Ginsenoside Rh2 enhances the apoptotic effect ofcisplatin on PC-3M prostatic cancer cells

AIM:To study the inducing apoptotic effect of Cisplatin (DDP) combined with Ginsenoside Rh₂ on PC-3M prostatic cancer cells in vitro.METHODS:Cell viability was examined by MTT test and the apoptosis of PC-3M by electrophoresis of NP-40 extracting fragmental DNA and flowcytometry.RESULTS:Pc-3M cells were exposed to 15.0 mg/L Rh₂ combined with 0.4 mg/L DDP: ①Cell viabililty inhibitory rate was increased by 1.44 times from 25.0% (with 0.4 mg/L DDP) to 61.03%. ②Characteristic apoptotic DNA laddering was found more obviously than using either agent alone. ③Apoptotic rate of PC-3M was increased from 0.32% (with 0.4 mg/L DDP) to 52.2%, which showed similar effect with 4.0 mg/L DDP, it was much higher than that of 15.0 mg/L Rh₂ alone (13.6%) (P＜0.01).CONCLUSION:Rh₂ greatly enhances the apoptotic effect of DDP on PC-3M prostatic cancer cells.     

Influence of Weimaining on the cell cycle of murine Lewis lung carcinoma

AIM:To investigate the anti-tumor effect of Weimaining (WMN) on a murine Lewis lung carcinoma (3LL) and the influence on the cell cycle. METHODS:The inhibitory rate of WMN in 3LL growth was detected by replicating the model of 3LL. The effect of the drug on 3LL cell cycle and the influence of the drug on the expression of cyclin D1 protein were investigated by flow cytometry and immunohistochemical staining. RESULTS:The results showed that the inhibitory rate of drug in 3LL is 19.14%, 33.59%, 40.63% and 51.56% respectively at dosage ranging from 100,150, 200 and 250 mg·kg^-1·d^-1. The drug inhibits tumor growth in a dose-dependent manner. The drug arrests 3LL cells in G₀-G₁ phase and decreases the expression of cyclin D1 protein. CONCLUSION:WMN inhibits the growth of 3LL cells in vivo by decreasing the expression of cyclin D1, blocking the cells in G₀-G₁ phase and preventing the cells transition from G₁ to S phase while DNA is replicated.     

Effect of pioglitazone on glucose metabolism and PPAR-γ expression in lipid-infused rats

AIM: To investigate the effect of pioglitazone (Pio) on glucose metabolism and peroxisome proliferators-activated receptor (PPAR)-γ expression in free fatty acid (FFA) -induced insulin resistance in rats. METHODS: A hyperinsulinaemic-euglycaemic clamp and ［3-3H］-glucose tracing technique were used in awake rats. Glucose metabolism in vivo and PPAR-γ in adipose tissue expression were assessed with elevation FFA by lipid infusion over 4 h in rats pretreated with or without Pio.RESULTS: During steady-state of clamp, there was a significant increase in plasma FFA in two lipid-infused groups, compared to control rats (P<0.01). The glucose infusion rates (GIR) in Pio-treated rats (P/L group), compared with controls, were significantly reduced ［(20.6±0.4) mg·kg^-1·min^-1 vs (33.6±0.6)mg·kg^-1· min^-1, P<0.01］, whereas the GIR was lower in the lipid group (L group) than that in the P/L group［(12.6±0.8) mg·kg^-1·min^-1 vs (20.6±0.4) mg·kg^-1·min^-1, P<0.01］. The hepatic glucose production (HGP) was significantly suppressed (85%) ［(18.3±2.1)mg· kg^-1·min^-1 (basal) vs (2.7±2.4)mg· kg^-1·min^-1, and (17.5±2.6) mg· kg^-1·min^-1 vs (2.6±1.0)mg· kg^-1·min^-1］, all P<0.01 during clamp in control and P/L groups. The suppressive effect of insulin on HGP was significantly blunted in L group［(17.3±2.1)mg· kg^-1·min^-1 vs (15.8±1.5)mg· kg^-1·min^-1］. The rate of glucose disappearance (GRd) was significantly reduced in two lipid-infused rats compared with controls［(26.6±1.6)mg· kg^-1·min^-1 and (23.2±0.9)mg· kg^-1·min^-1 vs (37.7±2.6)mg·kg^-1·min^-1,P<0.01］. The PPAR-γ expression of adipose tissue in P/L group was significantly upregulated. CONCLUSION: Lipid-infusion induces an acute insulin-resistance in vivo. Pio treatment upregulates the PPAR-γ of adipose tissue and suppresses HGP. Pio can protect partly against lipid-induced insulin resistance.     

