ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of ginsenosides from stems and leaves of ginseng on ethanol-induced lipid deposition in human L02 hepatocytes.</p><p><b>METHODS</b>L02 cells were exposed to ethanol for 36 h and treated with or without ginsenosides. The viability of L02 cells was evaluated by methylthiazolyldiphenyl-tetrazolium bromide assay and the triglyceride (TG) content was detected. Lipid droplets were determined by oil red O staining. Intracellular reactive oxygen species (ROS) production and the mitochondrial membrane potential were tested by flow cytometry. The ATP level was measured by reverse phase high performance liquid chromatography. The expression of cytochrome p450 2E1 (CYP2E1) and peroxisome proliferator-activated receptor α (PPARα) was detected by reverse transcriptase-polymerase chain reaction and Western blotting, respectively.</p><p><b>RESULTS</b>Ethanol exposure resulted in the increase of TG level, lipid accumulation and ROS generation, and the decrease of mitochondrial membrane potential and ATP production in the cells. However, ginsenosides significantly reduced TG content (9.69±0.22 μg/mg protein vs. 4.93±0.49 μg/mg protein, P<0.01), and ROS formation (7254.8±385.7 vs. 5825.2±375.9, P<0.01). Meanwhile, improvements in mitochondrial membrane potential (10655.33±331.34 vs. 11129.52±262.35, P<0.05) and ATP level (1.20±0.18 nmol/mg protein vs. 2.53±0.25 nmol/mg protein, P<0.01) were observed by treatment with ginsenosides. Furthermore, ginsenosides could down-regulate CYP2E1 expression (P<0.01) and upregulate PPARα expression (P<0.01) in ethanol-treated cells.</p><p><b>CONCLUSIONS</b>Ginsenosides could prevent ethanol-induced hepatocyte steatosis in vitro related to the inhibition of oxidative stress and the improvement of mitochondrial function. In addition, the modulation of CYP2E1 and PPARα expression may also play an important role in the protective effect of ginsenosides against lipid accumulation.</p>
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<p><b>OBJECTIVE</b>To determine the effect of berberine (Ber) on norepinephrine (NE)-induced apoptosis in neonatal rat cardiomyocytes.</p><p><b>METHODS</b>The cultured neonatal rat cardiomyocytes were treated with NE in the presence or absence of Ber. The activity of lactate dehydrogenase (LDH) in the culture medium was examined, and apoptosis of cardiomyocytes was assessed by Hoechst 33258, isothiocyanate (FITC)-conjugated annexin-V, and propidine iodide (PI) staining. In addition, the activities of caspases-2 and-3 were measured by a fluorescent assay kit. The level of secreted tumor necrosis factor α (TNF-α) and production of intracellular reactive oxygen species (ROS) were also determined.</p><p><b>RESULTS</b>NE at a concentration of 50 μ mol/L induced an obvious increase in the activity of LDH in the culture medium (P<0.05), which was inhibited by coincubation with 0.5, 1.0, or 2.0 μ mol/L Ber (P<0.05). Ber also significantly attenuated NE-induced apoptosis in a dose-dependent manner (P<0.01). Moreover, Ber at a dose of 2 μ mol/L markedly decreased the ROS and TNF-α productions (P <0.05) and inhibited the activation of caspases-2 and -3 in cardiomyocytes exposed to NE (P<0.05)h.</p><p><b>CONCLUSION</b>The present study suggested that Ber could reduce NE-induced apoptosis in neonatal rat cardiomyocytes through inhibiting the ROS-TNF-α-caspase signaling pathway.</p>
Subject(s)
Animals , Rats , Animals, Newborn , Annexin A5 , Metabolism , Apoptosis , Berberine , Pharmacology , Caspase 2 , Metabolism , Caspase 3 , Metabolism , Caspases , Metabolism , Cell Nucleus , Metabolism , Cell Shape , DNA , Metabolism , Enzyme Activation , Flow Cytometry , Fluorescein-5-isothiocyanate , Metabolism , Immunohistochemistry , L-Lactate Dehydrogenase , Metabolism , Myocytes, Cardiac , Pathology , Norepinephrine , Pharmacology , Rats, Sprague-Dawley , Reactive Oxygen Species , Metabolism , Signal Transduction , Tumor Necrosis Factor-alpha , MetabolismABSTRACT
To explore the anti-HSV-1 effect of silencing gD gene expression by RNA interference, five 21-nucleotide duplex small interfering RNAs(siRNAs) targeting the HSV1 gD sequence were designed and the gD-EGFP fusion gene expression vector was constructed, then co-transfected into Vero cell, and screened the effective siRNA through analyzing the intensity of the EGFP fluorescence. Finally, the anti-HSV1 effect was confirmed by plaque reduction assay, real-time PCR and daughter virus titration of HSV1 infected Vero cells transfected with siRNAs. The study demonstrated that siRNAs could effectively and specifically inhibit gD gene expression in HSV1-infected cells, but only had a little effect on HSV1 infection, so taking gD as the target of siRNA against HSV1 needs further study.
Subject(s)
Animals , Humans , Chlorocebus aethiops , Genetic Vectors , Genetics , Green Fluorescent Proteins , Genetics , Metabolism , Herpesvirus 1, Human , Genetics , Polymerase Chain Reaction , RNA Interference , RNA, Small Interfering , Genetics , Recombinant Fusion Proteins , Genetics , Metabolism , Transfection , Vero Cells , Viral Envelope Proteins , Genetics , MetabolismABSTRACT
Objective To evaluate the relationship between children interleukin-1 receptor antagonist gene(IL-1RN) polymorphisms and susceptibility to febrile seizures(FS).Methods Matching case-control study was carried out,blood samples from 52 patients with FS and 53 healthy children were collected.Genomic DNA was extracted and examined by polymerase chain reaction(PCR) amplification for judging IL-1RN genetype.Results Only Ⅰ/Ⅰ,Ⅰ/Ⅱ,Ⅱ/Ⅳ genetypes of IL-1RN were found in the surveyed Chinese population of Han nationality in Guangdong Province.No significant differences of these genotypes were observed in patients and controls.But the allele frequencise of Ⅰ,Ⅱand Ⅳ in patients were 95.2%,3.8% and 1.0%,respectively,while these frequencise in controls were 84.9%,10.4% and 4.7%,respectively.The allele frequency of IL-1RNⅠin patients was obviously higher than that in controls(P
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<p><b>OBJECTIVE</b>To establish a new method on stratification analysis when the stratification limits of confounding factors was not clear or contradictory.</p><p><b>METHOD</b>Data on a study of diabetes mellitus in Guangdong province collected in the year of 1997 and 1998 was analyzed using cluster-stratification analysis.</p><p><b>RESULTS</b>The efficiency of stratification analysis was improved and the confounding bias was effectively controlled with information bias avoided when the clusters-stratification analysis was applied.</p><p><b>CONCLUSION</b>The problem was logically solved using cluster analysis as an assistant stratification means.</p>