冠心病患者血小板活化指标分析及高脂大鼠血小板活化指标评价与药物干预研究
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摘要
(一)冠心病患者血小板活化指标分析
[目的]评价冠心病(CAD)患者阿司匹林抵抗(AR)、P选择素(P-Selectin)、11脱氢血栓素B_2(11-dh-TXB_2)、6酮前列腺素F1a(6-k-PGF1a)、11-dh-TXB_2/6-k-PGF1a(dTP)比值的影响因素及与CAD严重程度、心血管事件(MACE)的相关性。
[方法]203名住院患者规范应用阿司匹林、氯比格雷1周,记录临床特征、血液生化指标、冠状动脉造影(CAG)检查结果及1年MACE随访结果。应用二磷酸腺苷(ADP)、花生四烯酸(AA)诱导血小板最大聚集率(MPAR)评价AR,应用酶联免疫法测定血浆P-Selectin、11-dh-TXB_2和6-k-PGF1a水平,计算的dTP比值。
[结果]1.203例患者中CAG正常16例(7.9%)、稳定性心绞痛(SAP)22例(10.8%)、不稳定性心绞痛(UAP)115例(56.7%)、心肌梗死(AMI)50例(24.6%)。各组患者AR、P-Selectin、11-dh-TXB_2、6-k-PGF1a、dTP比值差别显著,结果见表1。2.各种血小板活化指标及CAG严重病变数的相关因素分析,结果见表2。3.随访1年55例(27.1%)患者发生MACE,11-dh-TXB_2、6-k-PGF1a、dTP值及P-Selectin水平与患者MACE发生相关,结果见表3。4。dTP比值与P-Selectin的相关系数最大(0.766)。
[结论]1.不同类型CAD患者AR、P-Selectin、11-dh-TXB_2、6-k-PGF1a、dTP比值差别显著。2.AR与临床合并糖尿病、hs-CRP及LDL-C水平相关。P-Selectin、年龄、hs-CRP水平与患者严重病变数相关。3.P-Selectin、11-dh-TXB_2、6-k-PGF1a、dTP比值与MACE发生相关。4.除了P-Selectin之外,TP比值能够较其它指标更好的反映CAD患者血小板活化的程度。
(二)高脂大鼠血小板活化指标评价与药物干预研究
[目的]分析高脂血症大鼠各种血小板活化指标与血小板表面CD62P表达的相关性,评价高脂血症大鼠血小板活化状态及阿司匹林、阿托伐他汀、氯沙坦、复方丹参滴丸、前列腺素E_1对于高脂血症大鼠血小板功能的影响。
[方法]Wistar大鼠75只随机分为正常对照组、高脂血症组、阿司匹林组、阿托伐他汀组、氯沙坦组、复方丹参滴丸组、前列腺素E_1注射液组。分组干预后检测各组大鼠的血脂水平.应用HE染色制作肝脏、心肌、主动脉病理切片。应用ADP、AA诱导检测MPAR,酶联免疫法测定血浆中P-Selectin、TXB_2、6-k-PGF1a浓度,计算的TP比值。应用流式细胞术检测血小板表面CD62P的表达。
[结果]1.Wistar大鼠分组喂养4周后,正常对照组与高脂饲料组TG、TC、LDL-C分别是0.35 vs 0.88mmol/L、1.92 vs 10.72mmol/L、0.32 vs 8.66mmol/L。2.P-Selectin与血小板表面CD62P表达水平相关系数最大(0.847)。3.药物干预4周后各组大鼠ADP、AA诱导的MPAR,血浆TXB_2、6-k-PGF1_a、TP比值、P-Selectin水平及血小板表面CD62P水平差别显著(P＜0.001),结果见表1。
[结论]1.Wistar大鼠血浆P-Selectin水平与血小板表面CD62P表达密切相关,能够较好的反映血小板活化程度。2.高脂血症能够促使血小板活化,阿司匹林能够明显抑制血小板活化。3.阿托伐他汀在调脂作用的同时显著抑制血小板活化。
4.科素亚、复方丹参滴丸、前列腺素E_1具有不同程度的抑制血小板活化作用。
PartⅠThe study of platelet activity inpatients with coronary artery disease
[objective]To evaluate the influential factors of aspirin resistance (AR),P-Selectin,11-dehydro-thromboxane B_2(11-dh-TXB_2),6-keto-prostag-landin F1 alpha(6-k-PGF1a) and the ratio of 11-dh-TXB_2 over 6-k-PGF1a (dTP) in patients with coronary artery disease(CAD),and to estimate the relationship of the platelet activity indices with the CAD severity and with the major adverse cardiovascular events(MACE).
