2012年1月19日,，據(jù)《每日科學(xué)》報道,，一種在血小板上發(fā)現(xiàn)的受體,，其作為潛在藥物靶標(biāo)的重要性一直飽受質(zhì)疑,，實際上在藥物測試中可能是卓有成就的，根據(jù)美國密歇根州立大學(xué)的一項新研究,。
一個由密歇根州立大學(xué)化學(xué)系Dana Spence領(lǐng)導(dǎo)的團隊揭示了一種分離及測試P2X1受體的方法,。通過創(chuàng)建一種新的、簡單的方法來研究它,，研究團隊已解鎖了一個許多影響紅細(xì)胞的疾?。ㄈ缣悄虿?，高血壓和囊性纖維變性）潛在的新藥物靶標(biāo),。
研究人員不僅可以從開發(fā)新藥物方面評估受體,，從重新測試現(xiàn)有藥物附加到受體起作用方面也可以對受體進行評估。
"科學(xué)家們一直在人體內(nèi)尋找新的'可藥性（druggable）'受體,，"Spence說,。"P2X1受體,，長久以來一直被視為在血小板中不重要；我們的研究表明,，這不一定是對的。受體是非?；钴S的,，你需要小心的使用它。"
這項研究發(fā)表在本期的《分析方法》（Analytical Methods）上,。
血小板的主要工作是通過凝結(jié)幫助防止出血,，Spence說,。它們通過增加血液的粘稠,、阻止正常的血流及阻塞血管來工作，但有些疾病如糖尿病或鐮狀細(xì)胞貧血的問題是血小板變得粘稠--甚至在它們不該粘稠的時候,。
當(dāng)血小板的受體打開時,，血小板就被激活了，目前研究人員一直將研究重點放在P2Y受體上,，這很容易研究,。另一方面，并沒有想到P2X1受體在血小板激活中發(fā)揮了重要的作用,，而且它已被證明非常麻煩,，因為一旦血液從身體抽出它就變得不再敏感了。
盡管科學(xué)家嘗試了一對方法來解決這個問題--通過使用不同的添加劑或酶，但結(jié)果在研究受體中并沒有卓越的成效,。
Spence和他的團隊發(fā)現(xiàn)的是,，通過添加一種簡單的被稱為NF449（最初認(rèn)為能夠阻斷受體）的分子，他們在抽血后能夠激活血小板中的P2X1受體,。
"我們已經(jīng)發(fā)現(xiàn)了一種制備和處理血小板的方法,，使我們能夠真實可靠地研究受體，"他說,。"這項研究開辟了新的研究途徑,，將允許研究人員和制藥公司重新評估這種受體作為'可藥性'的靶標(biāo),。"（生物谷bioon.com）
doi:10.1039/C1AY05530E
PMC:
PMID:
Measuring P2X1 receptor activity in washed platelets in the absence of exogenous apyrase
Kari B. Anderson, Welivitiya Karunarathne, Dana M. Spence
Abstract： Purinergic receptor signaling events in platelets are a major determinant in platelet function. However, investigating the ATP-sensitive P2X1 platelet receptor is difficult due to its rapid desensitization in the washed platelet sample matrix. To minimize desensitization, most studies involving P2X1 activity in washed platelets require apyrase in the sample to reduce matrix ATP levels. Unfortunately, the apyrase will also rapidly degrade any ATP added exogenously during the studies. Here, we describe a method that employs the reported P2X1 inhibitor NF449 to sensitize washed platelets in the absence of any added apyrase. Sensitization is verified by spectrofluorometric determination of Ca2+ entry into the platelets after stimulation with concentrations of ATP ranging from 0.625 μM to 5 μM. Results suggest that sensitization of the P2X1 receptor by NF449 is not necessarily dependent upon the inhibitor concentration, but rather the ratio of the inhibitor to exogenously-added ATP concentrations. With a ratio of ATP agonist to NF440 concentration of 5 : 1, the resulting percent change in fluorescence due to Ca2+ entry into the platelet is 39.3 ± 0.8%; however, at a ratio of 1 : 8 ATP to NF449 the percent change is reduced to 13.1 ± 2.2%. The sensitizing effect is also investigated as a function of time. The results obtained verify that NF449 can behave as a concentration-dependent inhibitor and sensitizer of the platelet P2X1 receptor in washed platelet samples, depending on the ATP concentration in the matrix.
