夏至草醇提物干預(yù)實(shí)驗(yàn)性彌散性血管內(nèi)凝血大鼠肺損傷的實(shí)驗(yàn)研究

1、夏至草醇提物干預(yù)實(shí)驗(yàn)性彌散性血管內(nèi)凝血大鼠肺損傷的實(shí)驗(yàn)研究         09-09-07 14:27:00     編輯：studa20                   作者：晁懷宇，張玉平，劉正泉，雷慧，杜舒婷，姜華【摘要】  目的觀察夏至草醇提物對(duì)高分子右旋糖酐（Dextran

2、 500）致彌散性血管內(nèi)凝血（DIC）大鼠肺損傷的干預(yù)作用。方法Wistar雄性大鼠20只，隨機(jī)分為夏至草組（n=8）、模型組（n=6）和對(duì)照組（n=6）。靜脈推注10%Dextran 500（10 ml/kg·體重）復(fù)制DIC模型（對(duì)照組以等量生理鹽水代替）。6 min后，夏至草組自頸靜脈緩慢推注夏至草醇提物（5 g/ml，6 g/kg·體重），其它兩組以等量生理鹽水代替。40 min后，制備肺組織勻漿，觀察肺勻漿自由基指標(biāo)MDA含量及SOD活性、NO含量及NOS活性的變化。結(jié)果與對(duì)照組相比，模型組肺組織勻漿MDA、NO含量及NOS活性顯著升高，SOD活性降低；夏至草組肺

3、組織勻漿MDA、NO含量及NOS活性顯著低于模型組，SOD活性升高，各指標(biāo)與對(duì)照組比較無(wú)統(tǒng)計(jì)學(xué)意義。結(jié)論夏至草醇提物減輕Dextran 500致DIC大鼠肺損傷的機(jī)制與降低自由基損傷、降低NO的生成與釋放有關(guān)。 【關(guān)鍵詞】  夏至草；彌散性血管內(nèi)凝血；肺損傷；自由基；一氧Abstract：ObjectiveTo observe the effects of ethanol extract from Mrrubium incisum against lung injury of disseminated intravascuclar coagulation (DIC) rats ind

4、uced by Dextran 500.Methods20 male Wistar rats were randomly divided into three groups: Mrrubium incisum group (n=8), model group (n=6) and control group (n=6). The DIC model was established by intravenous injection of Dextran 500 (10 ml/kg·bw), and Dextran 500 was replaced by equivalent normal

5、 saline in control group. 6 minutes later, ethanol extract from Mrrubium incisum was infused  (5 g/ml, 6g/kg·bw); in other groups ethanol extract from Mrrubium incisum was replaced by normal saline. After 40 min, the lung was taken out, the 10% tissue homogenate was prepared, and the free

6、radical indexes of MDA content and SOD activity, NO content and NOS activity were determined.ResultsThe contents of MDA and NO and NOS activity of lung homogenate in model group were significantly increased, and SOD activity were significantly lower than that in control group. The contents of MDA an

7、d NO and NOS activity of lung homogenate in Mrrubium incisum group were significantly lowered, and SOD activity were significantly increased as compared with model group. There was no statistical difference between Mrrubium incisum group and control group. ConclusionEthanol extract from Mrrubium inc

8、isum can markedly abate lung injury of DIC rats by Dextran 500 and its mechanism may be related to scavenging free radical and decreasing NO release.Key words：Mrrubium incisum;  Disseminated intravascuclar coagulation (DIC);  Dextran 500;  Lung injury;  Free radical;  Nitric

