Lysosomal membrane protein Sidt2 knockout induces apoptosis of human hepatocytes in vitro independent of the autophagy-lysosomal pathway / 南方医科大学学报

OBJECTIVE@#To explore the regulatory mechanism of human hepatocyte apoptosis induced by lysosomal membrane protein Sidt2 knockout.@*METHODS@#The Sidt2 knockout (Sidt2-/-) cell model was constructed in human hepatocyte HL7702 cells using Crispr-Cas9 technology.The protein levels of Sidt2 and key autophagy proteins LC3-II/I and P62 in the cell model were detected using Western blotting, and the formation of autophagosomes was observed with MDC staining.EdU incorporation assay and flow cytometry were performed to observe the effect of Sidt2 knockout on cell proliferation and apoptosis.The effect of chloroquine at the saturating concentration on autophagic flux, proliferation and apoptosis of Sidt2 knockout cells were observed.@*RESULTS@#Sidt2-/- HL7702 cells were successfully constructed.Sidt2 knockout significantly inhibited the proliferation and increased apoptosis of the cells, causing also increased protein expressions of LC3-II/I and P62(P < 0.05) and increased number of autophagosomes.Autophagy of the cells reached a saturated state following treatment with 50 μmol/L chloroquine, and at this concentration, chloroquine significantly increased the expressions of LC3B and P62 in Sidt2-/- HL7702 cells.@*CONCLUSION@#Sidt2 gene knockout causes dysregulation of the autophagy pathway and induces apoptosis of HL7702 cells, and the latter effect is not mediated by inhibiting the autophagy-lysosomal pathway.

Effect and mechanism of matrine on steatosis Chang Liver cells induced by oleic acid / 中国药房

OBJECTIVE To investigate the effects of matrine （MT） on steatosis Chang Liver cell model induced by oleic acid （OA） and its possible mechanism. METHODS Chang Liver cells were divided into blank group， model group and MT low-dose， medium-dose group and high-dose groups （0.1， 0.5， 1.0 mmol/L）. Except for blank group， the other groups were treated with 1.0 mmol/L OA for 24 h to establish steatosis model， and MT groups were given corresponding concentrations of drugs for 24 h. The activities of steatosis Chang Liver cells were observed； the morphologies of intracellular lipid droplets were observed and lipid content was also determined. The contents of liver function indexes [alanine transaminase （ALT）， aspartate transaminase （AST）， total bilirubin （TBIL）， alkaline phosphatase （ALP）]， as well as mRNA and protein expressions of farnesoid X receptor （FXR）， cytochrome P450 7A1 （CYP7A1） and fibroblast growth factor 19 （FGF19） were all detected. RESULTS OA and MT had no significant effect on the activity of Chang Liver cells. After OA treatment， orange lipid droplets formed in cytoplasm； compared with blank group， relative lipid content and the levels of liver function indexes were increased significantly， while the mRNA and protein expressions of FXR， CYP7A1 and FGF19 were down-regulated significantly （P＜0.05）. After treatment of low， medium and high concentrations of MT， above indexes were all reversed significantly （P＜0.05）. CONCLUSIONS MT could significantly improve the lipid content and liver function indexes of steatosis Chang Liver cells induced by OA though regulating FXR/CYP7A1/ FGF19 signaling pathway.

Improvement effect and mechanism of different extracts from Tylophora yunnanensis on non-alcoholic steato-hepatitis / 中国药房

OBJECTIVE To investigate the improvement effect and mechanism of different extracts from Tylophora yunnanensis on non-alcoholic steatohepatitis （NASH）. METHODS Normal human liver LO2 cells were induced to steatosis by free fatty acid， then were divided into normal group， model group， silybin group （100 μmol/mL）， T. yunnanensis ethanol extracts （TYS） group （50 μg/mL）， T. yunnanensis ethyl acetate extracts （TYSA） group （50 μg/mL）， and T. yunnanensis n-butanol extracts （TYSB） group （50 μg/mL）. After 24 hours of drug intervention， the deposition of lipid droplets was observed in LO2 cells in each group. The contents of total cholesterol （TC）， triacylglycerol （TG）， malondialdehyde （MDA） and glutathione （GSH）， the activities of aspartate transaminase （AST）， alanine transaminase （ALT） and superoxide dismutase （SOD）， the mRNA expressions of Kelch-like ECH-associated protein 1（ Keap1）， nuclear factor E2-related factor 2（ Nrf2） and heme oxygenase 1（ HO- 1） were detected. NASH rat model was induced by a high-fat diet， and then divided into normal group， model group， silybin group （12.6 mg/kg）， TYS group （80 mg/kg）， TYSA group （80 mg/kg） and TYSB group （80 mg/kg）， with six rats in each group. The liver indexes of rats in each group were calculated after 6 weeks of drug intervention. The liver histopathological changes were observed， and the contents of TC， TG， HDL-C and LDL-C， AST and ALT activities in serum， the contents of MDA and GSH， SOD activities in liver tissue were detected. RESULTS Compared with model group， TYS， TYSA and TYSB could reduce lipid droplet deposition， intracellular TC， TG and MDA contents， AST and ALT activities， and increase SOD activity， GSH content， and Keap1， Nrf2， HO-1 mRNA expression levels in LO2 cells after steatosis to varying degrees， with some differences being statistically significant （P＜0.05）. They also significantly improved liver injury in NASH model rats， reduced their liver indexes， TC， TG， LDL-C and MDA contents， AST and ALT 1-042） activities， and increased HDL-C （except for TYS and TYSB）， GSH contents and SOD activity， with TYSA having the most significant effect （P＜0.05）. CONCLUSIONS TYS， TYSA and TYSB have a certain improvement effect on NASH， among which TYSA has the most obvious effect. Its mechanism of action may be related to upregulating the Keap1/Nrf2/HO-1 signaling pathway and inhibiting oxidative stress

