原花青素对急性呼吸窘迫综合征模型大鼠治疗作用的实验研究

目的探讨原花青素(Procyanidolic,pc)对急性呼吸窘迫综合征(Acute respiratory distess syndrome,ARDS)大鼠疾病模型的治疗作用.方法大鼠用百草枯(Paraquat,PQ)一次性灌胃,剂量250 mg/kg,治疗组分别于染毒6、24、48 h各给予PC治疗剂量500 mg/kg,染毒组与治疗组分别于24、48、72 h,取大鼠血浆和支气管肺泡液(BALF),测试其血浆和BALF中超氧化物歧化酶(SOD)、丙二醛(MDA)谷胱甘肽过氧化物酶(GSH-Px)的活力,各组取肺病理组织做病理组织观察研究.结果治疗组与染毒组比较,血浆和BALF中GSH-P\-X、SOD活力逐渐升高,差异有显著性 P <0.05,MDA含量下降,差异有显著性 P <0.05,染毒与对照组比较,血浆和BALF中MDA含量高于对照组,差异有显著性 P <0.05,SOD和GSH-P\-X活力下降,差异有显著性 P <0.05,肺组织病理学观察,ARDS组肺泡壁充血,炎症细胞浸润,Ⅰ型与Ⅱ型细胞破坏,肺泡内有炎症渗出物,对照组肺泡壁完好,肺泡内没有炎症渗出物,没有炎症细胞浸润;给予PC治疗后,肺泡壁充血、出血减轻,炎症渗出物减少;结论 PC对ARDS模型大鼠有一定的治疗效果.     

黄磷及其化合物急性吸入致大鼠急性肺损伤或急性呼吸窘迫综合征模型的制作

目的建立黄磷及其化合物急性吸入致大鼠急性肺损伤（ALI）/急性呼吸窘迫综合征（ARDS）的模型。方法健康SD大鼠48只随机分为对照组以及实验组（0、4、12、24、48 h时间点处死）。采用自制染毒装置,间歇染毒形成ALI/ARDS模型。观察ALI/ARDS大鼠动脉血气分析以及肺系数和肺组织病理变化。结果肺损伤后大鼠动脉血气分析以及肺组织病理改变明显恶化,肺系数较对照组明显增大。结论成功地建立了黄磷及其化合物急性吸入致大鼠ALI/ARDS的模型,为黄磷及其化合物吸入中毒的防治研究提供良好实验基础,同时也适用于其他气体吸入致ARDS的实验研究。     

硫化氢在急性百草枯中毒致大鼠肺损伤中的作用

目的阐明百草枯中毒致大鼠肺损伤时机体内源性H2S的变化,探讨硫化氢在急性百草枯中毒致大鼠肺损伤中的作用。方法按时间点将50只大鼠分为5组,同时染毒;选择对应时间点50只大鼠为对照组。分组检测肺组织中内源性H2S的含量,并及时处死大鼠,行肺组织损伤病理学评分。另外取大鼠40只分为4组,即空白对照组、染毒组、染毒+外源性H2S组、外源性H2S组,于12h后,检测肺组织中内源性H2S的含量,并及时处死大鼠,行肺组织损伤病理学评分。结果百草枯中毒致大鼠肺损伤在不同时间范围内,机体内源性H2S的含量差异有显著统计学意义(P0.01);与染毒组比较,染毒组+外源性H2S组肺损伤程度评分显著降低,差异具有显著统计学意义(P0.01)。结论百草枯致大鼠肺损伤过程中,内源性H2S的含量与肺损伤程度呈负相关;外源性H2S通过增加体内肺组织H2S的含量,抑制百草枯致肺损伤。。     

