OBJECTIVE: To investigate the salidroside on proliferation and apoptosis of human hepatocellular carcinoma HepG2 cells and its possible mechanisms of anti-tumor. METHODS: Cultured HepG2 cells in vitro were used as the research object and their cell morphology were observed by inverted microscope. The inhibitory rate of cell proliferation was measured by MTS assay. The cell apoptosis was determined by flow cytometry. The protein expressions of Bcl-2, Bax, caspase-3 and caspase-8 were detected by Western blot analysis. RESULTS: Salidroside could inhibit the proliferation of HepG2 cells and promote its apoptosis. The Bax, caspase-3 and caspase-8 protein expression were increased while Bcl-2 protein expression was decreased in HepG2 cells in a concentration-dependent manner. CONCLUSION: Salidroside can inhibit the proliferation and induce the apoptosis of HepG2 cells, and the mechanism may be associated with regulating protein expression of Bcl-2, Bax, caspase-3 and caspase-8.

OBJECTIVETo discuss the protective effect of Mailuoning injection on ischemia/reperfusion (I/R) injury in rats and its mechanism.

METHODHealthy male adult Sprague-Dawley (SD) rats were randomly divided into the sham operation group, the model group, the edaravone (3 mg x kg(-1)) control group, and Mailuoning high, middle and low-dose groups (4, 2, 1 mL x kg(-1)), with 10 rats in each group, and administered with drugs through tail intravenous injection. The middle cerebral artery occlusion (MCAO) was adopted to establish the rat ischemia/reperfusion model. After the ischemia for 2 h and reperfusion for 24 h, the pathological changes in neurovascular units (NVU) of brain tissues at the ischemia side was observed by HE staining. The expressions of glialfibrillary acidic protein (GFAP) and ionized calcium-binding adaptor molecule 1 (Ibal) were detected by the immunohistochemical method. The expressions of tumor necrosis factor-alpha (TNF-alpha), interleukin 1beta (IL-1beta), vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) were detected by the western blotting technique.

RESULTMailuoning injection could significantly improve the pathological changes in cortical penumbra brain tissue UVN of (I/R) rats, reduce the number of GFAP and Ibal positive cells, and significantly decrease the expressions of TNF-alpha, IL-1beta, VCAM-1 and ICAM-1 of brain tissues of I/R rats.

CONCLUSIONMailuoning injection shows an obvious protective effect on UVN of I/R rats. Its mechanism may involve the inhibition of the activation of astrocyte and microglia and the secretion and expression of various inflammatory factors.

Animals ; Brain ; drug effects ; metabolism ; Brain Ischemia ; surgery ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Infarction, Middle Cerebral Artery ; genetics ; metabolism ; Intercellular Adhesion Molecule-1 ; genetics ; metabolism ; Male ; Protective Agents ; administration & dosage ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; genetics ; metabolism ; prevention & control ; Tumor Necrosis Factor-alpha ; genetics ; metabolism ; Vascular Cell Adhesion Molecule-1 ; genetics ; metabolism

ObjectiveTo explore the effect of estrogen on osteoblast apoptosis induced by serum hungry in vitro.MethodsOsteoblasts of second or third generation from newly born SD rats calvaria were divided randomly into the control group, serum hungry group and serum hungry with estrogen group. Cells of each group were incubated for 24 h, 48 h, 72 h, 5 d, 7 d and 14 d, then labeled using TUNEL staining and examined for morphological characteristics of apoptotic cell under light microscopy after incubated for 72 h. The rates of apoptotic cells of each group were examined with flow cytometry.ResultsThe cells of the control group showed normal appears, the serum hungry group had many cells with purple and blue particles in nuclei, but serum hungry with estrogen group had less such cells. The rate of apoptotic cell significantly increased in serum hungry group and decreased in serum hungry with estrogen group compared with the control group examined with flow cytometry (P<0.05).ConclusionEstrogen can repress osteoblasts apoptosis of rats induced by serum hungry.

OBJECTIVEIn this study, we aimed to investigate the expressions of adhesion molecules on human bronchial epithelial cells and neutrophils in co-culture system, assess the effects of puerarin on suppressing these adhesion molecules expressions, and explore the roles of two crucial signal-transduction elements p38 mitogen-activated protein kinase (p38 MAPK) and nuclear factor kappa B (NF-κB) in modulating adhesion molecules expressions.

METHODSNeutrophils and BEAS-2B cells (one human bronchial epithelial cell line) were co-cultured, and adhesion molecules expressions on cell surface were detected using flow cytometry. The mRNA levels of adhesion molecules were assessed by real-time quantitative polymerase chain reaction (real-time qPCR). Phosphorylated p38 MAPK and inhibitor κB were analyzed by Western blot.

