[1]姬改娜,王彩琳.乌司他丁通过AMPK/Nrf2通路对急性肺损伤新生大鼠肺组织保护作用的实验研究[J].陕西医学杂志,2022,51(11):1323-1327.[doi:DOI:10.3969/j.issn.1000-7377.2022.11.001]
　JI Gaina,WANG Cailin.Protective effect of ulinastatin on lung tissue of neonatal rats with acute lung injury via AMPK/Nrf2 pathway[J].,2022,51(11):1323-1327.[doi:DOI:10.3969/j.issn.1000-7377.2022.11.001]
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乌司他丁通过AMPK/Nrf2通路对急性肺损伤新生大鼠肺组织保护作用的实验研究
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
51
期数:
2022年11期
页码:
1323-1327
栏目:
基础研究
出版日期:
2022-11-05

文章信息/Info

Title:
Protective effect of ulinastatin on lung tissue of neonatal rats with acute lung injury via AMPK/Nrf2 pathway
作者:
姬改娜; 王彩琳
(榆林市第二医院,陕西 榆林 719000)
Author(s):
JI Gaina; WANG Cailin
(Yulin No.2 Hospital,Yulin 719000,China)
关键词:
乌司他丁;  AMPK/Nrf2通路;  高氧;  急性肺损伤;  腺苷酸活化蛋白激酶;  核因子E2相关因子2
Keywords:
Ulinastatin;  AMPK/Nrf2 pathway;  Hyperoxia;  Acute lung injury;  AMP-activated kinase;  Nuclear factor erythroid-2 related factor 2
分类号:
R 563.9
DOI:
DOI:10.3969/j.issn.1000-7377.2022.11.001
文献标志码:
A
摘要:
目的:观察乌司他丁(UTI)通过腺苷酸活化蛋白激酶(AMPK)/核因子E2相关因子2(Nrf2)通路对高氧诱导的急性肺损伤(ALI)新生鼠的肺组织保护作用,并探究其可能的作用机制。方法:选取SPF级足月新生SD大鼠30只,随机分为对照组、ALI组和ALI+UTI组,每组10只。ALI组和ALI+UTI组大鼠置于高氧中建立ALI模型,ALI+UTI组大鼠腹腔注射10 万U/kg的UTI 0.9%氯化钠溶液,对照组和ALI组腹腔注射等体积的0.9%氯化钠溶液。处死各组大鼠,苏木精-伊红(HE)染色观察肺组织病理学变化,计算肺组织湿干(W/D)比,采集支气管肺泡灌洗液(BALF),检测其中炎症因子的表达水平,检测肺组织中活性氧(ROS)、超氧化物歧化酶(SOD)和丙二醛(MDA)的表达水平,RT-PCR法检测肺组织中相关基因mRNA的表达,Western blot检测其中相关蛋白的表达。结果:HE染色可见ALI组大鼠肺组织结构紊乱无规则,存在大量纤维渗出和炎性细胞浸润,而ALI+UTI组大鼠肺组织损伤程度明显改善。ALI组和ALI+UTI组大鼠肺W/D比高于对照组(均P<0.05),ALI+UTI组大鼠的肺W/D比低于ALI组(P<0.05)。ALI组和ALI+UTI大鼠BALF中肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)和白介素-6(IL-6)的表达水平均高于对照组(均P<0.05),ALI+UTI大鼠BALF中TNF-α、IL-1β和IL-6的表达水平低于ALI组(均P<0.05)。与对照组相比,ALI组和ALI+UTI组大鼠肺组织中ROS和MDA的表达显著升高、SOD的表达显著降低(均P<0.05); 与ALI组相比,ALI+UTI组大鼠肺组织中ROS和MDA的表达显著降低、SOD的表达显著升高(均P<0.05)。ALI+UTI组大鼠肺组织中Nrf2及其下游基因血红素加氧酶1(HO-1)和醌氧化还原酶1(NQO-1)mRNA的相对表达均高于对照组和ALI组(均P<0.05)。对照组和ALI组大鼠肺组织中pAMPK/AMPK、Nrf2蛋白表达比较无统计学差异(均P>0.05); ALI+UTI组大鼠肺组织中pAMPK/AMPK、Nrf2蛋白表达高于对照组和ALI组(均P<0.05)。结论:UTI可通过AMPK/Nrf2通路,激活AMPK磷酸化而增强Nrf2及其下游基因的活性,有效缓解高氧诱导的ALI,UTI可作为临床治疗ALI的备选药物之一。
Abstract:
Objective:To observe the protective effect of ulinastatin(UTI)on lung tissue of neonatal rats with hyperoxia-induced acute lung injury(ALI)through AMPK/Nrf2 pathway,and to explore its possible mechanism.Methods:A total of 30 SPF grade SD neonatal rats were randomly divided into control group,ALI group and ALI+UTI group,with 10 rats in each group.The ALI group and ALI+UTI group were placed into a high oxygen culture to establish the ALI model,and the ALI+UTI group was intraperitoneal injected with 100000 U/kg UTI solution,while the control group and ALI group was intraperitoneal injected with isovolumetric 0.9% sodium chloride solution.All rats were sacrificed,the pulmonary pathology were observed by HE staining.The lung wet/dry(W/D)ratio was calculated.The broncho alveolar lavage fluid(BALF)was collected and inflammatory factors were dectected.The expression levels of oxygen species(ROS),superoxide dismutase(SOD)and malondialdehyde(MDA)in lung tissues were detected.The mRNA expression of related genes in lung tissue was detected by RT-PCR,and the expression of related proteins was detected by Western blot.Results:HE staining showed that the lung tissue of ALI group was irregular and disorder,much fiberours and inflammatory cell infiltration were observed,while the damage degree of ALI+UTI group was amendatory.The lung W/D ratio in ALI group and ALI+UTI group was higher than that in control group,while the lung W/D ratio in ALI+UTI group was lower than that in ALI group(all P<0.05).The expression levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)in BALF of ALI and ALI+UTI rats were higher than those of control group,while the expression levels of TNF-α,IL-1β and IL-6 in BALF of ALI+UTI rats were lower than those of ALI group(all P<0.05).Compared with the control group,the expression levels of ROS and MDA in lung tissue of ALI group and ALI+UTI group were significantly increased,and the expression level of SOD was significantly decreased(all P<0.05).Compared with ALI group,the expression levels of ROS and MDA in lung tissue of ALI+UTI group were significantly decreased,and the expression level of SOD was significantly increased(all P<0.05).The relative mRNA expression of nuclear factor E2-related factor 2(Nrf2)and its downstream genes heme oxygenase-1(HO-1)and quinone oxidoreductase 1(NQO-1)in lung tissue of ALI+UTI group were higher than those in control group and ALI group(all P<0.05).There was no significant difference in the expression of pAMPK/AMPK and Nrf2 protein between control group and ALI group(all P>0.05).The expression of pAMPK/AMPK and Nrf2 protein in ALI group was higher than that in control group and ALI group(all P<0.05).Conclusion:UTI can activate AMPK phosphorylation and enhance the activity of Nrf2 and its downstream genes through the AMPK/Nrf2 pathway,effectively alleviating hyperoxia-induced ALI.UTI can be used as one of the alternative drugs for clinical treatment of ALI.

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备注/Memo

备注/Memo:
基金项目:陕西省榆林市科技计划项目(YF2020051)
更新日期/Last Update: 2022-11-09