Adult ; Altitude ; Arteries ; metabolism ; Calcitonin Gene-Related Peptide ; blood ; Humans ; beta-Endorphin ; blood

The purpose of the study is to construct R8 peptide (RRRRRRRR) and pH sensitive polyethylene glycols (PEG) co-modified liposomes (Cl-Lip) and utilize them in breast cancer treatment. The co-modified liposomes were prepared with soybean phospholipid, cholesterol, DSPE-PEG2K-R8 and PEG5K-Hz-PE (pH sensitive PEG). The size and zeta potential of Cl-Lip were also characterized. The in vitro experiment demonstrated that the Cl-Lip had high serum stability in 50% fetal bovine serum. The cellular uptake of Cl-Lip under different pre-incubated conditions was evaluated on 4T1 cells. And the endocytosis pathway, lysosome escape ability and tumor spheroid penetration ability were also evaluated. The results showed the particle size of the Cl-Lip was (110.4 ± 5.2) nm, PDI of the Cl-Lip was 0.207 ± 0.039 and zeta potential of the Cl-Lip was (-3.46 ± 0.05) mV. The cellular uptake of Cl-Lip on 4T1 cells was pH sensitive, as the cellular uptake of Cl-Lip pre-incubated in pH 6.0 was higher than that of pH 7.4 under each time point. The main endocytosis pathways of Cl-Lip under pH 6.0 were micropinocytosis and energy-dependent pathway. At the same time, the Cl-Lip with pre-incubation in pH 6.0 had high lysosome escape ability and high tumor spheroid penetration ability. All the above results demonstrated that the Cl-Lip we constructed had high pH sensitivity and is a promising drug delivery system.
Animals ; Cell Line, Tumor ; Cell-Penetrating Peptides ; chemical synthesis ; chemistry ; Cholesterol ; chemistry ; Drug Delivery Systems ; Liposomes ; Mice ; Oligopeptides ; chemical synthesis ; chemistry ; Particle Size ; Phospholipids ; chemistry ; Polyethylene Glycols

AIM: This study was designed to investigate the inhibition of tanshinone ⅡA on C6 glioma cell line and its mechanism. METHODS: MTT was used to measure the levels of the proliferation of C6 cultured with tanshinone ⅡA at different concentrations. The effects of tanshinone ⅡA on cell cycle of C6 were observed by FCM. The change of DNA was observed by Sepharose electrophoresis. The expression of proto-oncogenes c-myc was measured by RT-PCR. RESULTS: The proliferation of C6 was obviously inhibited by tanshinone ⅡA in a dose-dependent manner. The outcome of FCM showed that the apoptotic cell rate was 7.7%, when cultured with tanshinone ⅡA at 1.0 mg/L for 3 days. The apoptotic cell rate was 21.6%, when cultured with tanshinone ⅡA at 2.0 mg/L in 3 days. CONCLUSION: Tanshinone ⅡA inhibits the proliferation of C6 cells, induces apoptosis and inhibits the expression of proto-oncogene c-myc.

AIM To observe the effects of Wuwei Wentong Chubi Capsules (Cinnamomi Ramulus,Poria,Epimedii Folium,etc.) on autophagy proteins of Beclin-1 and LC3-Ⅱ in adjuvant-induced arthritis rats and to explore the possible mechanism of action.METHODS Sixty SD rats were randomized into six groups:normal group,model group,Wuwei Wentong Chubi Capsules (0.8,1.6,3.2 g/kg) groups and tripterygium glycosides (TPT,40 mg/kg) group.In addition to the normal group,adjuvant arthritis (AA) was induced with Freund's complete adjuvant.From the 2th day after injection of FCA,Wuwei Wentong Chubi Capsules with different doses and TPT were given by gavage once a day for 12 days.At the end of the experiment,ankle-joint samples were taken to examine the degree of AA by HE.Beclin-1 and LC3-Ⅱ mRNA were determined by real-time fluorescent quantitative PCR.Meanwhile,the protein expression levels of Beclin-1 and LC3-Ⅱ were determined by immunofluorescence histochemical staining and Western blot.RESULTS As compared with the model group,Wuwei Wentong Chubi Capsules (1.60,3.20 g/kg) not only significantly reduced histopathological injuries,but also effectively up-regulated mRNA and protein expressions of Beclin-1 and LC3-Ⅱ.CONCLUSION Wuwei Wentong Chubi Capsules has a therapeutic action on AA in rats,which might be partly associated with promoting autophagy,decreasing excessive proliferation of synovial cells,leading to the reduction of damage to articular cartilage.

