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1.
Article in Chinese | WPRIM | ID: wpr-906391

ABSTRACT

Objective:To observe the effect of total flavonoids from Epimedii Folium (TEF) on the angiogenesis of ischemic myocardium in rats after acute myocardial infarction (AMI) and discuss its molecular biological mechanism of attenuating myocardial ischemia and improving cardiac function. Method:AMI in rats was induced through the ligation of left anterior descending coronary artery. All male SD rats were randomized into sham-operated group, model group, diltiazem group (10 mg·kg<sup>-1</sup>·d<sup>-1</sup>), and TEF low-dose and high-dose groups (100 and 200 mg·kg<sup>-1</sup>·d<sup>-1</sup>), with 8 rats in each group. After modeling, rats in the diltiazem group and TEF groups were given corresponding doses of diltiazem and TEF, respectively, and those in the model group and sham-operated group received normal saline of equivalent volume, once a day for 7 days. After the administration, VisualSonics Vevo2100 imaging system was used to detect the cardiac structure and function and hematoxylin-eosin (HE) staining to observe the histomorphological changes in myocardial ischemic area. Immunohistochemistry was employed to analyze the expression of CD31 and <italic>α</italic>-smooth muscle actin (<italic>α</italic>-SMA) in ischemic myocardium and Western blot to detect the expression of vascular endothelial growth factor-receptor 2 (VEGF-R2) and phosphorylation of protein kinase B (Akt) in ischemic myocardium. Real-time PCR was applied to quantify the mRNA levels of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). Result:Compared with the sham-operated group, the model group demonstrated significant increase in left ventricular systolic diameter (LVIDs), left ventricular internal diameter at end-diastole (LVIDd), left ventricular end-systolic volume (LVEVs), and left ventricular end-diastolic volume (LVEVd), significant decrease in End-systolic thickness of left ventricular anterior wall (LVAWs), end-diastolic thickness of left ventricle anterior wall (LVAWd), end systolic thickness of left ventricular posterior wall (LVPWs), stroke volume (SV), ejection fraction (EF), fractional shortening (FS), and cardiac output (CO), obvious pathological changes in the ischemic myocardium, and plummet of the expression of CD31 and <italic>α</italic>-SMA (<italic>P</italic><0.01), Akt phosphorylation level, protein level of VEGF-R2, and mRNA levels of VEGF and bFGF (<italic>P</italic><0.05, <italic>P</italic><0.01). High-dose TEF significantly alleviated the pathological changes of ischemic myocardium as compared with the model group. Moreover, TEF high-dose group showed significantly lower levels of LVIDs, LVIDd, LVEVs, and LVEVd, significantly higher levels of LVAWs, LVAWd, LVPWs, SV, EF, FS, and CO, higher expression of CD31 and <italic>α</italic>-SMA (<italic>P</italic><0.05, <italic>P</italic><0.01), and higher levels of VEGF-R2 protein, phosphorylated Akt, and VEGF and bFGF mRNA than the model group (<italic>P</italic><0.05, <italic>P</italic><0.01). Conclusion:TEF can effectively improve myocardial perfusion in peri-myocardial infarction area and attenuate ventricular remodeling and heart failure after AMI by up-regulating the expression of bFGF, VEGF, and VEGF-R2 in ischemic myocardium following AMI and activating phosphatidylinositol 3-kinases (PI3K)/Akt/VEGF signaling transduction pathway which can promote angiogenesis in ischemic myocardium.

