鲨鱼肝刺激物对大鼠急性肝损伤和肝脏线粒体功能的影响

目的:研究鲨鱼肝刺激物(sHSS)对硫代乙酰胺(TAA)所致大鼠急性肝损伤和肝线粒体功能的影响.方法:雄性SD大鼠,体质量(200±20) g,随机分3组:对照组、模型组、治疗组,每组8只.以400 mg/kg TAA 2次腹腔注射建立大鼠急性肝损伤模型,治疗组在注射TAA前 1 h腹腔注射80 mg/kg sHSS,对照组注射等体积的生理盐水,24 h后观察大鼠血清中丙氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST)活性和肝中丙二醛(MDA)含量的变化,以及TAA和sHSS对肝线粒体呼吸功能、线粒体肿胀和膜电位的影响.结果:治疗组大鼠血清中ALT、AST的水平明显低于模型组,而模型组MDA含量明显高于治疗组和正常组(P<0.05);治疗组大鼠肝脏线粒体ADP诱导的3态氧消耗、呼吸控制率(RCR)、氧化磷酸化率(OPR)明显高于TAA模型组(P<0.05);注射TAA和sHSS后,线粒体肿胀和跨膜电位没有明显的变化.结论:sHSS能明显抑制TAA造成的急性肝损伤和脂质过氧化,改善因TAA而受损的线粒体呼吸功能.     

大鼠烫伤早期肝线粒体氧化磷酸化偶联增强与体内儿茶酚胺类…

ＳＤ大鼠皮下注射肾上腺素或去甲肾上腺素，３０ｍｉｎ后发现鼠肝线粒体琥珀酸呼吸链呼吸控制率、３态耗氧率和ＡＴＰ形成率均增高，注射肾上腺素组和注射去甲肾上腺组的４态耗氧率增高和有增高趋势。皮下注射肾上腺素受体阻断剂噻吗心安和酚苄明后，对烫伤后３０ｍｉｎ大鼠肝线粒体琥珀酸呼吸链活性各息起部分抑制作用。提示烫伤早期大鼠肝线粒体氧化磷酸化偶联增强与体内儿茶酚胺类释放有关。     

牛磺酸对大鼠肝线粒体氧自由基损伤的保护作用

目的 观察氧自由基对线粒体的损伤以及牛磺酸的保护作用。方法 采用氧自由基发生系统ＦｅＳＯ４／ＶｉｔＣ损伤线粒体,分离的肝线粒体分为３组：对照组（Ａ组）,损伤组（Ｂ组）,牛磺酸保护组（Ｃ组）。分别测定线粒体呼吸功能、３种氧化酶活性、丙二醛含量。结果 Ｂ组线粒体状态３速率、呼吸控制率、磷／氧比、氧化磷酸化效率降低,说明线粒体呼吸功能和氧化磷酸化功能受损;Ｂ组线粒体电子传递链的琥珀酸氧化酶１、ＮＡＤＨ氧     

氯胺酮对腹腔感染脓毒症大鼠肝线粒体能量代谢的影响

目的 探讨氯胺酮对脓毒症大鼠肝线粒体能量代谢的影响以及氯胺酮对脓毒症肝损伤的保护机制。方法 采用盲肠结扎穿孔（CLP）制作脓毒症模型，将大鼠随机分为假手术组（C组）、盐水组（NS组）、氯胺酮组（K组），模型制作12h后开始分别给予生理盐水和氯胺酮，观察4h后制备肝线粒体和组织匀浆，并采用Clark氧电极技术测定线粒体呼吸功能和定磷法测定ATP酶活性。结果 盐水组的呼吸控制率（respiratory control rate，RCR）和ADP／O明显低于假手术组，而态3呼吸速率和态4呼吸速率明显高于假手术组，且ATP酶各值均较假手术组明显降低；氯胺酮组能明显提高RCR、ADP／O和ATP酶的活性，降低态3呼吸速率态和4呼吸速率。结论 脓毒症大鼠肝损伤可能与ATP酶活性被抑制有关，而氯胺酮通过提高ATP酶的活力，减轻细胞和线粒体钙超载，改善细胞能量代谢功能，从而有效地保护了肝脏的结构和功能。     