Effects of bisphenols on proliferation and oxidative stress of BRL 3A rat liver cells and their mutagenicities / 预防医学

Objective@#To examine the effects of bisphenol A (BPA), bisphenol S ( BPS ), bisphenol F ( BPF ) and bisphenol AF ( BPAF ) on the proliferation and oxidative stress of BRL 3A rat liver cells, and to preliminarily evaluate their mutagenicities.@*Methods@#In vitro cultured BRL 3A rat liver cells were treated with BPA, BPS, BPF and BPAF at concentrations of 0, 5, 10, 25, 50, 100, 150 and 200 μmol/L for 48 h, respectively. Then, the cell viability was determined using the CCK-8 assay, and the half maximal inhibitory concentration ( IC50 ) was calculated. The minimum inhibitory concentration for BRL 3A cell proliferation was screened, and the intracellular reactive oxygen species ( ROS ) was measured in BRL 3A cells using the 2',7'-dichlorodihydrofluorescein diacetate ( DCFH-DA ) assay. In addition, the effects of BPA, BPS, BPF and BPAF at concentrations of 1 000, 200, 40, 8 and 1.6 μg/plate on the mutant colonies of histidine-deficient Salmonella typhimurium ( TA1535, TA97a, TA98, TA100 and TA102 ) were tested using the Ames test.@*Results@#Treatment with BPA and BPF at concentrations of 100 to 200 μmol/L and with BPAF at concentrations of 25 to 200 μmol/L inhibited BRL 3A cell survival at a concentration-dependent manner, while exposure to BPS at concentrations of 5 to 200 μmol/L resulted in no changes in BRL 3A cell survival. The IC50 values of BPA, BPS, BPF and BPAF were 131.7, >200, 187.5 and 21.6 μmol/L against BRL 3A cells, respectively. Treatment with BPS at 100 μmol/L or BPAF at 25 μmol/L caused no significant changes in the ROS level; however, exposure to BPA at 100 μmol/L and BPF at 100 μmol/L significantly increased the ROS level. Ames test showed that BPA, BPS, BPF and BPAF did not induce mutagenicity in TA1535, TA97a, TA98, TA100 or TA102 strains.@*Conclusions@#BPAF shows the highest cytotoxicity to BRL 3A cells, and low-concentration exposure to BPS has few effects on BRL 3A cells. The cytotoxicity of bisphenols against BRL 3A cells may be associated with the induction of oxidative stress. None of the four bisphenols show mutagenic effects under the present experimental conditions.

Toxicity of N-Succinyl-Chitosan to Bovine Hemoglobin and Human Liver Cells / 中国药学杂志

OBJECTIVE: To investigate the toxicity of N-succinyl-chitosan (NSCS) to bovine hemoglobin (BHb) and human liver cells (HL-7702). METHODS: BHb was used as a research object and the toxic effect of NSCS was investigated by UV-Vis absorption spectroscopy, fluorescence spectroscopy and synchrotron spectroscopy under the simulative human physiological condition. At the same time, human HL-7702 cells was used as a research object and methyl thiazolyl tetrazolium (MTT) assay was employed to examine the cytotoxicity of NSCS. RESULTS: The results of UV-Vis absorption spectroscopy and MTT showed that the toxicity of NSCS was weak, and substitution degree had little effect on it. The result of fluorescence spectroscopy demonstrated that the intrinsic fluorescence of BHb was quenched by NSCS and the quenching effect slightly increased with the increase of substitution degree. The quenching mechanism was mainly dynamic quenching, and the major driving forces were hydrophobic and electrostatic force. CONCLUSION: The result of synchronous fluorescence spectroscopy reveals that NSCS has almost no influence on the conformation of BHb. The toxicity of NSCS to BHb and HL-7702 is weak.

