燒傷早期注射胰島素對(duì)大鼠心、腎損害的影響*＊?

袁新章，王　偉，李偉人＊

(貴州醫(yī)科大學(xué)附院燒傷整形科，貴州貴陽　550004)

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燒傷早期注射胰島素對(duì)大鼠心、腎損害的影響*＊?

袁新章＊＊，王偉，李偉人＊＊＊

(貴州醫(yī)科大學(xué)附院燒傷整形科，貴州貴陽550004)

［摘要］目的:探討燒傷早期皮下注射精蛋白生物合成胰島素對(duì)延遲復(fù)蘇大鼠心、腎損害的影響。方法: 32 只SPF級(jí)SD大鼠隨機(jī)均分為假傷組、即時(shí)復(fù)蘇組、延遲復(fù)蘇組與延遲復(fù)蘇治療組4組;即時(shí)復(fù)蘇組、延遲復(fù)蘇組與延遲復(fù)蘇治療組大鼠置于(95.0±0.5)℃熱水中15 s，制作30%燒傷總面積(TBSA)的Ⅲ°燙傷動(dòng)物模型，假傷組置37℃溫水中15 s模擬燙傷;即時(shí)復(fù)蘇組和延遲復(fù)蘇組傷后立即腹腔注射生理鹽水(40 mL/kg)，延遲復(fù)蘇治療組傷后立即在右后肢腹側(cè)面皮下注射精蛋白生物合成胰島素(2.0 U/kg)，延遲復(fù)蘇組與延遲復(fù)蘇治療組傷后6 h時(shí)腹腔注射生理鹽水(40 mL/kg) ;各組均于傷后24 h時(shí)采集腹主動(dòng)脈血，采用分光光度法測(cè)定血糖(BG)、乳酸脫氫酶(LDH)、肌酸激酶(CK)、尿素氮(BUN)及肌酐(Cr)。結(jié)果:即時(shí)復(fù)蘇組、延遲復(fù)蘇組與延遲復(fù)蘇治療組BG、LDH、CK、BUN、Cr均顯著高于假傷組(P＜0.05)，延遲復(fù)蘇組BG、LDH、CK、BUN、Cr顯著高于即時(shí)復(fù)蘇組(P＜0.05)，延遲復(fù)蘇治療組BG、LDH、CK、BUN、Cr顯著低于延遲復(fù)蘇組(P＜0.05)，延遲復(fù)蘇治療組LDH、CK顯著高于即時(shí)復(fù)蘇組(P＜0.05)，延遲復(fù)蘇治療組BG、Cr顯著低于即時(shí)復(fù)蘇組(P＜0.05)，延遲復(fù)蘇治療組BUN與即時(shí)復(fù)蘇組比較無顯著差異(P＞0.05)。結(jié)論:燒傷早期皮下注射精蛋白生物合成胰島素可減輕延遲復(fù)蘇大鼠的心臟及腎臟損害。

［關(guān)鍵詞］燒傷;復(fù)蘇;胰島素;心臟;腎

＊＊貴州醫(yī)科大學(xué)2013級(jí)外科學(xué)燒傷整形科碩士研究生

＊＊＊通信作者E-mail: gz_lwr@163.com

網(wǎng)絡(luò)出版時(shí)間: 2016－02－23網(wǎng)絡(luò)出版地址: http: / /www.cnki.net/kcms/detail/52.5012.R.20160223.2045.048.html

胰島素在嚴(yán)重?zé)齻菘藦?fù)蘇中具有潛在的應(yīng)用價(jià)值，休克期應(yīng)用普通胰島素可降低大面積燒傷患者復(fù)蘇所需膠－晶體溶液需要量，增加尿量，同時(shí)可減輕組織損傷程度和保護(hù)臟器功能［1］。胰島素有多種類型，不同類型胰島素的作用各有特點(diǎn)［2］。為探討燒傷早期皮下注射胰島素對(duì)延遲復(fù)蘇大鼠器官損害的影響，參照甘精胰島素治療Ⅱ型糖尿病時(shí)發(fā)揮抗氧化效應(yīng)的劑量［即＜0.4 U/(kg ·d)］［3］以及人與大鼠用藥劑量的換算關(guān)系［大鼠的劑量(mg/kg) = 6.25×人的劑量(mg/kg)］［4］，本實(shí)驗(yàn)觀察了燒傷早期皮下注射精蛋白生物合成胰島素(2.0 U/kg)對(duì)30%燒傷總面積(TBSA)延遲復(fù)蘇大鼠心臟及腎臟損傷的保護(hù)作用。

