective: TCM collateral disease theory as a guide to the high methionine diet rat model of endothelial dysfunction and as a reference to establish fatigue model exhaustive swimming, two superimposed create a composite model, by observing the vascular endothelial structure and function, renin - angiotensin system, oxidative stress-related indicators of change, explore the exhaustive swimming due to overwork on vascular endothelial function and mechanism of the intervention effect of Tongxinluo for the study of social and psychological factors in the role of vascular lesions provide experimental evidence for its prevention. Method: 1. Exhaustive Swimming on endothelial cell structure and function of the male Wistar rats were randomly divided into four groups: ① normal control group; ② homocysteine ??group; ③ exhaustive swimming group; ④ complex model group (high homocysteine ??exhaustive swimming). In general clinical and biological characterization of semi-quantitative scores, objective evaluation of rat hanging pole time fatigue; by light microscopy and transmission electron microscopy observation of morphological changes of the aorta, immunohistochemical detection of endothelin receptor (endothelin receptor, ET1) , endothelial nitric oxide synthase (endothelial nitric oxide synthase, eNOS) expression, as well as blood endothelin (endothelin, ET), nitric oxide (nitro oxygen, NO) detection and analysis of vascular endothelial function. (2) Exhaustive Swimming on the renin - angiotensin system modeling methods and animal grouping ibid., blood samples were collected after the experiment were separated plasma and serum, plasma renin activity (plasma renninactivity, PRA), plasma vascular angiotensin Ⅱ (angiotensin Ⅱ, Ang Ⅱ) (RIA); serum superoxide dismutase (superoxide dismutase, SOD) activity (hydroxylamine) and MDA (malondialdehyde, MDA) content (TBA France); aortic tissue Ang Ⅱ, Ang Ⅱ 1 receptor (Angiotensin Ⅱ receptor, AT1R), angiotensin-converting enzyme Ⅰ (angiotensin-convertion enzyme Ⅰ, ACE Ⅰ) and NADH / NADPH oxidase subunit p22phox expression (immunohistochemistry). 3 tongluo intervention on Exhaustive Swimming cause RAS activation and vascular endothelial cell injury induced into the following four groups: ① normal control group; ② complex model group; ③ Tongxinluo group; ④ benazepril group. Indicators and methods for detection with the above two parts. Discussion of Tongxinluo on exhaustive swimming rats induced fatigue state of RAS and vascular endothelial cell injury. Results: 1. Exhaustive Swimming on aortic endothelial structure and function 1.1 Results 1.1.1 aortic endothelial morphology by light microscopy results were normal control group: continuous endothelial cell integrity, closely connected with the subendothelial layer, the membrane, the outer membrane were normal. Homocysteine ??Group: endothelial cell swelling, loss, inflammatory cells attached to the wall, infiltration, subendothelial layer hyperplasia, smooth muscle cell swelling. Exhaustive Swimming Group: endothelial cell swelling, uneven distribution of lymphocytes attached to the wall, infiltration, lower proliferation of endothelial and smooth muscle cell disorders. Model group: endothelial cell swelling, uneven distribution, local endothelial cells disappear, some thickening of the intima, endometrial and inflammatory cell infiltration, internal elastic plates are broken; smooth muscle cell disorder, edema. 1.1.2 TEM results normal control group, tight connections between cells, nucleus and organelles exist, no abnormal mitochondrial structure. Homocysteine ??group, most of vascular endothelial cell mitochondria cristae and membrane fusion or disappeared, rough endoplasmic reticulum, degranulation, the outer nuclear membrane disappears vesicle reducing the number of parts perinuclear expansion. Exhaustive Swimming Group: Most endothelial cells mitochondria cristae and membrane fusion or disappeared, rough endoplasmic reticulum, degranulation, the outer nuclear membrane disappears vesicle reducing the number of parts perinuclear expansion. Model group: Vascular endothelial cell mitochondria most ridges and membrane fusion or disappeared, rough endoplasmic reticulum expansion, were round or oval, degranulation clear cytoplasm, nucleus edema, some membrane rupture, missing, contents spill. 