丹参提取物对CCl4和DMN诱导的大鼠肝纤维化的影响

分别采用四氯化碳和二甲基亚硝胺诱导的大鼠肝纤维化模型,以秋水仙碱和丹参作对照,通过肝组织病理学及Ⅰ、Ⅲ型胶原免疫组化观察,肝组织羟脯氨酸、丙二醛及血清肝功能部分指标检测,探讨丹提取物和治疗肝纤维化的效果及部分作用机理。     

Antioxidant activity and hepatoprotective effect of 10 medicinal herbs on CCl_4-induced liver injury in mice

BACKGROUND Many natural products confer health benefits against diverse diseases through their antioxidant activities. Carbon tetrachloride(CCl_4) is often used in animal experiments to study the effects of substances on liver injury and the related mechanisms of action, among which oxidative stress is a major pathogenic factor.AIM To compare antioxidant and hepatoprotective activities of ten herbs and identify and quantify phytochemicals for the one with strongest hepatoprotection.METHODS The antioxidant activity of ten medicinal herbs was determined by both ferricreducing antioxidant power and Trolox equivalent antioxidant capacity assays. The total phenolic and flavonoid contents were determined by Folin–Ciocalteu method and aluminum chloride colorimetry, respectively. Their effects on CCl_4-induced oxidative liver injury were evaluated and compared in a mouse model by administrating each water extract(0.15 g/mL, 10 mL/kg) once per day for seven consecutive days and a dose of CCl_4 solution in olive oil(8%, v/v, 10 mL/kg). The herb with the strongest hepatoprotective performance was analyzed for the detailed bioactive components by using high-performance liquid chromatography-electrospray ionization source-ion trap tandem mass spectrometry.RESULTS The results revealed that all tested herbs attenuated CCl_4-induced oxidative liver injury; each resulted in significant decreases in levels of serum alanine transaminase, aspartate transaminase, alkaline phosphatase, and triacylglycerols.In addition, most herbs restored hepatic superoxide dismutase and catalase activities, glutathione levels, and reduced malondialdehyde levels. Sanguisorba officinalis(S. officinalis) L., Coptis chinensis Franch., and Pueraria lobata(Willd.) Ohwi root were the three most effective herbs, and S. officinalis L. exhibited the strongest hepatoprotective effect. Nine active components were identified in S. officinalis L. Gallic acid and(+)-catechin were quantified(7.86 ± 0.45 mg/g and 8.19 ± 0.57 mg/g dried weight, respectively). Furthermore, the tested herbs displayed a range of in vitro antioxidant activities proportional to their phenolic content; the strongest activities were also found for S. officinalis L.CONCLUSION This study is of value to assist the selection of more effective natural products for direct consumption and the development of nutraceuticals or therapeutics to manage oxidative stress-related diseases.     

吴茱萸碱复合纳米粒对急性肝损伤模型小鼠的保护作用

目的：探讨吴茱萸碱复合纳米粒（evodiamine complex loaded nanoparticles，ECNP）对四氯化碳（CCl₄）诱导的急性肝损伤模型小鼠的保护作用。方法：采用离子凝胶法制备ECNP，测定其粒径和电位。腹腔注射CCl₄复制小鼠急性肝损伤模型。评估ECNP和EVO干预对肝损伤小鼠血清指标AST、ALT、TNF-α、IL-1β和肝脏指数的影响；测定肝脏GSH、CAT、SOD水平和MDA含量。光镜下观察肝脏病理改变。结果：ECNP的粒径和电位分别为（435.30±24.70）nm和（43.43±0.50）mV。ECNP和EVO干预能降低小鼠肝脏指数，改善小鼠肝组织病理损伤，降低血清ALT、AST水平，减少炎症因子IL-1β、TNF-α的产生，增强肝组织抗氧化物SOD、GSH、CAT的能力；ECNP调节血清转氨酶、降低炎症因子、增加机体抗氧化能力的作用较等剂量的游离EVO更强。结论：ECNP对CCl₄致肝损伤有一定的保护作用，且比EVO效果更好。     

The protective effect of cinnamon and ginger hydro-alcoholic extract on carbon tetrachloride-induced testicular damage in rats

