重组人红细胞生成素对新生大鼠高氧肺损伤炎症反应的影响

Q4 Medicine

中华实用儿科临床杂志 Pub Date : 2020-03-30 DOI:10.3760/CMA.J.CN101070-20190202-00084

Qilin Huang, Jing-jing Ren, Yanmei Xie, Ze-quan Ji, Haiyan Liu, Cui-wen Huang

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The morphological structure changes of lung tissues were observed by HE staining under light microscope, and the radial alveolar count(RAC) in lung tissues were detected.The expression of nuclear factor kappa B(NF-κB) was detected by immunofluorescence staining; Western blot was applied to determine the protein expression of phosphorylated NF-κB(pNF-κB), inhibitor protein(IκB) and Caspase-3 in lung tissues; and the expression of interleukin-1β(IL-1β) in bronchoalveolar lavage fluid was determined using enzyme linked immunosorbent assay (ELISA). \n \n \nResults \n(1) On the 14th day, the body weight of neonatal rats in the BPD group was lower than that in the control group [(18.97±3.21) g vs.(27.97±2.30) g], and the difference was statistically significant(P<0.01); however, the body weights of neonatal rats in the hyperoxia+ EPO(L)group and hyperoxia+ EPO(H)group[(24.16±2.15) g, (26.04±1.97) g] was much heavier than that in the BPD group, and the differences was statistically significant(all P<0.05). (2) The morphological structure of lung tissues which was observed by HE staining showed that in the BPD group, there were a few inflammatory cells infiltration in alveolar septum on the 3rd day, the inflammatory response was more evident on the 7th day, when exudation could be seen in the alveolar cavity; and on the 14th day, the number of pulmonary alveoli decreased, pulmonary bulla formed, and septa were thickened.Besides, it was also observed that compared with control group, RAC was significantly decreased in BPD group on the 14th day(5.50±1.29 vs.14.33±2.80), and the difference was statistically significant(P<0.01). Pathological changes were ameliorated and the infiltration of inflammatory reaction cells was reduced in the hyperoxia+ EPO(L)group and hyperoxia + EPO(H)group.RAC was remarkably higher in the hyperoxia+ EPO(L)group and hyperoxia+ EPO(H)group than that in the BPD group on the 14th day, and the differences were statistically significant (all P<0.05). (3)Immunofluorescence results showed that: the number of NF-κB p65 positive cells increased significantly and fluorescence intensity increased in the BPD group, while EPO could greatly reduce the number of NF-κB p65 positive cells and lower the fluorescence intensity.(4)Western blot results indicated that compared with the control group, the expressions of pNF-κB p65 and Cleaved Caspase-3 was significantly increased, and the differences were statistically significant (all P<0.05); and IκB was significantly lower, and the difference was statistically significant (P<0.05). The expression of NF-κB p65 and Cleaved Caspase-3 was significantly lower, but IκB was significantly higher in the hyperoxia+ EPO(L)group and the hyperoxia+ EPO(H)group than those in the BPD group, and the differences were statistically significant (all P<0.05). (5) ELISA results revealed that the expression of IL-1β in the BPD group was significantly higher than that in the control group, and the difference was statistically significant (P<0.05); Compared with BPD group, the expression of IL-1β was significantly lower in the hyperoxia+ EPO(L)group and hyperoxia+ EPO(H)group, and the differences were statistically significant (all P<0.05). \n \n \nConclusions \nEPO can reduce hyperoxia-induced lung tissue apoptosis and protect newborn rats against hyperoxic lung injury by decreasing the inflammatory response of cells through the NF-κB pathway on BPD. \n \n \nKey words: \nBronchopulmonary dysplasia; Human recombinant erythropoietin; Hyperoxia; Inflammatory reaction; Nuclear transcription factor kappa B","PeriodicalId":9843,"journal":{"name":"中华实用儿科临床杂志","volume":"35 1","pages":"383-388"},"PeriodicalIF":0.0000,"publicationDate":"2020-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Effect of human recombinant erythropoietin on inflammation of hyperoxic lung injury in neonatal rats\",\"authors\":\"Qilin Huang, Jing-jing Ren, Yanmei Xie, Ze-quan Ji, Haiyan Liu, Cui-wen