百草枯对小鼠神经行为发育及学习记忆功能的影响

目的观察百草枯(PQ)对发育期C57BL/6J小鼠神经发育的毒性作用,并探讨百草枯对小鼠学习记忆的影响。方法 80只出生21日龄的仔鼠分为对照组(生理盐水)、1.25、2.5、5、10 mg/(kg·d)五组,灌胃染毒百草枯,每天一次,连续30 d。观察小鼠的一般生理和神经行为发育情况,并在染毒结束后进行Morris水迷宫实验和避暗实验,测试小鼠的学习记忆功能。神经行为学测试结束后取小鼠大脑,称重并进行病理检查,同时利用透射电镜观察各组小鼠中脑黑质部超微结构。结果染毒期间小鼠一般状况没有明显变化,染毒结束后各组体重没有统计学差异;在Morris水迷宫测试中,各组差异没有统计学意义,而避暗实验中与对照组相比,高剂量组的避暗潜伏期延长,差异有显著性(P<0.05);在病理切片和透射电镜观察中,在高剂量组分别观察到黑质细胞减少和神经元细胞凋亡。结论百草枯暴露对发育期小鼠成年后神经行为有影响,同时会使小鼠成年后出现脑组织的病理变化,发生器质性的病变。     

大鼠对乙酰柠檬酸三丁酯的安全耐受值测定

按体重将80只SD大鼠随机分为对照组,低、中、高剂量组,每组雌雄各10只。持续静脉给药观察4周后处死,观察体重、脏器系数、血液及生化指标以及病理切片。与对照组比较,各组大鼠体重无显著性差异(p0.05);高剂量组雄鼠脑、肺、胸腺脏器系数差异显著(p0.05),雌鼠肝、卵巢脏器系数有差异显著(p0.05);中高剂量组雌、雄鼠部分血液及生化指标差异显著(p0.05);病理切片观察,各组大鼠肝脏均出现脂变,中高剂量组大鼠附睾、大肠出现炎症,其他器官未见明显病理改变。中高剂量组ATBC溶液对大鼠有一定毒性,参考低剂量组给药浓度,大鼠对其安全剂量为500 mg/kg体重/d。     

纳米氧化锌经口染毒对C57BL/6J小鼠外周脏器的影响*

目的:探讨纳米氧化锌经口染毒60 d对C57BL/6J小鼠多种外周脏器的损伤作用。方法:20只雄性C57BL/6J小鼠随机分为对照组和实验组,每组10只,实验组将纳米氧化锌溶液以20 mg/kg体重的剂量连续灌胃染毒60 d,对照组给予相应量的生理盐水;小鼠每周称重一次,染毒结束后,眼球取血,检测血糖、血脂、肝功能和肾功能相关指标,以及血清中炎症因子PAF、IL-6和TNF-α含量;取心脏、肝脏、脾脏、肺、肾脏和小肠组织制备病理切片,HE染色后,观察组织形态学变化。结果:实验组和对照组之间的体重无显著性差异;与正常对照组比较,实验组大鼠血中白蛋白(ALB)、白蛋白/球蛋白比值(A/G)、碱性磷酸酶(ALP)、谷草/谷丙转氨酶比值(S/L)、尿酸(UA)和尿素氮(BUN)含量明显升高(P＜0.05或 P＜0.01);两组间血清中炎症因子含量无显著差异。病理学检查发现,实验组心肌中部分区域出现浊肿,肝脏出现轻度炎性病变(灶性或小灶性坏死),脾脏色素沉着减少,肺部出现轻或中度间质性炎症,肾脏和小肠未见明显病理改变。结论:纳米氧化锌经口染毒60 d未引起C57BL/6J小鼠血液系统炎症,但可诱导心脏、肝脏、脾脏和肺脏出现轻度的病理变化,并导致肝脏和肾脏的功能异常。     

肺纤维化大鼠模型造模方法的优化

目的 比较气管内滴注、气管内雾化喷入博来霉素(5 mg/kg)两种给药方式,以及比较腹腔注射3%戊巴比妥钠麻醉、异氟烷呼吸麻醉(与氧气混合吸入时,异氟烷浓度为0.5%)两种麻醉方式对肺纤维化大鼠模型的影响,探讨更优的造模方法。方法 选择雄性SPF级SD大鼠50只,随机分为空白对照组、腹腔麻醉气管内滴注组、腹腔麻醉气管内雾化喷入组、呼吸麻醉气管内滴注组和呼吸麻醉气管内雾化喷入组,每组各10只。观察给药后1、3、7、14、21 d,各组大鼠的生存状况及体重变化;给药3周后处死大鼠,取肺称重,计算肺系数;HE染色观察肺组织炎症变化;Masson染色观察肺组织中胶原增殖;Western Blot检测肺组织中转化生长因子-β1(TGF-β1)蛋白表达量;碱水解法检测肺组织中羟脯氨酸(HYP)的含量。结果 与空白对照组比,4种模型组的大鼠精神状态不佳、体重下降、肺指数上升;肺组织损伤明显,炎症水平增加,胶原增殖显著;肺组织中TGF-β1蛋白表达水平升高(P<0.001);发现仅气管内雾化喷入博来霉素组大鼠肺组织中羟脯氨酸的含量增加(P<0.05)。发现气管内滴注的建模方式其模型大鼠肺部...     

