Objective: To observe the effect of exogenous spermine on H/I-induced cardiomyocyte apoptosis in suckling rat, and to explore its mechanism.Methods: The hypoxia injury model was established on primarily cultured suckling rat cardiomyocytes (Hank's balanced salt solution (pH=6.8) was used as cell culture medium, the oxygen in culturing environment was discharged, and the suckling rat cardiomyocytes were cultured in an anaerobic incubator for 24 hours). Cultured suckling rat cardiomyocytes were randomly assigned into control, hypoxia and hypoxia+spermine (Hypoxia+Sp) groups. The protein expressions of key enzyme catalyzing polyamine metabolism (ODC, SSAT) in cardiomyocytes were detected by Western blot. The cell apoptosis was detected by using Cell Counting Kit-8 (CCK-8) and Hoechst 33342 dye. The activities of total-superoxide dismutase (SOD), caspase-3/-9 and the contents of malondialdehyde (MDA), glutathione (GSH) in cell medium were analyzed. Intracellular reactive oxygen species (ROS) generation was estimated by using DCFH-DA staining.Results: Compared with the control group, the expression of SSAT protein, apoptosis rate, MDA content, the activity of caspase-3/-9 and ROS generation were increased, but the expression of ODC protein, SOD activity and GSH content were decreased in hypoxia group. Compared with the hypoxia group, the changes of all index above-mentioned were weakened in the Hypoxia+Sp group.Conclusion: Exogenous spermine can attenuate cardiomyocyte injury and cell apoptosis of suckling rat induced by hypoxia,which mechanism is related with restoring polyamines homostasis and scavenging ROS.

Objective: To explore the therapeutic effect of treating with anti-CCL21 monoclonal antibody on left ventricular remodeling and cardiac function post-acute myocardial infarction in mice.Methods: C57BL/6 mice were randomly divided into sham-operated group, model group and anti-CCL21 monoclonal antibody intervention group, and further randomly divided into 1 d, 3 d,7 d and 21 d subgroups. A mouse model of acute myocardial infarction (AMI) was generated by left anterior descending coronary artery ligation in C57BL/6 mice. Mice in model group were intravenously given isotype-IgG 1.0 mg, mice in anti-CCL21 monoclonal antibody intervention group were intravenously given goat anti-mouse CCL21 monoclonal antibody 1.0 mg at 5 minute and on the 3rd day post-coronary artery ligation. After surgery, C-C motif chemokine receptor 7 (CCR7) expression in myocardium was detected on the 1st, 3rd, 7th day, and the expressions of matrix metalloproteinase-2 (MMP-2) and MMP-9 in myocardium were detected on the 7th day 8 mice per group using Western blot. Serum levels of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in each group were measured on the 1st, 3rd, 7th day post-surgery using ELISA, 8 mice per group. Transthoracic echocardiography was underwent in mice on the 7th and 21th day after surgery using an echocardiography system to evaluate left ventricular function.Results: Compared with sham-operated group, serum levels of CCL21, TNF-α, IL-6, and myocardiac expression of CCR7、MMP-2、MMP-9 in model group were significantly increased (P<0.05). Compared with model group, serum levels of TNF-α and IL-6 and myocardiac expression level of MMP-9 were significantly decreased in anti-CCL21 monoclonal antibody intervention group(P<0.05).Conclusion: Our study indicates that treating with anti-CCL21 monoclonal antibody is involved in preventing cardiac remodeling and protecting left ventricular function post AMI via inhibiting inflammatory response and MMP-9 expression level.

Objective: To investigate the protective effects of ginsenoside-Rg2 on ventricular remodeling induced by isoproterenol (ISO) in rats.Methods: Sixty male Wistar rats were randomly divided into six groups, i.e. control group, ventricular remodeling model group,ginsenoside-Rg2 (20,40,80 mg/kg) groups and propranolol (15 mg/kg) group, with 10 rats in each group.The ventricular remodeling model was established in Wistar rats by subcutaneous injection with isoproterenol for seven days; the control group was given the same volume of normal saline. The rats were administrated intragastrically with ginsenoside-Rg2 or propranolol according to the group protocol above-mentioned after isoproterenol injection, once a day for 6 weeks. Then, the hemodynamic parameters were tested by an eight channel physiological recorder, and the heart weight was weighed and the left ventricular remodeling index was calculated.Results: On the 7th week after ISO injection, systolic blood pressure (SBP), diastolic blood pressure (DBP), mean blood pressure (MBP), heat rate, left ventricular systolic pressure (LVSP), +dp/dt_max, -dp/dt_max in the model group were lower than those of control group (P<0.01), however, heart weight/body weight (HW/BW), left ventricular weight/body weight (LVW/BW) and left ventricular end-diastolic pressure (LVEDP) were higher significantly than those of control group (P<0.01). The levels of SBP, DBP, MBP, HR, LVSP, +dp/dt_max, -dp/dt_max in ginsenoside-Rg2 low, medium and high dose groups and propranolol group were higher than those of model group (P<0.05, P<0.01); while the levels of LVDP were lower than that of model group (P<0.05, P<0.01). The levels of HW/BW and LVW/BW in ginsenoside-Rg2 medium and high dose groups and propranolol group were lower than those of model group (P<0.01). Compared with propranolol group, the levels of SBP, DBP, MBP were significantly decreased in ginsenoside-Rg2 low dose group (P<0.05, P<0.01); the level of LVEDP in ginsenoside-Rg2 low dose group, the levels of LVSP and +dp/dt_max in ginsenoside-Rg2 high dose group were obviously increased (P<0.05, P<0.01).Conclusion: Ginsenoside-Rg2 can alleviate the ventricular remodeling induced by isoproterenol in rats.