AIM: To investigate the effect of 7-(4-methoxyphenyl)-5, 8a-diphenyl-1,2, 3, 7, 8, 8a-hexahydroimidazo[1,2-a] pyridine (TIP-6) on cell proliferation in human hepatoma cell line HepG2 and human normal hepatocyte cell line L02. METHODS: Typan blue assay was used to check the effect of TIP-6 on cell proliferation. The changes of cell morphology were observed by the phase contrast microscope. Flow cytometry (FCM) was used to check cell cycle. Autophagy and autophagic cell death were detected after acridine orange (AO) staining under fluorescent microscopy. Apoptosis was analyzed by Annexin V/7-AAD, DAPI staining and DNA ladder. NF-κB expression was detected with cellular immunochemistry. RESULTS: Cell proliferation inhibiting effect was appeared when treated with TIP6 from 60 μmol/L to 200 μmol/L, which was correlated with treated concentrations and time. The proliferation rates were just 12.10% and 18.75% (vs control) under 200 μmol/L 72 h in HepG2 and L02 respectively. Vacuolization were found more and more frequently with the increasing of TIP-6 concentrations and treated time prolonged. FCM results indicated that cells were blocked in G2/M phase, and more sensitive were found in HepG2 than L02. AO staining results indicated that the phenomenon of autophagy and autophagic cell death were occurred and appeared more potent with more TIP-6 and longer time treated. No apoptosis markers were found with Annexin V/7-AAD and DAPI staining, and no DNA ladders were found either, these indicated that TIP6 didnt induce apoptosis in these cells. NF-κB was found increased after treated with TIP-6, and the autophagic vacuole became more and more with the increasing of NF-κB protein, but the proliferation rates decreased at the same time. CONCLUSION: TIP-6 inhibited cell proliferation and induced autophagy and autophagic cell death in HepG2 and L02 cells. NF-κB activation may be involved in these effects.

OBJECTIVETo observe the inhibitory effect of gefitineb on the proliferation and its inducing effect on the apoptosis of mouse I-10 Leydig testicular cancer cells in vitro.

METHODSWe treated I-10 Leydig testicular cancer cells of mice with gefitineb at 0, 1.25, 2.5, 5, 10, 20, and 40 µmol/L. Then we determined the inhibitory effect of gefitineb on the growth of the cells by MTT, detected their early and late apoptosis by Annexin V-FITC/propidium iodide double staining and Hoechst 33258 nuclear staining, respectively, and observed the expressions of apoptosis-related proteins Bcl-2, Bax and caspase 3/9 by Western blot.

RESULTSCompared with the blank control group, gefitineb significantly inhibited the proliferation of the I-10 cells at 10 and 20 µmol/L (P < 0.05). The survival rate of the cells was (32.4 ± 2.8)% (P < 0.01) and their early and late apoptosis rates were (26.7 ± 4.2)% and (59.33 ± 10.2)% in the 40 µmol/L group, significantly different from those in the control (P < 0.05 and P <0.01). In comparison with the blank control group, gefitineb at 10, 20, and 40 µmol/L increased the expression of pro-apoptotic protein Bax by (41.9 ± 7.1), (60.1 ± 9.8), and (69.0 ± 11.3)% (all P < 0.05), decreased that of apoptosis-inhibitory protein Bcl-2 by (50.3 ± 8.9), (63.9 ± 6.9), and (88.7 ± 13.9)% (all P < 0.05), and elevated that of the cleft proteins caspase-3 by (69.0 ± 6.9)% (P < 0.05), (71.5 ± 8.1)% (P < 0.05), and (110.9 ± 14.2)% (P < 0.01) and caspase-9 by (51.8 ± 4.9), (54.7 ± 6.7), and (43.8 ± 11.8)% (all P < 0.05).

CONCLUSIONGefitineb can increase the cytotoxicity of I-10 Leydig testicular cancer cells of mice and induce their apoptosis via the mitochondria-mediated apoptosis signaling pathway.

Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; metabolism ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Proliferation ; drug effects ; Cell Survival ; Leydig Cell Tumor ; drug therapy ; metabolism ; pathology ; Male ; Mice ; Neoplasm Proteins ; metabolism ; Neoplasms, Germ Cell and Embryonal ; drug therapy ; metabolism ; pathology ; Quinazolines ; pharmacology ; Testicular Neoplasms ; drug therapy ; metabolism ; pathology ; bcl-2-Associated X Protein ; metabolism

The purpose of this study is to prepare T7 and TAT dual modified liposomes (T7-TAT-LIP) to penetrate through blood brain barrier and target to brain tumor cells. The liposomes were prepared with CFPE, T7 modified PEG-DSPE, TAT modified PEG-DSPE, soybean phospholipid, PEG-DSPE and cholesterol. The CFPE was used to track the cellular uptake efficiency. The density of T7 and TAT and the length of PEG were optimized, and then the liposomes were characterized by particle size, zeta potential, morphology and stability. Afterwards, the cellular uptake by bEnd.3 and C6 cells were evaluated. The results showed that the optimized parameters were 6% of T7, 0.5% of TAT, the molecular weight of PEG for T7 was 2000 and the molecular weight of PEG for TAT was 1000. After optimization, the particle size of T7-TAT-LIP was 118 nm, the zeta potential was -6.32 mV and the particles were spherical. The turbidity and particle size of liposomes were not obviously changed after 24 h incubation in PBS at 37 °C. The particle size and polydispersity index were also stable during 1 month incubation at 4-8 °C. The cellular uptake by both bEnd.3 and C6 cells of T7-TAT-LIP was higher than that of T7 or TAT modified liposomes, suggesting dual modified liposomes possessed better blood brain barrier targeting ability and brain tumor targeting ability than the single ligand modified liposomes.
Biological Transport ; Blood-Brain Barrier ; Brain Neoplasms ; drug therapy ; Cell-Penetrating Peptides ; pharmacology ; Cholesterol ; Liposomes ; Particle Size ; Phosphatidylethanolamines ; Polyethylene Glycols