1　材料與方法

1.1實(shí)驗(yàn)動(dòng)物

SPF級(jí)成年雄性SD大鼠32只，由第三軍醫(yī)大學(xué)大坪醫(yī)院野戰(zhàn)外科研究所醫(yī)學(xué)實(shí)驗(yàn)動(dòng)物中心提供(scxk渝，2007－0005)，體重200～250 g，清潔環(huán)境飼養(yǎng)、萬級(jí)屏障系統(tǒng)，實(shí)驗(yàn)前適應(yīng)性馴養(yǎng)1周。

1.2主要儀器和試劑

721型分光光度儀購自山東高密彩虹分析儀器有限公司，TDL80－2B臺(tái)式低速離心機(jī)上海滬譽(yù)儀器有限公司，微量可調(diào)加樣器購自德國(guó)Eppendorf公司，長(zhǎng)效精蛋白生物合成胰島素購自丹麥諾和諾德公司，血糖(blood glucose，BG)、乳酸脫氫酶(lactate dehydrogenase，LDH)、肌酸激酶(creatinekinase，CK)、尿素氮(blood urea nitrogen，BUN)及肌酐(creatinine，Cr)測(cè)定試劑盒均購自中國(guó)南京建成生物工程研究所。

1.3方法

32只SD大鼠按隨機(jī)數(shù)字表法分為假傷組、即時(shí)復(fù)蘇組、延遲復(fù)蘇組與延遲復(fù)蘇治療組，每組8只。大鼠背部脫毛后，腹腔注射2.5 %戊巴比妥鈉(35 mg/kg)麻醉;參考文獻(xiàn)［5］的方法，即時(shí)復(fù)蘇組、延遲復(fù)蘇組與延遲治療組大鼠置于(95.0± 0.5)℃熱水中15 s，制作30% TBSA、Ⅲ°燙傷模型;假傷組僅置37℃溫水中15 s模擬燙傷。假傷組和即時(shí)復(fù)蘇組傷后立即腹腔注射生理鹽水(40 mL/kg)，延遲復(fù)蘇治療組傷后立即在右后肢腹側(cè)面皮下注射精蛋白生物合成胰島素(2.0 U/ kg)，延遲復(fù)蘇組與延遲復(fù)蘇治療組傷后6 h時(shí)腹腔注射生理鹽水(40 mL/kg)。各組大鼠分籠飼養(yǎng)，實(shí)驗(yàn)期間無死亡。各組大鼠傷后24 h開腹采集腹主動(dòng)脈血，2 000 r/min離心10 min，取上清液，采用分光光度法測(cè)定血糖(BG)、乳酸脫氫酶(LDH)、肌酸激酶(CK)、尿素氮(BUN)及肌酐(Cr)，按試劑盒說明書要求測(cè)定。

1.4統(tǒng)計(jì)學(xué)方法

采用SPSS 17.0軟件進(jìn)行統(tǒng)計(jì)學(xué)分析。數(shù)據(jù)以均值±標(biāo)準(zhǔn)差(±s)表示，組間比較采用單因素方差分析，P＜0.05表示差異有統(tǒng)計(jì)學(xué)意義。

2　結(jié)果

即時(shí)復(fù)蘇組、延遲復(fù)蘇組與延遲復(fù)蘇治療組BG、LDH、CK、BUN及Cr顯著高于假傷組，延遲復(fù)蘇組BG、LDH、CK、BUN及Cr顯著高于即時(shí)復(fù)蘇組，延遲復(fù)蘇治療組BG、LDH、CK、BUN及Cr顯著低于延遲復(fù)蘇組，延遲復(fù)蘇治療組LDH、CK顯著高于即時(shí)復(fù)蘇組，延遲復(fù)蘇治療組BG、Cr顯著低于即時(shí)復(fù)蘇組，差異均有統(tǒng)計(jì)學(xué)意義(P＜0.05) ;延遲復(fù)蘇治療組BUN與即時(shí)復(fù)蘇組比較差異無統(tǒng)計(jì)學(xué)意義(P＞0.05)。見表1。

