Blood Ning Granule On Atherosclerosis Model Rabbit Cholesterol Reverse Operations In Mechanism Research

Atherosclerosis (atherosclerosis，AS) refers to an event in the arteriesof the non inflammatory，degenerative，hyperplastic lesions，eventually leadingto arterial wall thickening and hardening，loss of flexibility and luminalnarrowing．AS often involving the big，medium-sized arteries，intimal lesions from thestart，local lipid accumulation and complex carbohydrates，bleeding andthrombosis，fibrous tissue hyperplasia and calcinosis，and arterial medialgradual degeneration and calcification．AS is a long course, life in slowformation and development，pathogenesis is very complex，lesions in thedevelopment process can be seen in the activity，progress，static and subsidesin different periods corresponding changes．Clinical AS divided into early andlate two categories of lesions．Early lesion is characterized by a significantlipid stripe，namely fatty streaks，also somebody puts forward micro thrombusand gelatinous lesions is AS early lesions，also called striated．Late lesionis characterized by a significant atheromatous plaque or fibrous plaqueformation，and thus cause a variety of diseases including cardiovascular diseaseor cerebrovascular accident．Atherosclerosis disease belongs to the modern medicine，Chinese medicineand disease related records scattered in“phlegm”，“chest”，“stroke”，“vertigo”，“forget”，“the itch”，“dementia”，“true heart”，“diabetes”，“kidney paralysis”，“gangrene” diseases such as therecord．AS is the leading cause of heart，brain and other important organs ofvascular disease is the major cause for the incidence of cardiovascular andcerebrovascular disease，clinical disease，can also be found in the section ofperipheral vascular diseases．The patient manifested as dizziness，palpitations，shortness of breath，limb numbness， headache， chest tightness， negativelanguage， body movement disorder，neck discomfort，paroxysmal pain，precordial pain，lower limb claudication，red tongue，fur more greasy side stereotypies，Mai Shen fine tart，tension and other symptoms. Clinical manifestations ofvarious forms，various stages of syndrome，the symptoms are not the same．Itspathogenesis is complex， in medicine，AS occurs and the infirm，eat rich foodis delicious， internal causes，pathogenic cold invasion and other factors，Qi deficiency and blood stasis syndrome of spleen deficiency and dampness，Sheng，phlegm，phlegm and blood stasis of detoxification，liver and kidneydeficiency is the major pathogenesis of this disease．Modern medicine for atherosclerotic disease research in detail，reversecholesterol transport in atherosclerosis development plays a vital role，because the study found，reverse cholesterol transport cholesterol excretionis the only effective way，therefore, for the disease pathogenesis and as a targetfor the development of new drug research process，many involved in reversecholesterol transportation research．Over the years，tutor to traditional Chinese medicine theory，combined withmodern medical cutting-edge research results，as well as the rich treatment ofatherosclerotic disease experience， explored a compound Chinese herbal Xue MaiNing granule，the compound preparation for the treatment and prevention ofatherosclerosis have achieved good clinical curative effect. We think，atherosclerosis is mainly because of frail，the deficiency of vital qi，eatingdisorders，eat fatness，which leads to the spleen fails，stagnation of Qi andblood，phlegm and blood stasis syndrome，depression and of heat，heat injuryof vein，resulting in the sputum， blood stasis， poison， heat， etc. in theveins. Therefore，the disease pathogenesis is deficiency of Qi and blood stasis，vein blockage. The “Buqi Huoxue Tongluo，lipid-lowering treatment” as thefundamental law. Party in Astragalus， Gynostemma pentaphyllum as monarch drug，line Qi lipid-lowering of reactive power， supplemented with Danshen， RhizomaChuanxiong， three seven， to help blood circulation power． With hawthorn，tuckahoe， Alisma， Quzhi desiccant. The drug supplement each other， playeda total of Buqi Huoxue Tongluo， lipid-lowering． The study of atherosclerosis in the pathogenesis of Chinese medicine underthe guidance，combined with modern medical detection means，in atheroscleroticrabbits as