Sexual dysfunction of men is one of the most serious problems in human society. This study aimed to evaluate the protective effect of cinnamon and ginger extract on testicular damages induced by carbon tetrachloride (CCl4). Thirty-six male Wistar rats were randomly divided into 6 groups (n = 6): 1. Normal control; 2. Carbon tetrachloride (CCl4); 3. CCl4 + Cinnamon; 4. CCl4 + Ginger; 5. CCl4 + Cinnamon and Ginger; and 6. Cinnamon + Ginger. CCl4 (1 ml/kg) was injected intraperitoneally on the 14th day, and cinnamon (50 mg/kg, orally) and ginger (250 mg/kg, orally) were administered daily for 14 days. Fifty hours after the CCl4 injection, the testicles and epididymis were separated and examined as to histological alterations and oxidative stress markers. CCl4 significantly increased malondialdehyde level and decreased total antioxidant capacity when compared to the normal control group (p < .05). In addition, degenerative alterations in the testicular and epididymal tissue were observed in CCl4 group. The pre-treatment with ginger and cinnamon extract significantly improved these parameters when compared to the CCl4 group (p < .05). The results of this study indicated that co-treatment of ginger and cinnamon reduces the damages induced by CCl4 in testicular tissue by increasing antioxidant capacity and reducing lipid peroxidation.     

CCl4,黄磷,As2O3对大鼠肝脏,血液巯基含量的影响

CCl_4、黄磷可降低肝脏巯基含量,而As_2O_3对血液巯基含量降低更显著。这可能是由于CCl_4、黄磷在肝内分布多于血液,而As_2O_3在血液中的储存多于肝脏之故。     

板蓝根对小鼠实验性肝损伤的保护作用及机制

目的探讨板蓝根对四氯化碳(carbon tetrachloride,CCl4)致小鼠肝损伤的保护作用和机制。方法 2013年1—4月选取健康小鼠40只随机分为生理盐水组(A组)、CCl4组(B组)、低剂量板蓝根组(C组)和高剂量板蓝根组(D组)各10只,A、B两组腹腔注射生理盐水10 ml/kg,1次/d;C组腹腔注射板蓝根剂20 ml/kg,1次/d;D组腹腔注射板蓝根剂80 ml/kg,1次/d,四组均注射10 d。末次给药2 h后,B、C、D三组均腹腔注射1 ml/L的CCl4花生油溶液10 ml/kg建立急性CCl4小鼠肝损伤模型。比较四组小鼠肝脏病理组织学变化、肝匀浆中超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)水平、血清ALT、AST水平。计量资料采用t检验,P<0.05为差异有统计学意义。结果空泡变性、肝细胞坏死、炎性细胞浸润情况A组分别为0、0、0只,B组分别为8、10、10只,C组分别为7、6、7只,D组分别为5、4、5只。SOD、MDA水平A组分别为(36.24±3.12)U/mg、(5.20±1.93)nmol/mg,B组分别为(19.21±5.67)U/mg、(8.96±2.65)nmol/mg,C组分别为(25.72±4.35)U/mg、(8.65±2.22)nmol/mg,D组分别为(34.61±4.66)U/mg、(5.98±2.15)nmol/mg。A、B两组比较差异均有统计学意义(均P<0.05);C、D两组与B组比较差异均有统计学意义(均P<0.05)。ALT、AST水平A组分别为(39.06±15.42)、(42.34±16.27)U/L,B组分别为(129.46±39.01)、(144.14±30.51)U/L,C组分别为(100.68±29.36)、(106.54±30.36)U/L,D组分别为(74.25±25.87)、(81.73±20.21)U/L。A、B两组比较差异均有统计学意义(均P<0.05);C、D两组ALT、AST的水平与B组比较差异均有统计学意义(均P<0.05)。结论板蓝根对CCl4诱导的急性肝损伤具有明显的保护作用。     

五味子对CCl4中毒大鼠肝细胞核基质蛋白和结构的保护作用

目的探讨五味子对中毒大鼠肝细胞核基质蛋白和结构的保护作用.方法将Wistar大鼠随机分为对照组、中毒组和治疗组.中毒组和治疗组大鼠经腹腔一次性注射200 mL/L CCl4 (0.2 mL/100 g体质量)致毒.治疗组以五味子提取液[0.5 mL/(100 g·d)]灌胃,中毒组以同量生理盐水灌胃,不同时间处死大鼠,光镜、电镜下观察肝脏细胞结构,并用免疫组化方法检测Lamin B蛋白的表达和电泳分析核基质蛋白成分.结果光镜和电镜下,五味子提取物治疗后大鼠肝细胞病理改变明显减轻.五味子提取物促进了中毒大鼠肝细胞Lamin B蛋白的表达、并有调节大鼠肝细胞核基质蛋白代谢的作用.结论五味子可能通过促进CCl4中毒大鼠肝细胞Lamin B蛋白的表达和影响核基质蛋白的代谢而促进损伤肝细胞修复,对于CCl4中毒大鼠肝细胞核基质蛋白和结构具有保护作用.     

