[Mechanism of Calculus Bovis Sativus in inhibiting hepatocyte lipid deposition based on serum pharmacology].

The aim of this paper was to investigate the molecular mechanism of Calculus Bovis Sativus( CBS) in alleviating lipid accumulation in vitro by serum pharmacology. The CBS-containing serum of mice was obtained by serum pharmacology method to evaluate its effect on the proliferation of LO2 hepatocytes. The lipid reducing effects of CBS-containing serum through Nrf2 was evaluated by fructose-induced LO2 hepatocyte steatosis model,nuclear factor erythroid 2 related factor 2( Nrf2) agonist oltipraz combined intervention,cell oil red O staining and intracellular triglyceride( TG) content. The effects of CBS-containing serum on lipid peroxidation and hepatocytes apoptosis were evaluated by reactive oxygen species( ROS) and apoptosis assay,respectively. Real-time quantitative polymerase chain reaction( PCR) was used to detect the relative expression of lipid synthesis-related genes and apoptosis-related genes.RESULTS:: showed that CBS drug-containing serum had no significant effect on LO2 hepatocyte proliferation. As compared with the model group,CBS-containing serum could effectively reduce the formation of lipid droplets in fructose-induced LO2 hepatocytes,significantly reduce intracellular TG and ROS levels,and significantly reduce hepatocyte apoptosis rate( P < 0. 05). As compared with the model group,carbohydrate responsive element binding protein( ChREBP),sterol regulatory element binding protein-1 c( SREBP-1 c),fatty acid synthase( FAS),acetyl-CoA carboxylase 1( ACC1),stearoyl-CoA desaturase 1( SCD1),Bax and caspase-3 mRNA levels were significantly reduced in CBS drug-containing serum treatment group( P<0. 05). All of the above effects could be reversed by oltipraz.In conclusion,CBS-containing serum can significantly inhibit the fructose-induced LO2 liver fat deposition,and the mechanism may be related to reducing intracellular ROS level through the Nrf2 pathway and improving intracellular peroxidation state to reduce apoptosis.

Investigation of the synergistic effects of haloperidol combined with Calculus Bovis Sativus in treating MK-801-induced schizophrenia in rats.

Clinical studies that focused on treating schizophrenia showed that Calculus Bovis Sativus (CBS), a substitute of Calculus Bovis, when used in combination with haloperidol could significantly lower the dosage of haloperidol compared with treatment with haloperidol alone, whereas efficacy was maintained. The aim of this study was to investigate the synergetic anti-schizophrenia effects in rats using CBS in combination with haloperidol. An open field test was conducted to verify the pharmacodynamic effects of a combination treatment of CBS and haloperidol on MK-801-induced schizophrenic rats. Rat plasma concentrations of intragastric haloperidol and intravenous haloperidol were determined after oral administration of a single dose or 1-week of pretreatment with CBS (50 mg/kg). The pharmacodynamic data showed a significant decrease in locomotor activity and an increase in the percentage of the central distance when haloperidol was concomitantly administered with CBS compared with haloperidol administration alone. The AUC0-∞ and Cmax of haloperidol in the orally coadministered groups were significantly higher compared with the oral treatment with haloperidol alone. In conclusion, oral coadministration of CBS with haloperidol resulted in a synergistic effect in rats. The enhanced oral bioavailability of haloperidol when combined with CBS might be attributed to the interaction between them.

ADAM10 mediates the cell invasion and metastasis of human esophageal squamous cell carcinoma via regulation of E-cadherin activity.

A disintegrin and metalloprotease 10 (ADAM10) is involved in the tumorigenesis, invasion and metastasis of several types of solid tumors. However, the potential role of ADAM10 in human esophageal squamous cell carcinoma (ESCC) is not yet well understood. The present study showed that ADAM10 was overexpressed in human ESCC tissues in vivo, and positively associated with depth of tumor invasion, lymph node metastasis and TNM stage, contributing to tumor carcinogenesis, invasion and metastasis. Additionally, ADAM10 was overexpressed in 3 types of ESCC cell lines in vitro, as compared to that in normal esophageal epithelial cells (NEECs); and moreover, ESCC cells with high ADAM10 expression obtained enhanced invasion and migration ability. Subsequently, ADAM10 silencing by small interfering (si) RNA in ESCC cell line, EC-1, reduced cell invasion, migration and proliferation in vitro. Finally, ADAM10 negatively regulated E-cadherin in ESCC in vivo and in vitro. In conclusion, active ADAM10 promotes the carcinogenesis, invasion, metastasis and proliferation of ESCC and controls invasion and metastasis at least in part through the shedding of E-cadherin activity, which makes it a potential biomarker and a useful therapeutic target for ESCC.

Endoplasmic reticulum stress-mediated signaling pathway of gastric cancer apoptosis.

BACKGROUND/AIMS: To investigate ER stress-mediated CHOP-signaling pathway of gastric cancer apoptosis in vitro and in vivo. METHODOLOGY: Based on the dose-and time-response experiments about tunicamycin (TM),gastric cancer cell line BGC823 was treated with 10tg/mL of TM for 24h. BGC823 apoptosis was detected with TUNEL assay and ultrastructural changes in BGC823 cells under ER stress were observed with transmission electron microscopy (TEM). RT-PCR and western blot-ting were used to determine the expression of ERS-related proteins, glucose-regulated protein 78 (GRP78) and CHOP and apoptosis-associated protein B-cell lympho-ma 2 (Bcl-2). After the knockdown of CHOP, the changes were also observed in vitro and in vivo. RESULTS: TEM assay showed that after treatment with TM, BGC823 cell size became smaller with ER dilation, vacuolization and karyopyknosis. RT-PCR and western blotting indicated that TM up-regulated GRP78 and CHOP expression and down-regulated Bcl-2 expression. The knock-down of CHOP could convert Bcl-2 expression and reduce BGC823 apoptosis caused by ERS in vitro and in vivo, but failed to influence GRP78. CONCLUSIONS: TM can induce ESR and regulate downstream molecules CHOP up-regulation and Bcl-2 down-regulation which lead to BGC823 apoptosis. This study may provide a new theoretical basis for the pathogenesis of gastric cancer.

