Development of PEGylated serum albumin with multiple reduced thiols as a long-circulating scavenger of reactive oxygen species for the treatment of fulminant hepatic failure in mice

Reactive oxygen species (ROS) are involved in the pathophysiology of fulminant hepatic failure. Therefore, we developed polyethylene glycol-conjugated bovine serum albumin with multiple reduced thiols (PEG-BSA-SH) for the treatment of fulminant hepatic failure. As a long-circulating ROS scavenger, PEG-BSA-SH effectively scavenged highly reactive oxygen species and hydrogen peroxide in buffer solution. PEG-BSA-SH showed a long circulation time in the plasma after intravenous injection into mice. Fulminant hepatic failure was induced by intraperitoneal injection of lipopolysaccharide and d-galactosamine (LPS/d-GalN) into mice. The LPS/d-GalN-induced elevation of plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels was significantly inhibited by a bolus intravenous injection of PEG-BSA-SH. Furthermore, the changes in hepatic lipid peroxide and hepatic blood flow were effectively suppressed by PEG-BSA-SH. In contrast, l-cysteine, glutathione, and dithiothreitol, three traditional reduced thiols, had no statistically significant effects on the serum levels of ALT or AST. These findings indicate that PEG-BSA-SH is a promising ROS scavenger and useful in the treatment of fulminant hepatic failure.     

Highly purified eicosapentaenoic acid prevents the progression of hepatic steatosis by repressing monounsaturated fatty acid synthesis in high-fat/high-sucrose diet-fed mice

Eicosapentaenoic acid (EPA) is a member of the family of n-3 polyunsaturated fatty acids (PUFAs) that are clinically used to treat hypertriglyceridemia. The triglyceride (TG) lowering effect is likely due to an alteration in lipid metabolism in the liver, but details have not been fully elucidated. To assess the effects of EPA on hepatic TG metabolism, mice were fed a high-fat and high-sucrose diet (HFHSD) for 2 weeks and were given highly purified EPA ethyl ester (EPA-E) daily by gavage. The HFHSD diet increased the hepatic TG content and the composition of monounsaturated fatty acids (MUFAs). EPA significantly suppressed the hepatic TG content that was increased by the HFHSD diet. EPA also altered the composition of fatty acids by lowering the MUFAs C16:1 and C18:1 and increasing n-3 PUFAs, including EPA and docosahexaenoic acid (DHA). Linear regression analysis revealed that hepatic TG content was significantly correlated with the ratios of C16:1/C16:0, C18:1/C18:0, and MUFA/n-3 PUFA, but was not correlated with the n-6/n-3 PUFA ratio. EPA also decreased the hepatic mRNA expression and nuclear protein level of sterol regulatory element binding protein-1c (SREBP-1c). This was reflected in the levels of lipogenic genes, such as acetyl-CoA carboxylase α (ACCα), fatty acid synthase, stearoyl-CoA desaturase 1 (SCD1), and glycerol-3-phosphate acyltransferase (GPAT), which are regulated by SREBP-1c. In conclusion, oral administration of EPA-E ameliorates hepatic fat accumulation by suppressing TG synthesis enzymes regulated by SREBP-1 and decreases hepatic MUFAs accumulation by SCD1.     

T-5224, a selective inhibitor of c-Fos/activator protein-1, attenuates lipopolysaccharide-induced liver injury in mice

The effect of T-5224, a selective inhibitor of c-Fos/activator protein (AP)-1, on lipopolysaccharide (LPS) induced liver injury was examined in mice. Administration of LPS (10?mg?kg^?1, i.p.) markedly increased serum levels of tumor necrosis factor-alpha (TNFα), high mobility group box 1 (HMGB1), alanine aminotransferase/aspartate aminotransferase (ALT/AST), liver tissue levels of macrophage-inflammatory protein-1 alpha (MIP-1α) and monocyte chemoattractant protein-1 (MCP-1), as well as hepatic necrosis and inflammation, leading to 67?% lethality. Administration of T-5224 (300?mg?kg^?1, p.o.) after intraperitoneal injection of LPS imparted appreciable protection against acute elevations in serum levels of TNFα, HMGB1, ALT/AST as well as in liver tissue levels of MIP-1α and MCP-1, and reduced the lethality (27?%). These data indicate that T-5224 ameliorates liver injury and improves survival through decreasing production of proinflammatory cytokines and chemokines in endotoxemic mice.     