2.
Article in Chinese | WPRIM | ID: wpr-906023

ABSTRACT

Objective:To observe the clinical efficacy of modified Buyang Huanwutang combined with electroacupuncture (EA) in the treatment of traumatic spinal cord injury (TSCI) due to Qi deficiency and blood stasis. Method:Eighty-seven TSCI patients who met the inclusion requirements were randomly divided into an observation group (<italic>n</italic>=44) and a control group (<italic>n</italic>=43). On the basis of comprehensive western medical treatments, patients in the control group were further provided with Wuwei Tongshuan oral liquid,10 mL per time,three times per day, while those in the observation group received modified Buyang Huanwutang,one bag per day,for 12 consecutive weeks. Besides, EA was performed in both groups in the same way, once per day, six times per week, for six weeks in total. The American Spinal Injury Association (ASIA) motor score, modified Barthel index (MBI),visual analog scale (VAS) pain score,Berg balance scale (BBS) score,modified Ashworth scale (MAS) score, spinal cord independence measure-Ⅲ(SCIM-Ⅲ) score, lower limb range of motion (ROM), and Qi deficiency and blood stasis syndrome score before and after treatment were evaluated, followed by the recording of the occurrence of complications during treatment. The brain-derived nerve growth factor (BDNF), nerve growth factor (NGF), vascular endothelial growth factor (VEGF), neurotrophic factor-3 (NT-3), malondialdehyde (MDA) and superoxide dismutase (SOD) levels before and after treatment were determined. Result:The motor, light touch, needling sensation, MBI, and BBS scores of the observation group were higher than those of the control group (<italic>P</italic><0.01), while the AS and MAS scores were lower(<italic>P</italic><0.01). The angles of adductor and straight leg raising in the observation group were greater than those of the control group (<italic>P</italic><0.01),but the Qi deficiency and blood stasis syndrome score was lower(<italic>P</italic><0.01). Both the scores of self-care, respiration, and sphincter management in SCIM-Ⅲ and the total score in the observation group were elevated as compared with those of the control group (<italic>P</italic><0.01). The cumulative incidence of complications in the observation group was 34.09%,significantly lower than 55.81% in the control group (<italic>χ</italic><sup>2</sup>=4.149,<italic>P</italic><0.05). Compared with the control group, the observation group exhibited remarkably increased BDNF, NGF, VEGF, NT-3, and SOD (<italic>P</italic><0.01) and decreased MDA (<italic>P</italic><0.01). Conclusion:Modified Buyang Huanwutang combined with EA is effective in alleviating spinal cord injury, promoting neural functional recovery, improving independence in activities of daily living, reducing the incidence of complications of patients with TSCI, which may be related to the amelioration of ischemia and hypoxia, inhibition of lipid peroxidation, and acceleration of nerve cell repair and regeneration.

3.
Article in Chinese | WPRIM | ID: wpr-888151

ABSTRACT

This study aimed to observe the inhibitory effect of icariin against oxidative stress-induced calcification in aortic vascular smooth muscle cells(VSMCs) and elucidate the molecular mechanism of icariin in inhibiting endoplasmic reticulum stress(ERS)-mediated atherosclerotic calcification, so as to provide new ideas for exploring the anti-atherosclerotic mechanism of Epimedii Folium. The VSMCs in rat thoracic aorta were subjected to adherent culture and then treated with the complete calcification DMEM containing high glucose and hydrogen peroxide(H_2O_2) for three weeks. The resulting calcified VSMCs were divided into different treatment groups. Icariin was added one week after calcification induction for protecting the VSMCs, whose viability was then detected using cell counting kit-8(CCK-8). Alizarin red-S staining was conducted to observe the calcification degree. The activity of alkaline phosphatase(ALP) in VSMCs was measured using the disodium phenyl phosphate substrate and the calcium content was measured by arsenazo Ⅲ method. The mRNA expression levels of ossification-related factors including osteocalcin(OC), osteopontin(OPN), Runt-related transcription factor 2(Runx2), and type Ⅰ collagen(Col Ⅰa) were detected by real-time PCR. Western blot was carried out to determine the protein expression levels of α-smooth muscle actin(α-SMA), Runx2, activating transcription factor 4(ATF4), and eukaryotic translation initiation factor(eIF)-2α. The results showed that H_2O_2 significantly induced the calcification of VSMCs, increased the ALP activity and calcium content in VSMCs, promoted OC, OPN, Runx2, and Col Ⅰa mRNA expression and Runx2 protein expression, and reduced α-SMA protein expression. The ATF4 protein expression and eIF2α phosphorylation were also elevated significantly. Icariin reversed the calcification of VSMCs induced by H_2O_2, inhibited ALP activity and calcium content in VSMCs, down-regulated the mRNA expression levels of OC, OPN, Runx2 and Col Ⅰa and Runx2 protein expression, and relatively up-regulated the expression of α-SMA. The expression of ATF4 and phosphorylation of eIF2α also declined significantly. All these have demonstrated that icariin inhibited VSMCs calcification by down-regulating the ossification-related factors and lowering ALP activity and calcium content in VSMCs. Besides, the down-regulation of Runx2 expression and the inhibition of ATF4 and eIF2α-mediated cellular calcification pathway in ERS might also be involved in such calcification-suppressing process.