[Method]203 patients were given one week regulationly treatment of aspirin and clopidogrel.Clinical features,blood biochemical indicators and coronary arteriography(CAG) were recorded,and the MACEs of these patients were followed up for one year.Maximum platelet agglutination rate(MPAR) induced by both ADP and AA was used to estimate AR.Enzyme linked immunosorbent assay was used to detect the serum levels of P-Selectin,TXB_2 and 6-k-PGF1a,then dTP ratio was calculated.
[Results]1.16 cases(7.9%) with normal CAG group were found in all 203 cases,22 cases(10.8%) for SAP group,115 cases(56.7%) for UAP group, 50 cases(24.6%) for AMI group.The diferences of AR,P-Selectin,11-dh-TXB_2, 6-k-PGF1a and dTP ratio between each groups were significant and resules were showed in table 1.2.Correlative factors of platelet activity indices and the number of severe affected(CAG stenosis≥75%) coronary artery(CA) were showed in table 2.3.After one-year follow-up,55 patients(27.1%) had MACEs.P-Selectin,11-dh-TXB_2,6-k-PGF1a and dTP ratio correlated to MACEs and resules were showed in table 3.4.The correlated coefficient of dTP to P-Selectin was the largest(0.766) among those indices.
[Conclusion]1.The differences of AR,P-Selectin,11-dh-TXB_2,6-k-PGF1a and dTP ratio in different type of CAD patients were significant.2. Diabetes,high level of hs-CRP and LDL-C were risk factors of AR.P-Selectin level,age,and hs-CRP level positively correlated to the number of severe affected CA.3.P-Selection,11-dh-TXB_2,6-k-PGFla and diP ratio correlated with MACEs.4.ThedTPratiocouldreflect theplateletactivation better than other indices except for P-Selection in CAD patients.
PartⅡThe evaluation of platelet activity indices and the medicinal intervention study in hyperlipidemic rats
[objective]To analyze the association between various kind of platelet activation indices with the expression of CD62P on the platelet surface of hyperlipidemic rats,evaluate the change of platelet function in those rats.The effects of aspirin,atorvastatin,losartan,Fufang Danshen Diwan and Drostaglandin E_1 intervention on the platelet function were also investigated.
[Method]Seventy-five Wistar rats were divided into control group, hyperlipidemic group,aspirin treated group,atorvastatin treated group, losartan treated group,Fufang Danshen Diwan treated group and prostaglandin E_1,treated group.After four weeks treatment,the serum levels of blood lipid were measured.HE dyeing was used for the pathological section of liver,heart and aorta.ADP and AA induced MPAR were detected and Enzyme linked immunosorbent assay was used to detect the serum levels of P-Selectin,TXB_2,6-k-PGF1a.Flow cytometric analysis was applied to detect the expression of CD62P on the surface of platelet.
[Results]1.After four weeks,the serum TG,TC and LDL-C in the control group compared with those in the high fat feeding group was 0.35 vs 0.88mmol/L,1.92 vs 10.72mmol/L and 0.32 vs 8.66mmol/L respectively.2.The correlated coefficient of P-Selectin to CD62P was the largest(0.847) among all indices.3.After four weeks drug intervention,the levels of ADP and AA induced MPAR,TXB_2,6-k-PGF1a,TP ratio,P-Selectin and CD62P in each group were significantly different and results were showed in table 1.
[Conclusion]1.P-Selectin level could reflect the platelet activation better than than other indices except for CD62P in Wistar rats.2. Hyperlipidemia could increase platelet activation which could be significantly inhibited by aspirin.3.Atorvastatin can not only regulate dyslipidemia but also inhibit platelet activation remarkably.4.Losartan, Fufang Danshen Diwan and prostaglandin E_1 could inhibit platelet activation to some extend.
