彭燕
摘要:目的? 評(píng)價(jià)?;撬釋?duì)LPS誘導(dǎo)急性肺損傷小鼠的治療作用。方法? 將BALB/c小鼠隨機(jī)分為正常對(duì)照組、LPS模型組、?;撬岬?、中、高劑量組,正常對(duì)照組小鼠經(jīng)鼻吸入50 μl生理鹽水,LPS模型組小鼠經(jīng)鼻吸入8 mg/ml的LPS溶液50 μl,?;撬岬?、中、高劑量組分別經(jīng)腹腔注射100、200、400 mg/kg?;撬?,監(jiān)測(cè)并比較5組小鼠肺功能、肺指數(shù)及髓過(guò)氧化物(MPO)酶活性。結(jié)果? ①LPS模型組呼氣末期停頓(EEP)、最大呼氣流量(PEF)、支氣管收縮的程度(PENH)、吸氣時(shí)間(TI)及呼氣時(shí)間(TE)均高于正常對(duì)照組及?;撬岬汀⒅?、高劑量組;?;撬岬?、中、高劑量組EEP、PEF、PENH、TI及TE均高于正常對(duì)照組,且依次降低,差異有統(tǒng)計(jì)學(xué)意義(P<0.05)。LPS模型組呼吸頻率(RR)、呼吸松弛時(shí)間(RT)、每分鐘呼氣量(MV)、吸氣末期停頓時(shí)間(EIP)、最大吸氣流量(PIF)、潮氣量(TV)及呼氣量(EV)均低于正常對(duì)照組及牛磺酸低、中、高劑量組;?;撬岬?、中、高劑量組RR、RT、MV、EIP、PIF、TV及EV均低于正常對(duì)照組,且依次升高,差異有統(tǒng)計(jì)學(xué)意義(P<0.05)。②LPS模型組肺濕/干重比、MPO酶活性高于正常對(duì)照組及?;撬岬?、中、高劑量組;?;撬岬?、中、高劑量組肺濕/干重比、MPO酶活性高于正常對(duì)照組,且依次降低,差異有統(tǒng)計(jì)學(xué)意義(P<0.05)。結(jié)論? ?;撬釋?duì)LPS誘導(dǎo)的小鼠急性肺損傷具有一定的保護(hù)作用,可減輕小鼠肺損傷,且其呼吸功能改善程度與劑量有一定聯(lián)系。
關(guān)鍵詞:急性肺損傷;?;撬?LPS;小鼠
中圖分類號(hào):R965? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?文獻(xiàn)標(biāo)識(shí)碼:A? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ? ?DOI:10.3969/j.issn.1006-1959.2020.03.023
文章編號(hào):1006-1959(2020)03-0079-03
Therapeutic Effect of Taurine on LPS-induced Acute Lung Injury in Mice
PENG Yan
(ICU of Tianjin TEDA Hospital,Tianjin 300457,China)
Abstract:Objective? To evaluate the therapeutic effect of taurine on LPS-induced acute lung injury in mice. Methods? BALB/c mice were randomly divided into normal control group, LPS model group, taurine low, medium and high dose groups. Normal control group mice inhaled 50 μl physiological saline,mice in the LPS model group inhaled 50 μl of 8 mg / ml LPS solution nasally, and the low, medium, and high dose groups of taurine were injected intraperitoneally with 100, 200, and 400 mg / kg taurine. Rat lung function, lung index, and myeloperoxidase (MPO) enzyme activity.Results? ①The end-expiratory pause (EEP), the maximum expiratory flow (PEF), the degree of bronchoconstriction (PENH), the inspiratory time (TI) and the expiratory time (TE) in the LPS model group were higher than those in the normal control group and taurine. The low, medium, and high dose groups; the low, medium, and high dose taurine EEP, PEF, PENH, TI, and TE were all higher than the normal control group, and they decreased in order, the difference was statistically significant(P<0.05). LPS model group breathing frequency (RR), breathing relaxation time (RT), expiratory volume per minute (MV), end of inhalation time (EIP), maximum inspiratory flow (PIF), tidal volume (TV) and exhalation amount (EV) is lower than the normal control group and low, medium and high dose of taurine; RR, RT, MV, EIP, PIF, TV and EV of the low, medium and high dose of taurine are lower than the normal control group, and increased in order, the difference was statistically significant(P<0.05). ②The lung wet / dry weight ratio and MPO enzyme activity of the LPS model group were higher than those of the normal control group and low, medium, and high dose taurine; lung wet/dry weight ratio and MPO enzyme activity of the low, medium, and high dose taurine group was higher than the normal control group and decreased in order,the difference was statistically significant (P<0.05).Conclusion? Taurine has a certain protective effect on acute lung injury induced by LPS in mice, which can alleviate lung injury in mice, and its degree of improvement in respiratory function is related to the dose.
