CMS Rats And Offspring Model And N-acetylcysteine,Dracocephalum Moldavica L Drug Intervention Study

Objective:This study used drug intervention by giving N-acetylcysteine(DA) and Dracocephalum moldavica L(DML)on the basis of establishing CMS, the model groups of rats and rats offspring.We discussed the study of CMS (Chronic mountain sickness)in rats and the impacts of DA and DML on CMS high altitude heart disease in rats by using cardiac ultrasound、ECG、pathology、pharmacology and metabolomics, explaining the CMS high altitude heart disease pathogenesis in rats and drug treatment at the same time stated that high altitude、low pressure、low oxygen environment caused changes on myocardium、blood inflammatory mediators and blood gass analysis as well as learning the difference between that CMS rats and CMS rats offspring in the above aspects and providing the theoretical base in finding an efective preventive measurements.This study included:(1)effect of DA on CMS rat model (2)meehanism and role of acetyl-l-cysteine on the CMS rats using NMR-based metabolomism research (3)effect of DML on CMS rats model (4)establishment and study of CMS rats model and rats offspring. Methods:Part one120Sprague-Dawley rats were randomly divided into6groups, BG、MGNE、LDA、 MDA、HDA and each group has20rats. Serum markers such as IL-6、CPR、ET-1、 VEGF、HCY、NO were measured in each group.we did analysis and determination of ECG、cardiac ultrasound、pathology、routine blood test、blood gas analysis in each group’ s rats. Part two32rats were divided into four groups, BG、MG、Plain acetylcysteine medication group、plateau acetylcysteine medication group respectively and each group has8rats. We used metabolomics approach for the analysis. Part three100Sprague-Dawley rats were randomly divided into5groups, BG、MG、NE、DMLWG、 DMLAG respectively, and each group has20rats.The serum markers (IL-6, CPR、ET-1、 VEGF、HCY、NO) and ECG、cardiac ultrasound、pathology、routine blood test、blood gas analysis were measured and analyzed in each group. Part four This part of the study is divided into3experimental groups, BG、Mg、P.O. Serum markers (IL-6, CPR、ET-1、 VEGF、HCY、NO)ECG、echocardiography、routine blood test and blood gas analysis were measured and analyzed in group’s rats. Result:Part one Pathological examination results observed under low magnification showed the mild congestion in epicardial arterial vessels rats with,myocardial interstitia vascular congestion the significant congestion in myocardial interstitial vessels and unclear striated muscles in MG rats. At high magnification, there were partial heart muscle swelling and cytoptasmic granular degeneration,and there also was eosinophilic Infiltration in partial myocardial fibrosisin with inflammatory cell infiltration. Cardiac muscles were arranged neatly with normal structure and occasionally partial myocardial interstitium was mildly congested in SD rats in HDA。Neither significant Eosinophilic diseased cells nor significant inflammatory cells infiltration were seen under high magnification. Serum markers revealed that the CRP、ET-1、IL-6、HCY、VEGF (vascular endothelial growth factor)、pulmonary arterial pressure、aortic pressure、Hb、Hct significantly increased in normal control group compared with the high altitude model group while NO、SaO2and PaO2significantly decreased with a statistical significant difference(P<0.05). Drug interventions can effectively improve the indicators; nifedipine can effectively improve all other indicators except for Hb while acetylcysteine can effectively improve all other indicators except for PaO2, and both with a significant statistical differences(P<0.05). Results in different dosage group of acetylcysteine showed that LDA group has the best improving effect on ET-land NO,medium and high dose group could extremely increase this indicator,while HDA has the best effect on other indicators. Echocardiogram showed that left atrial diameter change was not obvious and right atrial diameter change was significant in the comparison of plateau model group with plain model group.Again by comparing plateau model group and plain model group we found that left ventricular both systolic and diastolic diameter enlarged, right ventricular diameter increased, right ventricular anterior wall thickened, ventricular septal broadened, right ventricular outflow tract broadened, cardiac ejection fraction decreased, the above description showed that the high altitude disease modeling was successful.Left ventricular systolic and diastolic diameter decreased both in high dose and medium dose acetylcysteine medication group, but not obvious in the low dose group. Right ventricle shrinks in the high dose acetylcysteine group but it was not obvious both in the medium dose and low dose group. Ⅱ leads in electrocardiogram in experimental rats of each group showed that plain control group rats had normal ECG with normal P wave, QRS wave and ST segment. ECG of MG rats showed tall P wave and cyrta melichar ST elevation. ECG of high dose acetylcysteine group rats showed normal P wave、QRS complex and ST segment. Part two The blood metabolic components have significant difference in each group’s rats; comparing with the CMS model group, the positive correlation coefficient metabolites are the increased metabolites in plain model group, while the negative correlation coefficient are the decreased metabolites in plain model group. Comparing with the CMS model group, the positive correlation coefficient metabolites in those have significant difference are the increased metabolites in acetylcysteine medication group, while the negative correlation coefficient are the decreased metabolites in acetylcysteine medication group. Comparing with the CMS model group, the positive correlation coefficient metabolites in those have significant difference are the increased metabolites in plateau acetylcysteine medication group, while the negative correlation coefficient are the decreased metabolites in plateau