玉郎伞多糖对四氯化碳诱导大鼠原代肝细胞损伤的保护作用

目的:研究玉郎伞多糖(YLS)对大鼠原代肝细胞损伤的保护作用及其机制。方法:采用IV型胶原酶灌流法分离大鼠肝细胞进行原代培养,用四氯化碳(CCl4)体外诱导肝细胞损伤,检测培养上清液中天门冬氨酸转换酶(AST)和丙氨酸氨基转换酶(ALT)水平,测定肝细胞中丙二醛(MDA)和谷胱甘肽(GSH)含量,MTT法检测细胞存活和增殖活性。结果:YLS(0.125~1.000g.L-1)可明显降低由CCl4升高的肝细胞培养上清液中AST和ALT水平及肝细胞MDA含量,还可提高CCl4降低的肝细胞存活率和GSH含量。结论:提示YLS对大鼠原代培养肝细胞损伤有直接保护作用,该作用可能与其抗氧化作用有关。     

南沙参多糖对四氯化碳损伤原代培养大鼠肝细胞保护作用的研究

目的:探讨南沙参多糖(RAPS)对四氯化碳(CCl4)损伤原代培养大鼠肝细胞的保护作用及其机制。方法:通过原位灌流法分离大鼠肝细胞,培养36h后加入RAPS,同时造成CCl4损伤,分别于损伤24h和48h时检测培养液中丙二醛(MDA)的含量、丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)的活性,48h后用MTT法测定肝细胞存活率。结果:RAPS对CCl4引起的ALT、AST活力的升高有抑制作用,同时抑制MDA的产生及肝细胞损伤造成的SOD活性及GSH-PX活性的降低。而且能明显改善肝细胞存活率。结论:RAPS能有效抑制CCl4造成的原代培养大鼠肝细胞损伤,机制可能与其抗氧化作用有关。     

金丝桃苷对四氯化碳诱导肝细胞损伤的保护作用

目的研究金丝桃苷(hyperfine,HP)对四氯化碳(CCl4)诱导肝细胞损伤的保护作用及其机制。方法用CCl4体外诱导人正常肝细胞L-02损伤,检测培养上清液中天门冬酸氨基转移酶(AST)和丙氨酸氨基转移酶(ALT)水平,测定肝细胞中丙二醛(MDA)和谷胱甘肽(GSH)的含量,四甲基偶氮唑盐微量酶反应比色(MTT)法检测肝细胞存活率。结果金丝桃苷可明显降低由CCl4升高的肝细胞培养上清液中ALT和AST水平及肝细胞MDA含量,还可提高由CCl4降低的肝细胞存活率和GSH含量。结论金丝桃苷对人肝细胞氧化性损伤有直接保护作用,这可能与金丝桃苷抑制抗氧化酶活性和抗自由基活性有关。     

黄芪总提物对体外肝细胞损伤的保护作用

目的研究黄芪总提物 (TEA)对体外肝细胞损伤的保护作用及其机理。方法采用Ⅳ型胶原酶灌流法分离大鼠肝细胞进行原代培养 ,用CCl4 和H2O2 体外分别诱导肝细胞损伤 ,检测肝细胞丙二醛 (MDA)、谷胱甘肽 (GSH)含量和谷胱甘肽过氧化物酶 (GSHpx)活性以及培养上清液中天门冬氨酸转换酶 (AST)和/或丙氨酸氨基转换酶 (ALT)水平。结果 (1)TEA(5～80mg·L-1)可明显降低或恢复由CCl4 升高的肝细胞MDA含量及肝细胞培养上清液中AST水平 ,还可使CCl4 降低的肝细胞GSH含量和GSHpx活性升高或恢复 ;(2)TEA(5～80mg·L-1)可使H2O2升高的肝细胞培养上清液中ALT水平和肝细胞MDA含量明显降低或恢复 ,还可使H2O2降低的肝细胞GHS含量和GSHpx活性明显升高或恢复。结论提示TEA对体外肝细胞损伤有直接保护作用 ,该作用可能与其抗氧化作用有关     