The regulation effect of emodin on human embryonic hepatocyte L02 cell strain Farnesoid X receptor / 中华实用儿科临床杂志

Objective To investigate the regulation effect of emodin on human embryonic liver L02 cells strain farnesoid X receptor (FXR) pathways.Methods By using Guggulsterones,FXR genes were intervened with in L02 cells as model group,in three different concentrations of emodin (50.0 μ mol/L,25.0 μmol/L,12.5 μmol/L) of emodin in the model group cells,FXR,small heterodimer parter (SHP),UDP-glucuronosyltransferase 2B4 (UGT2 B4),bile salt export pump(BSEP) mRNA and protein expressions were detected by real-time fluorescent quantitative PCR and Western blot test.Results (1) The relative expressions of FXR mRNA and protein in the model group (0.240 ± 0.021,0.385 ±0.119) decreased significantly than those of control group (1.000 ± 0.088,1.000 ± 0.223),the differences were statistically significant (t =14.62,4.21,all P ＜ 0.01).Compared with the model group,the relative expressions of FXR mRNA in the high,the middle and the low-dose emodin groups (0.755 ±0.083,0.817 ±0.097,0.547 ± 0.080) were significantly higher (t =10.42,10.03,6.39,all P ＜ 0.01).The relative expressions of FXR protein in the medium-dose group (0.865 ± 0.203) increased significantly (t =3.53,P ＜ 0.01).The relative expressions of FXR mRNA in the high and the medium-dose groups (0.755 ± 0.083,0.817 ± 0.097) were higher than those in the low-dose group (0.547 ± 0.080),the differences were statistically significant (t =3.11,3.70,all P ＜ 0.01).(2)The relative expressions of SHP,UGT2B4,BSEP mRNA and protein in the model group (0.148 ±0.025,0.205 ± 0.039,0.184 ± 0.020;0.458 ± 0.130,0.255 ± 0.170,0.303 ± 0.100) were significantly lower than those in the control group (1.000 ±.0.099,1.000 ±0.104,1.000 ±0.125;1.000 ±0.129,1.000 ±0.157,1.000 ±0.162),the differences were statistically significant (t =14.50,12.44,11.19,5.13,5.57,6.33,all P ＜ 0.01).The relative expressions of SHP,UGT2B4 and BSEP mRNA in the high,the middle and the low-dose groups (0.610 ± 0.058,0.514 ± 0.041,0.707 ± 0.062;0.755 ± 0.108,0.800 ± 0.086,0.727 ± 0.076;0.470 ± 0.070,0.582 ± 0.050,0.500±0.108) were significantly lower than those in the model group (0.148 ± 0.025,0.205 ± 0.039,0.184 ± 0.020),the differences were statistically significant (t =12.75,9.38,13.94,9.46,10.90,11.96,7.53,10.31,5.00,all P ＜0.01).The relative expressions of SHP,UGT2B4 and BSEP mRNA in the high and the middle-dose emodin group (0.658 ±0.091,0.624 ±0.113,0.607 ±0.097;0.868 ±0.194,0.883 ±0.099,0.913 ±0.131) were significantly higher than those in the low-dose group (0.458 ±0.130,0.255 ±0.170,0.303 ±0.100),the differences were statistically significant (t =2.18,3.13,3.78,3.05,5.53,6.41,all P ＜ 0.01).The relative expression of SHP and BSEP protein in the low-dose group (0.645 ±0.135,0.572 ±0.076) increased,the differences were statistically significant (t =1.73,P ＜ 0.05,t =3.72,P ＜ 0.01).The relative expression of BSEP protein in the high and the medium-dose groups (0.607 ±0.097,0.913 ± 0.131) was significantly higher than those in the low-dose group (0.572 ± 0.076),the differences were statistically significant (t =1.99,3.90,all P ＜ 0.01).The relative expressions of SHP and UGT2B4 mRNA in the high and the low-dose group (0.610 ±0.058,0.470 ±0.070;0.514 ± 0.041,0.582 ± 0.050) were significantly lower than those in the medium-dose group (0.800 ± 0.086),the differences were statistically significant (t =3.75,6.47,3.83,3.42,all P ＜ 0.01).The expression levels of UGT2B4 and BSEP in the high and the low-dose groups (0.624 ± 0.113,0.644 ± 0.097;0.607 ± 0.097,0.572 ± 0.076) were significantly lower than those in the medium-dose group (0.883 ± 0.099,0.913 ± 0.131),the differences were statistically significant (t =4.27,2.98,6.30,3.90,all P ＜ 0.01).Conclusions Guggulsterones can inhibit FXR and downstream genes SHP,UGT2B4,BSEP expressions in L02,and emodin can enhance FXR gene expression,promote SHP,UGT2B4,BSEP gene expression,inhibit cholestasis pathway,protection of liver cell,which shows a dosage discreapancy.