Huang\",\"doi\":\"10.3760/CMA.J.CN101070-20190202-00084\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Objective \\nTo explore the effects of human recombinant erythropoietin (rhEPO) on inflammation of hyperoxic lung injury in neonatal rats. \\n \\n \\nMethods \\nSeventy-two neonatal rats were randomly divided into 4 groups: control group, BPD group, hyperoxia + low-dose recombinant erythropoietin [EPO(L)]group, and hyperoxia + high-dose recombinant erythropoietin [EPO(H)]group.The neonatal rats in BPD group, hyperoxia + EPO(L)group and hyperoxia + EPO(H)group were exposed to 850 mL/L oxygen.Then the neonatal rats in hyperoxia + EPO(L)group and hyperoxia + EPO(H)group were given 800 IU/kg and 2 000 IU/kg rhEPO by subcutaneous injection respectively at 1 d, 3 d and 7 d, while the control group and BPD group were given the same amount of 9 g/L saline water.Initially, the body weight of each group was recorded at 3 d, 7 d and 14 d. 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(2) The morphological structure of lung tissues which was observed by HE staining showed that in the BPD group, there were a few inflammatory cells infiltration in alveolar septum on the 3rd day, the inflammatory response was more evident on the 7th day, when exudation could be seen in the alveolar cavity; and on the 14th day, the number of pulmonary alveoli decreased, pulmonary bulla formed, and septa were thickened.Besides, it was also observed that compared with control group, RAC was significantly decreased in BPD group on the 14th day(5.50±1.29 vs.14.33±2.80), and the difference was statistically significant(P<0.01). Pathological changes were ameliorated and the infiltration of inflammatory reaction cells was reduced in the hyperoxia+ EPO(L)group and hyperoxia + EPO(H)group.RAC was remarkably higher in the hyperoxia+ EPO(L)group and hyperoxia+ EPO(H)group than that in the BPD group on the 14th day, and the differences were statistically significant (all P<0.05). (3)Immunofluorescence results showed that: the number of NF-κB p65 positive cells increased significantly and fluorescence intensity increased in the BPD group, while EPO could greatly reduce the number of NF-κB p65 positive cells and lower the fluorescence intensity.(4)Western blot results indicated that compared with the control group, the expressions of pNF-κB p65 and Cleaved Caspase-3 was significantly increased, and the differences were statistically significant (all P<0.05); and IκB was significantly lower, and the difference was statistically significant (P<0.05). The expression of NF-κB p65 and Cleaved Caspase-3 was significantly lower, but IκB was significantly higher in the hyperoxia+ EPO(L)group and the hyperoxia+ EPO(H)group than those in the BPD group, and the differences were statistically significant (all P<0.05). (5) ELISA results revealed that the expression of IL-1β in the BPD group was significantly higher than that in the control group, and the difference was statistically significant (P<0.05); Compared with BPD group, the expression of IL-1β was significantly lower in the hyperoxia+ EPO(L)group and hyperoxia+ EPO(H)group, and the differences were statistically significant (all P<0.05). \\n \\n \\nConclusions \\nEPO can reduce hyperoxia-induced lung tissue apoptosis and protect newborn rats against hyperoxic lung injury by decreasing the inflammatory response of cells through the NF-κB pathway on BPD. \\n \\n \\nKey words: \\nBronchopulmonary dysplasia; Human recombinant erythropoietin; Hyperoxia; Inflammatory reaction; Nuclear transcription factor kappa B\",\"PeriodicalId\":9843,\"journal\":{\"name\":\"中华实用儿科临床杂志\",\"volume\":\"35 1\",\"pages\":\"383-388\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2020-03-30\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"中华实用儿科临床杂志\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.3760/CMA.J.CN101070-20190202-00084\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"Medicine\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"中华实用儿科临床杂志","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.3760/CMA.J.CN101070-20190202-00084","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Medicine","Score":null,"Total":0}