Objective: To observe the effects of A_2aR on pulmonary fibrosis induced by bleomycin (BLM) in wild type and A_2aR gene knockout mice.Methods: Thirty male BALB/c wild type (WT) mice and twenty male A_2aR gene knockout (A_2aR KO) mice, were randomly divided into 5 groups (n=10 each):WT control group (A), WT fibrosis group (B), A_2aR KO control group(C), A_2aR KO fibrosis group(D), WT fibrosis+A_2aR agonist (CGS21680) group (E). For the induction of pulmonary fibrosis, mice were intratracheally injected with a single dose of bleomycin (5.0 mg/kg body weight), while the control groups with an equal volume of NS. After injection, the mice were vertically rotated immediately for 3 or 5 minutes in order to make the liquor evenly distributed in lung. The mice in groups A to D received intraperitoneal injection of 0.5 ml NS, those in E group received 0.5 ml CGS-21680 (0.25 mg/kg body weight) daily for a period of 4 weeks. On the 29th day, the blood and tissue samples were taken. The chloramine T method was used to detect the content of hydroxyproline (Hyp) in lung. ELISA was used to detect transforming growth factor-β1 (TGF-β1) in sera. The immunohistochemical technique and Western blot were used to detect the protein expression of TGF-β1 and A_2aR. And the mRNA expression of TGF-β1 and A_2aR were measured by in situ hybridization. The microstructure, ultrastructure and fiber staining of lung tissue in mice were observed in each group.Results: ①Thickened and destroyed alveolar walls, disordered, stenosed or partial collapsed alveolar spaces, fibrous proliferation and infiltration of inflammation cells were observed in lung of mice in group B under microscope. The described performance was even more serious in group D, and that the intervention in group E, could significantly ameliorate the pathological changes. The vacuolation in epithelial cells of type I and Ⅱ as well as the lamellar body, were observed on the visualizing of ultrastructure of lung tissue in group B. More severe pathological lesion in lung described above were presented in Group D than those in group B, obviously lessen pathological damage were presented in group E. The lung positive region in mice of group B and D was increased significantly with a fusion of patchy distribution by masson staining of fibrous tissue, the lung positive region in group D was higher than that in group B. However, the lung positive area in group E was significantly decresded compared with that in group B and D. The results above indicated A_2aR gene knockout exacerbated the pulmonary fibrosis. ②The content of TGF-β1 in sera and the content of Hyp in lung, the expression of A_2aR protein and TGF-β1 protein, the A_2aR mRNA and TGF-β1 mRNA were significantly increased in group B compared with group A(P <0.01,P<0.05). The content of TGF-β1 in sera, the content of Hyp, the expression of TGF-β1 protein and the TGF-β1 mRNA in lung were significantly increased in group D compared with group C(P <0.01,P<0.05). Those results indicated that content of Hyp and expression of TGF-β1 in mice with lung fibrosis were increased. ③Compared to group B, the content of TGF-β1 in sera and the content of Hyp in lung, the expression of TGF-β1 protein and TGF-β1 mRNA were significantly increased in group D(P<0.01). The expression of A_2aR protein and A_2aR mRNA in group E was higher than those in group B (P<0.01), the content of TGF-β1 in sera, Hyp in lung, the expression of TGF-β1 protein and TGF-β1 mRNA were much less than those in group E (P<0.01). The results above indicated that the content of Hyp and expression of TGF-β1 were increased in lung of A_2aR gene knockout mice, A_2aR agonist might upregulate the expression of A_2aR and deregulate the expression of TGF-β1.Conclusion: Pulmonary fibrosis is significantly increased in A_2aR gene knockout mice. A_2aR response to the pulmonary fibrosis is increased in wild type mice, the effect of A_2aR on anti-inflammatory in BLM-induced pulmonary fibrosis is acted by inhibiting TGF-β1 expression.