人脐带间充质干细胞对百草枯中毒肺损伤小鼠模型的治疗作用

目的:探讨脐带间充质干细胞(UCMSC)治疗百草枯中毒引起的小鼠肺损伤的可行性。方法:小鼠腹腔一次性注射百草枯制备百草枯中毒小鼠模型,24 h后尾静脉注射UCMSC,分别于治疗后7和21 d取材,观察UCMSC对急性肺损伤和慢性肺纤维化的治疗作用。结果:UCMSC移植对40和50 mg/kg百草枯染毒组急性肺损伤有效,动物死亡率显著降低,但对60 mg/kg百草枯染毒组动物无效。UCMSC治疗对慢性肺纤维化有显著治疗作用,治疗组动物体重恢复早,死亡率降低,肺纤维化评分降低。RT-PCR结果显示,UCMSC移植3 h有人特异性线粒体基因的表达,但24 h后未检测到。结论:UCMSC对百草枯中毒性急慢性肺损伤有一定的治疗作用,这种作用可能是通过旁分泌机制实现的。     

共聚焦激光扫描对呼吸窘迫综合症大鼠肺组织的观察

目的：为了更直观地观察和显示呼吸窘迫综合症(acute respiratory distress syndrom,ARDS)典型的病理变化(肺泡内形成一层蛋白质透明膜)。方法：利用百草枯(Paraqual)染毒SD大鼠复制ARDS实验动物模型,取肺病理组织,切片,试剂Goat—Anti-Rat—FITC IgM IgG染色,共聚焦激光扫描显微镜(confocal laser scarming microscope,CLSM)观察。结果：CLSM能清晰到样品内不同层面的病理变化。结论：共聚焦激光扫描显微镜能清晰观察样品内不同层面的结构,相比于传统的光学显微镜,其观察到的图像更直观、更具立体感,能更好表达ARDS的病理变化特征。     

百草枯中毒大鼠TNF-α、IL-10 的表达及大黄保护作用的研究

目的:观察大黄对急性百草枯中毒大鼠TNF-α、IL-10的干预作用,探讨其可能的作用机制。方法:90只SD大鼠随机分为生理盐水对照组(A组)、PQ(60 mg/kg)灌胃染毒组(B组)、生大黄(300mg/kg.d)干预组(C组),每组30只。中毒后6h、24h、72h分批处死存活的大鼠,并且检测大鼠血浆TNF-α、IL-10水平。结果:B组、C组TNF-α、IL-10水平在染毒后6h开始升高,72h达到高峰,与A组相比,差异有统计学意义(P<0.05、P<0.01),在相同时间点C组TNF-α和IL-10的表达低于B组,差异均有统计学意义(P<0.01)。B组、C组血浆TNF-α、IL-10水平与中毒时间呈显著正性相关关系(r=0.849,P<0.01;r=0.790,P<0.01;r=0.0.943,P<0.01;r=0.892,P<0.01)。结论:大黄能够通过降低百草枯中毒大鼠体内的TNF-α、IL-10水平,减轻百草枯对大鼠的损伤作用。     