acetylcysteine medication group, comparing with the plain acetylcysteine medication group, the positive correlation coefficient metabolites in those have significant difference are the increased metabolites in plain model group, while the negative correlation coefficient are the decreased metabolites in plain model group. Part three Pathological result showed that there was no epicardial congestion but there was significant congestion in myocardial interstitial vessels under low magnification in DMLWG group. Under high magnification there was significant cellular edema in epicardial myocardium as well as scattered acidophilic degeneration. There was nor significant epicardial congestion or significant myocardial interstitial congestion under low magnification in DMLWG group. There was several cellular edema of myocardium with rarely seen acidophilic degeneration under high magnification. Blood serum revealed that comparing with MG group, IL-6、CRP、Hcy、ET-1、VEGF significantly decreased (P<0.05) in control group while IL-6、CRP、Hcy、ET-1、VEGF decreased (P<0.05) in DMLAG group and DMLWG group; B ultrasound result:two-dimensionalultrasound of the successfully modeled SD rats which accepted the drug intervention by DMLAG group showed that comparing with the model group, ejection fraction increased with narrowing of the right ventricular outflow tract; ultrasound also showed that left ventricular systolic and diastolic diameter greatly reduced and ventricular septum and left ventricular posterior wall was in same directional motion, indicating that left ventricular systolic and diastolic function was severely damaged. In DMLWG medicational intervention group right atrial and ventricular diameter increased with little obvious increasing in ejection fraction. Ultrasound showed systolic snd diastolic diameter decreased which indicated that the left ventricular systolic and diastolic function failed furthurly. ECG results showed:P wave,QRS waves and ST segment were all normal in DMLWG group while it showed there was tall P wave in DMLAG group. Part four pathological result showed:the change in Plateau model group was the same with the first part.High land descend not congested epicardial vessels with rarely seen congested myocardial interstitial vessels and unclear striated muscle with neatly arranged myocardial fibers were seen in P.O under low magnification. Under high magnification there were mild edema of myocardial cell and acidophilic degeneration in myocardial cells and inflammatory cells infiltration. Comparing with MG group, IL-6、CRP、Hcy、ET-1、VEGF、pulmonary and carotid arterial pressure、Hct、Hb significantly decreased (P<0.05) in BG group while IL-6、CRP、 Hcy、ET-1、VEGF significantly while NO、SaO2and PaO2significantly decreased in plateau model group with a significant statistical difference(P<0.05). Comparing with plateau model group, there was a obvious improvement on above indicators in P.O with a significant statistical difference (P<0.05). Cardiac ultrasound showed:by comparing P.O with MG group it was seen that the right ventricule was enlarged,and right ventricular anterior wall was thickened,ventricular septum was broadened,ventricular ejection fraction was increased,right ventricular outflow tract was broadened was increased.The ultrasound showed that:by comparing the P.O with BG, left ventricular systolic and diastolic diameter significantly reduced and there was untypical same directional motion of left ventricular anterior wall and ventricular septum, indicating that environment during pregnancy affects embryo development resulting in left ventricular systolic and diastolic dysfunction, particularly in diastolic dysfunction. ECG result:P wave、QRS wave and ST segment were normal in BG group and in MG group there was tall P wave and elevated St segment while there was normal P wave and ST segment with little change.Conclusions: according to this experiment we can conclude that (1) DA has protective role on the hearts of CMS rats and has significant improvement on myocardial tissue damage. DA has a role on the reduction of the pulmonary artery and carotid artery pressure. After performing the B ultrasound we can conclude that DA has protective function both on the ventricule and atrium of CMS rats. DA has a role on the reduction of these indicators as (CRP、ET-1、 IL-6、HCY、VEGF、Hb、Hct) only except for PaO2.(2) Valine、tyrosine、1-methylHistidine、Leucine、phenylalanine、methionine、lacticacid、glucose、lipids、 proteins、acetone(VLDL)creatine,LDLand beta-hydroxybutyric acid were detected in decreasing rate while B-glucose and a-glucose were detected in decreasing rate in CMS rats blood by doing metabolomics investigation. Acetic acid、lactic acid、 1-methyl-Histidine,creatine level all decreased in the blood after giving DA intervention,indicating that as being a powerful exogenous antioxidant,DA has a role on the improvement of metabolism of CMS rats.(3)We can conclude from the DML intervention studies on CMS rats that DML has an improving effect on the myocardial tissue damage.DML has a reduction effect on the pulmonary and carotid artery. DML has an effective reduction role on CRP、ET-1、IL-6、HCY、VEGF、Hb、Hct、PaO2in CMS rats blood. DML has improving effect on ventricular size、motion and myocardial ischemia, in dicating that DML can be used as a drug for treatment and prevenion of CMS with its little side effects.(4)By comparing with the MG rats,the P.O rats have a degree of adaptive ability to the high altitude environment but comparing with the BG rats all physiological parameters that we detected had changes both in P.O and MG rats. Comparing to the MG, the physiological indexes of P.O rats moved to the direction of indexes of BG rats, and this indicated that P.O rats has a certain degree of adaptability and acclimatization. We also found that MG pregnant mice have low farrowing rate and the rat pups have a low survival rate afterbirth, which may be related to high altitude hypoxia environment.