Ectopic over-expression of oncogene Pim-2 induce malignant transformation of LO2 / 中国现代普通外科进展

Objective:To research whether ectopic over-expression of Pim-2 could cause chang-liver cell (LO2) malignant transformation,to explore the relationship between Pim-2 protein and hepatocellular carcinoma.Methods:Three groups of cells were arranged including human chang liver cell line LO2 (group C),LO2 cells transfected with empty-vector (group B) and LO2 cells transfected with Pim-2 gene (group A).Pim-2 expression levels were detected.The morphology,proliferation level,apoptosis rate and migration ability of the cells were detected respectively.The cells were subcutaneously inoculated into athymic mice and the microstructures of the neoplasm were observed by optical and electron microscopy.Results:Compared with group B,Pim-2 expression levels were significantly higher in group A (P＜0.05),and their morphology had obvious malignant changes.They also showed a significantly increased proliferation rate (P＜0.05) and migration capacity (P＜0.05),as well as a significantly decreased apoptosis rate (P＜0.05).Only the athymic mice inoculated with group A could generate neoplasm,and the morphology of the neoplasm coincided with that of the hepatoma.Conclusion:Both the morphological and biological changes of LO2/Pim-2 cells indicate the trend of malignant transformation,which could generate hepatoma in athymic mice.Pim-2 could induce malignant transformation of human liver.

An investigation of trichloroethylene-induced effects on histone methylation in L-02 hepatic cells / 中华预防医学杂志

Objective@#To further explore TCE-induced hepatotoxicity and its mechanisms by identification of trichloroethylene (TCE) induced abnormal histone methylation in human liver cells.@*Methods@#L-02 cells were treated with 0 and 8 mmol/L TCE for 24 h. Histones were extracted by acid. Liquid chromatography electrospray ionization tandem mass spectrometry (ESI-LC-MS/MS) were used to identify and quantify TCE related histone methylations. TCE induced abnormal methylation of H3K79 me2 and H3K79 me3 were validated by Western blot analysis. The further analysis of the function of histone abnormal methylation modifications were done by single cell gel electrophoresis (SCGE) and Western blot analysis of p53 and ɤH2AX.@*Results@#After treatment with TCE for 24 h in L-02 cells, the 36 TCE related histone methylation sites in 28 peptide segments were identified by MS. After treatment with TCE in concentrations of 0 and 8.0 mmol/L in L-02 cells for 24 h, the relative expression level of histone H3K79 me3 were 1.00±0.06, 0.70±0.09 (t=15.01, P=0.015); the relative expression level of histone H3K79 me2 were 1.00±0.05, 0.74±0.07 (t=16.69, P=0.018); the Olive Tail Moment about DNA damage were 1.46±0.28, 3.12± 0.68 (t=15.22, P=0.018); the relative expression levels of p53 were 1.00±0.04, 1.24±0.04 (t=18.71, P= 0.012); and the relative expression levels of ɤH2AX were 1.00 ± 0.03, 1.56 ± 0.11 (t=8.32, P=0 045).@*Conclusion@#TCE can induce changes in the relative expression level of H3K79 me2 and H3K79 me3 in L-02 cell, and induce DNA damage, suggesting that TCE may induce changes in the relative expression level of H3K79 me2 and H3K79 me3 by DNA damage.

Anti-liver fibrosis activities of human insulin-like growth factor-1 in vitro / 中国药科大学学报

This study was focus on investigating the anti-liver fibrosis effects of insulin-like growth factor-1 (IGF-1) in vitro.The effects of IGF-1 on human liver L-02 cell viability and cell cycle were observed.CC14-induced L-02 cell injury was set up to detect the anti-apoptotic activity of insulin-like growth factor-1 (IGF-1).Transforming growth factor β1 (TGF-β1) induced hepatic stellate cell line (HSC-T6) were used as a liver fibrosis model in vitro to analyze the effects of IGF-1 on the expression of liver fibrosis proteins and intracellular TGF-β1/Smad signaling pathway in HSC-T6 cells.The results showed that IGF-1 could relieve the growth inhibition effects of TGF-β1 on L-02 cells,increase the viability of L-02 cells injured by CCl4,decrease the expression of liver fibrosis proteins,and inhibit the TGF-β1/Smad signaling pathway by inhibiting the phosphorylation of Smad3.Our study suggested that IGF-1 exerted anti-liver fibrosis effects by stimulating L-02 cells proliferation,reducing cell damage and inhibiting ECM accumulation via interfering TGF-β1/Smad signaling pathway.

Experimental study on acute hepatotoxicity in beagles induced by Amanita exitialis / 中华急诊医学杂志