腹腔注射百草枯构建小鼠肺纤维化模型

目的:探讨百草枯一次性腹腔注射致小鼠肺纤维化的病理改变及半数致死剂量(LD50),进而制备肺纤维化病理改变稳定的百草枯中毒小鼠肺纤维化模型。方法:60只正常雌性C57BL/6J小鼠被随机分为6组,10只/组,分别为正常对照组及百草枯给药30、40、50、60和80 mg/kg组,所有小鼠于造模后28 d处死,取其左肺用于病理观察(HE染色),并计算LD50及各组肺纤维化Ashcroft评级。结果:至观察期28 d,小鼠一次性腹腔注射百草枯溶液的LD50为55.1923 mg/kg;各染毒组均出现不同程度的肺纤维化改变,且注射剂量越高,肺纤维化病变越严重,但早期死亡率亦越高。结论:一次性腹腔注射百草枯可制备小鼠肺纤维化模型,40和50 mg/kg为较合适的造模剂量。     

Using Bosentan to Treat Paraquat Poisoning-Induced Acute Lung Injury in Rats

Background

Paraquat poisoning is well known for causing multiple organ function failure (MODS) and high mortality. Acute lung injury and advanced pulmonary fibrosis are the most serious complications. Bosentan is a dual endothelin receptor antagonist. It plays an important role in treating PF. There is no related literature on the use of bosentan therapy for paraquat poisoning.

Objective

To study the use of bosentan to treat acute lung injury and pulmonary fibrosis as induced by paraquat.

Method

A total of 120 adult Wister male rats were randomly assigned to three groups: the paraquat poisoning group (rats were intragastrically administered with paraquat at 50 mg/kg body weight once at the beginning); the bosentan therapy group (rats were administered bosentan at 100 mg/kg body weight by intragastric administration half an hour after paraquat was administered, then the same dose was administered once a day); and a control group (rats were administered intragastric physiological saline). On the 3rd, 7th, 14th, and 21st days following paraquat exposure, rats were sacrificed, and samples of lung tissue and venous blood were collected. The levels of transforming growth factor-β1 (TGF-β1), endothelin-1 (ET-1), and hydroxyproline (HYP) in the plasma and lung homogenate were determined. Optical and electronic microscopes were used to examine pathological changes.

Result

The TGF-β1, ET-1, and HYP of the paraquat poisoning group were significantly higher than in the control group, and they were significantly lower in the 21st day therapy group than in the paraquat poisoning group on the same day. Under the optical and electronic microscopes, lung tissue damage was observed to be more severe but was then reduced after bosentan was administered.

Conclusion

Bosentan can reduce inflammation factor release. It has a therapeutic effect on acute lung injury as induced by paraquat.     

内质网应激及自噬参与百草枯中毒所致肺损伤

目的:探讨内质网应激及自噬在百草枯中毒所致大鼠肺脏损伤中的作用。方法:选取Wistar大鼠腹80只,腹腔注射百草枯(15 mg/kg)建立百草枯中毒肺脏损伤的动物模型。染毒后1、3、7、14、21 d处死动物取肺组织,采用HE染色和Van Gieson(V.G)染色观察大鼠肺脏损伤及纤维化情况,电镜观察Wistar大鼠肺脏胞浆空泡变、自噬体的形成以及肺脏损伤。Western-blot方法观察Wistar大鼠内质网应激相关蛋白(GRP94、Caspase-12和CHOP)和自噬相关蛋白(LC3-II、Beclin-1)的表达。结果:HE及V.G染色结果显示随中毒时间延长,百草枯中毒肺损伤及肺纤维化逐渐加重;电镜结果显示百草枯中毒肺脏发生胞浆空泡变、自噬体形成。与对照组比较,在百草枯中毒组内质网应激相关蛋白GRP94在3 d表达达到峰值(P0.001),7 d表达开始降低(P0.05),CHOP蛋白表达3 d开始增加(P0.001),cleaved caspase-12蛋白表达7 d开始增加(P0.001),并逐渐加强,自噬相关蛋白LC3-II和Beclin-1表达3 d开始增加(P0.001),14 d表达最高(P0.001)。结论:内质网应激以及细胞自噬共同参与百草枯中毒所致肺脏损伤。     