Objective: To investigate whether chronic renal insufficiency is an independent risk factor or not for all-cause death, cardiac/cerebral disease death and thromboembolic events in patients with non-vavular atrial fibrillation aged over 45 years.Methods: Eight hundred and twenty-five non-vavular atrial fibrillation patients without anticoagulation therapy aged over 45 years were enrolled in this study. The average age of the patients was 76.52±11.80 years old, 18.91% of the patients was female, while 64.73% of the patients was over 75 years old. The baseline characteristics and adverse events during the observation period were recorded. Multivariate Cox regression analysis was adopted to calculate the adjusted hazard ratio (HR) of renal insufficiency for clinical adverse events.Results: During a median of 33.5 months' follow-up, 209 patients died of all-cause death, and 61 patients died of cardiac/cerebral disease death. A total of 139 patients had thromboembolic events (stroke or transient ischemic attack (TIA), or peripheral arterial thromboembolism). All of the adverse events were higher in atrial fibrillation patients with chronic renal insufficiency than those in control group (P<0.05). After adjustment for CHA2DS2-VASc score and other traditional risk factors, chronic renal insufficiency was also an independent predictor of the all-cause death, cardiac/cerebral disease death, thromboembolic events (P<0.05).Conclusion: After adjustment for CHA₂DS₂-VASc score, chronic renal insufficiency was one of the independent risk factors for all-cause death, cardiac/cerebral disease death, thromboembolic events in non-vavular atrial fibrillation patients aged over 45 years without anticoagulation therapy.

Objective: To observe the effect of extracellular Ba²⁺ on the recording L-type calcium channel of rat ventricular cardiocytes.Methods: Whole-cell patch clamp technique was used to record the currents of L-type calcium channels (I_Ca,L) in a single ventricular cardiocyte isolated from a rat heart by acute enzymatic digestion combined with mechanical method. Ba²⁺ was used to replace Ca²⁺ in Tyrode's solution, and Ba²⁺ (0~0.8 mmol/L) was directly added to Tyrode's solution to observe the change of the peak current within 15 minutes. The data were repeated with 5 cells or more.Results: ①The inactivation rate of I_Ca,L was slowed down significantly (P<0.01) with Ba²⁺ instead of Ca²⁺ gradually in Tyrode's solution. There was no change in the inactivation rate of I_Ca,L when the Ba²⁺ (0.2, 0.4 mmol/L) was added in the extracellular solution, but when 0.8 mmol/L Ba²⁺ was added, the inactivation rate would slow down significantly (P<0.01). ②Ba²⁺(0.2, 0.4 mmol/L) added in the extracellular solution could decrease the peak current attenuation. Compared with the group in Tyrode's solution, the peak current attenuation rate of the group in Ba²⁺ (0.2, 0.4 mmol/L) added in the extracellular solution was decreased significantly at the two time points (10 min, 15 min, P<0.01). ③The extracellular Ba²⁺ (0.4 mmol/L) could move the current-voltage curve down, change the reversal potential, weaken I_Ca,L inhibition induced by salvianolic acid A and right-shift concentration-response curve.Conclusion: The addition of Ba²⁺ to extracellular solution can attenuate the peak current decay, change the voltage-dependent characteristics of calcium channels and influence drug dose-effect relationship when whole-cell patch clamp technique is used to record L-type calcium channel of rat ventricular cardiocytes.

Objective: To observe the effect of 4 weeks eccentric endurance exercise on metabolic disturbance and muscle atrophy in type 2 diabetic rats, and to investigate the role of myostatin/Smad3/atrogin-1 signaling pathway in muscle atrophy.Methods: Type 2 diabetic rats were established by high fat diet combined with streptozotocin(STZ) injection for 9 weeks. The normal chow diet rats were randomly divided into control group (C, n=6) and exercise group (E, n=9), the type 2 diabetic mellitus rats were randomly divided into diabetic control group (D, n=8) and diabetic exercise group (DE, n=12). Exercise program:slope of -5°, speed of 16 m/min, 60 min each time, 1 time a day, 5 d per week for 4 weeks. Fasting blood glucose (FBG), fasting insulin (FINS), insulin resistance index (homeostasis model assessment insulin resistance index, HOMA-IR), insulin sensitivity index (insulin sensitivity index, ISI) and glucose tolerance test were measured on fasting 12 hours after exercise. Soleus muscle was drawed after exercise and the degree of muscle atrophy was detected. The expressions of myostatin, phospho-Smad3 (p-Smad3), drosophila mothers against decapentaplegic protein 3 (Smad3) and atrogin-1 were detected.Results: ①Compared with control group, body weight, soleus mass/tibia length, muscle fiber cross-sectional area, FINS and ISI of diabetic group rat were decreased significantly (P<0.01), FBG, HOMA-IR, area under the curve of blood glucose (AUC_BG), the expression of all myostatin, Smad3, p-Smad3 and atrogin-1 was increased significantly (P<0.01). ②After 4 weeks of eccentric exercise, compared with diabetic group, muscle fiber cross-sectional area of diabetic exercise group rat was increased significantly(P<0.01). AUC_BG, HOMA-IR, the expression of myostatin, p-Smad3 and atrogin-1 were decreased significantly (P<0.05, P<0.01).Conclusion: The up-regulation of myostatin/Smad3/atrogin-1 signaling pathway is an important cause of muscle atrophy in type 2 diabetes. The down-regulation of myostatin, p-Smad3, and atrogin-1 after 4 weeks of eccentric endurance exercise may inhibit muscle atrophy, and then ameliorate metabolic disorders and increase insulin sensitivity in type 2 diabetes.

