OBJECTIVETo investigate the changes of sugar chain structures of alkaline phosphatase (ALP) in hepatoma tissue and its relation to the invasiveness of hepatocellular carcinoma (HCC).

METHODSThe binding ratios of ALP from 9 normal liver tissues, 16 hepatoma tissues and 16 noncancerous tissues surrounding hepatoma were analysed by affinity chromatography on various lectin columns including leukoagglutinating phytohemagglutinin (L-PHA), lentil lectin (LCA), Datura stramonium agglutinin (DSA), erythroagglutinating phytohemagglutinin (E-PHA) and Sambucus nigra bark agglutinin (SNA).

RESULTSThe binding ratios of ALP on L-PHA (22.94%+/-5.30%), DSA (55.97%+/-13.72%), LCA (38.16%+/-8.87%), E-PHA (11.56%+/-4.81%) and SNA (69.80%+/-13.71%) in HCC tissues were significantly increased (P<0.01) compared with that in normal liver tissues (L-PHA 5.89%+/-2.75%, DSA 36.20%+/-11.58%, LCA 17.90%+/-6.71%, E-PHA 5.38%+/-2.20%, SNA 57.32%+/-11.27%), respectively. t values between the two groups were 8.94, 3.64, 5.94, 3.62 and 2.32, respectively. L-PHA-binding ratio (25.84%+/-4.67%) of ALP in HCC with invasiveness was significantly higher than that (18.10%+/-3.64%) without invasiveness (t=3.71, P<0.01).

CONCLUSIONThe changes of ALP sugar chain structures occur in HCC tissue. b1-6 branching sugar chain structure of ALP is related to the invasiveness of HCC.

Alkaline Phosphatase ; chemistry ; Carbohydrates ; chemistry ; Carcinoma, Hepatocellular ; enzymology ; pathology ; Chromatography, Affinity ; Humans ; Lectins ; metabolism ; Liver Neoplasms ; enzymology ; pathology ; Neoplasm Invasiveness

OBJECTIVETo investigate the intervention effect of aqueous fractions from Boschniakia rossica (BRAF) on hepatic oxidative stress in mice with liver injury induced by carbon tetrachloride (CCl4).

METHODThe experimental mice were randomly assigned into the normal control group, the model group, the silymarin (positive control) group, as well as high and low dose BRAF groups. Mice were treated intragastrically with silymarin or BRAF once every day for 7 days. At the end of the experiment, CCl4 was injected intraperitoneally into the mice to establish the acute liver injury model. The pathological changes was detected with hematoxylin and eosin (HE) staining, and the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), superoxide dismutase (SOD) , catalase (CAT), glutathione peroxidase (GPx), Na+ -K+ -ATPase, Ca2+ -Mg2+ -ATPase, and the contents of reduced glutathione (GSH) and malondialdehyde (MDA) were detected by the colorimetric method.

RESULTBRAF significantly reduced ALT, AST and ALP activities in serum, alleviated hepatic injury induced by CCl4, increased SOD, CAT, GPx and GSH levels in liver, and SOD, Na + -K + -ATPase and Ca2+ -Mg2 + -ATPase activities in liver mitochondria, and decreased the MDA content in liver and liver mitochondria.

CONCLUSIONBRAF reduces hepatic oxidative stress in mice with acute liver injury induced by CCl4, thereby showing the protective effect on mice with acute liver injury induced by CCl4.

Animals ; Carbon Tetrachloride ; toxicity ; Chemical and Drug Induced Liver Injury ; enzymology ; metabolism ; pathology ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Liver ; drug effects ; enzymology ; metabolism ; pathology ; Male ; Mice ; Mitochondria ; drug effects ; metabolism ; Orobanchaceae ; chemistry ; Oxidative Stress ; drug effects ; Solubility ; Water ; chemistry

OBJECTIVETo study on the time-toxicity and dose-toxicity relationships caused by multiple dose water extraction components of Evodia Fructus to mice.

METHODMice were grouped according to different time or dose points, to observe the death condition and toxicity of mice. The changes of the activity of ALT, AST and liver, kidney index were detected, and the morphological changes of liver tissue were observed under light microscope.

RESULTOn the first day after administration the hepatotoxicity which displayed with obvious increase of ALT, AST activity in serum and liver tissue and hepatic injury appeared. On the third day the hepatotoxicity kept a higher level that the active units in serum ALT, AST were significantly higher than the normal group. On the 7th day after administration ALT, AST level in serum are restored near normality. Compared with the normal group, within 7 days after the administration, water extracted components in 0.63-5.0 g x kg(-1) dose scope could cause significant damage to liver, the activity of ALT, AST, AKP, TBI elevated, while ALB reduced, and liver ratio increased, and under light microscope, the different doses' liver tissue of mice all had different degree's edema, fatty degeneration in liver cells and interstitial congestion. There were certain time-toxicity and dose-toxicity relationships. The above-mentioned change gradually aggravated with dose increasing, and it was the obvious discrepancy compared with distilled water control group.

