On Vascular Endothelial Cells. Goose Ice Shuguan, Dripping Pills Lipid Oxidative Damage Protective Effect Study

Part 1.The observation of the Ebingxuguan Drop Pill's treatment effects and the influence of superoxide of the coronary heart disease.The Ebingxuguan Drop Pill(SGDW) is a new medicine for the coronary heart disease patient of TCM(traditional Chinese medicine) which was developed by professor Chengchoufu,based on his distance clinic practices and the theory of TCM.SGDW has the efficacy of accelera the blood block,increase the Qi' function then resolve the phlegm obstruct,and lighten the pain.It should be recommended to the coronary angina that is QI weak and phlegm block.This research adopting the random compared,double blind trail to make sure the treatment effects and the affection to the coronary heart disease patient's superoxidize.Purpose:Make sure the clinical affection of the SGDW use for the patient with certain coronary heart disease,then get further definite evidence of the influence of the coronary heart disease patient's oxidize.Method:The research adopted random,double blind,double imitate,the masculine medicine parallelism trial,choused the patients which matched the diagnoses of coronary angina pectoris and TCM,and paroxysm of angina more then 2 times every week.The compare drug is Xexiangbaoxi Pill.Dose:the patients of test group use 5 pills/times Ebingshuguan Pill and 2 pills/times Xexiangbaoxi Pill,3 times everyday and half an hour after meal each time;the control group,2 pills/times Xexiangbaoxi Pill and 5 pills/ times Ebingshuguan Pill,3 times everyday and half an hour after meal each time.A month later,examine the ET-1,MDA and SOD' activity to make sure the effect. Result:The experiment observed 87 patients totally,the test group 45,the control group 42,the result showed that the two groups with good balanced before the experiment.The test group:The effect for the angina is 26 example (57.78%),valid 15,the total efficient is 91.11%;The electrocardiogram curative effect shows the effect is 14 example(32.56%),valid 20,the total efficient is 79.07%;for the Zheng of the TCM the effect 24 example (53.33%),valid 18,total efficient 93.33%,all were higher than the control group,but the difference did not show any statistic meaning(P＞0.05).And the result show a lot patients with Ebingshuguan Drop Pill stop or reduce the dose of nitric acid glycerin,the ratio is 69.25%and 58.87%,a little lighter then the compare patients,but with no statistics meaning(P=0.0708).With the better of the patient's condition,the concentration of the MDA which is the last outcome of the oxidize action of fat dropped lower,and the activity of the SOD which was the main elimination for oxygen-derived flee radicals increases a lot than before(P＜0.01);and ET-1,the impartment vein active substance which is excreted by endothelia cell,which can reflect the function of the endothelia cell's function,fall remarkable.Conclusion:The Ebingshuguan Drop Pill has good affection on treating the coronary angina pectoris,it's resisting of oxidize fat and the meliorate of the endothelia cell's function may be one of the mechanisms of the clinical curative effect.Part 2.The research of Ebingshuguan Drop Pill's affection on the damage of vessels endothelia cell which is caused by ox-LDLFor make sure the function of the SGDW's infection on the damage of vessels endothelia cell that which is caused by ox-LDL,adopted molecular biology's solution,take apoptosis,NO,ET,t- PA,PAI etc as the index sign,carry on the research of the protection of the damage of vessels endothelia cell that which is caused by ox-LDL.Purpose:Study the Ebingshuguan Drop Pill's protection of the damage of vessels ndothelia cell,which is caused by ox-LDL.Method:Adopted MTT method examine the SGDW' s toxicity to the vessels endothelia cell VEC—304 which is trained outside the human body,take the 1/2 concentration that the biggest density which uninfluenced the cell as the test dose.According to the report,take the model of the damage of vessels endothelia cell which is caused by 200ug/ml eventually ox-LDL,joining the solution of the SGDW before or build the mold as the treatment factor,imitate the clinical usage of the SGDW to treatment the damage of vessels endothelia cell that caused by the dangerous factor of coronary angina pectoris.Examine apoptosis,NO,ET,t- PA,PAI etc,working out the effect ion of the SGDW.Divide group:divided the cell grew in the 12 knotholes board into 4 parts:①:Bland group:Hatch 24 hours in DMEM developing liquid(contain 10% calf serum) at the temp at 37℃;②Model group:Hatch 24 hours in DMEM developing liquid which contains 200ug/ml ox-LDL at the temp at 37℃,resulting in the cell hurt;Ebingshuguan Drop Pill protection set:Each hatch 2 hours in DMEM developing liquid which contains 1000ug/ml and 500ug/ml Ebingshuguan Drop Pill solution,then hatch 24 hours in DMEM developing liquid which contains 200ug/ml ox-LDL;④Ebingshuguan Pill group:Each hatch 24 hours in DMEM developing liquid contained 200ug/ml ox-LDL and 1000ug/ml or 500ug/ml Ebingshuguan Drop Pill solution. The apoptosis examination adopted immunocytochemical method and flow cytometry techniques;The measurement of NO adopted the nitric acid revivification enzyme method;The ET measurement adopted radiation immunity method;the PAI-1 and the t-PA examination adoption chromomeric funds method.Result:The fluorescence microscope observed the bland group cell dyes distribute uniformity.After the ox-LDL function 24 hours,can observe various cell of typical model of early,medium or later period apoptosis,and can detected the condensation of apoptotic nuclear,overflowing into the cell and formatted small bodys,present the typical model of dyeing quality coagulate which used can be fund in apoptosis.The decrease of apoptosis in the test group is improved obviously.The result showed that the apoptosis of the bland group was 1.67%,the model group is 32.28%,the G1 apex(AP) appeared before the g1 period of time. Compared with the model group,the rate of cell demise treat group descend(13.82%～17.43%,P＜0.05) obviously,the protection group was more lower than the test group(P＜0.05),but no different in each subgroup. The examination of the active substance excreted by vessels endothelia cell in the cell development liquid showed the NO,ET-1,PAI of model group all increased,but the NO/ET-1,the t- PA descended,the Ebingshuguan Drop Pill can resist the increase of NO,ET-1,PAI and descend of NO/ET-1,t- PA, caused by ox- LDL,and under most circumstances the protect set's function more obviously.Conclusion:The Ebingshuguan Drop Pill can protect the vessels endothelia cell apoptosis caused by ox-LDL,and increase the excretion of the endothelia cell under the ox- LDL condition,lowering endothelia element,confirmed the Ebingshuguan Drop Pill's protection to the injury of endothelia cell caused by ox- LDL condition.