肝大部切除及Galactosamine肝中毒后肝脏再生中AFP的免疫细胞化学研究

肝脏有极大的再生能力,但其再生机制和过程至今仍不十分清楚.本实验取标记肝细胞分化的特异蛋白——AFP对肝再生进行研究.以112只成年Wistar雄性大鼠分为标准肝大部分(70％)切除术组(PH组)和galactosamine注射组(Gal组,一次性腹腔注射D-galaetosamine 1500g/kg b.w.).术后1、2、3、

Lycopene stabilizes lipoprotein levels during D-galactosamine/lipopolysaccharide induced hepatitis in experimental rats

Objective:To investigate the effect of lycopene on lipoprotein metabolism during D-galactosamine/lipopolysacchoride(D-Gal/LPS)induced hepatitis in experimentul rats.Methods:The efficacy of lycopene was validated during D-Gal/LPS induced hepatitis by analyzing the activity of lipid metabolizing enzymes such as lipoprotein lipase(LPL),lecithincholesterol acyl transferase(LCAT)and hepatic triglyceride lipase(HTCL).Lipo protein analyses were done by the estimation of very low density lipoprotein cholesterol(VLDL),low density lipoprotein cholesterol(LDL)and high density lipoprotein cholesterol(HDL).Remits:The toxic insult of D-galactosamine/lipopolysaccharide(D-Gal/LPS)in experimental group of animals reduces the normal values of lipid metabolizing enzymes due to liver injury.The significant drop in the levels of HDL and concomitant increase in the values of VLDL and LDL were observed.The pretreatment of lycopene restore these altered values to near normal level in experimental group of animals.Conclusions:In the light of results,it can be concluded that administration lycopene stabilizes the lipoprotein levels by regulating the lipid metabolizing enzymes through its antioxidant defense and helps to maintain the normal lipid metabolism during toxic injury in liver.

Arachidonic acid Metabolism in Galactosamine／Endotoxin Indudced Acute Liver injury in Rats

The changes of the levels of LTC4, PGI2 and TXA2 in the liver tissue in SD rats with GaIN/LPS-induced acute liver injury was studied with radioimmunoassay (RIA). As a result,12h after the administration of GaIN/LPS, serum AST (398±37u), ALT (565 ±43u) increased (P<0.001 ) and the concentration of TXA2 (12188±588pg/g· w· wt) in liver tissue increased sigiuficantly(p<0.001), while the content of LTC4 (9713± 3557ng/g·w·wt ) and PGI2 (1748±560 pg/g· w·wt) in liver tissue were not obviously changed(p>0.05) and the inflammatory changes of the pathological findings were observed. The improvement of serum ALT (300±168u)(p< 0.05) and AST(273±424 u) (P<0. 05) and histopathological damage was observed after the administration of diethylcarbamazine (DEC), a LTA4 synthesis inhibitor, the liver TXA2(12740±699) concentration significantly increased (P<0. 001), while the levels of LTC4 (8179±1653) and PGI2 (2320±630) were not obviously changed. Serum ALT (536±74u) and AST (416± 41u)(P> 0. 05) levels and histopathology did not change with administration of indomethacin, a cyclooxygenase inhibitor, but the liver LTC4 (12166±13027) contents increased (P<0.05 ) and TXA2 (1868±791) reduced significantly (P<0. 001). The present study suggests that arachidonic acid metabolism in rats with acute liver injury are significantly abnormal. Leukotrienes and thromboxane are important inflammatory mediators in the liver injury.

THE EFFECT OF SUPERANTIGEN STAPHYLOCOCCALENTEROTOXIN B AND D-GALACTOSAMINE ON BALB/CMOUSE HEPATOCYTES

Objective To observe the role of superantigen staphylococcal enterotoxin B(SEB) andD - galactosamine (D - GalN) on Balb/c mouse hepatocytes and its mechanism. Methods After Balb/c mice wereinjected intraperitoneally with SEB, D- GalN or both, blood samples were collected and livers were removed at 2,6, 12, 24h. Patterns of hepatocellular death were studied morphologically and biochemically, circulating cytokines(TNF, IFN-γ) were determined, and mice mortality within 24h was assessed. Results SEB could induce thetypical apoptotic changes of hepatocytes morphologically and biochemically. The mechanism is probably associatedwith the production and release of Cytokines (such as TNF, IFN- γ, etc).D - GalN could induce hepatocytesapoptosis and degeneration at the same time. Besides this, we confirmed hepatocytes of the mice which wereadministered SEB and D - GalN developing apoptosis at 2, 6h, but after 12h hepatocytes were characterized bysevere injury, the mice mortality within 24h is 50%. Conclusion SEB or D - GalN alone could induce the typicalapoptotic changes of hepatocytes. SEB+D-GalN developed hepatocytes apoptosis in the early stage and necrosisin the later. It suggests that there is some relationship between hepatic cell apoptosis and necrosis, and massivehepatocyte apoptosis is the probably initiating step of acute hepatic necrosis in mice.