大鼠烫伤早期肝线粒体氧化磷酸化偶联增强与体内儿茶酚胺类释放的关系

SD大鼠皮下注射肾上腺素或去甲肾上腺素,30min后发现鼠肝线粒体琥珀酸呼吸链呼吸控制率、3态耗氧率和ATP形成率均增高,注射肾上腺素组和注射去甲肾上腺素组的4态耗氧率增高和有增高趋势。皮下注射肾上腺素受体阻断剂噻吗心安和酚苄明后,对烫伤后30min大鼠肝线粒体琥珀酸呼吸链活性各自起部分抑制作用。提示烫伤早期大鼠肝线粒体氧化磷酸化偶联增强与体内儿茶酚胺类释放有关。     

心肺复苏后大鼠脑细胞线粒体呼吸功能及能量代谢的变化

目的研究心肺复苏后大鼠脑细胞线粒体呼吸功能及能量代谢的改变。方法采用窒息合并冰氯化钾停跳液致大鼠心跳骤停5min后开始心肺复苏的动物模型，SD大鼠48只，随机分为6组：对照组（假手术组）、复苏后3、12、24、48和72h组（每组8只）。各组取脑组织测定线粒体呼吸Ⅲ、Ⅳ态及呼吸控制率（RCR），磷氧比（P／O）；应用高效液相及紫外检测法测定脑细胞ATP、ADP和AMP含量及能荷值。结果心肺复苏后大鼠脑细胞线粒体功能明显受损，线粒体呼吸Ⅲ态速率下降，线粒体呼吸Ⅳ态速率升高，线粒体呼吸控制率（RCR）和P／O明显下降；随着复苏成功后时间的延长，脑细胞线粒体呼吸控制率（RCR）和P／O持续下降，其中，24h有所恢复，48h再次下降；心肺复苏后的脑细胞ATP含量明显下降，ADP和AMP相对增加。结论心肺复苏后大鼠脑细胞线粒体呼吸功能明显下降，能量代谢障碍。     

硒和锗对镉所致大鼠肝脏脂质过氧化的影响

采用给大鼠腹腔注射硒（０．４ｍｇ／ｋｇ）、锗（７５ｍｇ／ｋｇ）或镉（１ｍｇ／ｋｇ）的方法，研究硒和锗对镉所致大鼠肝脏脂质过氧化作用的影响。结果表明，给大鼠连续腹腔注射镉（１ｍｇ／ｋｇ）３ｄ，可显著增强大鼠肝脏脂质过氧化作用；注射硒、锗可分别拮抗镉的脂质过氧化作用，但锗对镉所致大鼠肝微粒体、线粒体谷胱甘肽过氧化物酶活性下降没有明显影响；将硒、锗剂量减半联合应用，也能拮抗镉所致大鼠肝脏的脂质过氧化作用，表明硒和锗具有有协同抗氧化作用。     

重度烧伤狗肝,肾线粒体呼吸功能及ATP含量的变化

本文用呼吸速率、呼吸控制率(RCR)、磷氧比值(ADP/O)和ATP含量为指标,观察了50％TBSA Ⅲ°烧伤狗肝、肾线粒体呼吸功能的变化。 结果发现,烧伤后肝、肾线粒体呼吸速率增强,而RCR、ADP/O和ATP含量有明显降低,肝分别降至正常对照组的43.1％、70.6％和40.1％;肾分别降至73.4％、76.2％和51.4％,差异非常显著(P<0.001)。伤后立即输液组肝RCR、ADP/O和ATP分别是正常对照组的96.5％、95.4％和89.8％;肾分别是88.7％、93.5％和85.0％,均较接近正常对照组,差异不显著(P>0.05)。结果提示,重度烧伤严重损害肝、肾线粒体正常生理功能,可致线粒体氧化磷酸化生成ATP能力降低。而伤后立即输液对肝、肾线粒体功能有明显保护作用。     