自噬抑制剂氯喹对急性酒精诱导小鼠肝损伤的影响

目的：探讨自噬抑制剂氯喹（CQ）对急性酒精诱导肝损伤的影响及其作用机制。方法：将雄性C57BL/6小鼠随机分为3组：正常对照组、酒精组、氯喹干预组（n=7）,其中酒精组按4.5 g/kg剂量给予33%（V/V）酒精灌胃。HE和油红O染色检测各组小鼠肝组织脂滴变化;检测肝组织甘油三酯（TG）含量变化;检测血清谷草转氨酶（AST）和谷丙转氨酶（ALT）活性;免疫荧光法检测微管相关蛋白轻链3（LC3）蛋白变化;Western blot法检测LC3蛋白和核蛋白P65表达的变化;ELISA法检测促炎因子TNF-α、IL-6的变化。结果：与对照组比较,酒精组脂滴形成、TG含量、血清AST和ALT活性明显增高。与对照组比较,酒精组LC3-Ⅱ蛋白表达明显增加;与酒精组比较,氯喹干预组使酒精诱导的LC3-Ⅱ蛋白表达增强进一步加剧,使酒精诱导的TG含量、血清AST和ALT活性进一步增高,同时增加了酒精诱导的p65入核及TNFα、IL-6释放。结论：急性酒精能引起小鼠肝脏脂肪变化及炎症,而自噬抑制剂氯喹抑制自噬进程,加剧酒精诱导的肝损伤,说明自噬在酒精诱导肝损伤中可能具有保护效应。     

Reduction of high-fat diet-induced liver proinflammatory state by eicosapentaenoic acid plus hydroxytyrosol supplementation: involvement of resolvins RvE1/2 and RvD1/2

High-fat diet (HFD)-fed mice show obesity with development of liver steatosis and a proinflammatory state without establishing an inflammatory reaction. The aim of this work was to assess the hypothesis that eicosapentaenoic acid (EPA) plus hydroxytyrosol (HT) supplementation prevents the inflammatory reaction through enhancement in the hepatic resolvin content in HFD-fed mice. Male C57BL/6J mice were fed an HFD or a control diet and supplemented with EPA (50 mg/kg/day) and HT (5 mg/kg/day) or their respective vehicles for 12 weeks. Measurements include liver levels of EPA, DHA and palmitate (gas chromatography), liver resolvins and triglyceride (TG) and serum aspartate transaminase (AST) (specific kits) and hepatic and serum inflammatory markers (quantitative polymerase chain reaction and enzyme-linked immunosorbent assay). Compared to CD, HFD induced body weight gain, liver steatosis and TG accumulation, with up-regulation of proinflammatory markers in the absence of histological inflammation or serum AST changes; these results were accompanied by higher hepatic levels of resolvins RvE1, RvE2, RvD1 and RvD2, with decreases in EPA and DHA contents. EPA+HT supplementation in HFD feeding synergistically reduced the steatosis score over individual treatments and increased the hepatic levels of EPA, DHA and resolvins, with attenuation of proinflammatory markers. Lack of progression of HFD-induced proinflammatory state into overt inflammation is associated with resolvin up-regulation, which is further increased by EPA+HT supplementation eliciting steatosis attenuation. These findings point to the importance of combined protocols in hepatoprotection due to the involvement of cross-talk mechanisms, which increase effectiveness and diminish dosages, avoiding undesirable effects.     

银杏叶提取物对对乙酰氨基酚诱导的小鼠急性肝损伤的保护作用

目的：探究银杏叶提取物（GBE）对对乙酰氨基酚（APAP）诱导的小鼠急性肝损伤的保护作用及其机制。方法：30只小鼠随机分为对照组、模型组、GBE低、中、高剂量组（50,100,and 200 mg·kg^-1）,每组6只。除对照组外,剩余小鼠腹腔注射APAP （300 mg/kg）一次,随后GBE低、中、高剂量组按照相应剂量灌胃给药,治疗2 d后取材。观察各组肝脏大体情况和肝组织的病理组织学变化;取血测定各组小鼠血清中ALT、AST的活性和TNF-α、IL-6的水平;取肝检测各组肝组织中SOD、MPO的活性和GSH、MDA的含量;通过Western blot检测各组肝组织中Nrf2、HO-1蛋白的表达量。结果：与对照组相比,模型组肝脏明显肿大,病理表现差,血清中ALT、AST、TNF-α、IL-6的水平显著升高（P<0.01）,肝组织中GSH的含量和SOD的活性显著降低（P<0.01）,MDA的含量和MPO的活性显著升高（P<0.01）,Nrf2、HO-1蛋白表达明显下调（P<0.01）。与模型组相比,GBE组肝脏肿大减轻,病理表现有所改善,血清中ALT、AST、TNF-α、IL-6的水平显著降低（P<0.01）,肝组织中GSH的含量和SOD的活性显著提高（P<0.01）,MDA的含量和MPO的活性显著降低（P<0.01）,Nrf2、HO-1蛋白表达上调（P<0.05）,其中高剂量GBE组治疗效果最明显。结论：GBE可对APAP诱导的小鼠急性肝损伤具有保护作用,其作用机制可能是通过Nrf2/HO-1抗氧化途径发挥作用。     