Rifampicin protects rats against rotenone-induced dopaminergic neurons damage

AIM:To investigate the protective effect of rifampicin on rotenone-induced apoptosis of dopaminergic neurons and expression of α-synuclein in rats. METHODS:Highly selective lesions and high expression of α-synuclein in nigrostriatal dopaminergic neurons in rats were induced by chronic subcutaneous exposure to rotenone at dose of 1.5 mg·kg^-1·d^-1 for 3 weeks. At the same time, rifampicin was administered at dose of 30 mg·kg^-1·d^-1 by intragastric administration for 3 weeks. The changes of behavior, pathology and immunoreactivity of TH and α-synuclein in SNc were observed. RESULTS:Obvious changes of behavior, pathology and TH immunoreactivity in SNc were observed in male SD rats injected subcutaneously with rotenone and rifampicin protected rats against these toxic effects induced by rotenone. CONCLUSION:Rifampicin has extensive protective effects against rotenone-induced neurotoxicity, which is related to inhibiting the expression and aggregation of α-synuclein.     

Effects of crocetin on the apoptosis and the changes of its related regulating proteins caspase-3 and Bcl-2 induced by H2O2 in myocardial cells

AIM: To observe the effect of crocetin on the apoptosis and the changes of its related regulating proteins caspase-3 and Bcl-2 expression induced by hydrogen peroxide (H₂O₂) in cultured cardiomyocytes. METHODS: Changes of cellular morphology were detected under microscope. Apoptosis rates of the cells were analyzed by PI staining with flow cytometry. Expressions of caspase-3 and Bcl-2 proteins in the cells were determined by immunofluorescence with flow cytometry. RESULTS: In the concentrations used, more severe morphological changes with higher apoptosis rate of the cultured myocardial cells were seen in each H₂O₂ group than that in control group. When treated with 1×10-4 mol·L^－1 H₂O₂, the caspase-3 was increased and Bcl-2 protein decreased remarkably in the cells. But each dosage of crocetin, especially the highest one (5×10^－5 mol·L^－1, P<005 compared with 5×10^－7 mol·L^－1 group), seemed efficient in maintaining the cell morphology, reducing the cell apoptosis rate and improving the changes in caspase-3 and Bcl-2 protein expression in the cells exposed to 1×10^－4 mol·L^－1 H₂O₂. CONCLUSION: Crocetin obviously inhibits the apoptosis induced by H₂O₂ in the cultured myocardial cells. The mechanisms may involve the balance of the functions of the apoptosis-related regulating proteins, caspase-3 and Bcl-2 protein.     

Protective effects of pioglitazone on endothelial function in rats with hypercholesterolemia

AIM: To investigate the effects of pioglitazone,a PPARγ agonist,on endothelial cell (EC) dysfunction in hypercholesterolemic rats.METHODS: 36 healthy male Wistar rats were assigned to one of the following groups randomly (six rats in each group): control,hypercholesterolemia (HC),and HC treated with pioglitazone 1.5 mg·kg^-1·d^-1,3 mg·kg^-1·d^-1,10 mg·kg^-1·d^-1 and 20 mg·kg^-1·d^-1 (HC＋PIO),respectively.EC function was determined by comparing vasorelaxation to ACh,an EC dependent vasodilator,and acidified NaNO₂,an EC-independent vasodilator.Maximal positive and negative values of the instantaneous first derivative of LVP (+dp/dt_max and dp/dt_max) were determined by MS2000 system.RESULTS: (1) Hypercholesterolemia caused a significant endothelial diastolic dysfunction (maximal relaxation to ACh: 50.51%±2.45% vs 99.78%±3.01% in control,P<0.01).(2) Treatment with pioglitazone relieved EC-dependent vasodilatation in a dose dependent manner,and 10 mg·kg^-1·d^-1 is the best dose.(3) Pioglitazone not only improved EC function,but also reduced cardiac functional injury induced by hypercholesterolemia.CONCLUSION: EC dysfunction induced by hypercholesterolemia can be directly extenuated by pioglitazone,which may effectively prevent from subsequent atherosclerosis and ischemic heart disease.     