衰老大鼠肝线粒体质与量的改变

目的：探究大鼠衰老时肝组织细胞线粒体的变化规律。方法：分别从４月龄和２４月龄大鼠的肝组织分离出线粒体，并用双缩脲法测定单位组织的线粒体蛋白；用Ｃｌａｒｋ氧电极极谱法分别测定线粒体呼吸链３个氧化酶活性：ＮＡＤＨ氧化酶、琥珀酸氧化酶和细胞色素氧化酶；用差光谱法（连二亚硫酸还原的消光值减去正铁氰化钾氧化的消光值）获得细胞色素ａ，ｂ，ｃ和ｃ１（Ｃｙｔａ，ｂ，ｃ和ｃ１）含量。结果：与４月龄大鼠相比，２４月龄大鼠的肝组织细胞线粒体含量有一定的下降，而３个氧化酶活性显著增强，同时细胞色素含量尤其是Ｃｙｔｂ和ｃ１也显著升高。结论：衰老大鼠肝组织线粒体呼吸链氧化功能增强可能是通过某种反馈机制使残存尚好的线粒体代偿性功能增强。     

养心通脉方对大鼠心肌缺血损伤模型心肌细胞线粒体功能的影响

目的：研究养心通脉方对异丙肾上腺素诱导的大鼠心肌缺血损伤模型心肌细胞线粒体呼吸功能的影响。方法实验大鼠被随机分为3组（每组15只）：对照组、模型组、养心通脉方组,实验结束后所有动物采用脱颈椎法处死,通过测定呼吸控制率(RCR)、二磷酸腺苷磷/氧(ADP/O)比值及 ATP 的浓度来评价线粒体的呼吸功能,并对线粒体内胆固醇、磷脂（PL）、甘油三酯（TG）和游离脂肪酸（FFAs）的含量进行测定。结果模型组中大鼠心肌细胞线粒体内RCR、ADP/O比值及ATP浓度均明显低于对照组（P＜0.001）,而在养心通脉方组中心肌细胞线粒体内RCR、ADP/O比值及ATP浓度均接近于对照组（P＞0.05）。与对照组相比,模型组中胆固醇、TG 和 FFAs 的水平均明显升高（P＜0.001）,同时磷脂（PL）水平明显降低（P＜0.001）。但在养心通脉方组中心肌细胞线粒体内胆固醇、TG、FFAs以及PL的水平均与对照组无明显差异（P＞0.05）。结论养心通脉方对异丙肾上腺素诱导的大鼠心肌缺血损伤模型心肌细胞线粒体的呼吸功能及其膜完整性具有保护作用。     

Antioxidant and Anti-aging Activities of Silybum Marianum Protein Hydrolysate in Mice Treated with D-galactose

Objective In the present study, we investigated the antioxidant and anti‐aging effects of Silybum marianum protein hydrolysate(SMPH) in D‐galactose‐treated mice. Methods D‐galactose(500 mg/kg body weight) was intraperitoneally injected daily for 7 weeks to accelerate aging, and SMPH(400, 800, 1,200 mg/kg body weight, respectively) was simultaneously administered orally. The antioxidant and anti‐aging effects of SMPH in the liver and brain were measured by biochemical assays. Transmission electron microscopy(TEM) was performed to study the ultrastructure of liver mitochondria. Results SMPH decreased triglyceride and cholesterol levels in the D‐galactose‐treated mice. It significantly elevated the activities of superoxide dismutase(SOD) and glutathione peroxidase(GSH‐Px), and total antioxidant capacity(T‐AOC), which were suppressed by D‐galactose. Monoamine oxidase(MAO) and malondialdehyde(MDA) levels as well as the concentrations of caspase‐3 and 8‐OHd G in the liver and brain were significantly reduced by SMPH. Moreover, it increased Bcl‐2 levels in the liver and brain. Furthermore, SMPH significantly attenuated D‐galactose‐induced liver mitochondrial dysfunction by improving the activities of Na+‐K+‐ATPase and Ca2+‐Mg2+‐ATPase as well as mitochondrial membrane potential(ΔΨm) and fluidity. TEM showed that the degree of liver mitochondrial damage was significantly decreased by SMPH. Conclusion The results indicated that SMPH protects against D‐galactose‐induced accelerated aging in mice through its antioxidant and anti‐aging activities.     

