夏至草醇提物干预实验性弥散性血管内凝血大鼠肺损伤的实验研究

1、夏至草醇提物干预实验性弥散性血管内凝血大鼠肺损伤的实验研究         09-09-07 14:27:00     编辑：studa20                   作者：晁怀宇，张玉平，刘正泉，雷慧，杜舒婷，姜华【摘要】  目的观察夏至草醇提物对高分子右旋糖酐（Dextran

2、 500）致弥散性血管内凝血（DIC）大鼠肺损伤的干预作用。方法Wistar雄性大鼠20只，随机分为夏至草组（n=8）、模型组（n=6）和对照组（n=6）。静脉推注10%Dextran 500（10 ml/kg·体重）复制DIC模型（对照组以等量生理盐水代替）。6 min后，夏至草组自颈静脉缓慢推注夏至草醇提物（5 g/ml，6 g/kg·体重），其它两组以等量生理盐水代替。40 min后，制备肺组织匀浆，观察肺匀浆自由基指标MDA含量及SOD活性、NO含量及NOS活性的变化。结果与对照组相比，模型组肺组织匀浆MDA、NO含量及NOS活性显著升高，SOD活性降低；夏至草组肺

3、组织匀浆MDA、NO含量及NOS活性显著低于模型组，SOD活性升高，各指标与对照组比较无统计学意义。结论夏至草醇提物减轻Dextran 500致DIC大鼠肺损伤的机制与降低自由基损伤、降低NO的生成与释放有关。 【关键词】  夏至草；弥散性血管内凝血；肺损伤；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    弥散性血管内凝血（disseminated intravascuclar coagulation, DIC）是临床常见的危重病理过程，在此病理过程中所形成的微血栓、以及出血后引起的低血容量、微循环障碍等因素，是导致危重患者出现肺损伤甚至多个器官功能障碍的主要因素之一。因此，寻求合理的治疗措施防止DIC引起的肺损伤成为临床研究的重点内容。研究表明，高分子右旋糖酐（Dextran 500）在导致大鼠DIC及血液流变性异常的同时，可引起大鼠肝、肾、肺等多个器官组织形态学损伤，组织学观察可见红细胞淤滞、微血栓及细胞坏死等1；夏至草可明显改善血淤大鼠的血

10、液和淋巴微循环障碍2，3，从而维持机体自稳态。为了进一步阐明夏至草醇提物干预DIC大鼠重要器官肺损伤的机制，本研究观察夏至草醇提物对Dextran 500致大鼠DIC肺组织自由基及一氧化氮（NO）的影响，为临床DIC致肺损伤的防治提供实验依据。1  器材与方法1.1  动物与分组Wistar大鼠20只，体重240300 g（中国医科院实验动物繁育中心提供），随机分为夏至草组（n=8）、模型组（n=6）和对照组（n=6）。前两组复制DIC模型。1.2  药品与仪器唇形科夏至草属植物夏至草mrrubium incisum Benth的全草（内蒙驻张家口医药站提供，经河

11、北北方学院中医系姜希强教授鉴定），按文献4报道方法，制备夏至草醇提物，经水煎醇沉，再经活性炭脱色、灭菌制成原生药含量为5g/ml的注射液（pH、澄清度、溶血试验检测均为合格）；戊巴比妥钠（北京化学试剂厂生产，批号20000919）；肝素钠（北京奥博星生物技术责任有限公司生产，批号001218）；高分子右旋糖酐（Dextran 500，瑞典Pharmacia Biotech）。WZF-50F2双道微量注射泵（浙江大学医学仪器有限公司）；FJ-200型高速分散均质机（上海标本模具厂）。1.3  DIC模型的复制及标本制备所有大鼠均以2%戊巴比妥钠溶液（50 mg/kg·体重）肌

12、注麻醉后，行颈部正中手术切口，剥离左颈静脉和右颈总动脉，插管，备注射、输液及放血用。稳定15 min后，夏至草组及模型组大鼠，以10%的Dextran 500（10 ml/kg·体重）经左颈静脉缓慢匀速推注，1min注完，复制急性微循环模型5，6。6 min后，夏至草组经微量注射泵由颈静脉缓慢推注夏至草醇提物（6 g/kg·体重，以等量生理盐水稀释）；模型组输入与夏至草醇提物等量的生理盐水；输液时间为5 min。对照组以等量生理盐水代替Dextran 500和夏至草醇提物。经夏至草醇提物治疗或相当于治疗40min后，分别处死动物。选择固定肺组织0.81.0 g，加9倍的4无菌生理盐水，应用高速分散均质机低温制备10%组织匀浆，应用高速低温离心机以2 500 r/min离心10min，取上清液置于冰箱内-80冷冻备测。1.4  指标检测应用改良硫代巴比妥酸（TBA）微量法测定肺组织匀浆丙二醛（MDA）水平、黄嘌