胰岛素对大鼠肝细胞损伤的保护作用

目的:探讨胰岛素对大鼠肝细胞损伤是否具有保护作用。方法:胶原酶原位灌流分离大鼠肝细胞并原代培养,分别用脂多糖(LPS)及胰岛素处理肝细胞,12 h后检测肝细胞损伤及存活率。结果:LPS可引起肝细胞损伤,使丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶(AST)增高;胰岛素可减轻上述肝细胞损伤(ALT活性降低55.5%、AST活性降低24.4%,P<0.01),提高肝细胞存活率(P<0.01)。结论:胰岛素可减少ALT和AST释放,促进肝细胞存活,减轻LPS直接诱导的肝细胞损伤,保护受损的肝细胞。     

玉郎伞多糖对抗结核药诱导肝细胞损伤的保护作用

目的 研究玉郎伞多糖(Y LSPS)对抗结核药诱导的肝细胞损伤的保护作用.方法 将不同浓度的利福平、异烟肼及吡嗪酰胺合用(RFP+ INH+ PZA)作用于人张氏肝细胞,用MTT法检测肝细胞的存活率,以80％ ～ 85％存活率的药物浓度为最佳浓度建立肝细胞损伤模型;给予YLSPS处理后,检测细胞培养上清液中丙氨酸氨基转移酶(ALT)、天门冬氨酸转移酶(AST)和乳酸脱氢酶(LDH)和细胞内丙二醛(MDA)和超氧化物歧化酶(SOD)的浓度.结果 与模型组比较,YLSPS各剂量可明显降低肝细胞培养上清液中的AST、ALT、LDH水平及肝细胞中MDA的含量,升高SOD的水平,且呈剂量依赖性.结论 YLSPS对RFP+ INH+ PZA合用诱导张氏肝细胞损伤具有保护作用,其机制可能与抗氧化和清除自由基的作用有关.     

牛磺酸对原代培养大鼠肝细胞的毒性作用

目的用原代培养正常大鼠肝细胞研究牛磺酸(β氨基酸)对肝细胞的毒性作用。方法用2步灌注法分离原代大鼠肝细胞;用MTT法测定细胞活力并计算IC50,观察药物作用后,培养液上清中AST、ALT和LDH活性及培养液中GSH和细胞内GSH的含量。结果细胞生长抑制率与剂量成正相关性,牛磺酸的IC50为24.23g.L-1。高浓度牛磺酸组培养液上清中AST、ALT和LDH活性显著升高,GSH含量显著减少。结论高浓度的牛磺酸对体外培养的肝细胞有一定损伤。     

异甘草素对大鼠急性化学性肝损伤的保护作用

目的研究异甘草素(ISL)对CCl4所致大鼠急性化学性肝损伤的保护作用及其机制.方法①在体实验选用♂Wistar大鼠48只,随机分6组,每组8只.ISL三剂量给药组分别灌服ISL 10,20,40 mg·kg-1·d-1;甘草酸二铵胶囊(DG)组灌服DG 500 mg·kg-1·d-1;正常对照组和模型组每日灌服等容量的溶媒.连续给药7 d,qd.以CCl4诱导大鼠急性肝损伤模型,酶学测定各组大鼠血清谷丙转氨酶(ALT)、谷草转氨酶(AST)和超氧化物歧化酶(SOD)活性,以及肝组织丙二醛(MDA)、谷胱甘肽(GSH)、谷胱甘肽过氧化物酶(GSH-Px)含量.②体外实验采用大鼠离体肝细胞原代培养,并建立CCl4诱导肝细胞损伤模型,检测ISL对其作用的影响.结果①lSL剂量依赖性降低大鼠血清中升高的ALT和AST活性,升高肝组织中降低的GSH含量、SOD和GSH-Px活性,同时降低过氧化物终产物含量.②ISL浓度(5.0～20.0 μmol·L-1)依赖性抑制CCl4引起的ALT和AST升高,ISL 20.0 μmol·L-1可阻断CCl4产生的肝细胞ALT和AST漏出.结论ISL对大鼠化学性肝损伤具有显著的保护作用.其机制与清除肝组织中的自由基和抗脂质过氧化等作用有关.     