OBJECTIVETo determine whether the cardioprotection of puerarin (Pue) against hypoxia/reoxygenation injury is mediated by mitochondrial ATP-sensitive potassium channel (mitoK(ATP)) and/or mitochondria calcium-activated potassium channel(mitoK(Ca)).

METHODSCardiomyocytes were isolated from male Sprague-Dawley rats and hypoxia/reoxygenation injury was induced by myocyte pelleting model. Cell viability was assessed by trypan blue exclusion and mitochondrial membrane potential was measured by loading with TMRE. The opening of mitochondrial permeability transition pore was determined spectrophotometrically.

RESULTSPretreatment with Pue at 0.24 mmol/L for 5 min increased the cell viability against hypoxia/reoxygenation injury, while mitochondrial ATP-sensitive potassium channel inhibitor 5-hydroxydecanoate (5-HD, 100 micromol/L, 20 min) or mitochondrial calcium-activated potassium channel blocker paxilline (Pax, 1 micromol/L, 5 min) attenuated the effect of puerarin. The pretreatment with Pue at 0.24 mmol/L for 5 min attenuated collapse of delta-psim induced by hypoxia/reoxygenation injury, 5-HD and Pax abrogated the effect of Pue. In mitochondria isolated from hearts pretreated with Pue, a significant inhibition of Ca(2+)-induced swelling was observed, and this inhibition was attenuated by 5-HD and Pax.

CONCLUSIONThese findings indicate that Pue protects cardiomyocytes against hypoxia/reoxygenation injury via activating mitoK(ATP) channel and mitoK(Ca) channel, and inhibiting mitochondrial permeability transition pore opening.

Androsterone ; pharmacology ; Animals ; Cell Hypoxia ; drug effects ; Cells, Cultured ; Indoles ; pharmacology ; Isoflavones ; pharmacology ; Male ; Mitochondrial Membrane Transport Proteins ; metabolism ; Myocardium ; cytology ; Myocytes, Cardiac ; drug effects ; metabolism ; Potassium Channels ; metabolism ; Potassium Channels, Calcium-Activated ; metabolism ; Rats ; Rats, Sprague-Dawley

AIM: This study was designed to investigate the effect of ginsenoside Rg1 on the amyloid ?-protein precursor (APP) and neprilysin (NEP)expression induced by lipopolysaccharide (LPS) in C6 cell line in order to discover effectual Alzheimer's disease (AD)-treated drugs. METHODS: MTT colorimetric analysis was used to measure the survival rate of C6 cultured with ginsenoside Rg1 at different concentrations (2 5, 5, 10 and 20 ?mol?L -1) and LPS (100 mg?L -1). The expression of APP and NEP mRNA was measured by RT-PCR. RESULTS: LPS decreased the survival rate of C6, furthermore, the increase in APP expression and the decrease in NEP expression were observed. On the other hand, the above alteration induced by LPS was reversed by ginsenoside Rg1. CONCLUSION: This study demonstrates that LPS can cause cell damage, the increase in APP expression and the decrease in NEP expression. Ginsenoside Rg1 can exert a neuroprotective action, protect C6 cells against LPS-induced injury via inhibiting APP expression and increasing NEP expression.

AIM: To investigate the inhibitory effect of TanshinoneⅡA on U251 glioma cell line and its mechanism. METHODS: MTT was used to measure the levels of the proliferation in U251 cultured with TanshinoneⅡA at different concentrations. The effects of TanshinoneⅡA on cell cycle of U251 were observed by FCM. The expression of proto-oncogene c-myc was measured by RT-PCR. RESULTS: The proliferation of U251 was obviously inhibited by TanshinoneⅡA in a dose dependent manner. The inhibitory rate came to the peak at (54 2?0 9)%, when cultured with TanshinoneⅡA at 0 10 g/L. The outcome of FCM showed that the proportion of G 0/G 1 phase cells was increased and the proportion of S phase cells was reduced obviously, when cultured with TanshinoneⅡA at 0 10 g/L for 3 days. The RT-PCR experiment showed that the expression of proto-oncogene c-myc was notably decreased, when the dose of TanshinoneⅡA was increased. CONCLUSION: TanshinoneⅡA inhibited the proliferation of U251 and the expression of proto-oncogene c-myc.