Objective: To investigate the protective effects of different dosage of L. meyennii on hypoglycemia of rats induced by exhaustive exercise.Methods: The exercise-induced hypoglycemia animal model was established using the increasing load to exhaustion swimming training. Fifty-five male Wistar rats aged six-week were randomly divided into five groups:①control group (C group), ②exercise control group (M group), ③~⑤exercise+ low dose or medium dose or high dose L. meyennii group (LM I, LM Ⅱ, LM Ⅲ group), 10 rats in each group (5 rats were excluded). L. meyennii was given to LM groups once a day, the dosage was 0.2,0.4,1.2 g/kg body weight respectively, the volume was 5 ml/kg body weight, C and M group were given the same volume of normal saline intragastrically. The exercise rats were drilled in an exhaustive swimming training for 42 d with an increasing load. After 42 d exhaustive training, the body weight, exhaustive swimming time were determined, and related biochemical indicators were detected from samples of plasma, liver and deep strands of quadriceps.Results: The body weight, the level of blood glucose, muscle glycogen, hepatic glycogen, expression of PEPCK in hepatocytes and hepatocyte positive staining accumulated absorbance of M group were lower than those of C group (P<0.05 or P<0.01), there was no obvious difference between C and M groups in the time of exhaustive swimming. Compared with the C group, the content of blood lactate of M group was increased significantly (P<0.01). Compared with M group, body weight, the level of blood glucose, muscle glycogen, hepatic glycogen, expression of PEPCK in hepatocytes and hepatocyte positive staining accumulated absorbance of LM groups were obviously increased (P<0.05 or P<0.01), the time of exhaustive swimming was prolonged (P<0.01), and the content of blood lactate was decreased significantly(P<0.01). There was no difference between LM Ⅰ and LM Ⅲ group in body weight. Compared with LM I group, the level of blood glucose, muscle glycogen, hepatic glycogen, expression of PEPCK in hepatocytes and hepatocyte positive staining accumulated absorbance of LM Ⅲ group was obviously increased (P<0.05), the time of exhaustive swimming of LM Ⅲ group was prolonged, and the content of blood lactate of LM Ⅲ group decreased significantly (P<0.05).Conclusion: High dosage of L. meyennii may effectively suppress and delay the occurrence and development of exercise-induced hypoglycemia and fatigue induced by the long-time and intensive training, and the mechanism may be related to optimize of muscle glycogen and hepatic glycogen reserve in one hand. It is also related to up-regulates the limited enzyme of PEPCK expression and activity in the other hand. Finally it promotes the impact of gluconeogenesis.

Objective: To investigate the effect of aerobic exercise on energy metabolism of skeletal muscle mitochondria in aging rats.Methods: Twenty female Wistar rats aging to 12-month old were randomly divided into two groups:aged control group (AC,n=10)and aged exercise group (AE,n=10), and another 10 female Wistar rats aging to 2-month old were set as the young control group(YC,n=10). The rats in YC and AC groups were reared normally, the rats in AE group were given treadmill exercise 6 days a week for 12 weeks (slope 5°, speed 15.2 m/min, 15 min on the 1st day, 30 min on the 2nd day, 45 min on the 3th day and beyond). After 12 weeks, all the rats were decapitated, the gastrocnemius samples were taken, the mitochondria were extracted by differential centrifugation and used for measuring the activities of superoxide dismutase(SOD), glutathione peroxidase(GSH-Px), tricarboxylic acid cycling rate-limiting enzymes citrate synthase(CS), isocitrate dehydrogenase(ICD) and α-ketoglutarate dehydrogenase(α-KGDHC), respiratory chain complexs(RCC)Ⅰ~Ⅳ and the content of malondialdehyde(MDA).Results: ①Compared with YC group, the SOD activity and the MDA content were significantly increased (P<0.05),the activities of CS and α-KGDHC were significantly decreased (P<0.05),the activities of RCCⅠ, RCCⅡ and RCC Ⅳ were significantly decreased (P<0.05), and the activity of RCCⅢ was significantly increased (P<0.05) in skeletal muscle mitochondria of AC group; the activities of SOD and GSH-Px and the MDA content were significantly increased (P<0.01), the activities of CS, ICD and α-KGDHC were significantly increased (P<0.01), the activities of RCCⅠ~Ⅳ were significantly increased (P<0.01) in skeletal muscle mitochondria of AE group.②Compared with AC group, the activities of SOD and GSH-Px were significantly increased (P<0.05), MDA content was significantly decreased (P<0.05), the activities of CS, ICD, α-KGDHC and RCCⅠ~Ⅳwere significantly increased (P<0.01) in AE group.Conclusion: Aerobic exercise can improve the antioxidant capacity of skeletal muscle mitochondria in aging rats, decrease the level of lipid peroxidation, improve the function of tricarboxylic acid cycle and respiratory chain, promote mitochondrial energy metabolism, and slow down the degenerative changes in mitochondria during aging.