Objective To establish acute hepatotoxic model induced by Amanita exitialis and to study the characteristics of acute toxic liver failure induced by mushrooms containing peptide toxins,in hope for providing some help to experimental research on poisoning induced by mushrooms containing peptide toxins.Methods UPLC-MS/MS (Ultra performance liquid chromatography-tandem mass spectrometry) method was used to detect peptide toxins in Amanita exitialis.To establish acute toxic liver hepatic failure model,the beagles were fed with 60 mg/kg of lyophilized powder of Amanita exitialis fungus which encapsulated in starch capsules.Toxic sighs were observed,coagulation function,hepatic and renal function,liver histopathological morphology,peptide toxin concentration in plasma and urine were detected during the experiment.Results Total peptide toxins in Amanita exitialis was (3 482.6 ± 124.94) mg/ kg.All the beagles had toxic signs including vomiting and diarrhea in 12-48 h after ingestion.On 24 h after ingestion,the beagles' ALT,AST,TBIL,ALP,PT and APTT levels increased obviously.On 36 h after ingestion,the beagles' ALT,AST,PT and APTT values reached their peaks (ALT:283.2 ± 112.9 Kallmann unit;AST:223.9 ±93.8 Kallmann units;PT:132.9 ± 152.6 s;APTT:131.4 ± 153.9 s).On 48 h after ingestion,the beagles' TBIL and ALP levels reached their peaks (TBIL:23.3 ± 14.6 mol/L;ALP:274.5 ± 115.5 U/L).The beagles' TBIL,TP and APTT returned to normal 1 week after ingestion,their ALT,AST and ALP levels returned to normal 3 weeks after ingestion.Three dogs died during 24-72 h after ingestion.Liver histopathological morphology study showed hemorrhagic necrosis of hepatocytes.Peptide toxins can be detected in plasma within 24 h after ingestion.Peptide toxins can be detected in urine within 96 h after ingestion.Conclusion Amanita peptide toxins can cause hemorrhagic necrosis of liver cells and lead to acute liver failure.This model is consistent with clinical pathophysiological process of acute toxic liver failure induced by mushrooms containing peptide toxins,and it can be applied to the study of diagnosis and treatment of poisoning induced by mushrooms containing peptide toxins.

Construction and in vitro functional evaluation on a new hybrid bioartificial liver / 中华肝胆外科杂志

Objective To design a new type of hybrid bioartificial liver (HBAL), evaluate its efficacy in vitro, and explore the feasibility in clinical application.Methods CL-1 human hepatocytes were cultured on microcarriers for 5 days, when cell count reached about 4.0 × 109 with cell density of about 4.0 × 107/ml.CL-1 cells cultured on microcarriers in home-made bioreactor constitute the biological part of the HBAL.The abiotic part included blood perfusion and bilirubin adsorption, and blood pump was employed as the circulation driver, which were parts of HBAL.The changes of the concentrations of indirect bilirubin (UBD), chenodeoxycholic acid (CDCD), cholic acid (CA), blood ammonia (AA), AST, ALT and LDH were observed under the condition of in vitro circulation.Meanwhile, the function, morphology and the cell activity of CL-1 cells were also observed.Results After in vitro circulation for 24 h, the concentrations of UBD, CDCD, CA and AA significantly decreased from (335.3 ± 6.0) μmol/L, (395.0 ± 5.6) μmol/L, (155.7 ± 4.5) μmol/L, (39.0 ± 2.6) μmol/L at 0 h to (106.0 ± 10.9) μmol/L, (131.8 ± 28.7) μmol/L, (42.2 ± 7.3) μmol/L, (3.5 ± 1.0) μmol/L, respectively.At 48 h, ALT, AST and LDH significantly increased from (25.9 ± 4.2) IU/L, (22.0 ± 3.6) IU/L, (0.28 ± 0.09) μmol/L to (31.0 ± 2.6) IU/L,(31.6 ± 8.0) IU/L, (0.41 ± 0.12) μmol/L, meanwhile the count and vitality of CL-1 cells were significant declined.Conclusions (1) In the new HBAL system, CL-1 cells can keep its viability and function in vitro;and (2) the HBAL appears to be effective in purifying the serum in liver failure simulation model by clearing out non-conjugated bilirubin, chenodeoxycholic acid, cholic acid and ammonium chloride, which seems to be a promising therapeutic option.

The clinical analysis of recombinant human endostatin hormone combined with transcatheter arterial chemoembolization of primary hepatocellular carcinoma for 52 patients / 中国医师进修杂志

Objective To study the effect of recombinant human endostatin hormone combined with transcatheter arterial chemoembolization (TACE) for the treatment of primary hepatocellular carcinoma (HCC).Methods Fifty-two primary HCC patients were divided into combined group (26 patients) and control group (26 patients) by random digits table method.The patients in combined group received TACE and recombinant human endostatin hormone added in embolism emulsion.The recent curative effect,level of serum vascular endothelial growth factor (VEGF),adverse reactions,recurrence rate and survival rate were compared between two groups.Results The response rate(RR) and clinical benefit rate(DCR) in combined group were 57.69％(15/26),84.62％(22/26),in control group were 34.62％(9/26),73.08％(19/26),there were significant differences (x2 =5.237,P ＜ 0.05 ; x2 =4.284,P ＜ 0.05).The 7th,14th,28th day after TACE,the level of VEGF in two groups was first increased and then a downward trend,the difference were statistically significant (P ＜ 0.05).The level of VEGF in combined group was significantly lower than that in control group (P＜ 0.05).The follow up rate was 94.23％(49/52),the 1-year and 2-year recurrence rate in combined group was significantly lower than that in control group (P＜ 0.05).The 2-year and 3-year survival rate in combined group was significantly higher than that in control group (P ＜ 0.05).Conclusion With TACE plus recombinant human endostatin hormone can effectively inhibit the increase of serum VEGF level,improve curative effect and disease control rate,reduce tumor recurrence and improve survival rate.