RESULTSIn co-culture system, adhesion molecules expressions on BEAS-2B cells and neutrophils were enhanced significantly (P<0.05). Correspondingly, the mRNA levels of adhesion molecules were also increased greatly. Moreover, the pretreatment of peurarin obviously suppressed adhesion molecules expressions on cell surface. Furthermore, phosphorylated p38 MAPK and inhibitor κB in BEAS-2B cells and neutrophils were elevated in co-culture system, but decreased significantly after upon the treatment of peurarin (P<0.05).

CONCLUSIONSCoculture boosted the interactions between human bronchial epithelial cells and neutrophils mimicking airway inflflammation, whereas peurarin decreased the expression of adhesion molecules on cell surface by suppressing the activities of p38 MAPK and NF-κB pathways, and exhibiting its anti-inflflammation activity.

Animals ; Base Sequence ; Bronchi ; cytology ; enzymology ; metabolism ; Cattle ; Cell Adhesion Molecules ; metabolism ; Cell Line ; Coculture Techniques ; DNA Primers ; Down-Regulation ; drug effects ; Epithelial Cells ; enzymology ; metabolism ; Isoflavones ; pharmacology ; NF-kappa B ; metabolism ; Neutrophils ; enzymology ; metabolism ; Phosphorylation ; Real-Time Polymerase Chain Reaction ; p38 Mitogen-Activated Protein Kinases ; metabolism

This study is to investigate the effect of the effective components group of Xiaoshuantongluo (XECG) on neuronal injury induced by oxygen-glucose deprivation (OGD) in primary cortical cultures isolated from SD rat cortex at day 3 and the possible mechanism. Cells were divided into control group, OGD model group and XECG group (1, 3 and 10 mg x L(-1)). The cell viability was assessed with MTT assay and the LDH release rate was measured by enzyme label kit. The cell apoptosis was analyzed using Hoechst staining. RT-PCR was applied to detect the mRNA levels of JAK2 and STAT3. Western blotting was used to detect the expressions of Bcl-2, Bax, p-JAK2 and p-STAT3 proteins. Results showed that XECG resulted in an obvious resistance to oxygen-glucose deprivation-induced cell apoptosis and decrement of cell viability, decrease the cell LDH release rate. XECG could adjust the expression of Bcl-2 and Bax proteins and increase Bcl-2/Bax ratio, up-regulate the expression of p-JAK2 and p-STAT3. In conclusion, XECG could protect against the neuronal injury cells exposed to OGD, which may be relevant to the promotion of JAK2/STAT3 signaling pathway, and impact the expression of Bax and Bcl-2.
Animals ; Apoptosis ; Cell Survival ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Glucose ; Janus Kinase 2 ; metabolism ; Neurons ; drug effects ; metabolism ; Neuroprotective Agents ; pharmacology ; Oxygen ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; STAT3 Transcription Factor ; metabolism ; Signal Transduction ; bcl-2-Associated X Protein ; metabolism

This study was conducted to investigate the paclitaxel loaded by hydrazone bonds in poly(ethylene glycol)-poly(caprolactone) micelles (mPEG-PCL-PTX) on proliferation and apoptosis of human lung cancer A549 cells and its possible mechanisms of anti-tumor activity. The cell proliferation was measured with MTT assay. Flow cytometry were used to analyze the cell cycle. The cell apoptosis was analyzed using Hoechst/P staining. The expression levels of apoptotic genes expression in the mitochondrial apoptosis pathway were detected by RT-PCR and Western blotting, respectively. The mPEG-PCL-PTX could inhibit the proliferation of A549 cells and promote the apoptosis. The Bax, caspase-3 protein expression were increased while Bcl-2 protein expression was decreased in A549 cells. Results showed that the polymer containing hydrazone bond is non-toxic in vitro, the mPEG-PCL-PTX micelles can inhibit the proliferation and induce the apoptosis of A549 cells. Key words: paclitaxel; micelle; A549 cell; proliferation; cell cycle; apoptosis
Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Micelles ; Paclitaxel ; pharmacology ; Polyesters ; Polyethylene Glycols ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

Objective To investigate the relationship of CAG repeat length polymorphism in the androgen receptor(AR)gene to the development of acne.Methods A total of 238 patients with ache vulgaris and 207 healthy human controls in Northeast China were included in this study.Genomic DNA was isolated and purified from the blood of these subjects.The CAG repeat lengths in the AR gene were analyzed by somatic microsatellites (STRs).Results A significant difference was found in the CAG repeat number between the male acne Patients(22.70±3.09)and male controls(23.48±2.83,P=0.046),but not between the female cases and controls(23.41±2.87 versus 23.85±0.21.P=0.12).In order to assess the risk associated with CAG repeats,the male and female subjects were dichotomized based on the median repeat length in the corresponding control group as the arbitrary cut-off point.Those men and women with a short CAG repeat length(＜23 in men,and＜24 in women)had a significantly increased risk for agne than those with a long CAG repeat length(men:95%confidence interval,1.21-3.54,OR=2.07,P=0.008;women:95%confidence interval.1.18-3.56,OR=2.05,P=0.01).Conclusions This study strongly indicates that the CAG repeat length in AR gene is associated with the development of acne in Northeast China,and those men with a short CAG repeat length seem to have a high risk for ague.Consequently,CAG repeat length may serve as a genetic susceptibility marker.