Effects of heroin at different concentrations on electrophysiological properties of rat hippocampal CA1 pyramidal neurons

AIM: To investigate changes of electrophysiological properties of rat hippocampal CA1 pyramidal neurons in perfusion with heroin at different concentrations. METHODS: The intracellular recordings were made from CA1 pyramidal neurons of isolated hippocampal slices in perfusion with 0.03-0.3 mmol·L-1 heroin. Compared with control parametes before herion perfusion, the electrophysiological data obtained from identical neurons at 15 min after acute perfusion of the different concentration heroin were analyzed. RESULTS: With the increase of the heroin concentrations in perfusion, the absolute values of neuron resting potential and threshold potential decreased (P<0.05). In 0.3 mmol·L-1 heroin perfusion group，the amplitude of action potential decreased （P<0.05）, 10%-90% risetime and 10%-90% decaytime of spike potential elongated （P<0.05， P<0.01）, and I-V relation curve indicated membran slope resistance increased in 0.1-0.3 mmol·L^-1 perfusion group （P<0.05，P<0.01）. I-F relation curve showed that neuron average firing frequency recored at 15 min after 0.03 mmol·L^-1 heroin infusion was similar to the control frequency when intracellular stimulus intensity was 0.1-1.8 nA, however, firing frequency kept the higher level （P<0.05，P<0.01） in 1.9-2.0 nA vs the control frequence. It was evident that EPSPs of pyramidal neurons were inhibited in perfusion of three concentration heroin, especially in 0.3 mmol·L^-1. CONCLUSION: These results indicate acute heroin perfusion has a significantly influence on electrical activities of hippocampus CA1 pyramidal neurons.     

Electrophysiological changes in rat ventricular myocardium of experimental diabetes and action of CVB-D

AIM: To investigate the electrophysiological changes of diabetic myocardium and effects of cyclovirobuxine D (CVB-D) on its electrophysiology. METHODS: Diabetes was induced in male SD rats, using a single injection of alloxan into tail vein. Untreated age-matched animals were used as controls. Animal electrocardiogram (ECG) was recorded by 2 weeks. Effects of CVB-D on isolated right ventricular papillary muscle from experimental diabetic rats and control group were observed by recording the transmembrane potentials with conventional glass microelectrodes. RESULTS: QT intervals in ECG and action potential duration (APD) at all levels were significantly lengthened in myocardium from week 2 of diabetes. Within the concentration of 13.3-63.3 μmol·L^-1, CVB-D prologated APD of diabetes in dose-dependent manner and more than that of control. Within the concentration of 33.3-63.3 μmol·L^-1, CVB-D depressed RP, APA, Vmax and OS of diabetes in dose-dependent way and more than that of control. In addition, CVB-D at concentration of 20 μmol·L^-1 prologated APD in a time-dependent manner. The most prologation of APD was attained about 40 min in control, while more than 40 min in diabetes. CONCLUSION: The results show that QT intervals in ECG and APD at all levels are significantly lengthened in myocardium from week 2 of diabetes. CVB-D prolongates APD and inhibits RP, APA, OS and Vmax more in diabetes than in control.     

Effect of vitamin K3 on apoptosis induced by androgen-independent prostate cancer cell PC-3M

AIM: To study the effect of vitamin K₃ (VK₃) on the induction of apoptosis in androgen-independent prostate cancer cell PC-3M in vitro.METHODS: Cell viability was estimated by MTT assay. AO/EB staining was performed to detect apoptotic cells. Apoptosis and the changes of cell cycle were detected by flow cytometry. NAC was used to observe the effect of growth inhibition by VK₃. RT-PCR was used to confirm the changes in gene expression. Levels of intracellular peroxides were estimated by using an oxidation-sensitive fluorescent probe DCFH-DA. RESULTS: PC-3M cells growth was significantly inhibited by VK₃ (≥60 μmol/L, P<0.05). The inhibitory effect was time and dosage dependent. The result of AO/EB staining showed that apoptosis of PC-3M cells were induced by VK₃. A typical subdiploid peak before G₀/G₁ phase was observed after treated for 12 h with VK₃ (60 μmol/L) by flow cytometry. The effect of growth inhibition treated with VK₃ was antagonized by antioxygen NAC (5, 10, 20, 40, 80 μmol/L). An increase in the level of DCF fluorescence after PC-3M cells were treated for 1-2 h with VK₃ was observed. Antioxidase GSH-Px and CAT were run-down after treated with VK₃. CONCLUSION: The results indicate that apoptosis in PC-3M cells is induced through oxidative stress by VK₃.     