Key words:Acute lung injury;Taurine;LPS;Mice
急性肺損傷(acute lung injury,ALI)是以急性炎癥和組織損傷為特征,影響肺部正常氣體交換,逐漸導(dǎo)致呼吸功能衰竭的急性呼吸綜合征[1]。歐美聯(lián)席會(huì)議在1994年將該綜合征重新命名為急性呼吸窘迫綜合征(acute respiratory distress syndrome,ARDS)[2],是監(jiān)護(hù)室患者病死率高的重要原因[3],因而ALI/ARDS被認(rèn)為是臨床常見(jiàn)危重癥。由LPS所導(dǎo)致的膿毒癥是引起ARDS的重要常見(jiàn)病因,大量研究表明,LPS引起ARDS的機(jī)制與自由基的產(chǎn)生、脂質(zhì)過(guò)氧化和炎癥反應(yīng)有重要關(guān)系[4,5]。?;撬幔╰aurine)大量存在于哺乳動(dòng)物體內(nèi),能夠維持機(jī)體的免疫功能,調(diào)節(jié)全身多個(gè)系統(tǒng)、器官[6]。在?;撬岬淖饔孟拢瑑?nèi)皮細(xì)胞的凋亡及由脂多糖、腫瘤壞死因子刺激的氧化應(yīng)激狀態(tài)明顯減輕,?;撬徇€可以降低中性粒細(xì)胞介導(dǎo)的內(nèi)皮細(xì)胞的壞死,可用于急性肺損傷的治療。基于此,本研究建立LPS誘導(dǎo)急性肺損傷小鼠動(dòng)物模型,旨在觀察?;撬釋?duì)急性肺損傷的治療作用。
1材料與方法
1.1材料與儀器? 雄性BALB/c小鼠共50只,體重20~24 g,購(gòu)自天津醫(yī)科大學(xué)實(shí)驗(yàn)動(dòng)物中心,許可證編號(hào):SYXK(津)2013-0001。脂多糖(LPS)和?;撬峋?gòu)自Sigma-Aldrich;髓過(guò)氧化物酶活性檢測(cè)試劑盒購(gòu)自Genmed Scientifics Inc;清醒動(dòng)物肺功能監(jiān)測(cè)系統(tǒng)購(gòu)自EMKA Technologies。
1.2方法? 將BALB/c小鼠按照隨機(jī)數(shù)字表法分為正常對(duì)照組、LPS模型組、?;撬岬?、中、高劑量組。LPS模型組及牛磺酸治療組小鼠經(jīng)鼻吸入8 mg/ml的LPS溶液50 μl,正常對(duì)照組小鼠經(jīng)鼻吸入同體積的生理鹽水。實(shí)驗(yàn)開(kāi)始0.5 h和12 h,?;撬岬?、中、高劑量組分別腹腔注射100、200、400 mg/kg?;撬帷?shí)驗(yàn)開(kāi)始24 h后,對(duì)全部BALB/c小鼠進(jìn)行肺功能監(jiān)測(cè),取血及肺組織。
1.3觀察指標(biāo)? 比較各組小鼠肺功能、肺指數(shù)及髓過(guò)氧化物(MPO)酶活性。①肺功能:實(shí)驗(yàn)開(kāi)始24 h后,監(jiān)測(cè)各組小鼠在清醒無(wú)束縛狀態(tài)下的肺呼吸情況,包括吸氣時(shí)間(TI)、呼氣時(shí)間(TE)、最大吸氣流量(PIF)、最大呼氣流量(PEF)、潮氣量(TV)、呼氣量(EV)、呼吸松弛時(shí)間(RT)、每分鐘呼氣量(MV)、呼吸頻率(RR)、吸氣末期停頓(EIP)、呼氣末期停頓(EEP)及支氣管收縮的程度(PENH)。②肺指數(shù)測(cè)定:取小鼠的肺臟,洗凈,記錄濕重;100℃干燥至恒重,稱干重,計(jì)算肺的濕干重比。③MPO酶活性測(cè)定:采用髓過(guò)氧化物酶活性測(cè)定試劑盒測(cè)定肺組織中MPO酶的活性,具體步驟嚴(yán)格參考試劑盒說(shuō)明書(shū)進(jìn)行。