experimental object，through to the rabbit serum cholesterol，triglyceride， high density lipoprotein，low density lipoprotein content in foamcells， lipid content， ABCA1gene and protein expression levels，the liver cellSR-BI gene and protein expression level monitoring， of Xue Mai Ning Granuleon blood lipid regulation role in atherosclerotic rabbits atherosclerosis modelof rabbit peritoneal macrophages， the surface of ABCA1gene and proteinexpression of Xue Mai Ning Granule on regulation and atherosclerosis model rabbitliver tissue in SR-BI mRNA and SR-BI protein expression influence. Thus fromthe blood rheology, gene expression，protein expression in different anglefurther proved the compound Chinese herbal Xue Mai Ning granule in the treatmentof atherosclerotic disease mechanisms， clinical atherosclerosis in patientswith treatment of Xue Mai Ning particles and clinical application and promotionto provide scientific experimental basis．PART ONEXue Mai Ning Granule on the experimental study of the effectsof AS rabbit lipidPurpose：Observation of Xue Mai Ning Granule on blood lipid regulation role inatherosclerotic rabbits，verify the clinical curative effect of Xue Mai Ninggranule．Materials and methods：This study in rabbits as experimental objects，40rabbits were in accordancewith the random number method， were randomly divided into four groups， respectively： blank control group， model group， Xue Mai Ning granule group，the control group of Western medicine．Adaptive feeding rabbits after a week，the model group， Xue Mai Ning granule group and Western medicine group weremodel replication. Using high fat diet feeding methods atherosclerosis modelreplication，model group， Xue Mai Ning granule group and Western medicine groupall the rabbits were fed high fat diet， and in experiment first day one-timeintraperitoneal injection of vitamin D3（600000IU/kg）. Blank control groupgiven normal granular feed， and in experiment first day was given the same amountof intraperitoneal injection of saline. Continuous with high fat diet for4weeks,beginning of Xue Mai Ning granule group and Western medicine group were treatedwith, by mouth filling feeding Xue Mai Ning granule and atorvastatin， once aday，after4weeks of continuous irrigation．To experiment eighth weekend，through ear vein collecting all rabbits venous blood4ml，isolated from theserum by enzyme assay，all rabbits serum triglyceride，cholesterol，high densitylipoprotein，low density lipoprotein in. The interception of the abdominal aortain rabbits of each group1.5～2CM, soaked in4%paraformaldehyde, embedded inparaffin, sections, HE staining was observed in the abdominal aorta,pathological changes．Using SPSS10.0software package for each of the groupsof experimental data for statistical analysis，measurement data to mean±SD(x±s)，multiple group were compared with single factor analysis of variance，with P＜0.05was considered statistically significant．Result：Blank control group rabbit abdominal aorta smooth muscle cells arrangedregularly, no lipid deposition and atherosclerotic plaque. Model group rabbitabdominal aorta showed obvious atherosclerotic plaque formation, and Xue MaiNing granule group and Western medicine group rabbit abdominal aortapathological change was significantly milder than that in the model group。Compared with the blank control group, model group，Xue Mai Ning granule group， the control group of Western medicine in rabbit serum cholesterol，triglyceride，high density lipoprotein，low density lipoprotein contentincreased significantly，with statistical significance (P＜0.01)．Compared with the model group， Xue Mai Ning granule group，the control groupof Western medicine in rabbit serum cholesterol，triglyceride，low densitylipoprotein content decreased，and high density lipoprotein content decreased，with statistical significance (P＜0.01)．The two treatment groups in rabbit serum cholesterol，triglyceride，highdensity lipoprotein，low density lipoprotein content， without statisticalsignificance (P＞0.05)．Conclusion：1．Simple high-fat feed continuous feeding rabbits after8weeks in suchlarge abdominal aortic artery formation was evident in the atheroscleroticplaque．2．Xue Mai Ning granule can effectively reduce hyperlipidemia andatherosclerosis model in rabbits serum triglyceride，total cholesterol，lowdensity lipoprotein；increase high density lipoprotein level．