抑制AKR1B1表达通过调控巨噬细胞极化改善大鼠脓毒症所致急性肺损伤  

Inhibition of AKR1B1 alleviates sepsis-induced acute lung injury in rats by regulating macrophage polarization

作　　者：董岩[1] 贾依娜尔[1] 吐尔逊古丽·麦麦提 阿尔申·木拉提 潘志威 杨立新[1] DONG Yan;JIA Yinaer;Tuersunguri Maimaiti(Department of Critical Care Medicine,the Third Affiliated Cancer Hospital of Xinjiang Medical University,Xinjiang,Urumqi 830011,China)

机构地区：[1]新疆医科大学第三附属肿瘤医院重症医学科,乌鲁木齐市830011

出　　处：《河北医药》2024年第15期2245-2250,共6页Hebei Medical Journal

基　　金：新疆维吾尔自治区自然科学基金项目(编号:2022D01C307)。

摘　　要：目的 探究抑制醛酮还原酶1成员B1(AKR1B1)对脓毒症所致急性肺损伤(ALI)模型大鼠的影响及作用机制。方法 将60只SD大鼠按随机数字法分为对照组、模型组、低剂量组、高剂量组,每组15只,除对照组的其余3组大鼠以盲肠结扎穿刺法制备脓毒症ALI模型,低、高剂量组大鼠分别按照50 mg/kg、100 mg/kg灌胃依帕司他,1次/d,连续6周。6周后,免疫组织化学染色检测对照组和模型组大鼠肺组织内AKR1B1表达;ELISA法测定4组大鼠支气管肺泡灌洗液(BALF)和血清中炎性细胞因子肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)及IL-6水平;天平称量4组大鼠左侧肺组织湿质量(WW)与干质量(DW),计算WW/DW比值;苏木精-伊红(HE)染色法观察4组大鼠肺组织病理形态变化,TUNEL染色检测4组大鼠肺组织细胞凋亡情况;流式细胞分析术检测4组大鼠BALF中M1型(F4/80^(+)CD86^(+))与M2型(F4/80^(+)CD206+)巨噬细胞表达情况;实时荧光定量PCR测定4组大鼠肺组织内M1型巨噬细胞标志基因与M2型巨噬细胞标志基因的表达水平。结果 模型组大鼠肺组织内AKR1B1表达较对照组增加(P<0.05);与模型组比较,低、高剂量组大鼠血清与BALF中TNF-α、IL-1β及IL-6含量均降低(P<0.05),右侧肺组织WW/DW值下降(P<0.05),肺组织病理损伤得到明显改善,炎症细胞浸润减少,TUNEL阳性细胞率减少(P<0.05),BALF中F4/80^(+)CD86^(+)阳性表达率降低、(F4/80^(+)CD206+)阳性表达率升高(P<0.05),肺组织内M1型巨噬细胞标志基因TNF-α、iNOS、IL-6的mRNA相对表达量均下调(P<0.05),而M2型巨噬细胞标志基因TGF-β、Arg-1、IL-10的mRNA相对表达量均上调(P<0.05)。结论 AKR1B1在脓毒症所致ALI大鼠肺组织中表达升高,而抑制AKR1B1通过抑制巨噬细胞向M1型转化并促进向M2型转化来减轻大鼠肺损伤。Objective To explore the effect of inhibition of aldo-ketone reductase 1 member B1(AKR1B1) on sepsis-induced acute lung injury(ALI) in rats and the underlying mechanism.Methods A total of 60 Sprague-Dawley(SD) rats were randomly assigned into control group,model group,low-dose group,and high-dose group,with 15 rats in each group.Except for the control group,rats in the other three groups were used to establish sepsis-induced ALI model by cecal ligation and puncture.Rats in the low-dose and high-dose groups were given epalrestat by gavage at 50mg/kg and 100mg/kg,respectively,once a day for 6 consecutive weeks.After 6 weeks,immunohistochemical staining was used to detect the expression of AKR1B1 in the lung tissue of the control and model groups.Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of inflammatory cytokines tumor necrosis factor α(TNF-α),interleukin 1β(IL-1β) and interleukin 6(IL-6) in bronchoalveolar lavage fluid(BALF) and serum.The wet weight(WW) and dry weight(DW) of the left lung tissue were weighed,and the WW/DW ratio was calculated.Hematoxylin-eosin(HE) staining was used to observe the pathological morphologic changes in lung tissue.The terminal deoxynucleotidyl transferase dUTP nick end-labeling(TUNEL) staining was used to detect the apoptosis of lung tissue cells.Flow cytometry was used to detect the expressions of M1(F4/80^(+)CD86^(+)) and M2(F4/80^(+)CD206+) macrophages in rat bronchoalveolar lavage fluid(BALF).Real-time fluorescence quantitative PCR was used to detect the expression levels of M1 and M2 markers in rat lung tissue.Results The expression of AKR1B1 in the lung tissue of the model group was significantly higher than that of the control group(P<0.05).Compared with those of the model group,TNF-α,IL-1β and IL-6 levels in serum and BALF were significantly lower in the low-dose and high-dose groups after epalrestat treatment(P<0.05).The WW/DW ratio of right lung tissue was lower in the low-dose and high-dose groups than that of control group(P<0.05).Low-dose

关 键 词：醛酮还原酶 脓毒症 急性肺损伤 巨噬细胞极化 

分 类 号：R631[医药卫生—外科学]