Effects of atorvastatin on the experimental autoimmune myocarditis in Lewis rats

AIM:To test the hypothesis that atorvastatin affects T cell-mediated autoimmunity through modulating the balance of Th1/Th2 and reduces the severity of EAM. METHODS:Myocarditis was induced in Lewis rats by injection of porcine cardiac myosin. High-dose (10 mg·kg^-1·d^-1) or low-dose (1 mg·kg^-1·d^-1) atorvastatin or vehicle was administered orally for 3 weeks. On day 21, echocardiography was examined and the severity of myocarditis was detected by histopathological evaluation. Levels of serum IFN-γ, IL-2, IL-4 and IL-10 were measured by ELISA. RESULTS:Cardiac function and histological severity of myocarditis were improved in the two atorvastatin-treated groups. Treatment with atorvastatin decreased the levels of Th1 cytokine (IFN-γ, IL-2) and increased the levels of Th2 cytokine (IL-4, IL-10). CONCLUSION:These results suggest that HMG-CoA reductase blockade may be a promising new strategy for the treatment of autoimmune myocarditis.     

Harmful factors affect glycine receptor α1 subunit mRNA expression in the neonatal rat cardiomyocytes

AIM: To investigate the effects of lipopolysaccharide, hypoxia/reoxygenation,isoproterenol and high concentration of glucose on glycine receptor α₁ subunit mRNA expression in the neonatal rat cardiomyocytes. METHODS: Isolation of cardiomyocytes from Sprague-Dawley rats aging 1～3 d were performed. Cardiomyocytes （1×10⁵～5×10⁵ cells·L^-1）were cultured in DMEM medium containing 15% fetal bovine serum at 37 ℃ in 5%CO₂ atmosphere for 72 h. Then, cultured rat cardiomyocytes were treated with lipopolysaccharide, isoproterenol or high concentration of glucose for 24 h, respectively, or were exposed to hypoxia for 3 h followed by reoxygenation for 3 h. Subsequently, the cell survival rate was measured using CCK-8 reactant and RT-PCR was applied to monitor the expression of glycine receptor α₁ subunit mRNA. RESULTS: Compared with the control group, no significant difference in the cell survival rate was observed （P＞0.05）. The expression of glycine receptor α₁ subunit mRNA was increased （P＜0.01） in lipopolysaccharide（5,10,20,40,80 mg/L）,isoproterenol（20,100,500 μmol/L） or hypoxia/reoxygenation, hypoxia groups, but decreased（P＜0.01）in the group treated with high concentration of glucose（25, 50 mmol/L）. CONCLUSION: Lipopolysaccharide, isoproterenol, hypoxia/reoxygenation or hypoxia upregulates the expression of glycine receptor α₁ subunit mRNA,but high concentration of glucose down-regulates the expression of glycine receptor α₁ subunit mRNA in cultured neonatal rat cardiomyocytes.     

Protective effects of tongxinluo,carvedilol and valsartan on microvascular endothelial function and integrity after late reperfused AMI in rabbits

AIM:To compare the protective effects of tongxinluo, a Chinese medicine, and carvedilol and valsartan on myocardium microvascular endothelial function and integrity after late reperfusion of acute myocardial infarction (AMI) in rabbits. METHODS:Forty-eight rabbits were randomly assigned to the following groups:(1) sham operated rabbits;(2) ischemia-reperfusion (I-R) controls;(3) tongxinluo (1.0 g·kg^-1·d^-1);(4) carvedilol (5 mg·kg^-1·d^-1);(5) valsartan (10 mg·kg^-1·d^-1) and (6) ticlopidine + aspirine (30 and 20 mg·kg^-1·d^-1, respectively) groups. After 3 d of drug treatment, the left coronary artery in the rabbit was ligated for 2 h and loosed subsequently for another 2 h. The serum levels of nitric oxide (NO₂^-/NO₃^-) and endothelin (ET) at baseline before AMI, 2 h after both AMI and reperfusion were examined. Also, the number of circulating endothelial cells (CEC), MI size and percentage myocardium focal bleeding incidence were determined 2 h after reperfusion. RESULTS:(1) The baseline level of NO₂^-/NO₃^- was significantly higher in tongxinluo group than that in other groups (all P<0.01), whereas the content of ET was not significantly different among the groups. 2 h after both AMI and reperfusion, NO₂^-/NO₃^- was significantly reduced (P<0.05, P<0.01) and ET was significantly increased in each group as compared with their baseline (P<0.05, P<0.01). Yet among the groups, NO₂^-/NO₃^- was still significantly higher and ET was significantly lower in tongxinluo-treated group than that in the other groups (P<0.05, P<0.01). (2) CEC number was significantly increased in I-R controls as compared with sham group (P<0.01), and was significantly reduced in the tongxinluo-treated groups as compared with I-R controls (P<0.05). (3) MI size was significantly reduced in the four treatment groups as compared with I-R controls (all P<0.01). (4) The percentage of myocardium focal bleeding incidence was significantly lower in tongxinluo and valsartan-treated groups than that in I-R controls (P<0.05, P<0.01). CONCLUSION:Tongxinluo as well as valsartan effectively protectsmyocardium endothelial function and integrity during AMI and late reperfusion,with the effects of tongxinluo being superior.     

