【摘要】：【背景】酒精飲品在日常生活中最流行的飲品之一。然而,酗酒已成為世界各國(guó)共同的社會(huì)問(wèn)題。酒精濫用導(dǎo)致嚴(yán)重的臨床事件的發(fā)生,如血小板減少,出血,但發(fā)病機(jī)制尚不完全清楚�！痉椒ā渴占�18-25歲、兩周內(nèi)未服用任何藥物的健康志愿者的空腹新鮮靜脈全血,分離得到洗滌血小板及富含血小板血漿(Platelet rich plasma,PRP)。洗滌血小板(3×108/ml)與不同濃度酒精或者陰性對(duì)照二甲基亞砜(Dimethyl sulfoxide,DMSO)于37℃孵育2小時(shí),使用流式細(xì)胞儀檢測(cè)血小板線粒體跨膜電位(Mitochondrial inner transmembrane potential,ΔΨm)變化、血小板活化指標(biāo)P-選擇素(P-Selectin)及血小板激活復(fù)合物(Patelet activated complex-1,PAC-1)的結(jié)合率。使用Western blot檢測(cè)酒精與正常人洗滌血小板孵育后血小板促凋亡蛋白Bax,抗凋亡蛋白Bcl-2的表達(dá)和caspase-3的活化情況。為了探明c-Jun氨基末端激酶(c-jun NH2-terminal kinase,JNK)蛋白家族在酒精誘導(dǎo)的血小板凋亡過(guò)程中的作用,我們使用Western blot檢測(cè)JNK蛋白家族磷酸化表達(dá)水平的變化。同時(shí)我們研究了酒精對(duì)不同致聚劑誘導(dǎo)血小板聚集功能的影響,探討酒精對(duì)血小板功能有無(wú)損傷,將PRP與酒精(200 mg/dl)或者陰性對(duì)照(DMSO)于37℃孵育2小時(shí),洗滌血小板與酒精(200 mg/dl)或陰性對(duì)照(DMSO)于37℃孵育2小時(shí)后,PRP中加入二磷酸腺苷(Adenosine diphosphate,ADP,10μmol/L),洗滌血小板中加入凝血酶(Thrombin,0.5 U/m L)或者膠原(collagen,5μg/m L),1000 rpm剪切力的情況下,使用血小板聚集儀檢測(cè)血小板聚集程度。在體內(nèi)實(shí)驗(yàn)中,對(duì)C57BL/6J小鼠進(jìn)行灌胃,之后使用Sysmex KX-21N血細(xì)胞計(jì)數(shù)儀測(cè)定小鼠血小板計(jì)數(shù),檢測(cè)尾出血時(shí)間以及胃粘膜出血情況�！窘Y(jié)果】乙醇呈劑量依賴性誘導(dǎo)線粒體內(nèi)跨膜電位的去極化,上調(diào)Bax蛋白的表達(dá)量和Bcl-2的表達(dá)量的下調(diào),和caspase-3的活化。乙醇不誘導(dǎo)P-選擇素或PAC-1的表達(dá),并明顯降低膠原,凝血酶,和ADP誘導(dǎo)的血小板聚集。此外,酒精誘導(dǎo)JNK凋亡通路的激活。在急性酒精中毒的體內(nèi)小鼠模型中,乙醇明顯降低循環(huán)血小板數(shù),延長(zhǎng)尾出血時(shí)間,并造成胃粘膜出血�！窘Y(jié)論】這些數(shù)據(jù)表明,乙醇導(dǎo)致線粒體介導(dǎo)的血小板的細(xì)胞凋亡,導(dǎo)致循環(huán)血小板數(shù)量的減少和體內(nèi)止血機(jī)制損害。這些發(fā)現(xiàn)揭示急性酒精中毒患者出血現(xiàn)象的可能發(fā)病機(jī)制。
[Abstract]:Background: alcohol is one of the most popular drinks in daily life. However, alcoholism has become a common social problem all over the world. Alcohol abuse leads to serious clinical events, such as thrombocytopenia and bleeding, but the pathogenesis is not entirely clear. [methods] fasting fresh venous whole blood was collected from healthy volunteers aged 18-25 years who had not taken any drugs within two weeks. Washing platelets and platelet-rich plasma (Platelet rich plasma,PRP were isolated. Washed platelets (3 脳 108/ml) were incubated with dimethyl sulfoxide (Dimethyl sulfoxide,DMSO) at 37 鈩，

本文編號(hào)：2387979