黄芪多糖对乙型肝炎小鼠肝脏脂肪变性的作用及机制

目的:探讨黄芪多糖对乙型肝炎相关的肝脏脂肪变性的治疗作用及机制。方法:选取5-6周龄的SPF雄性小鼠共20只作为研究对象,随机分为研究组和对照组,各10例,研究组小鼠进行黄芪多糖溶液灌胃,对照组小鼠采用同样剂量的生理盐水灌胃。分别于用药前后测量体重(BW)、甘油三酯(TG)、谷丙转氨酶(ALT)、谷草转氨酶(AST)。结果:研究组小鼠用药后BW和TG明显低于对照组,数据经统计学比较具有显著差异(P<0.05)。用药前两组小鼠ALT和AST比较无显著差异,用药后研究组ALT和AST明显下降,与对照组用药后比较具有显著差异(P<0.05)。结论:黄芪多糖可有效降低乙型肝炎小鼠BW和血脂TG,减少ALT和AST,促进肝脏的排毒与修复,从而改善肝脏脂肪变性,缓解和治疗HBV的毒性和侵害性,提高治疗效果。     

番茄皂苷A对小鼠血脂及肝脏脂肪的作用

该研究以ApoE基因缺陷小鼠和高脂饲料诱导的高血脂症模型小鼠为对象,采用药理学方法研究了番茄皂苷A对血脂及肝脏脂肪的调节作用。在ApoE基因缺陷小鼠和高脂饲料诱导的高血脂症模型小鼠中,通过灌胃给予番茄皂苷A:取血,测定血清中总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDLC)、低密度脂蛋白胆固醇(LDLC)、谷丙转氨酶(ALT)、谷草转氨酶(AST)、尿素氮(BUN)、肌酐(Cr)、葡萄糖(Glu)的含量和活性;处死小鼠后,取肝脏称重,计算肝脏指数;精确称取一部分肝脏,测定肝脏脂质的含量。结果表明:番茄皂苷A对ApoE基因缺陷小鼠可以降低血清TC、HDLC、LDLC的含量,对ALT、AST、BUN、Cr、Glu没有影响,说明番茄皂苷A可以降低ApoE基因缺陷小鼠血中胆固醇含量,对血糖没有影响,对肝肾功能无影响;对高脂饲料诱导的高血脂症模型小鼠,可以降低血清TC、HDLC的含量,可以降低肝脏TC的含量,对ALT、AST、BUN、Cr、Glu没有影响,说明番茄皂苷A可以改善高脂饲料诱导的高血脂症模型小鼠的脂质代谢,且对肝肾功能无影响。该研究结果表明番茄皂苷A具有一定的降低胆固醇的作用,且不影响肝肾功能。     

Liver iron overload induced by tamoxifen in diabetic and non-diabetic female Wistar rats