珠蚌多糖对四氯化碳诱导肝细胞损伤的保护作用

目的 研究珠蚌多糖(HCP)对肝细胞损伤的保护作用及其机制.方法 培养L-02型肝细胞,用四氯化碳(CCL)体外诱导肝细胞损伤,检测培养上清液中天门冬氨酸转挟酶(AST)和丙氨酸氨基转换酶(ALT)水平,测定上清液中丙二醛(MDA)的含量扣过氧化物岐化酶(SOD)活力,MTT法检测细胞存活和增殖活性.结果 珠蚌多糖(25,250及1000μg·L-1)剂量组均可明显降低由CCl4升高的肝细胞培养上清波中AST争ALT水平及MDA含量,还可提高CCl4降低的肝细胞存活率和SOD活力.结论 提示珠蚌多糖对肝细胞损伤有直接保护作用,该作用可能与其抗氧化作用有关.     

海兔素对大鼠原代肝细胞酒精性氧化损伤的保护作用

目的探讨海兔素对大鼠原代肝细胞酒精性氧化损伤保护作用。方法门静脉胶原酶Ⅳ原位灌注及密度梯度离心获得大鼠原代肝细胞。MTT实验检测乙醇和海兔素最佳作用剂量及肝细胞活力。酶学实验检测细胞AST、LDH、SOD、MDA、GSH水平;流式细胞术检测细胞凋亡情况;单细胞凝胶电泳观察细胞DNA损伤状况;JC-1荧光探针检测细胞线粒体膜电位水平;比色法及Western blot检测细胞CYP2E1活性及蛋白表达。结果经30 mg·L~(-1)。海兔素预作用2 h,再与300 mmol·L~(-1)。乙醇共作用8 h后,肝细胞活力较酒精模型组明显上升,AST和LDH释放也得到明显抑制;同时,肝细胞SOD和GSH水平明显升高,MDA含量则明显降低,差异均具有显著性(P<0.05)。海兔素干预后,肝细胞凋亡率明显降低,DNA损伤及线粒体膜电位水平明显得到改善。海兔素干预后,肝细胞CYP2E1活性及蛋白表达水平明显受到抑制(P<0.05)。结论海兔素对大鼠原代肝细胞酒精性氧化损伤具有保护作用,其作用机制可能与海兔素抑制酒精对CYP2E1的活化,缓解氧化应激,提高机体抗氧化能力有关。     

地塞米松对双氯芬酸钠肝损伤的保护作用

目的探讨地塞米松(dexamethasone,Dex)对双氯芬酸钠诱导的大鼠药物性肝损伤的保护作用及部分机制。方法大鼠随机分为正常对照组、模型对照组、Dex(10 mg.kg-1)给药组。Dex(10 mg.kg-1)腹腔注射,1 h后腹腔注射双氯芬酸钠100 mg.kg-1,24 h后检测ALT和AST活性、测定肝匀浆中MDA、GSH含量和GSH-Px、SOD活性,观察肝组织病理学变化,并测肝线粒体膜电位、线粒体肿胀度、NADH水平、SDH及ATPase活性。结果模型对照组血清ALT、AST升高,光镜下可见肝小叶内肝细胞片状坏死,肝匀浆MDA含量升高,GSH、GSH-Px和SOD含量降低,肝线粒体NADH含量、SDH及ATPase活性降低。Dex可明显降低ALT、AST活性(P<0.05),减轻肝脏炎症,降低肝匀浆中MDA含量(P<0.01),升高GSH含量、GSH-Px和SOD活性以及线粒体中NADH含量、SDH及ATPase活性(P<0.01)。结论 Dex对双氯芬酸钠诱导的大鼠药物性肝损伤有保护作用,作用机制可能与减轻线粒体损伤有关。     