Protective effect of Pisonia aculeata on thioacetamide induced hepatotoxicity in rats

Objective

To evaluate the protective effect of Pisonia aculeata (P. aculeata) on thioacetamide induced hepatotoxicity in rats.

Methods

Male Wistar rats were administered 250 or 500 mg/kg p.o. of P. aculeata extract for 21 days and simultaneously administered thioacetamide (TAA) 50 mg/kg bw s.c. 1 h after the respective assigned treatments every 72 h. At the end of all experimental methods, all the animals were sacrificed by cervical decapitation. Blood samples were collected. Serum was separated and analyzed for various biochemical parameters.

Results

TAA induced a significant rise in aspartate amino transferase (AST), alanine amino transferase (ALT), alkaline phosphatase (ALP), total bilirubin, gamma glutamate transpeptidase (GGTP), lipid peroxidase (LPO) with a reduction of total protein, superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx) and glutathione S-transferase (GST). Treatment of rats with different doses of plant extract (250 and 500 mg/kg) significantly (P<0.001) altered serum marker enzymes and antioxidant levels to near normal against TAA treated rats. The activity of the extract at a dose of 300 mg/kg was comparable to the standard drug, silymarin (50 mg/kg, p.o.).

Conclusions

It can be concluded that P. aculeata extract possesses a remarkable hepatoprotective and antioxidant activity against TAA induced hepatotoxicity. More research is required to derive an optimal therapeutic dose.     

Protective effect of Tetracera scandens L.leaf extract against CCl_4-induced acute liver injury in rats

Objective:To investigate the protective potential of ethanolic extracts of Tetracera scandens L.(T.scandens) against CCl_4 induced oxidative stress in liver tissues.Methods:Dried leaf powder of T.scandens was extracted with ethanol and concentrated to yield a dry residue.Rats were administered with 100 mg/kg of ethanolic extracts orally once daily for one week.Animals were subsequently administered with a single dose of CCl_4(I mL/kg body weight,intraperitoneal injection).Various assays,such as serum levels of alanine aminotransferase,aspartate aminotransferase,lipid peroxidation,protein oxidation(carbonyl protein group),tumor necrosis factor alpha,catalase,superoxide dismutase,and glutathione peroxidase,were used to assess damage caused by CCl_4 and the protective effects of the ethanol extract on liver tissues.Results:Hepatotoxicity induced by CCl_4 was evidenced by a significant increase in serum aspartate aminotransferase and alanine aminotransferase level,lipid peroxidation,protein carbonyl group,and tumor necrosis factor alpha,as well as decreased activity of the hepatic antioxidant enzymes(catalase.superoxide dismutase.and glutathione peroxidase).Treatment with ethanolic T.scandens extracts prevented all of these typically observed changes in CCl_4-treated rats.Conclusions:Our findings indicate that T.scandens has a significant protective effect against CCl_4 induced hepatotoxicity in rat.which may be due to its antioxidant properties.     

Effect of morin on the levels of circulatory liver markers and redox status in experimental chronic hyperammonaemic rats

INTRODUCTION: Hyperammonaemia is a major contributing factor to neurological abnormalities observed in hepatic encephalopathy and in congenital defects of ammonia detoxication. Ammonia toxicity results in free radical generation that leads to oxidative stress and tissue damage. Morin is a bioflavonoid, a constituent of many herbs and fruits that are used as herbal medicines and also several biological activities. Our aim was to investigate the effect of morin on circulatory liver markers, lipid peroxidation and antioxidant status in ammonium chloride (AC)-induced hyperammonaemic rats. METHODS: Male albino Wistar rats weighing 180-200 g were used for the study. The hyperammonaemia was induced by interaperitonial injection of AC (100 mg/kg body weight). Rats were treated with morin (30 mg/kg body weight) via oral administration. Administration of morin in hyperammonaemic rats reduced the levels of ammonia and urea. The antioxidant property of morin was studied by assessing the activities of thiobarbituric acid reactive substances (TBARS), hydroperoxides (HP) and liver markers (alanine transaminase, aspartate transaminase and alkaline phosphatase) and the levels of glutathione peroxidase, superoxide dismutase, catalase, reduced glutathione, vitamins A, C and E in AC-treated rats. RESULTS: Oxidative stress was effectively modulated by morin administration. Morin significantly improved the status of antioxidants and decreased the levels of ammonia, urea, TBARS, HP and liver markers enzymes, as compared to the AC-treated group. CONCLUSION: The study offers evidence for the antihyperammonaemic, hepatoprotective and antioxidant effects of morin against oxidative stress induced by AC.     

