Aqueous extract of Anoectochilus formosanus attenuate hepatic fibrosis induced by carbon tetrachloride in rats.

The aim of this study was to investigate the effects of aqueous extract of Anoectochilus formosanus (AFE) on liver fibrogenesis in carbon tetrachloride (CCl4)-induced cirrhosis. Fibrosis was induced in rats by oral administration of CCl4 (20%, 0.5 ml/rat, p.o.) twice a week for 8 weeks. AFE (0.5 and 2.0 g/kg, p.o., daily for 8 weeks) was administered to rats simultaneously. AFE showed reducing actions on the elevated levels of GOT and GPT caused by CCl4. Liver fibrosis in rats induced by CCl4 led to the drop of serum albumin concentration; the AFE increased the albumin concentration. The CCl4-induced liver fibrosis markedly caused liver atrophy and splenomegalia, while AFE increased the liver weight, and decreased the spleen weight. The CCl4-induced liver fibrosis decreased the protein content, and increased collagen contents in rat's liver. AFE significantly increased the contents of protein and reduced the amount of collagen in the liver. In CCl4-treated rats, glutathione concentrations of liver were not affected. AFE significantly increased liver glutathione concentrations. All these results clearly demonstrate that AFE can reduce the liver fibrogensis in rats induced by CCl4.     

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。     

Use of phospholipids to repair rat liver membranes during carbon tetrachloride poisoning

The effect of phospholipid multilayer liposomes on the properties of endoplasmic reticular membranes has been studied in hepatic cells damaged with CCl4. It has been shown that the repair effect of phosphatidylcholine liposomes was manifested in vivo by normalization of phospholipid membrane composition, reduction in the degree of cytochrome P-450 inactivation, partial normalization of its hydroxylase activity and recovery of glucose-6-phosphatase activity. The degree of phosphatidylcholine unsaturation, and the introduction of antioxidants--SH-compounds, sphingomyelin, phosphatidyl inositol--into liposomes did not influence the efficacy of liposomes.     

Cationic liposomes-mediated plasmid DNA delivery in murine hepatitis induced by carbon tetrachloride

In order to elucidate the influence of hepatic disease stage on cationic liposomes-mediated gene delivery, we investigated the cationic liposomes-mediated plasmid DNA delivery with time in murine hepatitis induced by subcutaneous injection of CCl₄. Liver injury after injection of CCl₄ was confirmed by the determination of serum aspartate aminotransferase and alanine aminotransferase activities. Two kinds of liposomes constructed with N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethlylammoniumchloride and dioleylphosphatidylethanolamine (DOTMA-DOPE) or DOTMA and cholesterol (DOTMA-CHOL) were used for the gene-delivery vector. We determined luciferase activities in various organs after the intravenous administration of the lipoplexes. The CCl₄-treated mice administered with DOTMA-DOPE lipoplexes showed the more significant decreases of transgene expression in the liver and spleen at 18 hours after CCl₄ injection. On the other hand, the CCl₄-treated mice administered with DOTMA-CHOL lipoplexes showed a significant increase in the liver at 48 hours. In conclusion, our findings demonstrate that murine hepatitis induced by CCl₄ can influence cationic liposomes-mediated plasmid DNA delivery. The extent of influences was also affected by lipid contents. These results indicate the necessity of considering the timing and the formulation for gene therapy according to the disease stage.     

Effective protection of Terminalia catappa L. leaves from damage induced by carbon tetrachloride in liver mitochondria

The protective effects of chloroform extracts of Terminalia catappa L. leaves (TCCE) on carbon tetrachloride (CCl4)-induced liver damage and the possible mechanisms involved in the protection were investigated in mice. We found that increases in the activity of serum aspartate aminotransferase and alanine aminotransferase and the level of liver lipid peroxidation (2.0-fold, 5.7-fold and 2.8-fold) induced by CCl4 were significantly inhibited by oral pretreatment with 20, 50 or 100 mg/kg of TCCE. Morphological observation further confirmed the hepatoprotective effects of TCCE. In addition, the disruption of mitochondrial membrane potential (14.8%), intramitochondrial Ca2+ overload (2.1-fold) and suppression of mitochondrial Ca2+-ATPase activity (42.0%) in the liver of CCl4-insulted mice were effectively prevented by pretreatment with TCCE. It can be concluded that TCCE have protective activities against liver mitochondrial damage induced by CCl4, which suggests a new mechanism of the hepatoprotective effects of TCCE.     

Systemic inflammatory response syndrome (SIRS) induced by carbon tetrachloride in rats

We have observed the symptoms of systemic inflammatory response syndrome (SIRS) in male rats intoxicated by carbon tetrachloride (CCl(4)). Severe hypothermia, tachypnoea and increase in the heart beat min were diagnosed. These symptoms developed in the first hour of intoxication. The hepatic dysfunction was characterized by elevated bilirubin levels. In the sera we have measured increases in the activity of secretable (group II) phospholipase A(2) sPLA(2) (2,8x) and 6-ketoprostaglandin F(1alpha) (KPGF) (1,44x). Supposedly the free radicals derived from CCl(4)-mainly trichloromethyl-could induce the prompt reaction of SIRS and the release of sPLA(2) as well as the formation of KPGF. Our findings show that in the early phase of CCl(4) intoxication the symptoms of SIRS can be related to elevation of sPLA(2) and the products of cyclooxygenase II.     

Potentiation by chlordecone of the defect in hepatic microsomal calcium sequestration induced by carbon tetrachloride

The effect of carbon tetrachloride (CCl4) on the capacity of hepatic microsomes to sequester calcium was studied following pretreatment of rats with chlordecone. Chlordecone pretreatment alone had no effect on the kinetics of calcium uptake by hepatic microsomes. It was found, however, that chlordecone pretreatment of rats potentiated by sixfold the potency of CCl4 to suppress microsomal calcium sequestration capacity when measured one hour after CCl4 administration.