Tamoxifen (TX), a drug used in the treatment of breast cancer, may cause hepatic changes in some patients. The consequences of its use on the liver tissues of rats with or without diabetes mellitus (DM) have not been fully explored. The purpose of this study was to evaluate the correlation between plasma hepatic enzyme levels and the presence of iron overload in the hepatic tissue of female Wistar rats with or without streptozotocin-induced DM and using TX. Female rats were studied in control groups: C-0 (non-drug users), C-V (sorbitol vehicle only) and C-TX (using TX). DM (diabetic non-drug users) and DM-TX (diabetics using TX) were the test groups. Sixty days after induced DM, blood samples were collected for glucose, alanine aminotransferase (ALT), aspartate aminotransferase (AST) alkaline phosphatase (ALP) and bilirubin measures. Hepatic fragments were processed and stained with hematoxylin and eosin, Masson’s trichrome, Perls. The hepatic iron content was quantified by atomic absorption spectrometry. AST, ALT and ALP levels were significantly elevated in the DM and DM-TX groups, with unchanged bilirubin levels. Liver iron overload using Perls stain and atomic absorption spectrometry were observed exclusively in groups C-TX and DM-TX. There was positive correlation between AST, ALT and ALP levels and microscopic hepatic siderosis intensity in group DM-TX. In conclusion, TX administration is associated with liver siderosis in diabetic and non-diabetic rats. In addition, TX induced liver iron overload with unaltered hepatic function in non-diabetic rats and may be a useful tool for investigating the biological control of iron metabolism.     

Protective Effect of a Fermented Substance from Saccharomyces cerevisiae on Liver Injury in Mice Caused by Acetaminophen

The protective effect of a fermented substance from Saccharomyces cerevisiae (FSSC) on liver injury caused by acetaminophen (AAP) was studied in mice. Mice were pretreated with FSSC (0.5–2.0 g/kg, p.o.) for 4 d, and on the fourth day, the mice received an overdose of AAP (500 mg/kg, i.p.). Subsequently, they were sacrificed at 7 h, and blood was drawn from the abdominal vein and liver samples were collected. Histological and biochemical examinations revealed that the administration of AAP caused liver injury in the mice, including increases in plasma alanine aminotransferase and asparate aminotransferase activities and decreases in the hepatic reduced form of glutathione (GSH) content and antioxidant enzyme activities. Prior to AAP treatment, the mice pretreated with FSSC showed significantly reduced levels of alanine aminotransferase (ALT) and aspirate aminotransferase (AST) activity. Liver histology in the FSSC-pretreated mice was significant. In these mice, pretreatment with FSSC also served to reduce hepatic GSH depletion and the inhibition of antioxidant enzyme activity caused by AAP overdose. In conclusion, oral administration of FSSC significantly reduced AAP-induced hepatic injury in the mice.     

Lysosomal unesterified cholesterol content correlates with liver cell death in murine Niemann-Pick type C disease

Niemann-Pick type C (NPC) disease is a multisystem disorder resulting from mutations in the NPC1 gene that encodes a protein involved in intracellular cholesterol trafficking. Significant liver dysfunction is frequently seen in patients with this disease. The current studies used npc1 mutant mice to investigate the association between liver dysfunction and unesterified cholesterol accumulation, a hallmark of NPC disease. Data from 92 npc1(-/-) mice (age range, 9-56 days) revealed a significant positive correlation between the plasma activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and whole liver cholesterol content. In 56 day old npc1(-/-) mice that had been fed from 35 days of age a rodent diet or the same diet containing either cholesterol (1.0%, w/w) or ezetimibe (a sterol absorption inhibitor; 0.0125%, w/w), whole liver cholesterol content averaged 33.5 +/- 1.1, 87.9 +/- 1.7, and 20.8 +/- 0.9 mg, respectively. Again, plasma ALT and AST activities were positively correlated with hepatic cholesterol content. In contrast, plasma transaminase levels remained in the normal range in npc1(+/+) mice, in which hepatic esterified cholesterol content had been increased by 72-fold by feeding a high-cholesterol, high-fat diet. These studies suggest that the late endosomal/lysosomal content of unesterified cholesterol correlates with cell damage in NPC disease.     

Dietary eicosapentaenoic acid supplementation accentuates hepatic triglyceride accumulation in mice with impaired fatty acid oxidation capacity