CONCLUSIONMultiple intragastric administrations of water extracted components of Evodia Fructus with certain dosage may induce acute hepatotoxical injury in mice and show certain "dosage-time-toxicity" relationship.

Alanine Transaminase ; blood ; metabolism ; Animals ; Aspartate Aminotransferases ; blood ; metabolism ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; metabolism ; toxicity ; Evodia ; chemistry ; Female ; Fruit ; chemistry ; Kidney ; drug effects ; enzymology ; metabolism ; Liver ; drug effects ; enzymology ; metabolism ; pathology ; Male ; Mice

Transferrin receptor (TR) performs the major function of binding and internalizing its specific iron-loaded ligand, transferrin, and its expression is closely linked to the proliferation status of the cell. This study was undertaken to elucidate TR expression in the hyperplastic lesion of hepatocyte in chemically induced hepatic carcinogenesis. The resistant hepatocyte model was chosen for a rat model of carcinogenesis and Sprague-Dawley rats were divided into the following groups: the control groups of normal diet and iron-rich diet with or without hydroxyquinoline and the groups of carcinogen alone and carcinogen plus iron-rich diet with or without administration of hydroxyquinoline. Microscopic changes in the liver, expression of transferrin receptor and glucose-6-phosphatase were studied. The hepatocyte of the control group showed both cytoplasmic and membranous expression of TR. The liver of rats fed on high iron diet accumulated iron and the expression of TR was down regulated by intrahepatic iron accumulation. In the carcinogen administered group the resistant hepatocyte of hyperplastic lesion revealed strong membranous expression of TR and failed to accumulate iron in spite of high iron diet but in contrast the surrounding non-resistant hepatocyte expressed TR in both the membrane and cytoplasm and stored iron when fed on high iron diet. The strong membranous expression of TR is one of the characteristics of the resistant hepatocyte of hyperplastic lesion and it seems to be related to the inability to accumulate iron in spite of a high iron diet.
Animal ; Glucose-6-Phosphatase/metabolism ; Immunohistochemistry ; Iron/analysis/pharmacology ; Liver/chemistry/enzymology/pathology ; Liver Neoplasms, Experimental/enzymology/*ultrastructure ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, Transferrin/*biosynthesis ; Support, Non-U.S. Gov't

OBJECTIVEThis study is to investigate the effect of tea polyphenols and tea pigments on telomerase activity of human liver cancer cell line, HepG2 cells.

METHODSTRAP-PCR-ELISA was applied to investigate the telomerase activity.

RESULTSTelomerase was positive in tea polyphenols treated groups, tea pigments treated groups and blank control group. Telomerase activities (A(450 approximately 690) values) were 1.56 and 1.46 in 50 mg/L and 100 mg/L tea polyphenols-treated groups, 1.55 and 1.49 in 50 mg/L and 100 mg/L tea pigments-treated groups, respectively. The results showed that telomerase activity was significantly inhibited by tea polyphenols and tea pigments treatment as compared with the blank control group (A(450 approximately 690) = 2.11).

CONCLUSIONSTea polyphenols and tea pigments could significantly inhibit telomerase activity of HepG2 cells, and telomerase activity may be a useful biomarker for cancer chemoprevention.

Biomarkers, Tumor ; metabolism ; Carcinoma, Hepatocellular ; enzymology ; pathology ; Flavonoids ; isolation & purification ; pharmacology ; Humans ; Liver Neoplasms ; enzymology ; pathology ; Phenols ; isolation & purification ; pharmacology ; Pigments, Biological ; isolation & purification ; pharmacology ; Polyphenols ; Tea ; chemistry ; Telomerase ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo research the treatment effect of complex prescription of Chinese crude drug in BALB/c athymic mice with human liver cancer, which were built by Bel-7402.

METHOD48 male BALB/c athymic mouse models were built by Bel-7402 with an indirect method. After 24 hours of postoperation, the 48 athymic mice were distributed randomly into 4 groups which were treated by intragastric administration with complex prescription of Chinese crude drug that had been deliquated into 3 groups by the different density as the low, middle, and high and FT207 (Tegafur) for 4 weeks. At last, athymic mice were put to death and PTEN was detected in hepatic tissue, latero-cancer tissue and cancer tissue by immunohistochemistry (PowerVision Two-Step Histostaining Reagent).