Protective effects of echinacoside on carbon tetrachloride-induced hepatotoxicity in rats

The aim of this study was to investigate the possible protective effects of echinacoside, one of the phenylethanoids isolated from the stems of Cistanches salsa, a Chinese herbal medicine, on the free radical damage of liver caused by carbon tetrachloride in rats. Treatment of rats with carbon tetrachloride produced severe liver injury, as demonstrated by dramatic elevation of serum ALT, AST levels and typical histopathological changes including hepatocyte necrosis or apoptosis, haemorrhage, fatty degeneration, etc. In addition, carbon tetrachloride administration caused oxidative stress in rats, as evidenced by increased reactive oxygen species (ROS) production and MDA concentrations in the liver of rats, along with a remarkable reduction in hepatic SOD activity and GSH content. However, simultaneous treatment with echinacoside (50mg/kg, intraperitoneally) significantly attenuated carbon tetrachloride-induced hepatotoxicity. The results showed that serum ALT, AST levels and hepatic MDA content as well as ROS production were reduced dramatically, and hepatic SOD activity and GSH content were restored remarkably by echinacoside administration, as compared to the carbon tetrachloride-treated rats. Moreover, the histopathological damage of liver and the number of apoptotic hepatocytes were also significantly ameliorated by echinacoside treatment. It is therefore suggested that echinacoside can provide a definite protective effect against acute hepatic injury caused by CCl(4) in rats, which may mainly be associated with its antioxidative effect.     

舒芬太尼减轻大鼠肝缺血再灌注损伤的机制研究

目的 探讨舒芬太尼预处理减轻大鼠肝缺血再灌注损伤(HIRI)的作用机制。方法 48只雄性SD大鼠按照随机数字表法分为：假手术组(S组)、HIRI组(IR组)、舒芬太尼预处理+HIRI组(SF组)、转化生长因子β₁(TGF-β₁)抑制剂+舒芬太尼预处理+HIRI组(SB组)、TGF-β₁激动剂+舒芬太尼预处理+HIRI组(SRI组)和二甲基亚砜(DMSO)+HIRI组(DMSO组),每组8只,于造模完成后4 h取材。HE染色观察肝组织的形态变化;收集腹主动脉血测定各组丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平;制备10%肝组织匀浆,测定各组丙二醛(MDA)及超氧化物歧化酶(SOD)水平;TUNEL法检测肝细胞凋亡率;Western blot法检测肝组织TGF-β₁及Smad2/3、p-Smad2/3、p38、p-p38蛋白表达水平。结果 与S组相比,其余各组损伤程度加重,ALT、AST、MDA水平及肝细胞凋亡率升高(P<0.05),SOD水平降低(P<0.05),IR组、SF组、...     