Tab.1 Results of BG，LDH，CK，BUN and Cr of rats in the 4 groups

(1)與假傷組比較P＜0.05;(2)與即時(shí)復(fù)蘇組比較P＜0.05;(3)與延遲復(fù)蘇組比較P＜0.05

3　討論

患者于燒傷6 h后或更晚開始復(fù)蘇治療，屬于延遲復(fù)蘇之列［6］。燒傷延遲復(fù)蘇將導(dǎo)致多臟器并發(fā)癥發(fā)生和病死率增加［7］。本實(shí)驗(yàn)結(jié)果顯示，延遲復(fù)蘇組燒傷后24 h時(shí)BG、LDH、CK和BUN、Cr均顯著高于即時(shí)復(fù)蘇組，提示延遲復(fù)蘇可加重?zé)齻笫笮呐K及腎臟的損害，使血糖升高。在燒傷休克期，高血糖很常見［8］。動(dòng)物實(shí)驗(yàn)結(jié)果也顯示，TBSA30%、Ⅲ°燙傷大鼠的血糖傷后3 h時(shí)即顯著升高，一直持續(xù)到傷后24 h時(shí)［9］。胰島素作為體內(nèi)重要的激素，調(diào)節(jié)糖、蛋白與脂肪的代謝，并具有器官保護(hù)、抗炎、抗凋亡效應(yīng)［10］。本實(shí)驗(yàn)結(jié)果顯示，延遲復(fù)蘇治療組心肌酶活力、腎功能均顯著低于延遲復(fù)蘇組，并且Cr顯著低于即時(shí)復(fù)蘇組，BUN與即時(shí)復(fù)蘇組比較無顯著差異，提示燒傷早期皮下注射精蛋白生物合成胰島素可減輕延遲復(fù)蘇大鼠的心臟及腎臟損害。但延遲復(fù)蘇治療組心肌酶活力仍顯著高于即時(shí)復(fù)蘇組，表明燒傷后即時(shí)復(fù)蘇對(duì)保護(hù)臟器組織尤為重要。本實(shí)驗(yàn)結(jié)果同時(shí)顯示，延遲復(fù)蘇治療組血糖水平較延遲復(fù)蘇組顯著降低但仍高于假傷組，這有助于減少低血糖發(fā)生風(fēng)險(xiǎn)。

綜上，燒傷早期皮下注射精蛋白生物合成胰島素可減輕延遲復(fù)蘇大鼠的心臟及腎臟損害。在后續(xù)研究中，將進(jìn)一步探討胰島素發(fā)揮器官保護(hù)效應(yīng)的類型差異及其可能機(jī)制。

4　參考文獻(xiàn)

［1］施燕，張勤，鄭捷新，等.胰島素治療在燒傷休克復(fù)蘇中的應(yīng)用初探［J］.中華損傷與修復(fù)雜志，2012(1) :62－66.

［2］Asamoah E.Variability of insulin action and clinical effects［J］.The Endocrinologist，2007(17) : 282－290.

［3］Monnier L，Colette C，Michel F，et al.Insulin therapy has a complex relationship with measure of oxidative stress in type 2 diabetes: a case for further study［J］.Diabetes Metab Res Rev，2011(4) : 348－353.

［4］施新酋.醫(yī)用實(shí)驗(yàn)動(dòng)物學(xué)［M］.西安:陜西科學(xué)技術(shù)出版社，1989: 421.

［5］馮世杰，花蘭女，金曙雯，等.大鼠燙傷模型的制作［J］.上海第二醫(yī)科大學(xué)學(xué)報(bào)，1995(2) : 195－197.