1.4統(tǒng)計(jì)學(xué)方法? 選用SPSS 20.0軟件處理實(shí)驗(yàn)數(shù)據(jù),計(jì)量資料以(x±s)表示,兩組間比較采用獨(dú)立樣本t檢驗(yàn),多組間比較采用單因素方差分析;采用Pearson 相關(guān)性分析?;撬釀┝颗c呼吸功能的相關(guān)性。P<0.05表示差異有統(tǒng)計(jì)學(xué)意義。
2結(jié)果
2.1各組小鼠肺功能比較? LPS模型組EEP、PEF、PENH、TI及TE均高于正常對(duì)照組及?;撬岬?、中、高劑量組;同時(shí)?;撬岬?、中、高劑量組EEP、PEF、PENH、TI及TE均高于正常對(duì)照組,且依次降低,差異有統(tǒng)計(jì)學(xué)意義(P<0.05)。LPS模型組RR、RT、MV、EIP、PIF、TV及EV均低于正常對(duì)照組及?;撬岬?、中、高劑量組;同時(shí)牛磺酸低、中、高劑量組RR、RT、MV、EIP、PIF、TV及EV均低于正常對(duì)照組,且依次升高,差異有統(tǒng)計(jì)學(xué)意義(P<0.05),見(jiàn)圖1。
2.2各組小鼠肺濕/干重比、MPO酶活性比較? LPS模型組肺濕/干重比、MPO酶活性高于正常對(duì)照組及?;撬岬?、中、高劑量組;?;撬岬?、中、高劑量組肺濕/干重比、MPO酶活性高于正常對(duì)照組,且依次降低,差異有統(tǒng)計(jì)學(xué)意義(P<0.05)。見(jiàn)圖2、圖3。
3討論
牛磺酸可以明顯減輕內(nèi)皮細(xì)胞的凋亡及由脂多糖、腫瘤壞死因子刺激下的氧化應(yīng)激狀態(tài),并可以降低中性粒細(xì)胞介導(dǎo)的內(nèi)皮細(xì)胞壞死[5]。本實(shí)驗(yàn)采用LPS誘導(dǎo)急性肺損傷小鼠模型,考察了?;撬釋?duì)小鼠呼吸功能保護(hù)作用及對(duì)肺損傷的治療作用。實(shí)驗(yàn)結(jié)果顯示,與正常對(duì)照組相比LPS模型組的各項(xiàng)呼吸功能均有受損,但?;撬峤M小鼠的呼氣末期停頓時(shí)間、單位體重的最大呼氣流量、支氣管收縮的程度和呼吸頻率等呼吸功能指標(biāo)優(yōu)于LPS模型組,呼吸功能改善程度與牛磺酸使用劑量密切相關(guān),而且牛磺酸組小鼠的肺濕/干重比低于LPS模型組,提示?;撬釋?duì)LPS誘導(dǎo)的小鼠急性肺損傷具有治療作用,能夠改善肺水腫程度。
髓過(guò)氧化物酶又稱過(guò)氧化物,是由中性粒細(xì)胞、單核細(xì)胞或巨噬細(xì)胞等炎性細(xì)胞分泌的含血紅素輔基的血紅素蛋白酶,也是中性粒細(xì)胞的功能標(biāo)識(shí)和激活標(biāo)志,其活性變化反映著嗜中性多形核細(xì)胞的功能和活性狀態(tài)[6]。本研究結(jié)果顯示,吸入400 μg的脂多糖24 h后,實(shí)驗(yàn)小鼠發(fā)生明顯的肺水腫,肺組織中MPO酶活性提高,但給予?;撬嶂委熀竽軌蚓徑庑∈蠓嗡[狀況,降低肺組織中MPO酶活性,提示牛磺酸有可能通過(guò)抗氧化作用調(diào)節(jié)細(xì)胞內(nèi)鈣離子濃度,進(jìn)而對(duì)急性肺損傷發(fā)揮一定的保護(hù)作用。
綜上所述,?;撬釋?duì)LPS誘導(dǎo)的小鼠急性肺損傷具有一定的肺臟保護(hù)作用,可減輕小鼠肺損傷,且其對(duì)呼吸功能的改善程度與應(yīng)用劑量密切相關(guān)。
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收稿日期:2019-11-21;修回日期:2019-12-13
編輯/錢洪飛