RESULTAll of the 48 athymic mice survived 12 to 28 days (Ms 24 days) and every mouse with liver cancer demonstrated by dissection. The result of immunohistochemistry represents that the intensity of PTEN in latero-cancer tissue is the highest, and then the hepatic tissue, the lowest is cancer tissue, P < 0.01. It also represents that the intensity of PTEN in treatment groups (A, B, C) is more higher than the control group (D), P < 0.05 or P < 0.01, and group B is the highest in the treatment groups, P < 0.05 or P < 0.01. However, there is no significant statistic difference between group A and group C.

CONCLUSIONThe higher expression of PTEN in the laterocancer tissue can represent the protective reaction of stress of the organism. And anticancer effect of this complex prescription of Chinese crude drug relates to an eligible density of it. Mechanisms of this complex prescription of Chinese crude drug healing HCC may partially be explained by enhancing the expression of PTEN in liver.

Animals ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Carcinoma, Hepatocellular ; drug therapy ; enzymology ; pathology ; Cell Line, Tumor ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Humans ; Immunohistochemistry ; Liver ; drug effects ; enzymology ; pathology ; Liver Neoplasms ; drug therapy ; enzymology ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; PTEN Phosphohydrolase ; metabolism ; Phytotherapy ; Plants, Medicinal ; chemistry ; Random Allocation ; Xenograft Model Antitumor Assays

BACKGROUND/AIMS: Cyclooxygenase-2 (COX-2) inhibitors reportedly inhibit the growth of hepatocellular carcinoma (HCC) via caspase-dependent or caspase-independent apoptosis, which is due to COX-2 being associated with hepatocarcinogenesis. Survivin is highly expressed in most human cancers, but the mechanism regulating survivin expression remains unclear. We investigated the regulatory expression of survivin in selective-COX-2-inhibitor-induced growth inhibition of hepatoma cells. METHODS: After treatment with NS-398 (a selective COX-2 inhibitor) at various concentrations (10, 50, 100, 150, and 200 micrometer), the growth inhibition of Hep3B hepatoma cells was assessed by an MTT cell-viability assay, DNA fragmentation gel analysis, and flow cytometry. The expression of survivin transcript was analyzed by reverse-transcription polymerase chain reactions. RESULTS: NS-398 inhibited the growth of hepatoma cells by an amount dependent on the concentration and the time since treatment. Apoptotic DNA ladder and flow-cytometry shifting to the sub-G1 phase were revealed in NS-398-induced growth inhibition of hepatoma cells. NS-398 suppressed the expression of the survivin gene in a concentration- and time-dependent manner. CONCLUSIONS: Survivin was down-regulated in the growth inhibition of hepatoma cells induced by a selective COX-2 inhibitor, NS-398, in a concentration- and time-dependent manner. These results suggest the therapeutic inhibition of COX-2 via suppression of survivin in HCC.
Carcinoma, Hepatocellular/enzymology/*metabolism/pathology ; Cell Line, Tumor ; Cell Proliferation/drug effects ; Cyclooxygenase 2 Inhibitors/chemistry/*pharmacology ; G1 Phase ; Humans ; Liver Neoplasms/enzymology/*metabolism/pathology ; Microtubule-Associated Proteins/*antagonists & inhibitors/metabolism ; Neoplasm Proteins/*antagonists & inhibitors/metabolism ; Nitrobenzenes/chemistry/*pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Sulfonamides/chemistry/*pharmacology ; Time Factors

OBJECTIVETo study the time-toxicity and dose-toxicity relationships caused by multiple dose of total Bupleurum saponin extracts to rats.

METHODRats were picked according to different time or dose points, and total Bupleurum saponin crude extracts were administered to rats. The death circumstance and toxicity of mice were observed, ALT and AST in serum were detected. Indice of the liver, index was calculated. And the morphological changes of liver tissue were observed under light microscope.

RESULTThe "time-toxicity" relationship study showed that ALT and AST in rats' serum began to increase after 7 days' administration, and with obvious hepatotoxicity after 15 days', then there was concurrent toxicity and death. The "dose-toxicity" relationship study showed that compared with the control group, the total Bupleurum saponin crude extracts between 51. 2 and 125.0 g x kg(-1) dose could cause the obvious hepatotoxicity to rats in 15 days' administration, which was represented by that ALT and AST in serum increased significantly with the dose increased, the ratio of liver to body increased, and under light microscope, the different doses' liver tissue of mice all had edema in different degree and fatty degeneration in liver cells, and the high-dose and long time administration group appeared to be necrosis, lobular structure unclear. The above-mentioned change gradually aggravated with dose increasing, and obviously diversity compared with distilled water control group.

CONCLUSIONWith administrated a multiple dose and long time to rats, the total Bupleurum saponins crude extracts could cause serious liver injury and even death, and there were certain time-toxicity and dose-toxicity relationships.