1.2 endothelial function related indicators of changes in 1.2.1 NO, ET Changes: Compared with normal control group, high homocysteine ??group, exhaustive swimming group, model group the serum NO levels were significantly lower in (P lt; 0.001), plasma ET levels were significantly increased (P lt; 0.05, P lt; 0.001). With high homocysteine ??group, exhaustive swimming changes of NO and ET was no significant difference (P gt; 0.05), model group was significantly higher ET, NO was significantly lower (P lt; 0.05). 1.2.2 aortic tissue ET1, eNOS expression in the normal control group, the positive aortic endothelial cells stained positive ET1 almost no signal; homocysteine ??group showed strong positive signals ET1 yellow dye; eNOS stained weakly positive signal ; exhaustive swimming group showed strong positive signals ET1 yellow dye and eNOS stained weakly positive signal; composite model group showed strong positive signals ET1 yellow dye, and stronger than homocysteine ??group, almost no positive signals eNOS yellow dye . (2) Exhaustive Swimming on the renin - angiotensin system RAS 2.1 pairs of cyclic effects compared with the normal control group, high homocysteine ??plasma PRA, Ang Ⅱ tended to increase, but no significant difference (P gt; 0.05); exhaustive swimming group, complex model of plasma PRA, Ang Ⅱ were significantly higher (P lt; 0.01, P lt; 0.05); with high homocysteine ??group, model group plasma PRA, Ang Ⅱ was significantly higher (P lt; 0.01). 2.2 pairs of tissue-type RAS arterial tissue in the normal control group no Ang Ⅱ, ACE, AT1R stained positive signal; homocysteine ??group Ang Ⅱ, ACE, AT1R expression is induced in arterial tissue seen in the cytoplasm yellow dye signal; exhaustive swimming group arterial tissue visible Ang Ⅱ, ACE, AT1R-positive yellow dye signal; composite model group arterial tissue visible Ang Ⅱ, ACE, AT1R strong positive yellow dye signal. 2.3 SOD, MDA, p22phox changes compared with normal control group, high homocysteine ??group, exhaustive swimming group, model group MDA levels were significantly higher (P lt; 0.05, P lt; 0.01), serum SOD activity were significantly lower (P lt; 0.05, P lt; 0.01), p22phox expression were enhanced to varying degrees, to the most complex model group expression. 3 tongluo intervention on Exhaustive Swimming cause RAS activation and vascular endothelial injury in 3.1. Tongxinluo on endothelial damage caused by exhaustive swimming Improvement of Tongxinluo on 3.1.1 Exhaustive Swimming morphological changes induced endothelial Light microscopy role ① normal control group: continuous endothelial cell integrity, closely connected with the subendothelial layer, the membrane, the outer membrane were normal; composite model group: endothelial cell swelling, uneven distribution, local endothelial cells disappear, some thickening of the intima , endometrial and inflammatory cell infiltration, internal elastic plates are broken; smooth muscle cell disorder, edema. Two treatment groups: endothelial cell swelling, loss level and eosinophilic material endometrial hyperplasia was significantly reduced compared with model group, tongxinluo considerable role with benazepril. ② TEM results of the normal control group: tight connection between endothelial cells, nucleus and organelles exist, no abnormal mitochondrial structure. Model group: endothelial cell mitochondria cristae and membrane fusion most or disappear, rough endoplasmic reticulum, were round or oval, degranulation clear cytoplasm, nucleus edema, some membrane rupture, missing, contents spill. Two treatment groups: the more complex model group were significantly reduced, tongxinluo with benazepril considerable improvement. 