Effect of In Vivo and in Vitro Treatment with Arsenite on Rat Hepatic Mitochondrial and Microsomal Enzymes

ＥｆｆｅｃｔｏｆＩｎＶｉｖｏａｎｄｉｎＶｉｔｒｏＴｒｅａｔｍｅｎｔｗｉｔｈＡｒｓｅｎｉｔｅｏｎＲａｔＨｅｐａｔｉｃＭｉｔｏｃｈｏｎｄｒｉａｌａｎｄＭｉｃｒｏｓｏｍａｌＥｎｚｙｍｅｓＣＨＥＮＧＪｉｚｈｏｎｇ（程继忠）；ＷＵＨｕｉｑｉｏｎｇ（邬惠琼）；Ｓ...     

金线莲乙醇提取物抗氧化作用的研究

目的 探讨金线莲(Anoectochilus Formosanus,AF) 乙醇提取物对自然衰老模型小鼠抗氧化作用及其体外抗氧化活性的影响.方法 取8月龄(老龄)KM小鼠75只,按体质量随机分为5组:自然衰老组、阳性对照组(维生素E组:0.1 g/kg)、AF乙醇提取物高剂量组(4 g/kg)、AF乙醇提取物中剂量组(2 g/kg)、AF乙醇提取物低剂量组(1 g/kg).各组动物每日给予相应剂量的药物,自然衰老组给予等体积的生理盐水,连续给药30 d.于给药前、给药14 d、给药28d测定小鼠体质量.实验结束时,测定受试动物血清与肝脏中MDA、GSH-PX、SOD水平.采用总抗氧化能力、羟基自由基(·OH)、超氧阴离子(O-·2)的反应体系,考察金线莲乙醇提取物的体外抗氧化能力.结果 与自然衰老组比较,给药前、给药14 d、28 d各组动物体质量差异无统计学意义(P>0.05);维生素E组和AF乙醇提取物高、中、低剂量组血中GSH-PX和T-SOD的活力明显升高(P<0.01或P<0.05);维生素E组和AF乙醇提取物高、中剂量组血清MDA的含量降低,差异有统计学意义(P<0.01或P<0.05);维生素E组和AF乙醇提取物高剂量组肝中GSH-PX和SOD的活力显著升高(P<0.01)、MDA含量明显降低(P<0.01),AF乙醇提取物中剂量组肝中SOD的活力升高(P<0.05)、MDA含量明显降低(P<0.01).金线莲乙醇提取物在一定浓度范围内清除·OH、O-·2的能力及总抗氧化能力均呈量效关系;金线莲乙醇提取物对·OH、O-·2的IC50分别为5.5 mg/mL、12.5 mg/mL.结论 金线莲乙醇提取物具有一定的体内外抗氧化作用.     

参附注射液对大鼠阿霉素心肌损伤的影响

目的:探讨参附注射液(shenfu injection,SFI)是否能够防治阿霉素(doxorubicin,DOX)所致大鼠心肌损伤。方法:将SD雄性大鼠随机分为空白(SAL)组、空白参附(SAL+SFI,5 mL·kg^-1)组、模型(DOX)组、参附高剂量(DOX+SFI,5 mL·kg^-1)组、参附低剂量组(DOX+SFI,1 mL·kg^-1)。除SAL和SAL+SFI组外,其余各组皮下注射阿霉素(2 mg·kg^-1),每周1次,共7周。加用SFI组在DOX损伤前一天开始腹腔注射SFI,SAL组给予等容积的无菌生理盐水,持续给药5 d。停止注射DOX后1周处死动物,测定体质量、心脏质量、线粒体呼吸功能、线粒体肿胀程度、心脏超氧化物岐化酶(superoxide dismutase,SOD)等指标。结果:DOX损伤后,大鼠体质量、心脏质量及心脏指数降低,心脏线粒体更容易肿胀,DOX抑制了大鼠心脏态Ⅲ呼吸,降低了线粒体呼吸控制率;给予SFI后,上述指标有一定程度改善,SFI有升高大鼠心肌CuZn-SOD的趋势。结论:SFI具有一定的防治阿霉素心肌损伤的作用,其机制可能与保护心肌线粒体、增强SOD的活性有关。     