Tubuloside B rescues the PC12 neuronal cells from H2O2-induced apoptosis

AIM:To investigate the neuroprotective effect of tubuloside B, one of the phenylethanoids isolated from the stems of Cistanche salsa, on H₂O₂-induced injury in PC12 cells.METHODS:PC12 cells were exposed to various doses of tubuloside B for 12 h, then treated with H₂O₂ at concentration of 100 μmol/L for 24 h. The cell viability was observed with MTT assay. Reactive oxygen species and the mitochondrial membrane potential were measured with laser scanning confocal microscopy (LSCM). The DNA content and percentage of apoptosis were assayed by DNA agarose gel electrophoresis and flow cytometry. The activation of caspase-3 was detected with the caspase-3 activity assay kit. RESULTS:Following treatment with H₂O₂ for 24 h, H₂O₂ induced a significant decrease in cell viability; DNA ladder was observed and apoptosis percentage was as high as 48.0%. Accumulation of intracellular ROS, increase in caspase-3 activity and the decrease in mitochondrial membrane potential as indicated with the decrease of red/green ratios (from 5.97 to 0.41) were detected. However, pretreatment with tubuloside B (1, 10 or 100 mg·L^-1) for 12 h exhibited cytoprotective effects in a dose-dependent manner. Tubuloside B obviously enhanced the cell viability, reduced formation of the DNA ladder, and significantly reduced the number of cells labeled with Annexin-V. The percentage of apoptosis/necrosis neurons was significantly decreased to 30.9%, 18.3% and 6.2%, respectively. LSCM showed that the tubuloside B attenuated the accumulation of ROS and the H₂O₂-induced collapse of mitochondrial membrane potential in PC12 cells. The significant decrease in caspase-3 activity was detected, compared to the H₂O₂-treated cells at the same time point. CONCLUSION:Tubuloside B has the neuroprotective capacity to antagonize H₂O₂-induced apoptosis and injury in PC12 cells, indicating it may be useful for treating some neurodegenerative diseases.     

Salinomycin inhibited proliferation and induced apoptosis of cisplatin-resistant human lung adenocarcinoma cell line A549/DDP

AIM: To investigate the effect of salinomycin on the proliferation and apoptosis of cisplatin-resistant human lung adenocarcinoma cell line A549/DDP. METHODS: The inhibitory effect of salinomycin on the growth of A549/DDP cells was tested by MTT method in vitro . The apoptosis and mitochondrial membrane potential (ΔΨ_m) of A549/DDP cells were assayed by flow cytometry. The activity of caspase-3, 8 and 9 was determined by the method of colorimetry. The levels of cytochrome C, Bcl- 2, Bax, β-catenin, and phosphorylated low-density lipoprotein receptor-related protein 6(p-LRP6) were measured by Western blotting. RESULTS: Salinomycin inhibited the growth of A549/DDP cells in a dose-dependent manner. Salinomycin at concentration of 0.2 μmol/L decreased ΔΨ_m level, and increased reactive oxygen species (ROS), cytochrome C and cytosolic Ca²⁺ release in the cells. Salinomycin also increased the acti-vity of caspase-3, 8, and 9 in the cells, reduced the ratio of Bcl-2/Bax, and decreased the levels of β-catenin and p-LRP6. CONCLUSION: Salinomycin depresses the cell growth by inhibiting Wnt signaling, and induces the apoptosis of cisplatin-resistant human lung adenocarcinoma cell line A549/DDP via mitochondria-dependent and Bcl-2/Bax pathways.     