Alanine Transaminase ; metabolism ; Animals ; Aspartate Aminotransferases ; metabolism ; Bupleurum ; chemistry ; Complex Mixtures ; administration & dosage ; toxicity ; Dose-Response Relationship, Drug ; Drug Administration Schedule ; Female ; Liver ; drug effects ; enzymology ; pathology ; Male ; Rats ; Rats, Wistar ; Saponins ; chemistry ; Time Factors ; Toxicity Tests ; methods

OBJECTIVETo study the role of changes of matrix metalloproteinase-2, 9 (MMP-2, 9) activity in the development of dimethylnitrosamine (DMN)-induced liver fibrosis in rats.

METHODSThe rat liver fibrosis model was established by peritoneal injection of DMN (at a dose of 10 mg/kg, 3 times a week, for 4 weeks). The dynamic changes of liver fibrosis were observed at different time points (1d, 2d, 3d, 1 week, 2 weeks, 4 weeks, 6 weeks and 8 weeks). The MMP-2, 9 activity was measured by zymogram method. Liver ultrastructure was observed by electron microscope. The expressions of type IV collagen (CIV), laminin (LN), type I collagen (CI) and alpha-smooth muscle actin (alpha-SMA) were examined by immunohistochemistry. The tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) content was measured by Western blot method.

RESULTSThe MMP-2, 9 activity (gray value) significantly increased in the 2d and 3d DMN model rats (2d: normal/model group, MMP-2: 54.72+/-4.56/70.76+/-7.63; F = 16.27, P < 0.05; MMP-9: 25.72+/-4.29/51.76+/-15.33, F=13.38, P < 0.05). The positive staining area percentage of CIV in the sinusoidal walls decreased in the 2d, 3d and 1 weeks model rats (2d: normal/model group, 6.06+/-1.35/2.86+/-0.63, F=69.12, P < 0.05), but significantly increased in the 4w model rats (normal/model group, 6.06+/-1.35/8.04+/-1.50, F=14.42, P < 0.05). There was a remarkable negative correlation between the MMP-9 activity and expression of CIV in the sinusoidal walls (r = -0.729, P < 0.05). Positive expressions of LN and CI increased, and the strongest positive staining of them displayed in the 4w model rats. The formation of basement membrane was also observed in the 4 weeks model rats. Expression of TIMP-2 significantly increased in the late stage of fibrosis.

CONCLUSIONSThe increase of MMPs activity, especially MMP-9 which degrades the CIV normally distributed under the sinusoidal endothelium is the important factor in the formation of sinusoidal capillarization. The deposition and reconstitution of LN and new synthetic CIV, adding the deposition of CI constitute the high density basement membrane. The increase of TIMP-2 expression in the late stage of the fibrosis may be one of reasons why natural resolution of DMN-induced liver fibrosis is difficult.

Animals ; Collagen Type IV ; analysis ; Laminin ; analysis ; Liver ; chemistry ; ultrastructure ; Liver Cirrhosis, Experimental ; enzymology ; pathology ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Rats ; Rats, Wistar ; Tissue Inhibitor of Metalloproteinase-2 ; analysis

OBJECTIVETo investigate the protective effect of the roots of F. hirta against the cocaine-induced hepatotoxicity and it's active components.

METHODCocaine hydrochloride was subcutaneously injected to make male ICR mice liver wounded. Male ICR mice were randomly ig administered with the F. hirta decoction. The dose groups are 100, 200, 300 g x kg(-1) herb materials per body weight. Cocaine hydrochloride was subcutaneously injected into the mice after the administration. The serum ALT, AST activity and the activity of CAT in liver homogenate were assayed, and liver change of pathomorphism was evaluated to prove the effect of the F. hirta decoction on cocaine-induced hepatotoxicity. And the activity of psoralean which was separated from the F. hirta decoction by bioassay-guided fractionation, was proofed in the same method.

RESULTWe find that the F. hirta decoction shows a distinct effect on reducing serum transferase. The serum transferase and the content CAT in liver homogenate were dose-related reduced, and the histopathological examination found a significantly change of the liver tissues. And the psoralean, qua the mainly component, shows the same effect.

CONCLUSIONF. hirta has the protective effect against the cocaine-induced hepatotoxicity. Psoralean is the basis.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Catalase ; metabolism ; Chemical and Drug Induced Liver Injury ; Cocaine ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Ficus ; chemistry ; Ficusin ; isolation & purification ; pharmacology ; Liver ; drug effects ; enzymology ; pathology ; Liver Diseases ; blood ; prevention & control ; Male ; Mice ; Mice, Inbred ICR ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Random Allocation