引用次数: 0

摘要

目的探讨重组人红细胞生成素（rhEPO）对新生大鼠高氧肺损伤炎症反应的影响。方法72只新生大鼠随机分为4组：对照组、BPD组、高氧+低剂量重组红细胞生成素[EPO（L）]组和高氧+高剂量重组红血球生成素[EEPO（H）]组。高氧+EPO（L）组和高氧+EPO（H）组新生大鼠分别于1d、3d和7d皮下注射rhEPO 800IU/kg和2000IU/kg，对照组和BPD组给予等量的9g/L盐水。最初，在第3天、第7天和第14天记录各组的体重。在光学显微镜下通过HE染色观察肺组织的形态结构变化，并检测肺组织中的径向肺泡计数（RAC）。免疫荧光染色检测核因子κB（NF-κB）的表达；Western blot检测肺组织中磷酸化NF-κB（pNF-κB）、抑制蛋白（IκB）和半胱氨酸蛋白酶-3的蛋白表达；应用酶联免疫吸附法（ELISA）检测支气管肺泡灌洗液中白细胞介素1β（IL-1β）的表达。结果（1）第14天，BPD组新生大鼠体重低于对照组[（18.97±3.21）gvs.（27.97±2.30）g]，差异有统计学意义（P<0.01）；高氧+EPO（L）组和高氧+EPO（H）组的新生大鼠体重[（24.16±2.15）g，（26.04±1.97）g]明显重于BPD组，差异有统计学意义（均P<0.05），肺泡隔第3天有少量炎性细胞浸润，第7天炎症反应更为明显，肺泡腔内可见渗出；第14天肺泡数量减少，肺大泡形成，间隔增厚。此外，与对照组相比，BPD组在第14天的RAC显著降低（5.50±1.29 vs.14.33±2.80），高氧+EPO（L）组和高氧+EPO（H）组病理改变明显改善，炎性反应细胞浸润减少，14天AC明显高于BPD组，（3）免疫荧光结果显示：BPD组NF-κB p65阳性细胞数显著增加，荧光强度增加，而EPO可显著减少NF-κBp65阳性的细胞数，降低荧光强度。（4） Western blot结果显示，与对照组相比，pNF-κB p65和Cleaved Caspase-3的表达显著增加，差异有统计学意义（均P<0.05）；NF-κBp65和Cleaved Caspase-3的表达在高氧+EPO（L）组和高氧+EPO（H）组明显低于BPD组，（5）ELISA结果显示，BPD组IL-1β的表达明显高于对照组，差异有统计学意义（P<0.05）；高氧+EPO（L）组和高氧+EPO（H）组IL-1β表达明显低于BPD组，结论EPO可通过抑制细胞对BPD的炎症反应，减少高氧诱导的肺组织凋亡，保护新生大鼠免受高氧肺损伤。关键词：支气管肺发育不良；人重组红细胞生成素；高氧血症；炎症反应；核转录因子κB

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Effect of human recombinant erythropoietin on inflammation of hyperoxic lung injury in neonatal rats