[目的]评价冠心病(CAD)患者阿司匹林抵抗(AR)、P选择素(P-Selectin)、11脱氢血栓素B_2(11-dh-TXB_2)、6酮前列腺素F1a(6-k-PGF1a)、11-dh-TXB_2/6-k-PGF1a(dTP)比值的影响因素及与CAD严重程度、心血管事件(MACE)的相关性。
[方法]203名住院患者规范应用阿司匹林、氯比格雷1周,记录临床特征、血液生化指标、冠状动脉造影(CAG)检查结果及1年MACE随访结果。应用二磷酸腺苷(ADP)、花生四烯酸(AA)诱导血小板最大聚集率(MPAR)评价AR,应用酶联免疫法测定血浆P-Selectin、11-dh-TXB_2和6-k-PGF1a水平,计算的dTP比值。
[结果]1.203例患者中CAG正常16例(7.9%)、稳定性心绞痛(SAP)22例(10.8%)、不稳定性心绞痛(UAP)115例(56.7%)、心肌梗死(AMI)50例(24.6%)。各组患者AR、P-Selectin、11-dh-TXB_2、6-k-PGF1a、dTP比值差别显著,结果见表1。2.各种血小板活化指标及CAG严重病变数的相关因素分析,结果见表2。3.随访1年55例(27.1%)患者发生MACE,11-dh-TXB_2、6-k-PGF1a、dTP值及P-Selectin水平与患者MACE发生相关,结果见表3。4。dTP比值与P-Selectin的相关系数最大(0.766)。
[结论]1.不同类型CAD患者AR、P-Selectin、11-dh-TXB_2、6-k-PGF1a、dTP比值差别显著。2.AR与临床合并糖尿病、hs-CRP及LDL-C水平相关。P-Selectin、年龄、hs-CRP水平与患者严重病变数相关。3.P-Selectin、11-dh-TXB_2、6-k-PGF1a、dTP比值与MACE发生相关。4.除了P-Selectin之外,TP比值能够较其它指标更好的反映CAD患者血小板活化的程度。
(二)高脂大鼠血小板活化指标评价与药物干预研究
[目的]分析高脂血症大鼠各种血小板活化指标与血小板表面CD62P表达的相关性,评价高脂血症大鼠血小板活化状态及阿司匹林、阿托伐他汀、氯沙坦、复方丹参滴丸、前列腺素E_1对于高脂血症大鼠血小板功能的影响。
[方法]Wistar大鼠75只随机分为正常对照组、高脂血症组、阿司匹林组、阿托伐他汀组、氯沙坦组、复方丹参滴丸组、前列腺素E_1注射液组。分组干预后检测各组大鼠的血脂水平.应用HE染色制作肝脏、心肌、主动脉病理切片。应用ADP、AA诱导检测MPAR,酶联免疫法测定血浆中P-Selectin、TXB_2、6-k-PGF1a浓度,计算的TP比值。应用流式细胞术检测血小板表面CD62P的表达。
[结果]1.Wistar大鼠分组喂养4周后,正常对照组与高脂饲料组TG、TC、LDL-C分别是0.35 vs 0.88mmol/L、1.92 vs 10.72mmol/L、0.32 vs 8.66mmol/L。2.P-Selectin与血小板表面CD62P表达水平相关系数最大(0.847)。3.药物干预4周后各组大鼠ADP、AA诱导的MPAR,血浆TXB_2、6-k-PGF1_a、TP比值、P-Selectin水平及血小板表面CD62P水平差别显著(P＜0.001),结果见表1。
[结论]1.Wistar大鼠血浆P-Selectin水平与血小板表面CD62P表达密切相关,能够较好的反映血小板活化程度。2.高脂血症能够促使血小板活化,阿司匹林能够明显抑制血小板活化。3.阿托伐他汀在调脂作用的同时显著抑制血小板活化。
4.科素亚、复方丹参滴丸、前列腺素E_1具有不同程度的抑制血小板活化作用。
PartⅠThe study of platelet activity inpatients with coronary artery disease
[objective]To evaluate the influential factors of aspirin resistance (AR),P-Selectin,11-dehydro-thromboxane B_2(11-dh-TXB_2),6-keto-prostag-landin F1 alpha(6-k-PGF1a) and the ratio of 11-dh-TXB_2 over 6-k-PGF1a (dTP) in patients with coronary artery disease(CAD),and to estimate the relationship of the platelet activity indices with the CAD severity and with the major adverse cardiovascular events(MACE).