百草枯致急性肺损伤大鼠模型的建立

目的建立一种百草枯诱导的急性肺损伤（ALI）大鼠模型。方法将20只SD大鼠随机分为正常对照组10只、实验组10只。实验组一次性口服灌胃百草枯（PQ）80mg／kg,于给药后1天处死大鼠,观察光镜下肺组织病理改变、肺动脉血氧分压（PaO2）、支气管肺泡灌洗液（BALF）蛋白含量等。结果给予百草枯1天后肺形态学出现显著异常,PaO2及BALF蛋白含量出现显著改变。结论一次性灌胃百草枯80mg／kg成功建立急性肺损伤动物模型。     

一种高尿酸血症大鼠模型诱导方法的改良和效果评价研究

目的: 探索短期内诱导高尿酸血症大鼠模型的有效方法,并对模型效果进行评价。方法: 雄性SD大鼠随机分为对照组(CT组,6只)和5个模型组(M1-M5组),每组8只;M1组(每天酵母膏10 g/kg+腺嘌呤100 mg/kg 2次灌胃,于模型诱导的第7日1次性腹腔注射氧嗪酸钾300 mg/kg)、M2组(每天酵母膏10 g/kg+腺嘌呤100 mg/kg灌胃2次,于模型诱导第1、3、7日每天腹腔注射1次氧嗪酸钾300 mg/kg)、M3组(每天酵母膏10 g/kg+腺嘌呤100 mg/kg灌胃 2次,每天腹腔注射1次氧嗪酸钾300 mg/kg)、M4组(每天酵母膏20 g/kg+腺嘌呤100 mg/kg灌胃 2次,每天腹腔注射1次氧嗪酸钾300 mg/kg)、M5组(每天酵母膏30 g/kg+腺嘌呤100 mg/kg灌胃2次,每天腹腔注射1次氧嗪酸钾300 mg/kg)、CT组(5个模型组按相同的时间、体重计算等体积灌胃和腹腔注射生理盐水),造模7 d;分别在造模结束时和2周后采集24 h尿样和血样检测尿酸、肌酐水平,取肾脏和胃称重,观察肾脏病理变化。结果: 与CT组相比,造模结束后,所有模型组大鼠体重均显著降低(P＜0.01);除M2组外,其他造模组大鼠均有亡,M4组和M5组因死亡率高未做后续分析,M1和M3组分别死亡4例和2例;造模结束后,模型大鼠血尿酸、尿尿酸水平明显升高(P＜0.01),并且M2组的血尿酸水平显著高于其他各组(P＜0.05);继续喂养2周后,各模型组的血尿酸和尿尿酸水平仍显著升高(P＜0.05);各模型组大鼠肾脏重量也明显增加(P＜0.01);病理检查显示,模型组大鼠肾脏出现明显炎症反应和结构破坏。结论: 采用酵母膏(10 g/kg)、腺嘌呤(100 mg/kg)联合氧嗪酸钾(300 mg/kg)间隔(第1、3、7日)注射的方案可在短期内安全地诱导高尿酸血症大鼠模型,模型效果持续时间较长,适合在相关研究中应用。     

不同剂量甲泼尼龙对大鼠急性百草枯中毒肾脏的影响

目的:观察不同剂量甲泼尼龙治疗大鼠百草枯中毒肾脏损伤的疗效。方法:将120只Wistar大鼠随机分为五组,空白组,染毒组和干预组(根据甲泼尼龙剂量不同分为三组),除空白组外,均予百草枯(22mg/kg)稀释后腹腔注射,2h后依照组别、体重注射甲泼尼龙,在第1、3、7天共3个时间点,按抽签法处死实验对象6只获取标本,观察肾功能和病理变化。结果:各组血尿素氮(P=0.001<0.05)和肌酐(P=0.01<0.05)差异有统计学意义,干预组中5mg/kg甲泼尼龙组同染毒组比较差异有统计学意义。不同时间点血尿素氮(P=0.007<0.05)和肌酐(P=0.016<0.05)差异有统计学意义,其中第七天明显低于第一、三天。病理评分各组(P=0.21>0.05)差异无统计学意义。讨论:早期应用糖皮质激素治疗PQ中毒大鼠,可以显著减轻PQ中毒所致的肾损伤程度,改善肾功能,尤其小剂量改善显著,传统的大剂量糖皮质激素冲击治疗不值的推崇。     

Attenuation of the extract from Moringa oleifera on monocrotaline-induced pulmonary hypertension in rats