9、 oxide    彌散性血管內(nèi)凝血（disseminated intravascuclar coagulation, DIC）是臨床常見(jiàn)的危重病理過(guò)程，在此病理過(guò)程中所形成的微血栓、以及出血后引起的低血容量、微循環(huán)障礙等因素，是導(dǎo)致危重患者出現(xiàn)肺損傷甚至多個(gè)器官功能障礙的主要因素之一。因此，尋求合理的治療措施防止DIC引起的肺損傷成為臨床研究的重點(diǎn)內(nèi)容。研究表明，高分子右旋糖酐（Dextran 500）在導(dǎo)致大鼠DIC及血液流變性異常的同時(shí)，可引起大鼠肝、腎、肺等多個(gè)器官組織形態(tài)學(xué)損傷，組織學(xué)觀察可見(jiàn)紅細(xì)胞淤滯、微血栓及細(xì)胞壞死等1；夏至草可明顯改善血淤大鼠的血

10、液和淋巴微循環(huán)障礙2，3，從而維持機(jī)體自穩(wěn)態(tài)。為了進(jìn)一步闡明夏至草醇提物干預(yù)DIC大鼠重要器官肺損傷的機(jī)制，本研究觀察夏至草醇提物對(duì)Dextran 500致大鼠DIC肺組織自由基及一氧化氮（NO）的影響，為臨床DIC致肺損傷的防治提供實(shí)驗(yàn)依據(jù)。1  器材與方法1.1  動(dòng)物與分組Wistar大鼠20只，體重240300 g（中國(guó)醫(yī)科院實(shí)驗(yàn)動(dòng)物繁育中心提供），隨機(jī)分為夏至草組（n=8）、模型組（n=6）和對(duì)照組（n=6）。前兩組復(fù)制DIC模型。1.2  藥品與儀器唇形科夏至草屬植物夏至草mrrubium incisum Benth的全草（內(nèi)蒙駐張家口醫(yī)藥站提供，經(jīng)河

11、北北方學(xué)院中醫(yī)系姜希強(qiáng)教授鑒定），按文獻(xiàn)4報(bào)道方法，制備夏至草醇提物，經(jīng)水煎醇沉，再經(jīng)活性炭脫色、滅菌制成原生藥含量為5g/ml的注射液（pH、澄清度、溶血試驗(yàn)檢測(cè)均為合格）；戊巴比妥鈉（北京化學(xué)試劑廠生產(chǎn)，批號(hào)20000919）；肝素鈉（北京奧博星生物技術(shù)責(zé)任有限公司生產(chǎn)，批號(hào)001218）；高分子右旋糖酐（Dextran 500，瑞典Pharmacia Biotech）。WZF-50F2雙道微量注射泵（浙江大學(xué)醫(yī)學(xué)儀器有限公司）；FJ-200型高速分散均質(zhì)機(jī)（上海標(biāo)本模具廠）。1.3  DIC模型的復(fù)制及標(biāo)本制備所有大鼠均以2%戊巴比妥鈉溶液（50 mg/kg·體重）肌

12、注麻醉后，行頸部正中手術(shù)切口，剝離左頸靜脈和右頸總動(dòng)脈，插管，備注射、輸液及放血用。穩(wěn)定15 min后，夏至草組及模型組大鼠，以10%的Dextran 500（10 ml/kg·體重）經(jīng)左頸靜脈緩慢勻速推注，1min注完，復(fù)制急性微循環(huán)模型5，6。6 min后，夏至草組經(jīng)微量注射泵由頸靜脈緩慢推注夏至草醇提物（6 g/kg·體重，以等量生理鹽水稀釋）；模型組輸入與夏至草醇提物等量的生理鹽水；輸液時(shí)間為5 min。對(duì)照組以等量生理鹽水代替Dextran 500和夏至草醇提物。經(jīng)夏至草醇提物治療或相當(dāng)于治療40min后，分別處死動(dòng)物。選擇固定肺組織0.81.0 g，加9倍的4無(wú)菌生理鹽水，應(yīng)用高速分散均質(zhì)機(jī)低溫制備10%組織勻漿，應(yīng)用高速低溫離心機(jī)以2 500 r/min離心10min，取上清液置于冰箱內(nèi)-80冷凍備測(cè)。1.4  指標(biāo)檢測(cè)應(yīng)用改良硫代巴比妥酸（TBA）微量法測(cè)定肺組織勻漿丙二醛（MDA）水平、黃嘌