Attenuation of Carbon Tetrachloride-lnduced Hepatotoxicity by Cow Urine Distillate in Rats

Objective To study the carbon tetrachloride-induced hepatoprotective activity in cow urine. Methods Effect of cow urine distillate on liver function was studied in vivo in rats intoxicated with carbon tetrachloride （CC14）. Hepatotoxicity was induced by a 1 ： 1 （v/v） mixture of CCl4 in olive oil （5 mL/kg i.p）. Protective effect of cow urine distillate （in three dose levels） and standard drug Silymarin （100 mg/kg, p.o） on liver function were studied in intoxicated rats. Parameters in the study included liver function tests and histological observations. Results The cow urine distillate decreased the levels of SGOT, SGPT, ALP, GGT, and total bilirubin in a dose-dependent manner （P〈0.05） as sylimarin. Conclusion The observed protective effects of cow urine distillate on liver function might be due to the presence of antioxidants in cow urine.     

PCNA在CCl4诱导的小鼠急性肝损伤组织中的表达

目的研究增殖细胞核抗原（PCNA）在CCl4诱导小鼠急性肝损伤过程中的动态变化,以了解小鼠急性肝损伤过程肝脏再生的规律。方法试验共分10组：CCl4腹腔注射（浓度0.1%溶于食用油中,0.1 mL/10 g）诱导小鼠急性肝损伤3、6、12、243、03、6、42、485、4 h及对照组小鼠,每组10只小鼠。使用赖氏法检测各组小鼠血清丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）水平的变化趋势,用Western blotting方法检测CCl4诱导的小鼠肝损伤过程中PCNA蛋白的表达。结果与对照组小鼠相比,CCl4诱导小鼠急性肝损伤之后,小鼠血清ALT、AST明显呈变化趋势（P〈0.05）,先逐渐升高,36 h达到高峰,后逐渐减低;与对照组小鼠相比,CCl4诱导小鼠急性肝损伤之后肝脏PCNA蛋白的表达明显呈变化趋势（P〈0.05）,PCNA在CCl4处理后36、1、22、4 h明显降低,30 h时升高,42 h至最高点,后又逐渐减低,54 h时恢复到正常水平。结论 PCNA在CCl4诱导小鼠急性肝损伤过程中有显著的变化,在CCl4诱导小鼠急性肝损伤的过程中肝脏再生具有明显的规律。     

四氯化碳诱导大鼠肝纤维化模型改良的研究

目的构建肝纤维化大鼠模型,并对经典四氯化碳(CCl4)复制模型方法进行适当改进。方法取Sprague-Dawley(SD)大鼠40只,雌雄各半,按随机数字表法分为A组10只、B组14只和C组16只。B组给予大鼠腹腔注射40%CCl4花生油溶液2 mL.kg-1,2次.周-1,同时给予普通全价饲料喂养(改良方法);A组为正常对照组,给予等量花生油腹腔注射,2次.周-1,同时给予普通全价饲料喂养;C组给予大鼠腹腔注射40%CCl4花生油溶液5 mL.kg-1,2次.周-1,同时给予添加10%猪油及2%胆固醇的饲料喂养(经典方法)。实验第8周末处死各组剩余大鼠:采用酶联免疫吸附法检测血清肝功能(ALT、AST、ALB、TP、A/G)水平;HE及Masson染色观察大鼠肝组织病理学变化。结果实验第8周B、C组均可见明显肝功能损害表现。B组可见部分标本肝脏假小叶形成,达到早期肝硬化,肝细胞少量脂肪变性,死亡率28.6%。C组均已形成明显肝硬化,弥漫性肝细胞脂肪变性,死亡率43.7%。结论低剂量CCl4诱导并给予普通饲料喂养可成功建立大鼠肝纤维化模型,并明显降低大鼠死亡率,提高了模型质量及实验效率。