Subject(s)
Animals , Cells, Cultured , Flavonoids/pharmacology , Muscle, Smooth, Vascular/metabolism , Myocytes, Smooth Muscle , Oxidative Stress , Rats
4.
Article in English | WPRIM | ID: wpr-880540

ABSTRACT

OBJECTIVE@#To evaluate the effect of Guilu Erxian Glue (, GEG) on cyclophosphamide (CTX)-induced bone marrow hematopoietic stem cells (HSCs) senescence in mice and explore the underlying mechanism.@*METHODS@#The H@*RESULTS@#Compared with the model group, GEG increased cell viability as well as proliferation (P<0.05 or P<0.01) and reduced β -gal expression. Furthermore, GEG significantly decreased the expressions of p16@*CONCLUSION@#GEG can alleviate CTX-induced HSCs senescence in mice, and the p16

5.
Article in Chinese | WPRIM | ID: wpr-700003

ABSTRACT

Revolution energy spectral CT had its structural innovation introduced from the aspects of detector,driving system and CT bulb as well as high voltage generator,technical principle analyzed from the aspects of SSF,multi-model iterative reconstruction, cardiac imaging unlimited as well as material separation and quantitative analysis, and clinical application described in diagnoses of liver cancer, cholecystolithiasis and kidney stone, coronary arteriongraphy and metal artifact elimination.It's pointed out Revolution energy spectral CT was a new method for identifying the focal nature,tumor homology and components of inorganic substance as well as analyzing multi material quantitatively and qualitatively.

6.
Article in Chinese | WPRIM | ID: wpr-360146

ABSTRACT

<p><b>OBJECTIVE</b>To examine whether transforming growth factor-β (TGF-β) pathway and adaptive T cell immunity play roles in the anti-atherosclerotic effects of pioglitazone (PIO) in ApoEmice.</p><p><b>METHODS</b>ApoEmice with atherosclerosis induced by high-fat feeding were treated daily with PIO (20 mg/kg) or vehicle for 8 weeks. The protein expressions of TGF-β pathway in the atheromatous lesions of the aorta and the percentages of IFN-γand Foxp3cells in the spleen of the mice were examined with immunohistochemical staining. In the in vitro experiment, primary cultured splenocytes were stimulated with oxidized low-density lipoproteins (oxLDL) and treated with PIO either alone or in combination with the PPARγ antagonist GW9662, after which the changes in percentages of CD4IFN-γcells and CD4CD25Foxp3cells were analyzed with flow cytometry.</p><p><b>RESULTS</b>PIO treatment of ApoEmice with high-fat feeding significantly attenuated the progression of atheromatous lesions (P<0.05) and resulted in increased expressions of TGFβ1 (P<0.01), TGFβRII (P<0.05), and p-Smad3 (P<0.05) and a decreased expression of Smad7 (P<0.05) in the lesions. PIO treatment also led to decreased percentage of IFN-γcells (P<0.05) and increased percentage of Foxp3cells (P<0.01) in the spleen of the mice. In primary cultured splenocytes, PIO treatment caused significant down-regulation of IFN-γ mRNA (P<0.05) and up-regulation of Foxp3 mRNA (P<0.05) and obviously increased the percentages of CD4IFN-γcells (P<0.05) and CD4CD25Foxp3(P<0.05); the effects of PIO on CD4IFN-γand CD4CD25Foxp3cells were abolished by treatment of the cells with GW9662.</p><p><b>CONCLUSION</b>The anti-atherosclerotic effect of PIO is probably mediated by the TGF-β/Smad signaling pathway and PPAR-γ-dependent modulation of Th1/Treg population.</p>

