子宫内膜再生细胞通过表达PD-L1对大鼠肠道缺血再灌注损伤的抑制作用观察  

Inhibition of endometrial regenerative cells on intestinal ischemia reperfusion injury in rats by expressing PD-L1

作　　者：叶奎[1] 彭秋凤[1] 史刚刚[2] 曲鑫[1] 王浩[3,4] YE Kui;PENG Qiufeng;SHI Ganggang;QU Xin;WANG Hao(Department of Vascular Surgery,The Fourth Central Hospital of Tianjin,Tianjin 300142,China;不详)

机构地区：[1]天津市第四中心医院血管外科,天津300142 [2]天津医科大学第二医院肛肠外科 [3]天津医科大学总医院普通外科 [4]天津普通外科研究所

出　　处：《山东医药》2022年第11期5-8,共4页Shandong Medical Journal

基　　金：国家自然科学基金面上项目(82071802);天津市自然科学基金重点项目(18JCZDJC35800);天津市卫生健康委员会科技项目(ZC20106,ZC20098,ZC20154);天津市教委科研计划项目(2020KJ175);天津市第四中心医院优秀青年基金资助项目(tjdszxyy20210009)。

摘　　要：目的观察子宫内膜再生细胞(ERC)通过表达程序性死亡配体1(PD-L1)对大鼠肠道缺血再灌注损伤(IRI)的抑制作用。方法月事杯收集健康育龄期女性志愿者的月经血,利用标准Ficoll密度梯度离心获取血中单个核细胞,培养后获得ERC。将50只Wistar大鼠随机分为假手术组、模型组、PD-L1过表达ERC组、ERC治疗组、PD-L1低表达ERC组,每组10只。除假手术组外,均利用肠系膜上动脉结扎30 min后恢复血供的方法建立肠道IRI大鼠模型,假手术组开腹后分离肠系膜上动脉但不结扎。建模后,PD-L1过表达ERC组、ERC治疗组和PD-L1低表达ERC组分别通过尾静脉注射的方法,给予5 ng/mL重组人干扰素γ预刺激72 h后的ERC、普通ERC、10μg/mL抗人PD-L1抗体作用72 h后的ERC各5×10^(6)个。72 h后处死各组大鼠,收集肠道组织和血清标本,HE染色后评价肠道组织损伤情况,ELISA法检测血清肠道屏障功能指标二胺氧化酶(DAO)、D-乳酸(D-Lac)及炎症相关因子白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)水平。结果假手术组、PD-L1过表达ERC组、ERC治疗组、PD-L1低表达ERC组、模型组肠道组织损伤评分分别为(0.500±0.534)、(2.000±0.535)、(2.875±0.641)、(3.750±0.463)、(4.125±0.641)分,组间两两比较P均<0.05。假手术组、PD-L1过表达ERC组、ERC治疗组、PD-L1低表达ERC组、模型组血清D-Lac、DAO、TNF-α、IL-6、IL-1β水平均依次升高,组间两两比较P均<0.05。结论ERC能够通过表达PDL1而减轻大鼠肠道IRI损伤,其机制可能与减轻肠道屏障功能损伤及炎症反应有关。Objective To observe the inhibitory effect of endometrial regenerative cells(ERC)on intestinal ischemia reperfusion injury(IRI)in rats by expressing programmed death ligand 1(PD-L1).Methods Menstrual blood was collected from healthy female volunteers using a menstrual cup,and mononuclear cells were obtained through standard Ficoll density gradient centrifugation,and cells were cultured to obtain the ERC.Fifty rats were randomly divided into five groups:sham group(we performed operation but not ligation),untreated group,ERC overexpressing PD-L1 treatment group(5 ng/mL IFN-γ stimulated ERC for 72 h),ERC treatment group(ERC),and ERC low expressing PD-L1 treatment group(10μg/mL anti-PD-L1 treated ERC for 72 h).Except for the sham group,the rats’superior mesenteric arteries were ligated for an hour and blood supply was restored to construct the experimental intestinal IRI models in the other groups.After 72 h,the rats in each group were sacrificed,and the intestinal tissues and serum samples were collected,and the intestinal tissue injury was evaluated after HE staining.ELISA was used to determine the levels of intestinal barrier function markers diamine oxidase(DAO)and D-lactate(D-Lac),and inflammatory cytokines interleukin 1β(IL-1β),tumor necrosis factorα(TNF-α)and interleukin 6(IL-6)in the serum.Results Histological scores in the sham group,ERC overexpressing PD-L1 treatment group,ERC treatment group,ERC low expressing PD-L1 treatment group,and untreated group were(0.500±0.534),(2.000±0.535),(2.875±0.641),(3.750±0.463),and(4.125±0.641)points,respectively,with statistically significant difference between any two groups(all P<0.05).Levels of D-Lac,DAO,TNF-α,IL-6 and IL-1β in the serum of the sham group,ERC overexpressing PD-L1 treatment group,ERC treatment group,ERC low expressing PD-L1 treatment group,and untreated group increased in sequence,with statistically significant difference between any two groups(all P<0.05).Conclusion ERC can alleviate intestinal IRI through PD-L1 expression probably by dec

关 键 词：子宫内膜再生细胞 肠道缺血再灌注损伤 程序性死亡配体1 肠道屏障功能 炎症反应 大鼠 

分 类 号：R459.9[医药卫生—治疗学]