Reduced mitochondrial fatty acid (FA) β-oxidation can cause accumulation of triglyceride in liver, while intake of eicosapentaenoic acid (EPA) has been recommended as a promising novel therapy to decrease hepatic triglyceride content. However, reduced mitochondrial FA β-oxidation also facilitates accumulation of EPA. To investigate the interplay between EPA administration, mitochondrial activity and hepatic triglyceride accumulation, we investigated the effects of EPA administration to carnitine-deficient mice with impaired mitochondrial FA β-oxidation. C57BL/6J mice received a high-fat diet supplemented or not with 3% EPA in the presence or absence of 500 mg mildronate/kg/day for 10 days. Liver mitochondrial and peroxisomal oxidation, lipid classes and FA composition were determined. Histological staining was performed and mRNA level of genes related to lipid metabolism and inflammation in liver and adipose tissue was determined. Levels of pro-inflammatory eicosanoids and cytokines were measured in plasma. The results showed that mildronate treatment decreased hepatic carnitine concentration and mitochondrial FA β-oxidation and induced severe triglyceride accumulation accompanied by elevated systemic inflammation. Surprisingly, inclusion of EPA in the diet exacerbated the mildronate-induced triglyceride accumulation. This was accompanied by a considerable increase of EPA accumulation while decreased total n-3/n-6 ratio in liver. However, inclusion of EPA in the diet attenuated the mildronate-induced mRNA expression of inflammatory genes in adipose tissue. Taken together, dietary supplementation with EPA exacerbated the triglyceride accumulation induced by impaired mitochondrial FA β-oxidation. Thus, further thorough evaluation of the potential risk of EPA supplementation as a therapy for NAFLD associated with impaired mitochondrial FA oxidation is warranted.     

The uncoordinated-5 homolog B receptor affects hepatic ischemia reperfusion injury

Recent evidence has demonstrated additional roles for the neuronal guidance protein receptor UNC5B outside the nervous system. Given the fact that ischemia reperfusion injury (IRI) of the liver is a common source of liver dysfunction and the role of UNC5B during an acute inflammatory response we investigated the role of UNC5B on acute hepatic IRI. We report here that UNC5B(+/-) mice display reduced hepatic IRI and neutrophil (PMN) infiltration compared to WT controls. This correlated with serum levels of lactate dehydrogenase (LDH), aspartate- (AST) and alanine- (ALT) aminotransferase, the presence of PMN within ischemic hepatic tissue, and serum levels of inflammatory cytokines. Moreover, injection of an anti-UNC5B antibody resulted in a significant reduction of hepatic IR injury. This was associated with reduced parameters of liver injury (LDH, ALT, AST) and accumulation of PMN within the injured hepatic tissue. In conclusion our studies demonstrate a significant role for UNC5B in the development of hepatic IRI and identified UNC5B as a potential drug target to prevent liver dysfunction in the future.     

ERCP术与腹腔镜手术治疗胆总管结石的疗效对比及对血清AST、ALT、ALP的影响

目的:探讨内镜逆行胰胆管造影术(ERCP)与腹腔镜手术治疗胆总管结石的疗效对比及对血清门冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、碱性磷酸酶(ALP)的影响。方法:选择2017年4月到2018年4月我院接诊的胆总管结石患者50例作为研究对象,以随机数表法分为观察组(n=26)和对照组(n=24),对照组使用腹腔镜手术治疗,观察组采用ERCP取石术治疗。比较两组手术时间、术中出血量、术后下床时间、住院时间、血清AST、ALT、ALP水平变化情况、术后胃肠道恢复时间及术后不良反应发生情况。结果:观察组患者手术时间高于对照组,(P0.05),术中出血量、术后下床时间及住院时间均显著低于对照组(P0.05);手术前,两组患者血清AST、ALT、ALP水平无明显差异;治疗后,两组患者血清AST、ALT、ALP水平均明显上升(P0.05),且观察组患者血清AST、ALT、ALP水平显著低于对照组(P0.05);观察组患者肠鸣音恢复时间、排气时间、排便时间均明显低于对照组(P0.05);治疗期间,观察组患者不良反应总发生率为7.69%,显著低于对照组的29.17%(P0.05)。结论:在胆总管结石患者中应用ERCP取石术效果显著,对患者血清血清AST、ALT、ALP影响较小,值得推广与运用。     

Traditional Chinese Medicine improves dysfunction of peroxisome proliferator-activated receptor alpha and microsomal triglyceride transfer protein on abnormalities in lipid metabolism in ethanol-fed rats