[Method]203 patients were given one week regulationly treatment of aspirin and clopidogrel.Clinical features,blood biochemical indicators and coronary arteriography(CAG) were recorded,and the MACEs of these patients were followed up for one year.Maximum platelet agglutination rate(MPAR) induced by both ADP and AA was used to estimate AR.Enzyme linked immunosorbent assay was used to detect the serum levels of P-Selectin,TXB_2 and 6-k-PGF1a,then dTP ratio was calculated.
[Results]1.16 cases(7.9%) with normal CAG group were found in all 203 cases,22 cases(10.8%) for SAP group,115 cases(56.7%) for UAP group, 50 cases(24.6%) for AMI group.The diferences of AR,P-Selectin,11-dh-TXB_2, 6-k-PGF1a and dTP ratio between each groups were significant and resules were showed in table 1.2.Correlative factors of platelet activity indices and the number of severe affected(CAG stenosis≥75%) coronary artery(CA) were showed in table 2.3.After one-year follow-up,55 patients(27.1%) had MACEs.P-Selectin,11-dh-TXB_2,6-k-PGF1a and dTP ratio correlated to MACEs and resules were showed in table 3.4.The correlated coefficient of dTP to P-Selectin was the largest(0.766) among those indices.
[Conclusion]1.The differences of AR,P-Selectin,11-dh-TXB_2,6-k-PGF1a and dTP ratio in different type of CAD patients were significant.2. Diabetes,high level of hs-CRP and LDL-C were risk factors of AR.P-Selectin level,age,and hs-CRP level positively correlated to the number of severe affected CA.3.P-Selection,11-dh-TXB_2,6-k-PGFla and diP ratio correlated with MACEs.4.ThedTPratiocouldreflect theplateletactivation better than other indices except for P-Selection in CAD patients.
PartⅡThe evaluation of platelet activity indices and the medicinal intervention study in hyperlipidemic rats
[objective]To analyze the association between various kind of platelet activation indices with the expression of CD62P on the platelet surface of hyperlipidemic rats,evaluate the change of platelet function in those rats.The effects of aspirin,atorvastatin,losartan,Fufang Danshen Diwan and Drostaglandin E_1 intervention on the platelet function were also investigated.
[Method]Seventy-five Wistar rats were divided into control group, hyperlipidemic group,aspirin treated group,atorvastatin treated group, losartan treated group,Fufang Danshen Diwan treated group and prostaglandin E_1,treated group.After four weeks treatment,the serum levels of blood lipid were measured.HE dyeing was used for the pathological section of liver,heart and aorta.ADP and AA induced MPAR were detected and Enzyme linked immunosorbent assay was used to detect the serum levels of P-Selectin,TXB_2,6-k-PGF1a.Flow cytometric analysis was applied to detect the expression of CD62P on the surface of platelet.
[Results]1.After four weeks,the serum TG,TC and LDL-C in the control group compared with those in the high fat feeding group was 0.35 vs 0.88mmol/L,1.92 vs 10.72mmol/L and 0.32 vs 8.66mmol/L respectively.2.The correlated coefficient of P-Selectin to CD62P was the largest(0.847) among all indices.3.After four weeks drug intervention,the levels of ADP and AA induced MPAR,TXB_2,6-k-PGF1a,TP ratio,P-Selectin and CD62P in each group were significantly different and results were showed in table 1.
[Conclusion]1.P-Selectin level could reflect the platelet activation better than than other indices except for CD62P in Wistar rats.2. Hyperlipidemia could increase platelet activation which could be significantly inhibited by aspirin.3.Atorvastatin can not only regulate dyslipidemia but also inhibit platelet activation remarkably.4.Losartan, Fufang Danshen Diwan and prostaglandin E_1 could inhibit platelet activation to some extend.
引文
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