Effects of bilirubin on oxygen free radicals and caspase-3 in acute lung injury rats

AIM:To investigate the mechanisms by which bilirubin inhibits acute lung injury (ALI). METHODS:30 female Wistar rats were divided into normal group, ALI group and bilirubin treatment group. ALI was induced by intravenous injection of LPS. The contents of OH^-, H₂O₂ and O₂^· in the lung as well as the expression of caspase-3 in the lungs were investigated. RESULTS:(1) The contents of OH^-, H₂O₂ and O₂^· in the lung homogenate and the expression of caspase-3 in the lungs in ALI group increased compared with those in normal group (P<0.05). (2) The contents of OH^-, H₂O₂ and O₂^· in the lung homogenate and the expression of caspase-3 in the lungs in bilirubin treatment group increased compared with those in normal group, but decreased compared with those in ALI models (P<0.05). CONCLUSION:(1) Bilirubin was shown to be able to ameliorate apoptosis in ALI rats. (2) The increase in the contents of OH^-, H₂O₂, O₂^· in ALI group indicated the development of oxidative lung injury, which was ameliorated by bilirubin. (3) Expression of caspase-3 confirmed that the model made by LPS was associated with apoptosis, which was reduced by bilirubin.     

Effect of rapamycin on proliferation,invasion, adhesion,apoptosis and autophagy of human lung adenocarcinoma A549 and resistant A549/DDP cells treated with cis-diamminedichloroplatinum

AIM: To study the effect of rapamycin (Rap) on the proliferation, invasion, adhesion, apoptosis and autophagy of human adenocarcinoma A549 and resistant A549/DDP cells treated with cis-diamminedichloroplatinum (DDP). METHODS: Human adenocarcinoma A549 and resistant A549/DDP cell lines were cultured. The inhibitory effects of Rap alone or combined with DDP on A549 and resistant A549/DDP cells were detected by MTT assay. The in vitroinvasion abilities of the 2 cell lines treated with Rap alone or combined with DDP were detected by Transwell methods. The in vitroadhesion abilities of the 2 cell lines treated with Rap alone or combined with DDP were detected by adhesion experiments. The apoptosis of A549 and resistant A549/DDP cells induced by Rap alone or combined with DDP was analyzed by flow cytometry. The cell autophagy marker proteins beclin-1 and LC3 in A549 and resistant A549/DDP cells treated with Rap alone or combined with DDP were detected by Western blotting.RESULTS: Compared with Rap or DDP alone group, the combination of Rap and DDP significantly inhibited the proliferation, invasion and adhesion of A549 and resistant A549/DDP cells in vitro, and promoted the cell apoptosis and autophagy marker proteins beclin-1 and LC3 expression (all P＜0.05).CONCLUSION: Rap enhances the effect of DDP through promoting the cell autophagy, thereby inhibiting the proliferation, invasion and adhesion of A549 and resistant A549/DDP cells and inducing the cell apoptosis with a synergistic effect.     

Preventive effect of 3,4-dihydroxyacetophenone on atherosclerosis and role of visfatin expression

AIM: To observe the preventive effect of 3,4-dihydroxyacetophenone (DHAP) on atherosclerosis (AS) and the role of visfatin expression in ApoE(-/-) mice.METHODS: Eight-week-old normal mice were used in normal control group (n=8). Eight-week-old male ApoE (-/-) mice were randomly divided into 3 groups: AS group (n= 8, im. NS), DHAP treatment group (n=8, im. DHAP 10 mg·kg^-1·d^-1) and simvastatin treatment group (n=8, im. simvastatin 10 mg·kg^-1·d^-1). All mice were fed with Western diet (21% fat, 0.15% cholesterol) for 12 weeks. The blood samples were collected and the concentrations of blood lipids and visfatin were detected. The frozen sections of aortic root were stained with oil red O. The visfatin in atherosclerotic plaques at aortic roots was examined by Western blotting. The structures of smooth muscle cells and endothelial cells were observed under electron microscope. RESULTS: In DHAP-treated mice, the concentrations of visfatin, TG and TC were decreased, the formation of AS plaque was reduced, the injuries of smooth muscle cells and endothelia cells were attenuated. Visfatin was also decreased at atherosclerosis plaque in DHAP-treated mice.CONCLUSION: DHAP effectively prevents and treats AS by inhibiting the production of visfatin and reducing lipid.     

