Adenosine triphosphate (ATP) is the most abundant and primary carrier of the required energy for various functions in cells. Prolonged ischemia, reperfusion, anaerobic metabolism and lactate accumulation can lead to a dramatic decrease of ATP, cell swelling, cell rupture, and finally cell death by necrotic, necroptotic, apoptotic, and autophagic mechanisms. Due to drastic hydrolysis of ATP in vivo by ectoenzymes and poor cellular penetration, the direct delivery of ATP to the ischemic tissues is difficult.
To increase delivery of ATP to the tissues and protect from enzymatic degradation, encapsulation in liposomes has been proposed and demonstrated in various models of ischemia [1,2]. Studies on myocardial [1,3,4], liver [5-8], retina [9] and wound healing [10-12] ischemia have shown the ability of liposomal encapsulated ATP to prevent cell death and tissue dysfunction following ischemic events.
The encapsulation of ATP in liposomes markedly promotes its effectiveness by preventing the hydrolysis by extracellular enzymes, increasing ATP circulation time and enhancing its intracellular penetration. Depending on the type of the cell line and the target organ various types of liposomes with different surface charges such as anionic, cationic and neutral has been studied by various groups. Moreover, ATP encapsulated liposomes has been demonstrated to improve energy state and function of the cold-stored liver [6,7,13].
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加尾法是采用加A酶先对mirna进行加尾,然后再用带oligodt的引物反转录,他的反转录引物是通用的,一次反转录可以获得所有miRNA的cdna,效率高。茎环法是采用特异性反转录引物序列+颈环结构作为反转录引物进行反转录的,一次反转录只能获得一种mirna的cdna,可能有几种一起反转录的,这个我不太清楚一起反转录的效果。然后茎环法的经典即ABI的探针法定量试剂盒。我用过茎环法反转录+染料法定量的,下游引物为通用引物,结果不太好,就现在用的这家的他采用的是双向引物都是特异性的,效果还可以。不知道我说的清楚不,希望能对你有帮助。
如果是用转录出的cDNA做那跟平时就一样了。不需要特别的试剂盒、