Partial protection by lipoic acid against carboplantin-induced ototoxicity in rats

To investigate the alterations in auditory brainstem evoked responses (ABRs) and the changes of carboplatin-induced ototoxicity in the cochlear oxidant/antioxidant systems and otoprotection by an antioxidant lipoate. Methods Male wistar rats were divided into four groups and treated as follows: 1) vehicle (saline) control, 2) carboplatin (256 mg/kg, i.p.), 3) lipoate (100 mg/kg, i.p.), 4) lipoate carboplatin. Post-treatment ABRs were performed after four days and rats were sacrificed with their cochleae harvested and analyzed. Results Carboplatin significantly elevated ABR threshold above the pretreatment thresholds. Lipoate carboplatin treated rats showed decreased elevation of hearing threshold. Carboplatin significantly depleted cochlear reduced to oxizized glutathione (GSH/GSSG) ratio, whereas lipoate carboplatin treatment increased GSH/GSSG ratio. Carboplatin significantly decreased cochlear copper zinc-superoxide dismutase (CuZn-SOD) catalase (CAT), glutathione peroxidase (GSH-Px), glutathione reductase (GR) and glutathione-S-transferase (GST) activities and enzyme protein expressions and a significant increase in Mn-SOD activity, protein expression and malondialdehyde (MDA) level. Cochlear antioxidant enzyme activities, enzyme protein expressions and MDA level were partially restored in lipoate carboplatin treated rats, compared to carboplatin alone. Conclusion Carboplatin-induced ototoxicity is related to impairment of cochlear antioxidant system and otoprotection conferred by lipoate is associated with partial sparing of the cochlear antioxidant defense system.     

Hepatoprotective effects of Ger-Gen-Chyn-Lian-Tang in thioacetamide-induced fibrosis in mice

BackgroundMany researchers have focused on developing traditional herbal medicines as pharmacological medicines to treat hepatic fibrosis. In this study, we evaluated the possible mechanism of Ger-Gen-Chyn-Lian-Tang (GGCLT) on thioacetamide (TAA)-induced hepatic injury in mice.MethodsHepatic fibrosis mice were established by intraperitoneal injection with TAA (100 mg/kg, 3 times/week), and treated with daily oral administration of 30 mg/kg, 100 mg/kg, and 300 mg/kg of GGCLT for 6 weeks. There were 40 mice randomly assigned to control, TAA and TAA+GGCLT groups. When the experiment was completed, Masson's trichrome staining was used to measure the degree of liver fibrosis. Hepatic fibrosis molecules were assessed by Western blot and real-time polymerase chain reaction. Hepatic glutathione levels, matrix metalloproteinase (MMP-2 and MMP-9), and hydroxyproline were also measured.ResultsTreatment with GGCLT significantly reduced the toxicity of TAA and exhibited effective hepatoprotective activity. The mechanism of the hepatoprotective effect of GGCLT is proposed to be by normalizing oxidative stress. Additionally, the data of fibrotic areas, expression of procollagen III, and MMP2 and 9 mRNA levels in the TAA+GGCLT group were much lower than those in the TAA group (p < 0.05). Furthermore, the upregulation of hepatic protein levels of nuclear factor-κB, transforming growth factor (TGF)-β receptor-1, and smooth muscle α-actin induced by TAA was significantly inhibited after GGCLT treatment.ConclusionGGCLT can efficiently ameliorate hepatic fibrosis by its inhibitory effects on the intrahepatic oxidative stress in TAA mice model. The antioxidant properties afforded by GGCLT may be attributed to its modulation on TGF-β/TGFβ receptor signaling through the downregulation of integrated signal pathways involving smooth muscle α-actin and lipid peroxidation.