Effects of thalidomide on the expression of NF-κB and TNF-α in experimental hepatic fibrotic rats

AIM: To study the effects of thalidomide on the expressions of nuclear factor κB (NF-κB) and tumor necrosis factor-α (TNF-α) in rat liver fibrosis.METHODS: The fibrosis of rat liver was induced by intraperitoneal injection of carbon tetrachloride thrice weekly.Meanwhile thalidomide (10 mg·kg^-1·d^-1 or 100 mg·kg^-1·d^-1) was given daily by the intragastric route for 8 weeks.Serum alanine aminotransferase (ALT),aspartate aminotransferase (AST),prealbumin (PA),hyaluronic acid (HA) and laminin (LN),and hydroxyproline (HYP) contents in the liver,NF-κB p65 and α-smooth muscle actin (α-SMA) protein in the liver,IκBα and TNF-α protein in cytoplasm and NF-κB p65 protein in nucleus and TNF-α mRNA levels in the liver were studied.RESULTS: Compared with the model group,the Knodell score,serum ALT,AST,HA,LN levels and HYP contents in liver,NF-κB p65 protein in nucleus and α-SMA protein in the liver,and TNF-α mRNA and protein in the liver of rats given high dose of thalidomide were decreased significantly (P<0.01).Meanwhile PA level and IκBα protein in cytoplasm were elevated significantly (P<0.01).CONCLUSION: Thalidomide exerts its effect on the down-regulation of NF-κB-induced TNF-α via inhibiting dissociation and degradation of IκB and prevents liver fibrosis in rats.     

Porcine surfactant in treatment of LPS-induced early-stage acute lung injury in rats

AIM: To evaluate the therapeutic efficacy of intratracheal instillation of porcine pulmonary surfactant (PPS) in rats with lipopolysaccharides (LPS)-induced early-stage ALI in this study.METHODS: SD rats weighing 200 g-300 g were randomly divided into 4 groups: LPS (1.5 mg·kg-1)+saline,LPS+PPS 100 mg·kg^-1,LPS+PPS 150 mg·kg^-1,LPS+PPS 200 mg·kg^-1.The PaO₂ and PaCO₂,as well as survival rate of rats were examined for 6 h after the start of PPS-instillation.Then,rats were killed and lungs were immediately removed for lung index (LI) and histological analysis.The bronchoalveolar lavage fluid (BALF) was collected for measurement of total protein (TP) contents,TNF-α level and white blood cell(WBC) numbers.RESULTS: Significantly increased PaO₂,reduced mortality rate,decreased total protein and TNF-α contents in BAL,as well as lung index and meliorated histological appearance were observed in three PPS-treated groups compared with group given saline after LPS (P<0.05).The therapeutic effect in PPS150 and PPS200 groups was better than that in PPS100 group.CONCLUSION: Intratracheal PPS instillation provides protective effect on acute lung injury in rats induced by LPS.     

Reversal effect of shikonin on cisplatin resistance of ovarian cancer SKOV3/DDP cells

AIM:To explore the reversal effect of shikonin on cisplatin resistance of ovarian cancer SKOV3/DDP cells and its potential mechanism. METHODS:The proper conditions of treatment with shikonin and cisplatin were determined by CCK-8 assay. The cell cycle and apoptotic rate were analyzed by flow cytometry. The protein levels of cell cycle-and apoptotic-related molecules, such as cyclin D1, cyclin-dependent kinases 2 (CDK2), P18, p-Rb, Bcl-2, Bax and cleaved caspase-3, were determined by Western blot. RESULTS:The results of CCK-8 assay showed that compared with cisplatin group, combined treatment with shikonin and cisplatin had a better inhibitory effect on the growth of cisplatin-resistant SKOV3/DDP cells. The cell cycle G₁/S transition was inhibited, while early apoptotic rate was increased after combined use of shikonin and cisplatin. The results of Western blot showed that compared with cisplatin group, the cells in combination group had lower protein levels of cyclin D1, CDK2, p-Rb and Bcl-2, accompanied with higher protein levels of P18, Bax and cleaved caspase-3. CONCLUSION:Shikonin reverses the cisplatin resistance of ovarian cancer SKOV3/DDP cells. The mechanism may be related to the regulation of cell cycle-and apoptotic-related molecules, and further inhibition of cell viability and promotion of cell apoptosis.