缺血预适应对肝脏热缺血再灌注损伤保护作用的机制研究

目的 探讨缺血预适应对肝脏热缺血再灌注损伤保护作用的分子机制。 方法 90 只成年清洁级 SD 大鼠随机均分为 3 组, A 组为假手术组, B、C 组建立 SD 大鼠肝脏热缺血再灌注损伤模型, 并对 C 组大鼠进行缺血预适应干预。 于缺血再灌注后 0、2、6、12、24 h 采集血标本测定丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）, 采用酶联免疫吸附（ELISA）技术检测肝细胞中肿瘤坏死因子（TNF）-α 及白细胞介素（IL）-1β 水平; 采集肝组织标本测定丙二醛（MDA）和超氧化物歧化酶（SOD）, 流式细胞仪测定肝细胞线粒体膜电位。 结果 B、C 组大鼠的血清转氨酶、TNF-α、 IL-1β 及 MDA 水平均明显高于 A 组（P＜ 0.05）; B、C 组大鼠的凝血酶原活动度和胆碱酯酶低于 A 组（P＜ 0.05）; B、C 组大鼠肝脏 SOD 水平明显低于 A 组;C 组大鼠各项指标均优于 B 组（P＜ 0.05）。 缺血再灌注发生后肝细胞线粒体膜电位水平在 0 h 后即达到最低值, 此后呈逐渐上升趋势（P＜ 0.05）, 并且 C 组的肝细胞线粒体膜电位水平在各时段均高于 B 组（P＜ 0.05）。 结论 缺血预适应对肝细胞热缺血再灌注损伤具有一定的保护作用, 缺血预适应可以通过减少炎症因子 TNF-α 及 IL-1β 的释放, 提高 SOD 拮抗自由基的活性, 减轻线粒体损伤等途径发挥其保护作用。     

软肝片抗肝纤维化作用的实验研究

目的探讨软肝片对四氯化碳中毒性肝纤维化的防治作用。方法用四氯化碳皮下注射造成大鼠肝纤维化模型 ,以联苯双酯作为阳性对照 ,测定血清丙氨酸氨基转移酶 (ALT)、天冬氨酸氨基转移酶 (AST)、玻璃酸(HA)、唾液酸及肝组织羟脯氨酸 (Hyp)、丙二醛 (MDA)、超氧化物歧化酶 (SOD)含量 ,以反映肝细胞损伤及肝纤维化程度。结果软肝片可明显降低肝纤维化大鼠血清ALT、AST、HA、唾液酸水平及肝组织Hyp和MDA水平 ,提高肝组织中SOD活力。结论软肝片具有一定的抗肝纤维化及抗脂质过氧化作用。     

Pomegranate peel extract prevents liver fibrosis in biliary-obstructed rats

Punica granatum L. (pomegranate) is a widely used plant that has high nutritional value. The aim of this study was to assess the effect of chronic administration of pomegranate peel extract (PPE) on liver fibrosis induced by bile duct ligation (BDL) in rats. PPE (50 mg kg(-1)) or saline was administered orally for 28 days. Serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) levels were determined to assess liver function and tissue damage. Proinflammatory cytokines (tumor necrosis factor-alpha and interleukin 1 beta) in the serum and antioxidant capacity (AOC) were measured in plasma samples. Samples of liver tissue were taken for measurement of hepatic malondialdehyde (MDA) and glutathione (GSH) levels, myeloperoxidase (MPO) activity and collagen content. Production of reactive oxidants was monitored by chemiluminescence assay. Serum AST, ALT, LDH and cytokines were elevated in the BDL group compared with the control group; this increase was significantly decreased by PPE treatment. Plasma AOC and hepatic GSH levels were significantly depressed by BDL but were increased back to control levels in the PPE-treated BDL group. Increases in tissue MDA levels and MPO activity due to BDL were reduced back to control levels by PPE treatment. Similarly, increased hepatic collagen content in the BDL rats was reduced to the level of the control group with PPE treatment. Thus, chronic PPE administration alleviated the BDL-induced oxidative injury of the liver and improved the hepatic structure and function. It therefore seems likely that PPE, with its antioxidant and antifibrotic properties, may be of potential therapeutic value in protecting the liver from fibrosis and oxidative injury due to biliary obstruction.     