Broccoli sprout extract induces detoxification-related gene expression and attenuates acute liver injury

AIM: To investigate the effects of broccoli sprout extract(BSEx) on liver gene expression and acute liver injury in the rat.METHODS: First, the effects of BSEx on liver gene expression were examined. Male rats were divided into two groups. The Control group was fed the AIN-76 diet, and the BSEx group was fed the AIN-76 diet containing BSEx. After a 10-d feeding period, rats were sacrificed and their livers were used for DNA microarray and realtime reverse transcription-polymerase chain reaction(RT-PCR) analyses. Next, the effects of BSEx on acute liver injury were examined. In experiments using acute liver injury models, 1000 mg/kg acetaminophen(APAP) or 350 mg/kg D-galactosamine(D-Gal N) was used to induce injury. These male rats were divided into four groups: Control, BSEx, Inducer(APAP or D-Gal N), and Inducer+BSEx. The feeding regimens were identical for the two analyses. Twenty-four hours following APAP administration via p.o. or D-Gal N administration via i.p., rats were sacrificed to determine serum aspartate transaminase(AST) and alanine transaminase(ALT) levels, hepatic glutathione(GSH) and thiobarbituric acid-reactive substances accumulation and glutathioneS-transferase(GST) activity. RESULTS: Microarray and real-time RT-PCR analyses revealed that BSEx upregulated the expression of genes related to detoxification and glutathione synthesis in normal rat liver. The levels of AST(70.91 ± 15.74 IU/m L vs 5614.41 ± 1997.83 IU/m L, P < 0.05) and ALT(11.78 ± 2.08 IU/m L vs 1297.71 ± 447.33 IU/m L, P < 0.05) were significantly suppressed in the APAP + BSEx group compared with the APAP group. The level of GSH(2.61 ± 0.75 nmol/g tissue vs 1.66 ± 0.59 nmol/g tissue, P < 0.05) and liver GST activity(93.19 ± 16.55 U/g tissue vs 51.90 ± 16.85 U/g tissue, P < 0.05) were significantly increased in the APAP + BSEx group compared with the APAP group. AST(4820.05 ± 3094.93 IU/m L vs 12465.63 ± 3223.97 IU/m L, P < 0.05) and ALT(1808.95 ± 1014.04 IU/m L vs 3936.46 ± 777.52 IU/m L, P < 0.05) levels were significantly suppressed in the D-Gal N + BSEx group compared with the D-Gal N group, but the levels of AST and ALT in the D-Gal N + BSEx group were higher than those in the APAP + BSEx group. The level of GST activity was significantly increased in the D-Gal N + BSEx group compared with the D-Gal N group(98.04 ± 15.75 U/g tissue vs 53.15 ± 8.14 U/g tissue, P < 0.05).CONCLUSION: We demonstrated that BSEx protected the liver from various types of xenobiotic substances through induction of detoxification enzymes and glutathione synthesis.

Impact of asialoglycoprotein receptor deficiency on the development of liver injury

The asialoglycoprotein （ASGP） receptor is a wellcharacterized hepatic receptor that is recycled via the common cellular process of receptor-mediated endocytosis （RME）. The RME process plays an integral part in the proper trafficking and routing of receptors and ligands in the healthy cell. Thus, the missorting or altered transport of proteins during RME is thought to play a role in several diseases associated with hepatocyte and liver dysfunction. Previously, we examined in detail alterations that occur in hepatocellular RME and associated receptor functions as a result of one particular liver injury, alcoholic liver disease （ALD）. The studies revealed profound ethanol- mediated impairments to the ASGP receptor and the RME process, indicating the importance of this receptor and the maintenance of proper endocytic events in normal tissue. To further clarify these observations, studies were performed utilizing knockout mice （lacking a functional ASGP receptor） to which were administered several liver toxicants. In addition to alcohol, we examined the effects following administration of anti- Fas （CD95） antibody, carbon tetrachloride （CCh） and lipopolysaccharide （LPS）/galactosamine. The results of these studies demonstrated that the knockout mice sustained enhanced liver injury in response to all of the treatments, as shown by increased indices of liver damage, such as enhancement of serum enzyme levels, histopathological scores, as well as hepatocellular death. Overall, the work completed to date suggests a possible link between hepatic receptors and liver injury. In particular, adequate function and content of the ASGP receptor may provide protection against various toxinmediated liver diseases.