We report the effects of Traditional Chinese Medicine (TCM) on alcohol-induced fatty liver in rats. TCM consists of Astragalus membranaceus, Morus alba, Crataegus pinnatifida, Alisma oriental, Salvia miltiorrhiza and Pueraria lobata. The rats were separated randomly into five groups; the CD group (n=10), which was fed a control diet for 10 weeks, the ED group (n=10), which was fed an isocaloric liquid diet containing ethanol for 10 weeks and given daily oral doses of TCM (0.222 g/kg/day; TCM222, 0.667 g/kg/day; TCM667, and 2.000 g/kg/day; TCM2000, n=10, respectively) over the last four weeks of the study. The ED group developed fatty livers, as determined by their lipid profiles and liver histological findings. Compared with the control group, liver/body weight, plasma triglyceride (TG) and total cholesterol (TC), liver TG and TC, plasma alanine aminotransferase (ALT) and aspartic aminotransferase (AST) significantly increased in the ED group. Also, free fatty acids (FFA) levels increased in both plasma and liver during the administration of ethanol. On the other hand, when rats were administrated with TCM, their liver/body weight, plasma TG, TC and FFA, liver TG, TC and FFA, plasma ALT and AST decreased significantly and the degree of hepatic lipid droplets was markedly improved compared with those in the ED group. Proper function of the peroxisome proliferator-activated receptor alpha (PPARalpha) is essential for the regulation of hepatic fatty acid metabolism. Microsomal triglyceride transfer protein (MTP) is essential for the secretion of triglycerides from the liver. mRNAs for PPARalpha and MTP were reduced in the livers of ethanol-fed rats. TCM restored the mRNA levels of PPARalpha and MTP, and prevented development of fatty livers in ethanol-fed rats. Impairment of PPARalpha and MTP function during ethanol consumption contributes to the development of alcohol-induced fatty liver, which can be overcome by TCM.     

吡非尼酮对四氯化碳诱导的小鼠肝纤维化的影响

目的：研究吡非尼酮对四氯化碳诱导的小鼠肝纤维化的影响。方法：选用8周健康雄性SPF级ICR小鼠40只,随机分为4组（n=10）：肝纤维化模型组（CCL₄组）、吡非尼酮低剂量组（PFD-L组）、吡非尼酮高剂量组（PFD-H组）及正常对照组。CCL₄肝纤维化模型采用0.4 ml/只10%的CCL₄大豆油溶液进行小鼠腹腔注射制备,低剂量、高剂量吡非尼酮干预组在造模后采用120 mg/kg、240 mg/kg吡非尼酮（PFD）灌胃,正常对照组采用与前三组等量的生理盐水腹腔注射的方法。全自动生化仪检测血清中谷丙转氨酶（ALT）、谷草转氨酶（AST）、碱性磷酸酶（ALP）;取肝脏组织HE染色观察组织的病理学变化;荧光定量PCR法测定肝脏中α-平滑肌肌动蛋白（α-SMA）相关基因的表达,酶联反应法检测肝纤维化指标透明质酸（HA）、层粘连蛋白（LN）、Ⅳ型胶原（IV-C）。结果：与正常组相比,CCL₄组小鼠肝小叶结构明显破坏,胶原纤维沉积明显,可见假小叶形成;血清ALT、AST、ALP均显著升高（P<0.05）;血清肝纤维化指标HA、LN、IV-C均显著升高（P<0.05）;α-SMA基因的表达水平也显著升高（P<0.05）。PFD-L组和PFD-H组小鼠AST、ALT、ALP相较于CCL₄组明显降低,PFD-L组和PFD-H组小鼠血清肝纤维化指标HA、LN、IV-C相较于CCL₄组明显降低,α-SMA基因的表达也受到抑制（P<0.05）。病理学观察发现,PFD-L组小鼠的肝纤维化程度减轻,胶原纤维减少,仅见少量假小叶;PFD-H组小鼠细胞排列恢复,小叶结构轻度紊乱,未见明显假小叶,恢复效果比PFD-L组好。结论：吡非尼酮对于肝纤维化有一定的治疗作用,其可成为肝纤维化早期干预的新药物。     