［6］黎鰲，楊宗城.黎鰲燒傷學(xué)［M］.上海:上?？茖W(xué)技術(shù)出版社，2001: 56－57.

［7］邱嘯臣，金以超，孫瑜，等.富氫鹽水對(duì)延遲復(fù)蘇燙傷大鼠血壓及肺組織抗氧化能力的影響［J］.中華燒傷雜志，2010(6) : 435－438.

［8］Holm C，H?rbrand F，Mayr M，et al.Acute hyperglycaemia following thermal injury: friend or foe［J］.Resuscitation，2004(1) : 71－77.

［9］王偉，李偉人，孔豫蘇，等.胰島素對(duì)嚴(yán)重燙傷后早期大鼠血乳酸濃度及乳酸脫氫酶活力的影響［J］.中國(guó)急救醫(yī)學(xué)，2013(8) : 746－748.

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(2015－08－30收稿，2015－12－20修回)

中文編輯:戚璐;英文編輯:劉華

Effect of Hypodemic Injection of Protamin Biosynthetic Insulin on the Damage of Heart and Kidney of Delayed Resuscitation Rats in the Early Stage of Burn Injury

YUAN Xinzhang，WANG Wei，LI Weiren
(Department of Burn and Plastic Surgery，the Affiliated Hospital of Guizhou Medical University，Guiyang 550004，Guizhou，China)

［Abstract］Objective: To investigate the effect of hypodemic injection of protamin biosynthetic insulin on the damage of hearts and kidneys of rats with delayed resuscitation in the early stage of burn injury.Method: Thirty-two specific-pathogen free Sprague-Dawley rats were randomly divided into 4 groups，namely，sham burn group，immediate resuscitation group，delayed resuscitation group and delayed resuscitation treatment group，with 8 rats in each group.The rats of immediate resuscitation group，delayed resuscitation group and delayed resuscitation treatment group were immersed in (95± 0.5)℃hot water for 15 seconds to make empyrosis rat model of 30% total burn surface area，Ⅲdegree burn injury.The sham burn group was immersed in 37℃warm water for 15 seconds to simulate burns.Immediate resuscitation group and delayed resuscitation group received an intraperitoneal injection of physiological saline(40 mL/kg) after burn injury immediately，while delayed resuscitation treatment group received subcutaneous injection of protamine biosynthetic insulin{ (2.0 U/kg) in the ventral side of the right hind limb after burn injury immediately.Delayed resuscitation group and de-book=180,ebook=61layed resuscitation treatment group received intraperitoneal injection of physiological saline (40 mL/ kg) 6 hours after burn injury.The abdominal aortic blood was collected 24 hours after burn injury in each group.The spectrophotometry was adopted for determination of blood glucose(BG)，lactate dehydrogenase (LDH)，creatine kinase(CK)，blood urea nitrogen(BUN) and creatinine(Cr).Results: BG，LDH，CK，BUN and Cr in immediate resuscitation group，delayed resuscitation group and delayed resuscitation treatment group were significantly higher than their counterparts of sham burn group (P＜0.05).BG，LDH，CK，BUN and Cr in delayed resuscitation group were totally higher than their counterparts of immediate resuscitation group (P＜0.05).BG，LDH，CK，BUN and Cr in delayed resuscitation treatment group were significantly lower than their counterparts of delayed resuscitation group (P＜0.05).LDH and CK in delayed resuscitation treatment group were significantly higher than their counterparts of immediate resuscitation group，while BG and Cr were significantly lower than their counterparts of immediate resuscitation group (P＜0.05).There was no significant difference of BUN between delayed resuscitation treatment group and immediate resuscitation group (P＞0.05).Conclusion: During the early stage of burn injury，hypodemic injection of protamine biosynthetic insulin can reduce the damage of heart and kidney of delayed resuscitation rats.

［Key words］burn; recovery; insulin; heart; kidney

*［基金項(xiàng)目］貴州省衛(wèi)生廳科學(xué)技術(shù)基金(gzwkj2011－1－022)

［中圖分類號(hào)］R644

［文獻(xiàn)標(biāo)識(shí)碼］A

［文章編號(hào)］1000-2707(2016) 02-0179-03