Tratamento minimamente invasivo do adenoma hepático em situações especiais / Minimally invasive treatment of hepatic adenoma in special cases

O adenoma hepatocelular é um tumor benigno raro, que apresentou aumento do número de diagnósticos nas décadas de 1980 e 1990, o que foi atribuído à difusão dos contraceptivos hormonais orais, e à melhor disponibilização e ao avanço dos exames radiológicos. Apresentamos aqui o relato de dois pacientes com grandes adenomas hepáticos submetidos ao tratamento minimamente invasivo por meio de embolização arterial. O primeiro caso foi submetido à embolização eletiva, por apresentar múltiplos adenomas, além de hemorragia recente de um desses nódulos. O segundo, caracterizado por vítima de trauma abdominal fechado e rotura de adenoma hepático, foi realizado em caráter de urgência, tendo a paciente sinais clínicos de choque hemodinâmico secundário à hemorragia intra-abdominal. O desenvolvimento de terapias minimamente invasivas locorregionais, como a embolização arterial, trouxe um novo horizonte para pacientes com adenomas hepáticos. Na ressecção emergencial de um adenoma hepático roto, as taxas de mortalidade são de 5 a 10%, enquanto que a ressecção eletiva reduz a 1% esse desfecho. A embolização arterial dos adenomas hepáticos na vigência de hemorragia é tema de debate. Essa constatação aponta para um possível papel da embolização transarterial desses tumores rotos e não rotos, visto que tal conduta poderá limitar a indicação cirúrgica em casos selecionados, resultando na redução da morbimortalidade. O seguimento das pacientes tratadas foi realizado por meio de ressonância magnética e, após 30 dias, já havia diminuição das lesões embolizadas bem como a presença de significativo componente avascular. O controle radiológico, após 12 meses, mostrou ausência de novas lesões e diminuição daquelas embolizadas.

Hepatocellular adenoma is a rare benign tumor that was increasingly diagnosed in the 1980s and 1990s. This increase has been attributed to the widespread use of oral hormonal contraceptives and the broader availability and advances of radiological tests. We report two cases of patients with large hepatic adenomas who were subjected to minimally invasive treatment using arterial embolization. One case underwent elective embolization due to the presence of multiple adenomas and recent bleeding in one of the nodules. The second case was a victim of blunt abdominal trauma with rupture of a hepatic adenoma and clinical signs of hemodynamic shock secondary to intra-abdominal hemorrhage, which required urgent treatment. The development of minimally invasive locoregional treatments, such as arterial embolization, introduced novel approaches for the treatment of individuals with hepatic adenoma. The mortality rate of emergency resection of ruptured hepatic adenomas varies from 5 to 10%, but this rate decreases to 1% when resection is elective. Arterial embolization of hepatic adenomas in the presence of bleeding is a subject of debate. This observation suggests a role for transarterial embolization in the treatment of ruptured and non-ruptured adenomas, which might reduce the indication for surgery in selected cases and decrease morbidity and mortality. Magnetic resonance imaging showed a reduction of the embolized lesions and significant avascular component 30 days after treatment in the two cases in this report. No novel lesions were observed, and a reduction in the embolized lesions was demonstrated upon radiological assessment at a 12-month follow-up examination.

δ-opioid receptor activation by protein kinase C pathway inhibits acid-induced liver cell apoptosis / 中华肝胆外科杂志

Objective To study the inhibitory effects of δ-opioid receptor activation in serumdeprivation induced apoptosis of human liver cells and the proposed protein kinase C(PKC)pathway mechanism.Methods MTT assay was used to detect the survival rate of human liver cells in vitro and Annexin V-FITC/PI double staining was used to detect the cell apoptosis rate.Flow cytometry was used to analyze cell cycle,RT PCR used to analyze the PKC mRNA and Western Blot analysis was used for detecting the protein expression of PKC and Caspase-3.Results After serum-deprivation for 48h of cultured human liver cells in vitro,significant liver cell apoptosis occurred.The apoptosis was suppressed by δ-opioid receptor activation,which manifested as a slower rate of apoptosis,decreased expression of Caspase-3and increased expression of PKC.After GF109203X was added,the inhibitory effects of DADLE decreased markedly.Conclusion Activation of δ-opioid receptor on the membrane of human liver cells has inhibitory effects on serum-deprivation induced apoptosis of liver cells.The underlying mechanism may be associated with PKC pathway activation.

The role of TSH in cholesterol synthesis / 中华内分泌代谢杂志

Thyroid diseases are often accompanied with dyslipidemia.In me past,low level of myroid hormone(TH) was considered a main factor in causing elevated level of blood cholesterol in hypothyroid patients.However,some clinical features can not be explained by this traditional theory. For example,in subclinical hypothyroid cases,both levels of thyroid-stimulating hormone (TSH) and serum total cholesterol (TC) increase,while TH level remains normal.From this phenomenon,we speculate that in case with hypothyroidism,not only there is a relationship between serum TC and TH levels,but also serum TC level change is related to TSH in some way,which has not been noticed.The purpose of this paper is to review the new and most advanced progress in the research of TSH and cholesterol.