Objective: To study the effect of ferulic acid on diabetic cardiomyopathy of type 2 diabetic mice induced by high fat diets combined with low dose streptozotocin (STZ), and to explore its possible mechanism.Methods: Twenty male ICR mice were fed with high fat diets for 6 weeks, and then were injected intraperitoneally with STZ (30 mg/kg) for 5 continuous days. Nine days later, the fasting blood glucose (FBG) was detected, mouse with FBG over 11.1 mmol/L was considered as diabetes mellitus. Twenty diabetic mice were randomly divided into two groups:model group and ferulic acid group (200 mg/kg, i.g.), 10 in each group. Taking another ten age and sex matched mice as a control group. After continuous administration for 8 weeks, FBG, body weight, heart weight, left ventricular weight, the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) were detected, the heart mass index (HMI) and left ventricular mass index (LVMI) were calculated. Masson staining was employed to observe pathological changes; immunohistochemistry was used to determine the expression of transforming growth factor-β1 (TGF-β1), collagen Ⅲ in the myocardial tissue.Results: Compared with the model group, HMI, LVMI, the content of MDA were significantly decreased (P<0.01, P<0.05) and the activity of SOD was notably increased (P<0.05) in the ferulic acid group. Collagen deposition and interstitial fibrosis in myocardial tissue were improved. Immunohistochemistry showed that expressions of TGF-β1 and type Ⅲ collagen remarkably decreased (P<0.01, P<0.05) in myocardial tissue.Conclusion: Ferulic acid has a protective effect on diabetic cardiomyopathy in diabetic mice, and its mechanism may be linked to anti-oxidation, inhibiting the expression of TGF-β1 protein and reducing the deposition of production oftypeⅢcollagen.

Objective: To investigate the improving effect of applephenon on the hypoxic-pulmonary hypertension by down-regulating the expression of calcium sensing receptor(CaSR).Methods: Thirty-six male SD rats were randomly divided into 3 groups(n=12):control group, hypoxia-exposure group and applephenon-intervention group.Rats in hypoxia-exposure group and applephenon-intervention group were intermittently exposed to hypoxia in a hypoxic-cabin (the O₂ concentration was 10%) for 8 hours everyday, while rats in control group were raised in normal environment, the experiments were conducted for 4 weeks.Meanwhile, rats in applephenon-intervention group were given intragastrically with applephenon according to 200 mg/kg per day, rats in control and hypoxia-exposure group were given intragastrically with the same amount of normal saline, it lasted for 4 weeks. Then the average pulmonary artery pressure(mPAP), pulmonary vascular resistance(PVR), right ventricular hypertrophy index(RVHI), medial thickness(MT), WA%, WT% and the expression of calcium sensitive receptor (CaSR) of rats in each group were measured.Results: mPAP, PVR, RVHI, MT, WA%, WT% and CaSR expression in hypoxia-exposure group were all significantly higher than those in control group(P<0.01), while applephenon could improved those obviously.Conclusion: Applephenon has a good prophylactic effect on hypoxic pulmonary hypertension, the mechanism may be involved in the down-regulation of CaSR expression.

Objective: To observe the effects of calcitonin gene-related peptide (CGRP) on epithelial-mesenchymal transition (EMT) of alveolar epithelial cells and its mechanism.Methods: The RLE-6TN cell were divided into 6 groups as follows:control group, cells were incubated with double distilled water (TGF-β1 solvent) for 48 h; transforming growth factor-β1(TGF-β1) group, cells were incubated with TGF-β1 (5 ng/ml) for 48 h; +CGRP (1, 10,100 nmol/L) group, cells were pre-treated with CGRP (1, 10,100 nmol/L) for 1 h, and then subjected to TGF-β1 for 48 h; CGRP 100 nmol/L + CGRP8-37 1 μmol/L group, cells were pre-treated with CGRP 100 nmol/L + CGRP8-37 1 μmol/L for 1 h, and then subjected to TGF-β1 for 48 h. RLE-6TN cell vitality was determined by MTT. The mRNA or protein expression levels of Notch1, eIF3a, α-SMA, E-cadherin and collagen Ⅲ were detected by real-time PCR or Western blot.Results: Compared with the TGF-β1 group, cell vitality and the expression of E-cadherin were significantly increased, and the expression of Notch 1, eIF3a, α-SMA and collagen Ⅲ were markedly decreased in CGRP (1, 10, 100 nmol/L) group. The effects of CGRP above mentioned could be abolished by CGRP 8-37.Conclusion: These results suggest that CGRP inhibits TGF-β1-induced EMT process, and down-regulation of the expressions of Notch1and eIF3a may be involved in its mechanisms.