Objective To explore the effects of human recombinant erythropoietin (rhEPO) on inflammation of hyperoxic lung injury in neonatal rats. Methods Seventy-two neonatal rats were randomly divided into 4 groups: control group, BPD group, hyperoxia + low-dose recombinant erythropoietin [EPO(L)]group, and hyperoxia + high-dose recombinant erythropoietin [EPO(H)]group.The neonatal rats in BPD group, hyperoxia + EPO(L)group and hyperoxia + EPO(H)group were exposed to 850 mL/L oxygen.Then the neonatal rats in hyperoxia + EPO(L)group and hyperoxia + EPO(H)group were given 800 IU/kg and 2 000 IU/kg rhEPO by subcutaneous injection respectively at 1 d, 3 d and 7 d, while the control group and BPD group were given the same amount of 9 g/L saline water.Initially, the body weight of each group was recorded at 3 d, 7 d and 14 d. The morphological structure changes of lung tissues were observed by HE staining under light microscope, and the radial alveolar count(RAC) in lung tissues were detected.The expression of nuclear factor kappa B(NF-κB) was detected by immunofluorescence staining; Western blot was applied to determine the protein expression of phosphorylated NF-κB(pNF-κB), inhibitor protein(IκB) and Caspase-3 in lung tissues; and the expression of interleukin-1β(IL-1β) in bronchoalveolar lavage fluid was determined using enzyme linked immunosorbent assay (ELISA). Results (1) On the 14th day, the body weight of neonatal rats in the BPD group was lower than that in the control group [(18.97±3.21) g vs.(27.97±2.30) g], and the difference was statistically significant(P<0.01); however, the body weights of neonatal rats in the hyperoxia+ EPO(L)group and hyperoxia+ EPO(H)group[(24.16±2.15) g, (26.04±1.97) g] was much heavier than that in the BPD group, and the differences was statistically significant(all P<0.05). (2) The morphological structure of lung tissues which was observed by HE staining showed that in the BPD group, there were a few inflammatory cells infiltration in alveolar septum on the 3rd day, the inflammatory response was more evident on the 7th day, when exudation could be seen in the alveolar cavity; and on the 14th day, the number of pulmonary alveoli decreased, pulmonary bulla formed, and septa were thickened.Besides, it was also observed that compared with control group, RAC was significantly decreased in BPD group on the 14th day(5.50±1.29 vs.14.33±2.80), and the difference was statistically significant(P<0.01). Pathological changes were ameliorated and the infiltration of inflammatory reaction cells was reduced in the hyperoxia+ EPO(L)group and hyperoxia + EPO(H)group.RAC was remarkably higher in the hyperoxia+ EPO(L)group and hyperoxia+ EPO(H)group than that in the BPD group on the 14th day, and the differences were statistically significant (all P<0.05). (3)Immunofluorescence results showed that: the number of NF-κB p65 positive cells increased significantly and fluorescence intensity increased in the BPD group, while EPO could greatly reduce the number of NF-κB p65 positive cells and lower the fluorescence intensity.(4)Western blot results indicated that compared with the control group, the expressions of pNF-κB p65 and Cleaved Caspase-3 was significantly increased, and the differences were statistically significant (all P<0.05); and IκB was significantly lower, and the difference was statistically significant (P<0.05). The expression of NF-κB p65 and Cleaved Caspase-3 was significantly lower, but IκB was significantly higher in the hyperoxia+ EPO(L)group and the hyperoxia+ EPO(H)group than those in the BPD group, and the differences were statistically significant (all P<0.05). (5) ELISA results revealed that the expression of IL-1β in the BPD group was significantly higher than that in the control group, and the difference was statistically significant (P<0.05); Compared with BPD group, the expression of IL-1β was significantly lower in the hyperoxia+ EPO(L)group and hyperoxia+ EPO(H)group, and the differences were statistically significant (all P<0.05). Conclusions EPO can reduce hyperoxia-induced lung tissue apoptosis and protect newborn rats against hyperoxic lung injury by decreasing the inflammatory response of cells through the NF-κB pathway on BPD. Key words: Bronchopulmonary dysplasia; Human recombinant erythropoietin; Hyperoxia; Inflammatory reaction; Nuclear transcription factor kappa B

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来源期刊

中华实用儿科临床杂志 Medicine-Pediatrics, Perinatology and Child Health

CiteScore

0.60

自引率

0.00%

发文量

14243

期刊介绍： Chinese Journal of Applied Clinical Pediatrics ( semi-monthly ) is a core journal of paediatrics under the supervision of China Association for Science and Technology, sponsored by Chinese Medical Association and undertaken by Xinxiang Medical College. Founded in 1986, it is openly circulated both at home and abroad. The journal has several columns, such as Expert Forum, Experimental Research and Paediatric Surgery, which are mainly for paediatric medical workers and medical researchers in hospitals. Its purpose is to reflect the new theories and technologies in paediatric medicine and scientific research at home and abroad, and to promote academic exchanges. Chinese Journal of Applied Clinical Pediatrics is a source journal of China Science Citation Database (CSCD), a core journal of Peking University, a source journal of Chinese science and technology paper statistics (China Science and Technology Core Journals), a core academic journal of RCCSE, a high-quality scientific and technical journal of China, a high-quality scientific and technical journal of China Association for Science and Technology, and a high-quality scientific and technical journal of China Biomedical Science and Technology Association. We have been published in China Biomedical Literature Database (SinoMed), China Knowledge Network, Wanfang Data Knowledge Service Platform, China Academic Journal Abstracts, Scopus Database, Chemical Abstracts (USA), Japan Science and Technology Agency (JSTA) Database, Copernicus Abstracts (Poland), Abstracts of the Centre for Agricultural and Biological Sciences (CABS) of the United Kingdom, Cambridge Scientific Abstracts ProQuest Database, WHO Medical Journal of the Western Pacific Region (WMPR), and WHO Medical Journal of the Western Pacific Region (WMPR) of the United States. We have been included in dozens of authoritative databases at home and abroad, such as WHO Western Pacific Region Index of Medicine (WPRIM), Ullrich's Guide to Periodicals, and so on.