Chronic Opium Treatment Can Differentially Induce Brain and Liver Cells Apoptosis in Diabetic and Non-diabetic Male and Female Rats

It has been shown that some opium derivatives promote cell death via apoptosis. This study was designed to examine the influence of opium addiction on brain and liver cells apoptosis in male and female diabetic and non-diabetic Wistar rats. This experimental study was performed on normal, opium-addicted, diabetic and diabetic opium-addicted male and female rats. Apoptosis was evaluated by TUNEL and DNA fragmentation assays. Results of this study showed that apoptosis in opium-addicted and diabetic opium-addicted brain and liver cells were significantly higher than the both normal and diabetic rats. In addition, we found that apoptosis in brain cells of opium-addicted and diabetic opium-addicted male rats were significantly higher than opium-addicted and diabetic opium-addicted female, whereas apoptosis in liver cells of opium-addicted and diabetic opium-addicted female rats were significantly higher than opium-addicted and diabetic opium-addicted male. Overall, these results indicate that opium probably plays an important role in brain and liver cells apoptosis, therefore, leading neurotoxicity and hepatotoxicity. These findings also in away possibly means that male brain cells are more susceptible than female and interestingly liver of females are more sensitive than males in induction of apoptosis by opium.

Radioprotective effects of nitroxides R-1 on human liver cells L-02 / 中华放射医学与防护杂志

Objcetive To investigate the protective effects of the nitroxides R-1 on human liver cells exposed to ionizing radiation.Methods Human liver cells L-02 were cultured and irradiated with 60Co γ-rays at the doses of 0,1,2,4,and 8 Gy,in order to screen the proper irradiation dose.WR2721 at the terminal concentration of 4 mmol/L was used as positive control.L-02 cells irradiated with 4 Gy were added with R-1 at the terminal concentration of 0.25 μmol/L at 30 min before irradiation or immediately after irradiation.MIT method was used to screen the proper conditions for follow-up experiment 72 h later.L-02 cell culture fluid was added with R-1 at the concentrations of 0,0.125,0.25,0.5,and 1 μmol/L,respectively for 30 min before irradiation at the doses of 0,1,2,4,and 8 Gy to ealculate clone formation rate at 10 d post-irradiation.L-02 cells were cultured and divided into 4 groups:control group without any treatment.drug group pretreated by 0.25 μmol/L R-1 only,irradiation group,irradiated at 4 Gy only,and drug + irradiation group with combination of 0.25 μmol/L R-01 and 4 Gy irradiation.The inverted microscopy and Hoechst 33258 staining and flow eytometry were used to observe the apoptosis of the cells at 24,48,and 72 h later.Results Nitroxides R-1 did not inhibit the viability of L-02 cell when its concentration was less than 1 μmol/L and it inhibited the L-02 cell growth when the concentration wu higher than 2 μmoL/L.The A value and colony formation rate of different concentration of R-1 groups were all higher than those of the irradiation group,and the effect of the 0.25 μmol/L drug concentration group was the most significant.Consequently,the concentration 0.25 μmoL/L was selected for follow-up experiment.Compared with the irradiation group,the L-02 cells of the pretreatment group showed solid adherence, increased refraction,clear outline,less apoptotic and dead cells at 4 Gy post-irradiation.Conclusions Nitroxides R-1 can protect the human liver cells from 60Coγ-ray induced injury effectively.The mechanism of its protective effect may be the reduction of apoptosis.

Immortalization of liver cells: experimental studies and application / 中华肝胆外科杂志

Hepatocyte is the core raw materials of bioartificial liver support system, primary hepatocyte is lim-ited to application because of short survival and difficult cul-ture in vitro. Porcine hepatocyte which has been used re-cently exist the risks of endogenous retrovirus transmission.With the development of molecular biology, it has been pos-sible that hepatocyte is immortalized recently. Immortalized hepatocytes have greatly significant to drug toxicology, bio-artificial liver support system and tissue engineering of liver.Therefore, we will review the prospects for research and ap-plication of immortalized hepatocytes.