The purpose of this study was to determine the effects of an extract from Moringa oleifera (MO) on the development of monocrotaline (MCT)-induced pulmonary hypertension (PH) in Wistar rats. An ethanol extraction was performed on dried MO leaves, and HPLC analysis identified niaziridin and niazirin in the extract. PH was induced with a single subcutaneous injection of MCT (60 mg/kg) which resulted in increases in pulmonary arterial blood pressure (Ppa) and in thickening of the pulmonary arterial medial layer in the rats. Three weeks after induction, acute administration of the MO extract to the rats decreased Ppa in a dose-dependent manner that reached statistical significance at a dose of 4.5 mg of freeze-dried extract per kg body weight. The reduction in Ppa suggested that the extract directly relaxed the pulmonary arteries. To assay the effects of chronic administration of the MO extract on PH, control, MCT and MCT+MO groups were designated. Rats in the control group received a saline injection; the MCT and MCT+MO groups received MCT to induce PH. During the third week after MCT treatment, the MCT+MO group received daily i.p. injections of the MO extract (4.5 mg of freeze-dried extract/kg of body weight). Compared to the control group, the MCT group had higher Ppa and thicker medial layers in the pulmonary arteries. Chronic treatments with the MO extract reversed the MCT-induced changes. Additionally, the MCT group had a significant elevation in superoxide dismutase activity when normalized by the MO extract treatments. In conclusion, the MO extract successfully attenuated the development of PH via direct vasodilatation and a potential increase in antioxidant activity.     

Effects of 2-azafluorenones on phosphatidyl-inositol specific phospholipase C activation in c6 glioma cells

The purpose of this study was to determine the effects of an extract from Moringa oleifera (MO) on the development of monocrotaline (MCT)-induced pulmonary hypertension (PH) in Wistar rats. An ethanol extraction was performed on dried MO leaves, and HPLC analysis identified niaziridin and niazirin in the extract. PH was induced with a single subcutaneous injection of MCT (60 mg/kg) which resulted in increases in pulmonary arterial blood pressure (Ppa) and in thickening of the pulmonary arterial medial layer in the rats. Three weeks after induction, acute administration of the MO extract to the rats decreased Ppa in a dose-dependent manner that reached statistical significance at a dose of 4.5 mg of freeze-dried extract per kg body weight. The reduction in Ppa suggested that the extract directly relaxed the pulmonary arteries. To assay the effects of chronic administration of the MO extract on PH, control, MCT and MCT+MO groups were designated. Rats in the control group received a saline injection; the MCT and MCT+MO groups received MCT to induce PH. During the third week after MCT treatment, the MCT+MO group received daily i.p. injections of the MO extract (4.5 mg of freeze-dried extract/kg of body weight). Compared to the control group, the MCT group had higher Ppa and thicker medial layers in the pulmonary arteries. Chronic treatments with the MO extract reversed the MCT-induced changes. Additionally, the MCT group had a significant elevation in superoxide dismutase activity when normalized by the MO extract treatments. In conclusion, the MO extract successfully attenuated the development of PH via direct vasodilatation and a potential increase in antioxidant activity.     

N-acetylcysteine pretreatment attenuates paraquat-induced lung leak in rats

We investigated the effects of N-acetylcysteine (NAC) pretreatment on paraquat-induced lung inflammation and leak. We found that administering a single intravenous dose (60 mg/kg) of paraquat rapidly (2 h) increased lung leak, lung lavage cytokine-induced neutrophil chemoattractant (CINC) levels, and lung myeloperoxidase (MPO) activity in rats. Rats pretreated with NAC (150 mg/kg, intraperitoneally) had increased lung tissue glutathione (GSH + GSSG) levels compared to saline-pretreated rats. In addition, rats pretreated with NAC and then given paraquat 2.5 h later had decreased lung leak compared to saline-pretreated rats given paraquat. In contrast, NAC pretreated rats given paraquat had the same lung lavage CINC levels and lung tissue MPO activity as saline-pretreated rats given paraquat. Our results indicate that paraquat causes an oxidative injury which may be decreased by the GSH-increasing or other properties of NAC.