红景天苷调节AMPK/mTOR/ULK1信号通路对结肠癌SW480细胞裸鼠的肝脏损伤的影响

摘要 目的：探讨红景天苷（Sal）调节单磷酸腺苷活化蛋白激酶（AMPK）/哺乳动物雷帕霉素靶蛋白（mTOR）/Unc51样激酶1（ULK1）信号通路对结肠癌SW480细胞裸鼠肝脏损伤的影响。方法：通过皮下注射SW480细胞悬浮液建立肝转移裸鼠模型,将造模后的裸鼠随机分为模型组、Sal低剂量（Sal-L,50 mg/kg Sal）组、Sal中剂量（Sal-M,100 mg/kg Sal）组、Sal高剂量（Sal-H,200 mg/kg Sal）组,Sal-H+AMPK抑制剂（Compound C,200 mg/kg Sal+10 mg/kg Compound C）组,以未接种SW480细胞悬液的裸鼠作为对照组。腹部主动脉取血,检测裸鼠血清中丙氨酸氨基转移酶（AST）、天冬氨酸氨基转移酶（ALT）水平;处死裸鼠,检测肝转移瘤数目及肝脏重量;HE染色观察肝脏组织病理变化;qRT-PCR检测肝脏组织中AMPK、mTOR、ULK1 mRNA表达水平;Western blot检测肝脏组织中自噬（Beclin1、p62）蛋白及通路相关蛋白表达。结果：与对照组相比,模型组裸鼠组织中出现肝转移瘤,肝脏重量、AST、ALT水平、mTORmRNA、ULK1 mRNA、p62表达显著增加（P<0.05）;Beclin1、AMPK mRNA及蛋白表达显著降低（P<0.05）;与模型组相比,Sal-L、Sal-M、Sal-H组肝转移瘤数目、肝脏重量、AST、ALT水平、mTORmRNA、ULK1 mRNA、p62表达显著降低（P<0.05）;Beclin1、AMPK mRNA及蛋白表达显著增加（P<0.05）;与Sal-H组相比,Sal-H+Compound C组肝转移瘤数目、肝脏重量、AST、ALT水平、mTORmRNA、ULK1 mRNA、p62表达显著增加（P<0.05）;Beclin1、AMPK mRNA及蛋白表达显著降低（P<0.05）。结论：Sal可通过减少裸鼠肝转移瘤形成,保护裸鼠肝脏,其机制可能与激活AMPK/mTOR/ULK1信号通路,促进肝脏自噬有关。     

紫檀芪调节Keap-1/Nrf2/HO-1信号通路对非酒精性脂肪肝大鼠氧化应激和细胞凋亡的影响

摘要 目的：研究紫檀芪调节Kelch样ECH关联蛋白1（Keap-1）/核因子E2相关因子2（Nrf2）/血红素加氧酶-1（HO-1）信号通路对非酒精性脂肪肝（NAFLD）大鼠氧化应激和细胞凋亡的影响。方法：将60只SD大鼠随机分为对照组、模型组、紫檀芪低剂量组（30 mg/kg）、紫檀芪高剂量组（60 mg/kg）、紫檀芪（60 mg/kg）+N-（4-（2,3-二氢-1-（2''-甲基苯甲酰）-1H-吲哚-5-基）-5-甲基-2-噻唑基）-1,3-苯并二氧唑-5-乙酰胺（ML385）（30 mg/kg）组，每组12只。模型组与药物干预组大鼠以高脂饲料饲养诱导NAFLD模型，对照组大鼠以普通饲料饲养，各组连续喂养12周。以紫檀芪和ML385分组处理14 d后（对照组以等剂量生理盐水处理），检测各组大鼠脂代谢指标[三酰甘油（TG）、总胆固醇（TC）及游离脂肪酸（FFA）水平]、肝指数、肝功能指标[谷丙转氨酶（ALT）及谷草转氨酶（AST）]水平、血清白细胞介素（IL）-17、IL-6、IL-10、氧化应激指标[丙二醛（MDA）、超氧化物歧化酶（SOD）及过氧化氢酶（CAT）]水平；原位末端标记法（TUNEL）染色检测各组大鼠肝细胞凋亡率；蛋白免疫印迹法检测各组大鼠肝组织凋亡相关蛋白及Keap-1/Nrf2/HO-1通路相关蛋白表达。结果：与对照组相比，模型组大鼠血清IL-10、SOD及CAT水平、肝组织Nrf2、HO-1、Bcl-2表达水平显著降低（P＜0.05），TG、TC及FFA水平、肝指数、ALT及AST水平、血清IL-17、IL-6、MDA水平、肝细胞凋亡率、肝组织Keap-1及Bax表达水平显著升高（P＜0.05）。与模型组相比，紫檀芪低、高剂量组大鼠血清IL-10、SOD及CAT水平、肝组织Nrf2、HO-1、Bcl-2表达水平均升高（P＜0.05），TG、TC及FFA水平、肝指数、ALT及AST水平、血清IL-17、IL-6、MDA水平、肝细胞凋亡率、肝组织Keap-1、Bax表达水平均降低（P＜0.05）；与紫檀芪低剂量组相比，紫檀芪高剂量组大鼠血清IL-10、SOD及CAT水平、肝组织Nrf2、HO-1、Bcl-2表达水平升高（P＜0.05），TG、TC及FFA水平、肝指数、ALT及AST水平、血清IL-17、IL-6、MDA水平、肝细胞凋亡率、肝组织Keap-1及Bax表达水平降低（P＜0.05）；与紫檀芪高剂量组相比，紫檀芪+ML385组大鼠血清IL-10、SOD及CAT水平、肝组织Nrf2、HO-1、Bcl-2表达水平降低（P＜0.05），TG、TC及FFA水平、肝指数、ALT及AST水平、血清IL-17、IL-6、MDA水平、肝细胞凋亡率、肝组织Bax表达水平升高（P＜0.05）。结论：紫檀芪可能通过激活Keap-1/Nrf2/HO-1信号通路，改善NAFLD大鼠脂代谢水平，调节炎症反应及氧化应激，减轻肝组织脂肪变性及细胞凋亡。     