3.1.2 Exhaustive Swimming Tongxinluo on endothelial function caused by changes in the role ① on NO, ET content in comparison with the normal control group, model group significantly decreased NO level (P lt; 0.01), ET was significantly higher (P lt; 0.01); and the composite model group, Tongxinluo group was significantly higher NO levels (P lt; 0.001); while Burnett Plymouth variation of NO was no significant difference (P gt; 0.05); than those without statistically significant difference (P gt; 0.05). ② The aortic tissue ET1, eNOS expression ET1: normal control group weak aortic endothelial cells ET1 expression; composite model group ET1 strong expression. Compared with the model group, the treatment group showed a weak positive expression of ET1. eNOS: normal control group, aortic endothelial cells were strongly positive eNOS expression; composite model group almost no expression of eNOS, two treatment group showed positive expression of eNOS, the yellow dye signal intensity compared with model group increased. 3.2 Tongxinluo on the impact of RAS 3.2.1 Tongxinluo on the impact of recycling RAS compared with normal control group, model group plasma PRA, Ang Ⅱ were higher (P lt; 0.01, P lt; 0.05); and compound model group, TXL group PRA, Ang Ⅱ levels were significantly lower (P lt; 0.05, P lt; 0.001); benazepril group Ang Ⅱ levels were significantly lower (P lt; 0.01). 3.2.2 Tongxinluo on the impact of tissue-type RAS control rats aorta endothelial cells almost no Ang Ⅱ, ACE, AT1R stained positive signal; composite model visible Ang Ⅱ, ACE, AT1R-positive yellow dye signal, endometrial Under thickening seen at ATR-1 stained weakly positive signal; and the composite model group, TXL group visible Ang Ⅱ, ACE, AT1R stained weakly positive signal, benazepril group visible ACE, AT1R weak positive signals. 3.2.3 Tongxinluo on SOD, MDA, p22phox change impacts ① SOD, MDA changes compared with normal control group, model group significantly increased MDA content, SOD activity was significantly decreased (P lt; 0.01); and complex model group, the two treatment groups decreased MDA content, SOD activity increased (P lt; 0.05). In Tongxinluo group effect is significant (P lt; 0.05). ② p22phox changes in the normal control group, no aortic tissue stained positive signals p22phox, p22phox complex model group showed a strong positive signal in yellow dye, two treatment group showed a weak positive signals p22phox yellow dye, and with Tongxinluo group changed significantly. Conclusion: this study, fatigue and other social psychological factors in the development of vascular lesions of the background of rats fed a high methionine diet cause vascular endothelial injury as a reference to explore Exhaustive Swimming on endothelial structure due to overwork and function, the results showed that rats can swim alone exhaustive structure and function of vascular endothelial cells appear obvious damage, caused by overwork display exhaustive swimming endothelial damage is caused by an important risk factor, with a high methionine diet exhaustive swimming overlay could be heavy homomethionin induced endothelial damage. (2) explore the exhaustive swimming on rat tissue circulation type RAS and RAS, and found that exhaustive swimming can activate rat circulation type RAS, but for local arterial tissue RAS is not obvious; high methionine diet rats can local tissue RAS activation, no significant effect on the cycle type RAS; combined cycle type rats and tissue type of RAS are activated. Reveals the exhaustive swimming can be induced by excessive activation of RAS rat recycling and to increase the high methionine diet-induced rat local RAS activation. 3 exhaustive swimming rats induced activation of RAS, while oxidation - antioxidant imbalance, this change further increase in the composite model, suggesting that travel-related exhaustion, which may be due to fatigue caused by exhaustive swimming vascular endothelial injury one of the mechanisms. 4 tongxinluo by reducing the complex model in rat plasma PRA, Ang Ⅱ levels excessive activation of RAS inhibition of cyclic and benazepril on recycling obvious effect of elevated PRA, but also inhibits tongxinluo tissue excessive activation of RAS, thereby inhibiting the activation of RAS resulting from oxidation, improve endothelial structure and function.
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