Objective: To investigate the effect of octreotide on the proliferation and apoptosis of the rat airway smooth muscles (ASMCs) induced by transforming growth factor β1 (TGF-β1), and whether the SHZ domain-containing protein tyrosine phosphatase-1(SHP-1) is involved in the effect mentioned above or not.Methods: ①The cultured ASMCs were divided into the following 4 groups:control group, TGF-β1 group, TGF-β1+octreotide group and TGF-β1+octreotide+NSC87877group. The proliferation of ASMCs was detected by MTT assay. The level of phospho-SHP-1(Tyr536) was examined by Western blot.②The cultured ASMCs were divided into 3 groups:control group, 0.01 nmol/ml octreotide group and 0.1 nmol/ml octreotide group. Apoptosis of ASMCs was detected by Annexin-V FITC/PI double staining.Results: ①The absorbance (A_{490 nm}) value of TGF-β1 group was higher than that of control group (P<0.01).The A_{490 nm} value of TGF-β1+octreotide group was lower than that of TGF-β1 group (P<0.05) but higher than that of TGF-β1+octreotide+NSC87877 group(P<0.01).②The apoptotic rate of ASMCs in 0.01 nmol/ml octreotide treated group was higher than that in control group (P<0.01). There was no significant difference of opoptosis rate between control and 0.1 nmol/ml octreotide treated group.③The level of p-SHP-1 in TGF-β1 group was lower than that in control group(P<0.01). There were no significant differences between TGF-β1 group and TGF-β1+octreotide group.The p-SHP-1 in TGF-β1+octreotide+NSC87877 group was the lowest in all the groups.Conclusion: ①Octreotide can inhibit rat ASMCs proliferation induced by TGF-β1. ②Low dose octreotide induces ASMCs apoptosis, but high dose octreotide has no significant effect on ASMCs apoptosis. ③The growth inhibitory effect of octreotide is independent of SHP-1 signal pathway.

Objective: To study the effects of nano-carbon black particles combined with cold exposure on lung tissue and its oxidative stress in mice.Methods: Seventy-two healthy male C57BL/6 mice were randomly divided into six groups:control group (Ctrl), cold exposure group (C), low dose exposure group (L), low dose exposure combined with cold exposure group (LC), high dose exposure group (H), high dose exposure combined with cold exposure group (HC). The suspensions of nanoscale carbon black particles were intratracheal instilled to mice once time, the exposure doses were 0.45 mg/ml (L) and 4.05 mg/ml (H). The mice in cold group and in nano-carbon black particles exposure combined with cold exposure groups had been intermittently exposed to 4℃ for 20 days (4 h/day). Then the mice were weighed and the lung were drawn. The activities of SOD (superoxide dismutase), GSH-Px (glutathione peroxidase) and the content of MDA (malondialdehyde) in lung were determined. The lung pathological slices were prepared to observe the structural changes of the lung.Results: All cold exposed mice were significantly lighter in body weight than mice in all non-cold-exposed groups(P<0.05),the body weight of the mice in control group and in simple nano-carbon black particles exposure group began to increase significantly after the 14th day of the experiment(P<0.05), the body weight of the mice in the C group, LC group and HC group tended to be stable after the 14th day of the experiment. The results of HE pathological examination showed that the alveolar spaces of mice in L group, LC group, H group and HC group were all deposited with black granules. The alveolar structure was disrupted, the arrangement was in disorder, and a large number of inflammatory cells infiltrated in lung of HC group. Compared with the control group,SOD activity of the other groups was significantly decreased(P<0.05). The GSH-Px activity of the high-dose exposure group and the high-dose exposure combined with cold exposure group was significantly lower than that of the control group (P<0.01). The MDA content in H group, LC group and HC group was significantly higher than that of the control group(P<0.01). The two factor variance analysis showed that as the administrated dose increased,SOD activity and GSH-Px activity were decreased significantly(P<0.05), and as the decrease of temperature, the MDA content of lung tissue was significantly increased(P<0.05). There was no interaction effect between 4℃ intermittent cold exposure and nano-carbon black particles exposure on the activity of SOD, GSH-Px and the content of MDA in lung tissue.Conclusion: Nano-carbon black particles combined with cold exposure might aggravate lung inflammation and increase the level of oxidative stress in mice.