Hepatoprotective effect of nitric oxide in experimental model of acute hepatic failure

AIM: To evaluate the effect of nitric oxide (NO) on the development and degree of liver failure in an animal model of acute hepatic failure (AHF).

Fulminant liver failure models with subsequent encephalopathy in the mouse

BACKGROUND:A reliable model of fulminant liver failure (FLF) is urgently required in this research field.This study aimed to develop a murine FLF model.METHODS:We used three groups of male C57BL/6 mice:control,with azoxymethane treatment (AOM group),and with galactosamine and tumor necrosis factor-alpha treatment (Gal+TNF-α group).The effects of body temperature (BT) control on survival in all three groups were investigated Using BT control,we compared the survival,histopathological findings and biochemical/coagulation profiles between the two experimental groups.The effects of hydration on international normalized ratios of prothrombin time (PT INRs) were also checked.Dose-dependent survival curves were constructed for both experimental groups.Neurological behavior was assessed using a coma scale.RESULTS:No unexpected BT effects were seen in the control group.The AOM group,but not the Gal+TNF-α group showed a significant difference in survival curves between those with and without BT care.Histopathological assessment showed consistent FLF findings in both experimental groups with BT care.There were significant differences between the experimental groups in aspartate aminotransferase levels and PT-INRs,and significant differences in PT-INRs between the sufficiently and insufficiently hydrated groups.There were significant differences between FLF models in the duration of each coma stage,with significant differences in stages 1 and 3 as percentages of the disease state (stages 1-4).The two FLF models with BT care showed different survival curves in the dose-dependent survival study.CONCLUSIONS:AOM provides a good FLF model,but requires a specialized environment and careful BT control.Other FLF models may also be useful,depending on the research purpose.Thoughtful attention to caregiving and close observation are indispensable for successful FLF models.

Study on liver injury models induced by CCl_(4)D-Gal and ANIT in mice

ＳｔｕｄｙｏｎｌｉｖｅｒｉｎｊｕｒｙｍｏｄｅｌｓｉｎｄｕｃｅｄｂｙＣＣｌ４ＤＧａｌａｎｄＡＮＩＴｉｎｍｉｃｅＹＡＮＧＸｉｎＢｏ１，ＨＵＡＮＧＺｈｅｎｇＭｉｎｇ１，ＣＡＯＷｅｎＢｉｎ１，ＺＨＥＮＧＭｉｎｇ１，ＣＨＥＮＨｏｎｇＹａｎ１，ＺＨＡＮ...

The Effects of Human Fetal Liver Cells on Experimental Fulminant Hepatic Failure in Rats

The beneficial dffects of human fetal liver cells on the D-galactosamine-inducedfulminant hepatic failure were observed in rats and the mechanisms of the effects investi-gated.The survival rate of the rats intraperitoneally injected with liver cell suspensionand cytosol was 47.37% and 42.11% respectively which was significantly higher than5.26% of the controls（P【0.05）.There was no statistical significance between the differ-ence of the survival rates due to two preparations.The administration of cytosol two hours before the intoduction of D-galactosamine re-sulted in a significant lowering of the plasma level of endotoxin and hepaticmalondialdehyde in rats and a marked increase of 3H-thymindinc incorporation intohepatic DNA(DPM/OD260)as compared with the parameters of the controls.These re-sults suggest that there might be some biological active substance in human fetal liver cellswhich is responsible for thc effects to increase the survival rate.

Isofuranodiene protect D-galactosamin/lipopolysacchride induced liver injury in rats

OBJECTIVE Isofuranodiene(ISO)is a natural product isolated in Chinese herb.Our recent results showed anticancer effect in vitro.In this study,we investigated its live protective effect with a Dgalactosamine/lipopolysacchride(GalN/LPS)induced rat model.METHODS SD rats were treated orally with or without ISO(20 and 50mg·kg-1,ig)for 3d and then treated with GalN/LPS for 8h.The serum were collected and the concentration of aspartate aminotransferase(AST),alanine aminotransferase(ALT),and malondialdehyde(MDA)were determined.The liver injury was examined by H&E staining.The mRNA expression of IL-1β,IL-6 and inducible nitric oxide synthase(iNOS)in liver tissues were determined by realtime PCR.RESULTS Oral administration of ISO(20 and 50mg·kg-1)dramatically inhibited GalN/LPS-induced serum elevation of AST,ALT,and MDA levels.The liver injury was also significantly ameliorated as evidenced by the histological improvement in H&E staining.Furthermore,ISO treatment significantly inhibited GalN/LPS-induced mRNA expression of IL-1β,IL-6,and inducible nitric oxide synthase(iNOS)in liver tissues.CONCLUSION This data showed that ISO has hepatoprotective effect in rats.