[Different doses transplanted microglia effect on ß-amyloid protein in rat Alzheimer's models].

OBJECTIVE: To explore the effects of different doses of transplanted microglia (MG) on ß-amyloid protein (Aß) in rat brain model of Alzheimer's disease (AD). METHODS: A total of 60 SD rats were randomly divided into experimental group and control group. The experimental group was injected with Aß-42 and the control group saline in hippocampus. At Day 3, different doses of continuously expressed enhanced green fluorescent protein (EGFP) microglial cells cultured in vitro were injected into the rats via carotid artery. At Day 3 post-injection, Aß was labeled by immunofluorescent method. And the expressions of EGFP microglia and Aß were detected in hippocampus by fluorescence. The relative brain expression of ß-amyloid precursor protein (APP) mRNA was analyzed by RT-PCR (reverse transcription-polymerase chain reaction). RESULTS: The mean fluorescent intensity of EGFP was significantly higher in each experimental group (5.62 ± 0.61, 8.85 ± 0.33, 10.24 ± 0.45, 11.26 ± 0.37, 12.75 ± 0.65) than the control group (2.22 ± 0.32, all P < 0.05); Fluorescent immunohistochemistry showed that the mean fluorescent intensity of Aß in the 4×10(6) dose group (9.53 ± 0.23) and the 12×10(6) dose group (8.32 ± 0.46) were significantly higher than that in the 8×10(6) dose group (4.56 ± 0.13, both P < 0.05); The fluorescent regions had partial consistence of EGFP and Aß; RT-PCR (reverse transcription-polymerase chain reaction) results showed that the relative expressions of APP mRNA in the 4×10(6) dose group (1.83 ± 0.22) and the 12×10(6) dose group (1.94 ± 0.28) were significantly higher than that in the 8×10(6) dose group (0.43 ± 0.12, both P < 0.05). CONCLUSION: Aß has the chemotactic role of MG. MG may swallow Aß protein partially. But a high dose of MG also accelerates the formation of Aß.

[Effects of estrogen on P-Tau, ChAT and nerve growth factor protein expressions in the brain tissue of rats with Alzheimer's disease].

OBJECTIVE: To examine the effect of estrogen on the expressions of phosphorylated Tau (P-Tau), ChAT and nerve growth factor (NGF) protein in the brain tissue of rat models of Alzheimer disease (AD). METHODS: Rat models of AD were established by injecting Aß1-42 protein fragments in the right lateral ventricle. Two weeks later, 17ß-estradiol tablets were implanted subcutaneously at the neck of the rats and maintained for 30 days. The pathological changes in the rats' brain neurons and alterations in the expressions of P-Tau, ChAT and NGF proteins were observed using HE staining and immunohistochemistry, respectively. RESULTS: In the AD rats, neurofibrillary tangles occurred in the brain tissue, and estrogen treatment significantly reduced the formation of neurofibrillary tangles. Estrogen treatment also resulted in lowered P-Tau expression and increased ChAT and NGF protein expressions in comparison with those in the AD model rats. CONCLUSION: Estrogen can up-regulate ChAT and NGF and down-regulate tau protein expression, thus producing obvious therapeutic effect on AD in rats.

[Effect of polyoxyl ether analogous surfactants on the activity of cytochromes P450 3A in rats in vivo].

To evaluate the effects of p-octyl polyethylene glycol phenyl ether (Triton X-100), polyoxyl 35 caster oil (EL35) and polyoxyl 40 hydrogenated caster oil (RH40) on the activity of Cytochrome P450 3A (CYP3 As) in vivo. Rats were administered with saline, ketoconazole (75 mg x kg(-1) x d(-1)), Triton X-100 (30 mg x kg(-1) x d(-1)), EL35 (150 mg x kg(-1) x d(-1)) and RH40 (150 mg x kg(-1) x d(-1)) intragastrically for 5 consecutive days, and then given midazolam 10 mg x kg(-1) 20 min after the last treatment of ketoconazole or three surfactants with the same dose through duodenal administration. Pharmacokinetics parameters for midazolam and its metabolite 1'-hydroxymidazolam were estimated from the plasma concentration-time data by a noncompartmental approach. The results showed that multiple dose administration of Triton X-100, EL35 and RH40 decreased the ratios of 1'-hydroxymidazolam and midazolam AUC0-infinity from 1.14 to 0.90, 1.03 and 0.64, respectively. In contrast, multiple dose administration of ketoconazole caused the ratios of 1'-hydroxymidazolam and midazolam a significant decrease to 0.50. This study indicated that Triton X-100 and EL35 would have no inhibition on CYP3A, while RH40 had significant inhibition on CYP3A. Therefore, RH40 might be used to prepare drug formulations in pharmaceutical industry and would increase the bioavailability of some drugs transformed by CYP3As and further lead to significant clinical pharmacologic effects.