Objective: 2minus 2-trinitrobenzene sulfonic acid (TNBS) copy ulcerative colitis model of rats, to explore purslane polysaccharide on ulcerative colitis rats colon organization IL6/STAT3 and the influence of the NF-kappa B, clear IL-6/STAT3 signaling pathway and the relationship of chronic inflammatory bowel disease, looking for new targets for the treatment of chronic ulcerative colitis.Methods: Fourty rats were randomly divided into control group, model group, mesalazine group, and purslane group (n=10). The colonic contents were weighed and weighed again after drying, and the colonic tissues were biopsies. The contents of serum interleukin-6 (IL-6), interleukin 1 β (IL-1 β), tumor necrosis factor α (TNF-α), nuclear transcription factor kappa B (NF-kappa B) were detected by ELISA kit. Immunohistochemical staining was used to determine the colonic MPO (myeloperoxidase). RT-PCR method was used to detect signal transduction and transcriptional activator 3 (STAT3), IL-6 mRNA.Results: Compared with model group and mesalazine group, defecation status of rats in purslane group were markedly improved, intestinal mucosa edema reduced, the serum IL-6, intestinal tissue MPO, the NF-kappa B and serum sIL-6Rα, gp130 content were decreased (P<0.01). Compared with the model group, the expression levels of STAT3 and IL-6 mRNA in the purslane group were significantly reduced (P<0.01). There was no significant difference of testing indicator mentioned above between control group and purslane group (P>0.05).Conclusion: By lowering the serum IL-6, sIL-6Rα and gp130 in rats, the purslane polysaccharides reduced the levels of MPO and NF-kappa B in the intestinal tissues, and further reduced the inflammatory response caused by the formation of sIL-6R alpha and IL-6. By using IL-6/STAT3 signaling pathway, the mRNA expression levels of STAT3 and IL-6 of rats intestinal tissues were down-regulated to inhibit the occurrence of inflammation.

Objective: To investigate the effect of white lentil polysaccharides on the apoptosis of human gastric cancer cells and its related mechanism.Methods: Gastric cancer cell lines HGC-27 and SGC-7901 were treated with 16, 8, 4, 2, 1 and 0 μg/ml white lentil polysaccharides for 24, 48 and 72 hours respectively, each experiment 3 wells. The proliferation activities of gastric cancer cells were detected with CellTiter 96^® AQueous Non-Radioactive Cell Proliferation Assay (MTS). HGC-27 and SGC-7901 cells were treated with 0 or 4 μg/ml white lentil polysaccharides for 24 hours respectively, each experiment 3 wells, then the changes of cell mitochondrial membrane potential were observed by JC-1 staining, cell cycle and cell apoptosis rates were evaluated with flow cytometry, the levels of mRNA expression of Bcl-2, caspase-3 and Bax genes were investigated with real-time quantitative PCR detecting system (QPCR).Results: After the gastric cancer cell lines HGC-27 and SGC-7901 were treated with white lentil polysaccharides, the mitochondrial membrane potential of the cells was significantly reduced, proliferation activities of the cells were inhibited obviously (P<0.01), and the inhibition degree was related with drug concentration and incubation time. Flow cytometry analysis showed that the apoptosis rates of HGC-27 and SGC-7901 cell lines were 53.15% and 38.77% respectively, which were significantly increased compared with PBS treatment groups (8.07% and 6.03%, P<0.01). And the cell cycle of these cells was not influenced significantly. The results of QPCR showed that Bcl-2 gene transcription level of HGC-27 and SGC-7901 cell lines was significantly down-regulated, and the transcription levels of Bax and caspase-3 genes were significantly increased (P<0.01).Conclusion: The white lentil polysaccharides can induce the apoptosis of gastric cancer cells lines HGC-27 and SGC-7901, and the mechanism of their apoptosis may be mediated by the regulation of Bax-Bcl-2-caspase3 pathway.

Objective: To investigate the therapeutic effects and mechanisms of Hedysari Polysaccharides (HPS) and Hedysari Polysaccharide HG-1 (HHG-1) on osteoarthritis (OA) of rats.Methods: Wistar rats were randomly divided into seven groups with eight in each as following:normal control group, model control group, positive control group(Sodium hyaluronate, SHA 1.00 mg/joint), HPS treated group (HPS 0.35 or 1.05 mg/join), and HHG-1 treated group(HHG-1 0.15 or 0.45 mg/joint). The rat's OA model was established with papain. The range of joint activity, synovial pathology and its glycosaminoglycan (GAG) content, whole blood viscosity, and serum antioxidant indexes were determined after treated with HPS and HHG-1.Results: HPS (0.35, 1.05 mg/joint) and HHG-1 (0.15, 0.45 mg/joint) could significantly enlarge the stretching angle of knee joint, relieve the pathological changes of articular cartilage, inhibit abnormal thickening of articular synovium, increase the content of GAG in synovi in the OA' rats. Meanwhile, HPS and HHG-1 could reduce the blood viscosity, enhance the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), decrease the serum level of malondialdehyde (MDA) (P<0.05, P<0.01).Conclusion: HPS and HHG-1 have some therapeutic effects on OA, but there is no significant difference between the two groups. Its possible mechanisms are improving hemorheology to alleviate abnormal cartilage metabolism and improving the activity of antioxidant enzymes to relieve cartilage injury.

