骨髓間充質干細胞對小鼠肺臟缺血再灌注損傷的影響
發(fā)布時間:2018-06-23 來源: 感悟愛情 點擊:
[摘要]目的 探討骨髓間充質干細胞對小鼠肺臟缺血再灌注損傷的影響。方法 采用全骨髓培養(yǎng)法分離純化小鼠骨髓間充質干細胞(BMSCs),結扎左側肺門60 min,再灌注240 min制備小鼠肺臟缺血再灌注損傷模型。70只小鼠隨機分為7組(n=10),包括:假手術組(Sham組)、缺血再灌注組(I/R組)、BMSCs 1組(尾靜脈注射1×106個BMSCs)、BMSCs 2組(尾靜脈注射2×106個BMSCs)、BMSCs 5組(尾靜脈注射5×106個BMSCs)、BMSCs 10組(尾靜脈注射10×106個BMSCs)、BMSCs it組(氣管內注射5×106個BMSCs)。再灌注結束時抽取小鼠股動脈血,行血氣分析,并計算氧合指數(shù)(PaO2/FiO2)。隨后處死小鼠,取肺組織,采用ELISA法檢測定肺組織中超氧化物歧化酶(SOD)的活性、白細胞介素(interleukin,IL)-8、腫瘤壞死因子(TNF-α)含量及髓過氧化物酶(MPO)的活性,光鏡下觀察肺組織損傷程度并進行病理學評分。結果 與Sham組比較,I/R組、BMSCs 1組、BMSCs 2組、BMSCs 5組、BMSCs 10組、BMSCs it組的PaO2/FiO2降低,肺組織病理學評分、MPO活性、IL-8、TNF-α含量升高,SOD活性降低,差異有統(tǒng)計學意義(P<0.05)。與I/R組比較,BMSCs 1組、BMSCs 2組、BMSCs 5組、BMSCs 10組、BMSCs it組的PaO2/FiO2升高,肺組織病理學評分、MPO、IL-8、TNF-α水平降低,SOD活性升高,差異有統(tǒng)計學意義(P<0.05);其中BMSCs 10組最明顯,差異有統(tǒng)計學意義(P<0.05),氣管內注射與血管內注射相同濃度BMSCs作用無明顯區(qū)別,差異無統(tǒng)計學意義(P>0.05)。結論 BMSCs能夠明顯減輕肺組織缺血再灌注損傷,其作用機制與調節(jié)肺組織炎癥反應有關,且具有明顯量效關系,氣管內注射與血管內注射相同濃度BMSCs作用無明顯區(qū)別。
[關鍵詞]骨髓間充質干細胞;肺;再灌注損傷;氧自由基
[中圖分類號] R332 [文獻標識碼] A [文章編號] 1674-4721(2018)2(b)-0018-04
Influence of bone marrow mesenchymal stem cells on lung ischemia-reperfusion injury in mice
SUN Chang-rong1,2 HOU Nian-guo1 LI Hui1 SHUAI Xun-jun1 SUN Ming-jie1 TANG Yu-ru1 XU Tang-wen1,2 AI Deng-bin1▲
1.Department of Anesthesiology,Weifang Medical College,Shandong Province,Weifang 261053,China;2.Department of Anesthesiology,Research Center of Clinical Anesthesiology of Qingdao City,Qingdao Municipal Hospital,Shandong Province,Qingdao 266011,China
[Abstract]Objective To investigate the influence of bone marrow mesenchymal stem cells (BMSCs) on lung ischemia-reperfusion injury (LIRI) in mice.Methods BMSCs from rats were isolated,cultured and purified by the whole bone marrow acherence method.LIRI was produced by occlusion of the left hilum of lungs for 60 min followed by 240 min reperfusion.70 mice were randomly divided into 7 groups (n=10),included the sham operation group (the Sham group),the ischemia-reperfusion group (the I/R group),the BMSCs 1 group (tail vein injections of 1×106 BMSCs),the BMSCs 2 group (tail vein injections of 2×106 BMSCs),the BMSCs 5 group (tail vein injection of 5×106 BMSCs),the BMSCs 10 group (tail vein injection of 10×106 BMSCs),the BMSCs it group (endotracheal injection of 5×106 BMSCs).At the end of reperfusion,blood samples were obtained for blood gas analysis and PaO2/FiO2 was calculated.Then,the animals were sacrificed and lung tissues were immediately removed for determination of superoxide dismutase (SOD),tumor necrosis factor-α (TNF-α),interleukin-8 (IL-8),and myeloperoxidase (MPO) and for microscopic examination of the pathological changes of lungs which were scored.Results Compared with Sham group,PaO2/FiO2 was significantly decreased,the pathological scores and the levels of TNF-α,IL-8,and MPO were increased,and SOD activity was decreased in I/R,BMSCs 1,BMSCs 2,BMSCs 5,BMSCs 10,BMSCs it group,the difference was statistically significant (P<0.05).Compared with I/R group,PaO2/FiO2 was significantly increased,the pathological scores and the levels of TNF-α,IL-8,and MPO in lung tissues were decreased,and SOD activity was increased in BMSCs 1,BMSCs 2,BMSCs 5,BMSCs 10,BMSCs it group,the difference was statistically significant (P<0.05).The most obvious group is BMSCs 10,the difference was statistically significant (P<0.05).There is no obvious difference between endotracheal injection and intravascular injection with the same concentration of BMSCs,the difference was not statistically significant (P>0.05).Conclusion BMCS can alleviate LIRI in mice,and the responsible mechanism is related to inhibition of inflammatory responses.The lung protective effect is dose dependent.There is no significant difference between endotracheal injection and intravascular injection with the same concentration of BMSCs.
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