發(fā)布時(shí)間：2019-05-27 02:42

【摘要】：目的： 探索microRNA-21在腎缺血再灌注損傷中作用機(jī)制,以期找到防治腎缺血再灌注損傷microRNA分子靶標(biāo)。 方法： 采用微型血管夾,同時(shí)夾閉雄性BALB/C小鼠雙側(cè)腎蒂構(gòu)建實(shí)驗(yàn)所需的腎缺血再灌注損傷模型，參照前期實(shí)驗(yàn)的結(jié)果和本次試驗(yàn)的目的將小鼠分為Pre-miR-21+IRI組（阻斷腎蒂血管45分鐘，，再灌注24h，于術(shù)前24小時(shí)和6小時(shí)分別按200ng/kg腹腔注射mir-21agomir）、PBS+IRI組（阻斷腎蒂血管45分鐘，再灌注24h于術(shù)前24小時(shí)和6小時(shí)分別腹腔注射等體積的PBS）、Pre-miR-21+shame組（僅打開腹腔不阻斷腎蒂血管，于術(shù)前24小時(shí)和6小時(shí)分別按200ng/kg腹腔注射mir-21agomir）、PBS+shame組（僅打開腹腔不阻斷腎蒂血管，于術(shù)前24小時(shí)和6小時(shí)分別腹腔注射等體積PBS），（n≥5）。四組小鼠缺血再灌注損傷后，收集小鼠血液檢測血清肌酐、尿素氮判斷腎功能。斷頸法處死小鼠，從小鼠背部取一手術(shù)切口獲取雙側(cè)模型腎臟。一側(cè)腎臟行HE染色，光學(xué)顯微鏡下觀察模型腎病理組織變化，另一側(cè)腎臟檢測模型腎中microRNA-21，PDCD4、PTEN基因的mRNA和caspase-3蛋白的表達(dá)。 結(jié)果： （1）Pre-miR-21+IRI、PBS+IRI組、Pre-miR-21+shame組和PBS+shame組組小鼠存活率分別為87%、75%、100%、100%。Pre-miR-21+IRI組中血清肌酐、尿素氮和腎組織病理評分明顯低于PBS+IRI組，但上述兩組均高于PBS+shame組和Pre-miR-21+shame（P＜0.05）；PBS+shame組與Pre-miR-21+shame組腎組織病理評分和血清肌酐、尿素氮無明顯差異(P＞0.05)。 （2）四組小鼠中miR-21的表達(dá)存在顯著差異，從高到低的排列順序?yàn)镻re-miR-21+IRI、Pre-miR-21+shame組、PBS+IRI組、PBS+shame組(P＜0.05)；Pre-miR-21+IRI組中靶基因PDCD4、PTEN的mRNA表達(dá)量低于PBS+IRI組（P＜0.05）；PBS+IRI組中靶基因PDCD4、PTEN的mRNA表達(dá)量低于PBS+shame組（P＜0.05）；PBS+shame組與Pre-miR-21+shame組兩組之間靶基因PDCD4、PTEN的mRNA表達(dá)量無明顯差異(P＞0.05)。 （3）、Pre-miR-21+IRI組中cleaved caspase-3表達(dá)明顯低于PBS+IRI組（P＜0.05），PBS+IRI組中cleaved caspase-3高于PBS+shame組和Pre-miR-21+shame組（P＜0.05），差異均有統(tǒng)計(jì)學(xué)意義。各組中未經(jīng)激活的pro-caspase-3無明顯差異，P＞0.05無統(tǒng)計(jì)學(xué)意義。 結(jié)論： （1）成功構(gòu)建了雄性BALB/c小鼠腎臟中mir-21過表達(dá)模型和腎缺血再灌注損傷模型； （2）mir-21能對其靶基因PDCD4和PTEN進(jìn)行負(fù)向調(diào)控； （3）在腎缺血再灌注損傷的環(huán)境中mir-21能通過對其靶基因PDCD4和PTEN的負(fù)向調(diào)控，抑制腎缺血再灌注損傷所造成凋亡，從而對腎臟起到一定的保護(hù)作用。
[Abstract]:Objective: to explore the mechanism of microRNA-21 in renal ischemia-reperfusion injury in order to find a molecular target for the prevention and treatment of renal ischemia-reperfusion injury. Methods: the model of renal ischemia-reperfusion injury was established by clamping bilateral renal pedicle of male BALB/C mice with microvascular clip. According to the results of the previous experiment and the purpose of this experiment, the mice were divided into Pre-miR-21 IRI group (blocking renal pedicle vessels for 45 minutes, reperfusing them for 24 hours, injecting mir-21agomir intraperitoneally according to 200ng/kg 24 hours and 6 hours before operation). PBS IRI group (blocking renal pedicle vessels for 45 minutes, 24 hours after reperfusion, 24 hours before operation and 6 hours before operation, respectively, intraabdominal injection of the same volume of PBS), Pre-miR-21 shame group (only open abdominal cavity did not block renal pedicle vessels, Mir-21agomir), PBS shame group was injected intraperitoneally with 200ng/kg 24 hours and 6 hours before operation (only open abdominal cavity did not block renal pedicle vessels, and PBS), (n 鈮

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