Objective: To investigate the changes of autophagy in cerebral cortex neurons at different times after cerebral ischemia and ischemia/reperfusion (I/R) in rats.Methods: Healthy male SD rats (n=60) were randomly assigned into sham-operation group (sham, n=10), cerebral ischemia and ischemia/reperfusion model group (I/R, n=50). In the model group, 10 rats were randomly sampled at five different time points:ischemia 30 min, 2 h, and reperfusion 1 h, 6 h and 24 h after ischemia 2 h. Cerebral infarction volume and cerebral water content were measured. Autophagy of cerebral cortical neurons was examined by transmission electron microscopy (TEM). The protein levels of microtubule associated protein light chain 3-Ⅱ (LC3-Ⅱ) in cerebral cortex of the rats in each group were determined by Western blot.Results: LC3-Ⅱ/Ⅰ ratio had no obvious rise at ischemia 30min,and it began to increase at ischemia 2 h point which was significantly higher than that in sham group (P<0.01).LC3-Ⅱ/Ⅰ ratio was still higher at ischemia/reperfusion 1h and 6h points than that in sham group (P<0.05),although it was decreased comparison with that at ischemia 2 h point. LC3-Ⅱ/Ⅰ ratio reached a peak at ischemia/reperfusion 24 h point, which was significantly higher than that in sham group (P<0.01). This phenomenon was further confirmed by the transmission electron microscopy. The infarction volume at ischemia/reperfusion 6h and 24 h points was increased significantly, and there was a statistical difference compared with sham group (P<0.01). The content of water in the brain tissue of rats at ischemia/reperfusion 24 h was increased obviously, which was significantly higher than that in sham group (P<0.05). Edema, rarefaction, some cell degeneration and apoptosis only in the cortical tissue of rats at ischemia/reperfusion 24 h were showed,and a large number of neurons nuclei in the hippocampus was shrunken,hyperchromatic showing degeneration and apoptosis by HE staining.Conclusion: Neurons autophagy in cerebral cortices of rats with focal cerebral ischemia and ischemia/reperfusion was obviously activated at ischemia 2 h point, it was increased continuously at ischemia/reperfusion 1 h and 6 h points and reached the peak at ischemia/reperfusion 24 h point.

Objective: To observe the effects of endothelin-1 (ET-1) on monocyte chemotactic protein-1 (MCP-1) generation and the primary mechanisms in rat vascular smooth muscle cells (VSMCs).Methods: Rats VSMCs were cultured and divided into two groups:ET-1 group and inhibitor group (the latter including BQ123+ET-1, BQ788+ET-1, NAC+ET-1, PD98059+ET-1, SB203580+ET-1, SP600125+ET-1 group). ①The ET-1 group was stimulated with different concentration of ET-1 for indicated time. The VSMCs in inhibitor groups were incubated with various inhibitor, including BQ123 and BQ788 (ET_A and ET_B receptor antagonist), antioxidant NAC (N-acetyl cysteine), PD98059 (ERK inhibitor), SB203580 (p38MAPK inhibitor), SP600125 (JNK inhibitor), and PDTC (NF-κB inhibitor) for 30 minutes, then incubated with ET-1 for 24 hours. At indicated time, concentration of MCP-1 protein and expression of MCP-1 mRNA were determined by ELISA and RT-PCR respectively. ②The VSMCs were incubated with inhibitors (including BQ123, BQ788, NAC, PD98059, SB203580, and SP600125 for 20 minutes, then incubated with ET-1 for 5 minutes. The levels of ERK, p38MAPK, JNK, and corresponding phosphorylated protein of them (i.e. p-ERK, p-p38MAPK, p-JNK) were determined by Western blot. Each assay was repeated by three independent experiments.Results: The results showed that ET-1 could stimulate MCP-1 generation in VSMCs both in protein and in mRNA levels in concentration-dependence manner(P<0.05, P<0.01). BQ123, NAC, PD98059, SB203580 and PDTC, but not BQ788 and SP600125, could inhibite ET-1-stimulated protein and mRNA expressions of MCP-1 in VSMCs (P<0.01). In addition, BQ123, NAC, and PD98059 or SB203580 could suppress ET-1 induced ERK and p38MAPK activation respectively (P<0.05, P<0.01), but there was no increase of phospho-JNK in ET-1 treated VSMCs.Conclusion: The results demonstrate that ET-1 induces MCP-1 production in VSMCs via ET_A receptor and subsequent ROS, ERK, p38MAPK, NF-κB signal pathway.