OBJECTIVETo analyze the change of the neuronal restricted silencing factor (NRSF) gene as well as the NRSF regulation genes in beta-mercaptoethanol induction of the marrow mesenchymal stem cells (MSCs) to neurons, and to discuss the function of NRSF in neural induction of the MSCs and the mechanism of the differentiation from MSCs to neurons.

METHODWe used beta-mercaptoethanol, serum-free DMEM, and dimethyl sulfoxide to induce rat MSCs to differentiate to neurons, and then analyzed the changes of the expressions of NRSF gene and NRSF-regulated genes through real-time PCR.

RESULTSThe rat MSCs were successfully induced to differentiate into neuron-like cells. The induced neuron marker, neuron-specific enolase, was positive. Real-time PCR showed that the expression of NRSF gene remarkably declined. The expressions of neurotrophic tyrosine kinase receptor, type 3, synaptosomal-associated protein 25, L1 cell adhesion molecular,neuronal pentraxin receptor in the NRSF-regulated genes also increased at varied extents.

CONCLUSIONSThe differentiation from MSCs to neurons is relevant with the decline of NRSF expression and the increase of the expressions of NRSF-regulated genes. The NRSF may be the key gene during the differentiation from MSCs to neurons.

Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; physiology ; Cells, Cultured ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Neurons ; cytology ; metabolism ; Phosphopyruvate Hydratase ; genetics ; metabolism ; Rats ; Rats, Wistar ; Repressor Proteins ; metabolism ; physiology ; Synaptosomal-Associated Protein 25 ; genetics ; metabolism

OBJECTIVETo investigate the effect of puerarin on interleukin (IL)-8 mRNA expression and the protein release in the co-culture of human bronchial epithelial (BEAS-2B) cells and human neutrophils.

METHODSBEAS-2B cells and neutrophills were cultured separately and co-cultured with puerarin (50, 100, and 200 μg/mL) for a predetermined time. Cytokines in culture supernatant were evaluated by protein array and IL-8 quantified by enzyme-linked immunosorbent assay (ELISA). IL-8 mRNA expression was evaluated by real-time quantitative polymerase chain reaction (real-time qPCR).

RESULTSThe co-culture of BEAS-2B cells and neutrophils exhibited synergistic effects on IL-8 mRNA expression in BEAS-2B cells, but not in neutrophils after 12 h incubation (P<0.01), as compared with that in BEAS-2B cells or neutrophils alone. IL-8 protein release in the culture supernatant was obviously elevated when BEAS-2B cells were co-cultured with human neutrophils as compared with that in the supernatant of BEAS-2B cells or neutrophils alone after incubated for 2, 6, 12, and 18 h (P<0.01). Treatment with puerarin could significantly down-regulate the expression of IL-8 mRNA in BEAS-2B cells and IL-8 release in the supernatant of the co-culture of BEAS-2B cells and neutrophils (P<0.01).

CONCLUSIONPuerarin could exhibit anti-inflammatory activity by suppressing IL-8 production from the co-culture of human bronchial epithelial cells and neutrophils.

Adult ; Bronchi ; cytology ; Cell Communication ; drug effects ; Cell Line ; Coculture Techniques ; Epithelial Cells ; cytology ; drug effects ; metabolism ; Fluorescence ; Gene Expression Regulation ; drug effects ; Humans ; Interleukin-8 ; genetics ; secretion ; Isoflavones ; pharmacology ; Neutrophils ; cytology ; drug effects ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction

To screen potential hamster chymase 2 inhibitors, a high-throughput screening (HTS) model was established. Recombinant hamster chymase 2 with active form was cloned and expressed in E. coli. The HTS model with total volume of 50 microL in 384-well microplate was based on fluorescence analysis and was proved sensitive as well as specific (Z' = 0.84). A total of 40 080 samples (including 28 060 compounds and 12 020 natural products) were screened, and 613 samples with inhibition greater than 90% were selected for further rescreening. Finally, compounds J16647 and J16648 were identified with high inhibitory activity on chymase 2, and whose IC50 values were 0.823 and 0.690 micromol x L(-1), respectively.
Animals ; Chymases ; analysis ; antagonists & inhibitors ; Cricetinae ; Enzyme Inhibitors ; analysis ; pharmacology ; Escherichia coli ; metabolism ; High-Throughput Screening Assays ; methods ; Inhibitory Concentration 50 ; Rats ; Structure-Activity Relationship