In vitro investigations of Cynara scolymus L. extract on cell physiology of HepG2 liver cells / Investigações in vitro de extrato de Cynara scolymus L. na fisiologia das células do fígado HepG2

The objective of this study was the investigation of a potential influence of artichoke leaf extract (ALE) on the cell physiology and gene expression of phase I/II enzymes of human liver cells HepG2 and investigation on potential cell protective effects against ethanol-induced cell toxicity against HepG2 cells. Cell biological assays under in vitro conditions using HepG2 liver cells and investigation of mitochondrial activity (MTT test), proliferation assay (BrdU incorporation ELISA), LDH as toxicity marker, gene expression analysis by RT-PCR and enzyme activity of glutationtransferase. Artichocke extract, containing 27 percent caffeoylquinic acids and 7 percent flavonoids induced mitochondrial activity, proliferation and total protein content under in vitro conditions in human liver cells HepG2. These effects could not be correlated to the well-known artichoke secondary compounds cynarin, caffeic acid, chlorogenic acid, luteolin and luteolin-7-O-glucoside. The flavones luteolin and luteolin-7-O-glucoside had inhibitory effects at 100 µg/mL level on HepG2 cells, with luteolin being a significant stronger inhibitor compared to the respective glucoside. Artichoke leaf extract had minor stimulating effect on gene expression of CYP1A2, while CYP3A4, GGT, GPX2, GSR and GST were slightly inhibited. GST inhibition under in vitro conditions was also shown by quantification of GST enzyme activity. Induction of gene expression of CYP1A2 was shown to be supraadditive after simultaneous application of ethanol plus artichoke extract. Artichoke leaf extract exhibited cell protective effects against ethanol-induced toxicity within cotreatment under in vitro conditions. Also H2O2 damage was significantly inhibited by simultaneous artichoke incubation. Pre- and posttreatments did not exert protective effects. DMSO-induced toxicity was significantly reduced by pre-, post- and cotreatment with artichoke extract and especially with luteolin-7-O-glucoside...

O objetivo deste estudo foi a investigação de uma potente influência do extrato das folhas da alcachofra (ALE) na fisiologia celular e na expressão gênica de enzimas de fase I/II de células hepáticas humanas HepG2 e investigação no potencial efeito protetor celular em células HepG2 contra toxicidade celular induzida por etanol. Ensaios biológicos de células em condições in vitro usando células de fígado HepG2 e investigação da atividade mitocondrial (teste MTT), ensaio de proliferação, LDH como marcador de toxicidade, análise de expressão gênica por RT-PCR e atividade da enzima glutationa transferase. O extrato da alcachofra, contendo 27 por cento de ácidos cafeoilquínico e 7 por cento de flavonóides, induzem a atividade mitocondrial, proliferação e o teor de proteína total em condições in vitro em células hepáticas humanas HepG2. Estes efeitos não podem ser correlacionados aos compostos secundários conhecidos da alcachofra, cinarina, ácido cafeico, ácido clorogênico, luteolina e luteolin-7-O-glicosídeo. As flavonas luteolina e luteolin-7-O-glicosídeo possuem efeitos inibitórios em nível de 100 µg/mL em células HepG2, com a luteonina sendo uma inibidora significativamente mais forte comparada com o respectivo glicosídeo. O extrato das folhas de alcachofra possui um efeito mínimo da estimulação na expressão gênica de CYP1A2, enquanto CYP3A4, GGT, GPX2, GSR e GST foram sutilmente inibidos. A inibição de GST em condições in vitro também foi mostrada pela quantificação da atividade da enzima GST. Indução da expressão gênica de CYP1A2 mostrou-se supraaditiva após aplicação simultânea do etanol mais o extrato de alcachofra. O extrato das folhas de alcachofra exibiu efeitos protetores celulares frente à toxicidade induzida por etanol em co-tratamento em condições in vitro. Além disso, danos por H2O2 foram significativamente inibidos pela incubação simultânea do extrato de alcachofra. Pré e pós-tratamento não exerceram efeitos protetores. Toxicidade...

The effect of cotransplantation of nonparenchymal liver cells on the survival of intraperitoneally transplanted hepatocytes / 대한내과학회지

BACKGROUND: Transplantation of primary hepatocytes (PH) has been shown to provide metabolic support during acute liver failure. However, PH are known to be subject to necrosis in the peritoneal cavity. This is because cell-cell interaction plays an important role in their survival, but the peritoneal cavity can not provide such an environment. We tried to improve the survival of PH by simultaneously transplanting nonparenchymal liver cells (NPL). METHODS: PH from normal Wistar rats, either alone (10(9) cells/kg, group 1, n=10) or mixed with NPL (5x10(8) cells/kg, group 2, n=10) were transplanted into the peritoneal cavity of hyperbilirubinemic Gunn rats which are congenitally devoid of bilirubin glucuronidation. Liver cells from Gunn rats were transplanted as a control. RESULTS: Bilirubin glucuronides (BG) were detected in the bile of both group 1 and 2 rats collected at 6 hours after transplantation, and reached peak levels in 4 days. However, in the third and fourth week, BG could be detected only in group 2 animals. The serum bilirubin levels were decreased by 12.1~18.9% of basal levels in the second and third week for group 2 rats, but decreased by 15.1% only in the second week for the group 1 rats. Using in situ hybridization, albumin mRNA positive cells could be detected until the fourth week for the group 2 animals, but only until the second week for the group 1 rats. CONCLUSIONS: PH start functioning in a short time after intraperitoneal transplantation and simultaneous transplantation of NPL with PH can prolong the survival and function of transplanted hepatocytes.