首页|参附注射液对失血性休克大鼠肺损伤的影响

参附注射液对失血性休克大鼠肺损伤的影响

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目的 观察参附注射液对大鼠失血性休克(HS)致肺损伤的影响并探讨潜在机制.方法 选择SPF级健康雄性SD大鼠36 只,16~17 周龄,体重400~600 g.采用随机数字表法将大鼠分为三组:假手术组(SH组)、失血性休克组(HS组)和参附注射液组(SF组),每组 12 只.SH组麻醉后仅分离出右股静脉和股动脉,不行动静脉置管,HS组和SF组制备HS大鼠模型.HS组经静脉导管进行液体复苏,复苏液为所失自体血加 1.5 倍失血量的复方氯化钠注射液与生理盐水 10 ml/kg,SF组复苏液为所失自体血加 1.5 倍失血量的复方氯化钠注射液与参附注射液10 ml/kg,灌注时间约 60 min.于术后24、48 h随机处死6 只大鼠,取肺组织检测湿重与干重比(W/D),采用ELISA法检测白细胞介素-6(IL-6)、IL-17、IL-10、转化生长因子-β(TGF-β)浓度,采用PCR法检测肺组织视黄酸相关孤核受体γt(RORγt)、叉头翼状螺旋转录因子3(Foxp3)、缺氧诱导因子1α(HIF-1α)mRNA表达量,采用Western blot法检测RORγt、Foxp3、HIF-1α、水通道蛋白 1(AQP1)和AQP5 蛋白含量,肺组织行HE染色后在光镜下观察病理学改变并行肺损伤评分.结果 与术后24h比较,术后48hSF组肺组织W/D、IL-6和IL-17 浓度、RORγt和HIF-1α mRNA表达量及蛋白含量、肺损伤评分均明显降低(P<0.05),IL-10 和TGF-β浓度、Foxp3 mRNA表达量及蛋白含量、AQP1 蛋白含量均明显升高(P<0.05).与SH组比较,术后 24、48 h HS组和SF组W/D、IL-6、IL-17、IL-10 和TGF-β浓度、RORγt、Foxp3 和HIF-1α mRNA表达量及蛋白含量、肺损伤评分均明显升高(P<0.05),AQP1、AQP5 蛋白含量明显降低(P<0.05),光镜下可见肺泡结构破坏,肺泡间质充满大量水肿液,其间浸润着大量炎性细胞.与HS组比较,术后 24、48 h SF组W/D、IL-6和IL-17 浓度、RORγt和HIF-1α mRNA表达量及蛋白含量、肺损伤评分明显降低(P<0.05),IL-10 和TGF-β浓度、Foxp3 mRNA表达量及蛋白含量、AQP1 和AQP5 蛋白含量均明显升高(P<0.05),光镜下可见肺泡结构紊乱改善,水肿程度减轻,炎性细胞数量减少.结论 参附注射液可调节促炎因子IL-6、IL-17 与抗炎因子IL-10、TGF-β平衡,升高肺组织AQP1、AQP5 蛋白含量,降低肺组织W/D和肺损伤评分,减轻HS大鼠肺损伤,机制可能与调节HIF-1α-RORγt/Foxp3 平衡有关.
Effect of Shenfu injection on acute lung injury in hemorrhagic shock rats
Objective To observe the effect of Shenfu injection on lung injury caused by hemor-rhagic shock(HS)in rats and explore the related potential mechanism.Methods Thirty-six SPF healthy male SD rats,aged 16-17 weeks,weighing 400-600 g,were randomly divided into three groups:sham op-eration group(group SH),HS group(group HS),and Shenfu injection group(group SF),12 rats in each group.In group SH,only the right femoral vein and femoral artery were separated after anesthesia,and ve-nous catheterization was not performed.HS model was established in groups SF and HS.In group HS,liquid resuscitation was performed through an intravenous catheter,and the resuscitation fluid consisted of the auto-blood lost and the compound sodium chloride injection equivalent to 1.5 times the blood loss and 10 ml/kg normal saline.In group SF,the resuscitation fluid consisted of the lost autoblood and the compound sodium chloride injection equivalent to 1.5 times the blood loss and Shenfu injection 10 ml/kg.The whole perfusion time was about 60 minutes.Six rats in the three groups were randomly anesthetized 24 and 48 hours after op-eration.The wet/dry weight ratio(W/D)of lung tissues was detected.The concentrations of interleukin-6(IL-6),IL-17,IL-10,and transforming growth factor-β(TGF-β)were detected by ELISA,the mRNA ex-pression of retinoic acid-related orphan nuclear receptor γt(RORγt),transcription factor forkhead box pro-tein 3(Foxp3),and hypoxia-inducible factor-1α(HIF-1α)in lung tissues were detected by PCR.The pro-tein contents of RORγt,Foxp3,HIF-1α,aquaporin 1(AQP1),and AQP5 in lung tissue were detected by Western blot.Pathological changesunder HE staining light microscope and lung injury scores were observed.Results Compared with 24 hours after operation,W/D,the concentrations of IL-6 and IL-17,mRNA ex-pression and protein content of RORγt and HIF-1α,and lung injury score were significantly decreased(P<0.05),the concentrations of IL-10,and TGF-β,Foxp3 mRNA expression and protein content,and AQP1 protein content were significantly increased in group SF 48 hours after operation(P<0.05).Compared with group SH,W/D,the concentrations of IL-6,IL-17,IL-10,and TGF-β,mRNA expression and protein content of RORγt,Foxp 3,and HIF-1α,and lung injury score were significantly increased(P<0.05),AQP1 and AQP5 protein contents were significantly decreased in groups HS and SF 24 and 48 hours after operation(P<0.05),and alveolar structure was damaged under light microscope and alveolar interstitium was filled with a large amount of edematous fluid,during which a large number of inflammatory cells infiltra-ted.Compared with group HS,W/D,the concentrations of IL-6 and IL-17,mRNA expression and protein content of RORγt and HIF-1α,and lung injury score were significantly decreased(P<0.05),the concen-trations of IL-10 and TGF-β,Foxp3 mRNA expression and protein content,AQP1 and AQP5 protein con-tents were significantly increased in group SF 24 and 48 hours after surgery(P<0.05),and the alveolar structure was improved under light microscope,and edema was reduced,and the number of inflammatory cells was reduced.Conclusion Shenfu injection can regulate the balance between pro-inflammatory factors IL-6 and IL-17,and anti-inflammatory factors IL-10 and TGF-β,increase the protein content of AQP1 and AQP5 in lung tissue,and decrease the W/D and injury score in lung tissue,thus alleviating lung injury in HS rats.The mechanism may be related to the regulation of HIF-1α-RORγt/Foxp3 balance.

Shenfu injectionHemorrhagic shockLung injuryHypoxia-inducible factor-1αRetinoic acid-related orphan nuclear receptor γtTranscription factor forkhead box protein 3

徐迎雪、迟永良、伊晓倩、刘改红、张栋斌、季加富、司尚坤、苏帆

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250355 济南市,山东中医药大学第一临床医学院

山东中医药大学附属医院麻醉科

参附注射液 失血性休克 肺损伤 缺氧诱导因子 视黄酸相关孤核受体γt 叉头翼状螺旋转录因子 3

山东省自然科学基金面上项目济南市科技创新发展计划

ZR2020MH391202134068

2024

临床麻醉学杂志
中华医学会南京分会

临床麻醉学杂志

CSTPCD北大核心
影响因子:2.225
ISSN:1004-5805
年,卷(期):2024.40(1)
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