3．Xue Mai Ning Granule on blood lipids has bidirectional regulation effect．PART TWOXue Mai Ning Granule on the experimental study of the effectsof AS rabbit macrophage cholesterol content and ABCA1expressionPurpose：Observation of Xue Mai Ning Granule on peritoneal macrophages inatherosclerotic rabbits lipid clearance effect of AS model； observation of rabbit peritoneal macrophages surface ABCA1gene and protein expressionregulating effect of compound traditional Chinese medicine，exploration of XueMai Ning granule in treating atherosclerosis mechanism．Materials and methods：This study in rabbits as experimental objects，40rabbits were in accordancewith the random number method，were randomly divided into four groups，respectively：blank control group， model group，Xue Mai Ning granule group，the control group of Western medicine. Adaptive feeding rabbits after a week，the model group，Xue Mai Ning granule group and Western medicine group were modelreplication. Using high fat diet feeding methods atherosclerosis modelreplication，model group， Xue Mai Ning granule group and Western medicine groupall the rabbits were fed high fat diet，and in experiment first day one-timeintraperitoneal injection of vitamin D3(600000IU/kg). Blank control groupgiven normal granular feed，and in experiment first day was given the same amountof intraperitoneal injection of saline. Continuous to high fat diet for4weeks，all rabbits through ear vein collecting venous blood3ml，isolated from the serumwere detected，the level of blood lipid of rabbits. By statistical analysis，compared with the blank control group，model group three rabbits blood lipidswere significantly higher than control group (P＜0.01)，that successfully mould，and the start of Xue Mai Ning granule group and the control group of Westernmedicine treatment，respectively，by mouth filling feeding Xue Mai Ning granuleand atorvastatin，once a day，after4weeks of continuous irrigation．In theexperiment until the end of the third day，every rabbits by intraperitonealinjection into the sterile starch solution2ml/only. After the end of theexperiment，put to death all rabbits，and to the peritoneal injection of PBSbuffer40ml，gently massage the abdomen for3minutes，then cut open the belly，collection of peritoneal fluid，separation of rabbit peritoneal macrophages wereobserved in each group，rabbit peritoneal macrophage lipid loaded condition， then the4%NP-40isopropanol solution elution staining of macrophages in the dye，492nm wavelength detection the eluate of the absorbance value，in response toall rabbits peritoneal macrophage lipid loaded weight；immunocytochemicaldetection of peritoneal macrophage surface expression of ABCA1protein；RT-PCRmethod for the detection of peritoneal macrophage surface ABCA1mRNA expression.Using SPSS10.0software package for each of the groups of experimental data forstatistical analysis，measurement data to mean±SD(x±s)，multiple group werecompared with single factor analysis of variance，with P＜0.05was consideredstatistically significant．Result：Model rabbits peritoneal macrophage lipid loaded weight was significantlyhigher than the other three groups，between the two treatment groups with nosignificant difference．Immunocytochemical detection results，AS model rabbits of group ABCA1ofmacrophages protein expression level was significantly higher，compared withthe blank control group was statistically significant (P＜0.01). Through thepositive cell percentage statistics study shows that，compared with the modelgroup，Xue Mai Ning granule could significantly increase the ABCA1proteinexpression，with statistical significance (P＜0.01)．The results of RT-PCR，AS ABCA1mRNA expression in rabbits were significantlyup-regulated，drug intervention，increase more significantly，indicating thatChinese medicine Xue Mai Ning granule for the ABCA1mRNA expression has someupregulate．The two treatment groups were comparable，rabbit peritoneal macrophageslipid loaded weight，cell surface ABCA1gene and protein expression levels werenot statistically significant (P＞0.05)． Conclusion：1．Xue Mai Ning granule can effectively remove foam intracellular lipidcontent，thereby effectively inhibit atherosclerosis disease．