【摘要】：目的:探究人工合成的小窩蛋白1(caveolin-1,Cav-1)腳手架區(qū)多肽cavtratin對血紅素加氧酶1(heme oxygenase-1,HO-1)活性及脂多糖(lipopolysaccharide,LPS)誘導(dǎo)的小鼠急性肺損傷的作用。方法:成年雄性BALB/c小鼠隨機分為6組,每組8~10只,實驗分為對照(control)組、觸足肽內(nèi)化序列(Antennapedia internalization sequence,AP)組、LPS組、LPS+血晶素(hemin)組、LPS+hemin+cavtratin組和LPS+hemin+cavtratin+鋅原卟啉(zinc protoporphyrin IX,Zn PP)組。小鼠氣管滴注LPS 24 h后,蘇木素-伊紅染色觀察肺組織病理形態(tài)變化;檢測肺組織濕/干重比、肺泡灌洗液中細胞數(shù)和血清中乳酸脫氫酶活性。免疫熒光觀察HO-1和Cav-1的結(jié)合情況并檢測HO-1活性;實時熒光定量PCR檢測炎癥因子(IL-1β、IL-6、TNF-α、MCP-1和i NOS)的mRNA水平。結(jié)果:組織免疫熒光以及HO-1活性檢測發(fā)現(xiàn),與LPS組比較,LPS+hemin+cavtratin組HO-1與細胞膜Cav-1的結(jié)合減少,HO-1的活性增高(P0.05);與LPS組比較,LPS+hemin+cavtratin組肺組織受損程度明顯減輕,肺濕/干重比、肺泡灌洗液細胞數(shù)和血清中乳酸脫氫酶活性顯著降低(P0.05);與LPS組比較,LPS+hemin+cavtratin組炎癥因子的mRNA表達降低(P0.05);HO-1活性抑制劑Zn PP可以消除cavtratin的保護作用。結(jié)論:Cavtratin可以通過減少HO-1與細胞膜Cav-1的結(jié)合使得HO-1活性增加,進而減輕LPS誘導(dǎo)的小鼠急性肺損傷。
[Abstract]:Aim: to investigate the effects of synthetic scaffold polypeptide cavtratin of caveolin-1,Cav-1 on heme oxygenase-1 (heme oxygenase-1,HO-1) activity and lipopolysaccharide (lipopolysaccharide,LPS) -induced acute lung injury in mice. Methods: adult male BALB/c mice were randomly divided into 6 groups (n = 10): control (control) group, (Antennapedia internalization sequence,AP group, LPS hemin cavtratin group and LPS hemin cavtratin zinc-protoporphyrin (zinc protoporphyrin IX,Zn PP group. 24 h after LPS was infused into mouse trachea, histopathological changes of lung tissue were observed by hematoxylin eosin staining, lung wet / dry weight ratio, cell number in alveolar lavage fluid and lactate dehydrogenase activity in serum were measured. The binding of HO-1 and Cav-1 and the activity of HO-1 were observed by immunofluorescence, and the levels of IL-1 尾, IL-6,TNF- 偽, MCP-1 and i NOS) mRNA were detected by real-time fluorescence quantitative PCR. Results: compared with LPS group, the binding of HO-1 to cell membrane Cav-1 decreased and the activity of HO-1 increased in, LPS hemin cavtratin group (P0.05), compared with LPS group, the damage degree of lung tissue in, LPS hemin cavtratin group was significantly reduced and lung wet / dry weight ratio was significantly decreased (P < 0.05). The number of alveolar lavage fluid cells and the activity of lactate dehydrogenase in serum decreased significantly (P0.05); compared with LPS group, the mRNA expression of inflammatory factors in, LPS hemin cavtratin group was decreased (P0.05); Zn PP, the inhibitor of HO-1 activity, could eliminate the protective effect of cavtratin. Conclusion: Cavtratin can increase the activity of HO-1 by reducing the binding of HO-1 to cell membrane Cav-1 and then attenuate the acute lung injury induced by LPS in mice.
【作者單位】： 江南大學(xué)無錫醫(yī)學(xué)院;無錫市第三人民醫(yī)院急救中心;
【基金】：國家自然科學(xué)基金資助項目(No.81270126) 中央高�；究蒲袠I(yè)務(wù)費專項資金資助(No.JUSRP51412B) 江蘇省研究生培養(yǎng)創(chuàng)新工程項目(No.KYZZ16_0312;No.KYLX15_1196) 國家級大學(xué)生創(chuàng)新創(chuàng)業(yè)訓(xùn)練計劃項目(No.201610295074) 江蘇省醫(yī)學(xué)會/康緣藥業(yè)臨床醫(yī)學(xué)科研專項基金(No.2015JZKY07)
【分類號】：R563

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