Orally administrated rare earth element cerium induces metallothionein synthesis and increases glutathione in the mouse liver

The influence of oral administration of rare earth element cerium (Ce) was studied in relation to metallothionein (MT) and glutathione (GSH) content in the organs of ICR mice, which were administered heavy metal cadmium (Cd) for comparison. Male ICR mice were divided into 9 groups: 1 control group, 4 cerium groups and 4 cadmium groups, each with 4 mice, for a total of 36 mice. Ce groups included a 20 ppm CeCl3 diet (Ce-low) group and a 200 ppm CeCl3 diet (Ce-high) group, as did Cd groups, i.e., a 20 ppm CdCl2 diet (Cd-low) group and a 200 ppm CdCl2 diet (Cd-high) group. Each group was subdivided in 2 groups except a control group: 6-week administration group and 12-week administration group. The level of plasma aspartate aminotransferase(AST) activity, plasma alanine aminotransferase(ALT) activity, plasma cholesterol and plasma triglyceride in the Ce-low, Cd-low, Ce-high, and Cd-high group were higher than that of control group, although there were no significant differences (p > 0.05). By contrast, both Ce and Cd groups had higher levels of MT and GSH in hepatic cells compared to the control group (p < 0.05) and decreased liver tissue level of lipoperoxide (p < 0.05). These groups also had decreased plasma superoxide dismutase (SOD) activity (p < 0.05), and increased plasma level of lipoperoxide (p > 0.05). In conclusion, it is suggested that orally administered Ce increases MT and GSH as an antioxidant in the mouse liver, and these reaction are probably caused by increases in the oxidative stress with Ce.     

乙酰辅酶A羧化酶抑制剂联合非诺贝特对小鼠非酒精性脂肪肝的治疗效果

摘要 目的：探讨乙酰辅酶A羧化酶抑制剂（MK-4074）联合非诺贝特对小鼠非酒精性脂肪肝（NAFLD）的脂质含量以及肝功能的改善效果。方法：20只C57BL/6小鼠给予60%高脂饲料连续喂养8周构建NAFLD小鼠模型后，随机分为安慰剂组、MK-4074组、非诺贝特组以及MK-4074联合非诺贝特治疗组，每组各5只，继续高脂喂养并分别给予安慰剂（Placebo）、MK-4074（10 mg/kg/天）、非诺贝特（30 mg/kg/天）、以及MK-4074（10 mg/kg/天）+ 非诺贝特（30 mg/kg/天）治疗持续8周。治疗结束后对小鼠体重、肝指数、肝脏脂质含量、肝功能以及肝脏病理和肝脏中性粒细胞和巨噬细胞浸润情况进行分析。结果：与安慰剂组相比，单用MK-4074治疗可显著降低肝指数、肝脏甘油三酯（TG）、胆固醇（TC）、非酯化脂肪酸（NEFA）的含量以及血清ALT和AST水平，而对小鼠体重和血清TC没有显著影响；单用非诺贝特可显著降低小鼠体重，肝脏TG、TC、NEFA以及血清TG、 ALT和AST水平，对小鼠的肝指数、血清TC没有显著影响；而MK-4074与非诺贝特联合治疗可显著降低小鼠体重、肝脏TG、TC、NEFA，以及血清TG、ALT和AST水平，降低肝脏脂质积累以及中性粒细胞与巨噬细胞浸润，效果优于MK-4074或非诺贝特单药治疗。结论：MK-4074联合非诺贝特可显著减少NAFLD小鼠肝脏的脂质含量，改善肝功能。