2．Xue Mai Ning granule through the up-regulation of ABCA1gene and proteinexpression level to achieve the removal of foam cells role of lipid．3．Xue Mai Ning granule upregulate ABCA1gene and protein expression，intracellular lipid removal of foam may be Xue Mai Ning granule in preventingand treating atherosclerosis is one of the mechanisms．PART THREEXue Mai Ning Granule on the experimental study of the effectsof AS rabbit liver tissue SR-BI expressionPurpose：Observation of Xue Mai Ning Granule on liver tissue in rabbit atherosclerosismodel of SR-BI mRNA and protein expression of SR-BI regulation．Exploration ofcompound Chinese herbal Xue Mai Ning granule in treating atherosclerosismechanism．Materials and methods：This study in rabbits as experimental objects,40rabbits were in accordancewith the random number method，were randomly divided into four groups，respectively：blank control group，model group，Xue Mai Ning granule group，the control group of Western medicine．Adaptive feeding rabbits after a week，the model group，Xue Mai Ning granule group and Western medicine group were modelreplication．Using high fat diet feeding methods atherosclerosis modelreplication，model group，Xue Mai Ning granule group and Western medicine groupall the rabbits were fed high fat diet，and in experiment first day one-timeintraperitoneal injection of vitamin D3(600000IU/kg). Blank control group given normal granular feed，and in experiment first day was given the same amountof intraperitoneal injection of saline. Continuous to high fat diet for4weeks，all rabbits through ear vein collecting venous blood3ml，isolated from the serumwere detected，the level of blood lipid of rabbits. By statistical analysis，compared with the blank control group，model group three rabbits blood lipidswere significantly higher than control group（P＜0.01），that successfully mould，and the start of Xue Mai Ning granule group and the control group of Westernmedicine treatment，respectively，by mouth filling feeding Xue Mai Ning granuleand atorvastatin，once a day，after4weeks of continuous irrigation．In theend of the experiment，through ear vein injection of air，cause an air embolism，put to death all rabbits，open abdominal，removal of liver caudate lobe，usingRT-PCR method to detect all rabbits liver tissues of SR-BI mRNA expression；using Western-blot for detecting clusters of rabbit liver tissue SR-BI proteinexpression level. Using SPSS10.0software package for each of the groups ofexperimental data for statistical analysis，measurement data to mean±SD(x±s)，multiple group were compared with single factor analysis of variance，with P＜0.05was considered statistically significant．Result：Western-blot results show，objective strap located approximately180kDa，integral optical density value of statistical results indicate that，comparedwith the blank control group，model group rabbit AS expression level of SR-BIwas significantly higher，with statistical significance (P＜0.01)；comparedwith the model group，treatment group two in rabbit SR-BI expression level wassignificantly higher，with statistical significance (P＜0.01)．RT-PCR results show，objective strap located approximately485bp，integraloptical density value of statistical results indicate that，compared with theblank control group，model group rabbit AS expression level of SR-BI wassignificantly higher，with statistical significance (P＜0.01)；compared with the model group，treatment group two in rabbit SR-BI expression level wassignificantly higher，with statistical significance (P＜0.01)．The two treatment groups were comparable，the rabbit liver tissue SR-BI geneand protein expression levels were not statistically significant (P＞0.05)．Conclusion：1．Xue Mai Ning granule can obviously increase in atherosclerotic rabbitsand SR-BI mRNA expression in liver cells．2．Xue Mai Ning granule can obviously increase in atherosclerotic rabbitsand expression of SR-BI protein in liver cells．3．Xue Mai Ning granule through upregulation of liver cell SR-BI gene andprotein expression and its effect on cholesterol reverse transport role．4．upregulation of liver cell SR-BI gene and protein expression，whichpromotes reverse cholesterol transport may be Xue Mai Ning granule in preventingand treating atherosclerosis and mechanism．...