8-甲氧补骨脂素对对乙酰氨基酚致小鼠急性肝损伤的保护作用

目的研究8-甲氧补骨脂素(8-methoxypsoralen,8-MOP)对对乙酰氨基酚(acetaminophen,APAP)致小鼠急性肝损伤的保护作用。方法采用对乙酰氨基酚所致小鼠急性肝损伤模型。24 h后,检测小鼠血清中丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)和乳酸脱氢酶(LDH);留取肝脏组织,常规石蜡包埋切片,HE染色,光镜观察肝脏组织病理变化;制备肝匀浆,测定肝中还原型谷胱甘肽(GSH)、氧化型谷胱甘肽(GSSG)和丙二醛(MDA)的含量。结果与正常对照组比较,模型组小鼠血清中ALT、AST和LDH活性明显升高,肝脏组织出现明显的肝细胞变性坏死;与模型组相比,8-甲氧补骨脂素可以明显降低小鼠血清中ALT、AST和LDH的活性,降低肝组织中MDA的含量,升高GSH/GSSG比值,肝组织病理损伤也明显减轻。结论 8-甲氧补骨脂素对对乙酰氨基酚致小鼠急性肝损伤具有明显的保护作用。     

甜菜碱对大鼠肝脏缺血-再灌注损伤的作用

目的观察甜菜碱对大鼠肝脏缺血-再灌注损伤的作用。方法 40只SD大鼠随机分为假手术组,模型组,甜菜碱200、400、800 mg·kg^-1组(均n=8)。甜菜碱各剂量组于造模前7 d灌胃给予相应剂量甜菜碱,假手术组和模型组给予生理盐水。夹闭门静脉、肝动脉分支40 min,再灌注24 h制大鼠部分肝脏缺血-再灌损伤模型。检测血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)水平和肝脏组织中丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性,流式细胞术检测细胞凋亡水平,Western blot法检测凋亡相关蛋白p53、cl-caspase-3和Bcl-2的表达,经HE染色观察肝脏组织病理学改变。结果与假手术组比较,模型组大鼠血清ALT和AST水平显著上升(P <0.01),肝脏组织中MDA含量显著增加、SOD活性降低(P <0.01),肝细胞凋亡率显著增高(P <0.01)。与模型组比较,甜菜碱400、800 mg·kg^-1组ALT和AST水平显著降低(P <0.05,P <0.01),肝组织中MDA含量显著降低、SOD活性显著增强(P <0.05),肝细胞凋亡率下降(P <0.05,P <0.01),p53、cl-caspase-3、Bax表达显著降低(P <0.05)。结论甜菜碱可改善肝脏缺血-再灌注损伤大鼠肝功能,可能与其抗氧化、抑制肝细胞凋亡有关。     

Alpha-lipoic acid protects against hepatic ischemia-reperfusion injury in rats

BACKGROUND AND AIM: To evaluate the protective effect of alpha-lipoic acid in reducing oxidative damage after severe hepatic ischemia/reperfusion (IR) injury. METHODS: Wistar albino rats were subjected to 45 min of hepatic ischemia, followed by 60 min reperfusion period. Lipoic acid (100 mg/kg i.p.) was administered 15 min prior to ischemia and immediately before reperfusion period. At the end of the reperfusion period aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH) activity, and cytokine, TNF-alpha and IL-1beta levels were determined in serum samples. Malondialdehyde (MDA), and glutathione (GSH) levels and myeloperoxidase (MPO) activity were determined in the liver tissue samples while formation of reactive oxygen species was monitored by using chemiluminescence (CL) technique with luminol and lucigenin probes. Tissues were also analyzed histologically. Results: Serum ALT, AST, and LDH activities and TNF-alpha and IL-1beta levels were elevated in the I/R group, while this increase was significantly lower in the group of animals treated concomitantly with lipoic acid. Hepatic GSH levels, significantly depressed by I/R, were elevated back to control levels in lipoic acid-treated I/R group. Furthermore, increases in tissue luminol and lucigenin CL, MDA levels and MPO activity due to I/R injury were reduced back to control levels with lipoic acid treatment. CONCLUSION: Since lipoic acid administration alleviated the I/R-induced liver injury and improved the hepatic structure and function, it seems likely that lipoic acid with its antioxidant and oxidant-scavenging properties may be of potential therapeutic value in protecting the liver against oxidative injury due to ischemia-reperfusion.