7.
Article in Chinese | WPRIM | ID: wpr-360142

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of postischemic treatment with endomorphin-1 (EM-1) against myocardial ischemia/reperfusion (IR) injury in rats and on extracellular signal regulated kinase 1/2 (Erk1/2)-dependent signaling pathway.</p><p><b>METHODS</b>Sprague-Dawley rats were randomly divided into 5 groups, namely the sham-operated group, IR group, EM-1 post-treatment group (EM50 group), EM-1 post-treatment group with PD98059 treatment (EM50+PD group), and PD98059 post-treatment group (PD group). The hemodynamic indexes of the rats were recorded. After reperfusion, CK-MB, LDH, CTnI, MDA, IL-6, TNF-α, and SOD activities or contents were measured, the infarct size was determined, and the expression levels of Erk1/2, P-Erk1/2 and cleaved caspase-3 were detected using Western blotting.</p><p><b>RESULTS</b>Compared with the sham group, the IR group showed significantly decreased heart rate and mean arterial pressure (P<0.05), which were increased obviously by EM-1 post-treatment (P<0.05). EM-1 post-treatment also resulted in significantly decreased LDH, CK-MB, CTnI, MDA, IL-6, and TNF-α activities or contents (P<0.05), increased SOD activity (P<0.05), reduced the infarct size (P<0.05), and increased the expression level of P-Erk protein (P<0.05). Compared with EM50 group, EM50+PD group showed significantly decreased heart rate and mean arterial pressure (P<0.05), increased LDH, CK-MB, CTnI, MDA, IL-6, and TNF-α activities or contents (P<0.05), decreased SOD activity, increased infarct size (P<0.05), and lowered expression of P-Erk protein (P<0.05).</p><p><b>CONCLUSION</b>Postischemic treatment with EM-1 protects the heart against IR injury by improving the cardiac function, inhibiting inflammation, and inhibiting oxidative stress and myocardial apoptosis, and Erk1/2 signaling pathway may be involved in this process.</p>

8.
Article in Chinese | WPRIM | ID: wpr-307146

ABSTRACT

To investigate the effects and mechanisms of total flavones of Epimedium (TFE) on oxidative stress induced by myocardial ischemia/reperfusion injury in rats, forty male SD rats were randomly divided into sham operated group, model group, diltiazem group and flavonoids of Epimedium low and high doses groups with 8 rats in each. Myocardial ischemia/reperfusion injury model was induced by ligaturing the left anterior descending artery for 30 min followed reperfusion for 4 h after TFE was taken by intragastric administration for 4 days. The degree of myocardial infarct was observed by N-BT staining. The concentrations of MDA and activities of SOD and T-AOC in cardiac tissue were measured by colorimetry. Serum TnI concentrations were checked by ELISA. HE stain was used to observe myocardium structure under light microscope. Expressions of SIRT1 and Nrf2 in cardiac tissue were evaluated by immunohistochemistry method and Western blot, respectively. Compared with the model group, the degree of myocardial infarct, MDA concentration in cardiac tissue and the levels of TnI in serum significantly decreased in the diltiazem group and flavonoids of Epimedium low and high doses groups (P<0.05 or P<0.01); flavonoids of Epimedium low and high doses groups and the diltiazem group also showed improvements in myocardium structure under ischemia/reperfusion injury. TFE significantly increased the activity of SOD and T-AOC and the expression of SIRT1 and Nrf2 in cardiac tissue when compared with the model group (P<0.05 or P<0.01). Therefore, TFE can increase anti-peroxidant capacity of myocardium tissue by using intrinsically anti-oxidant signaling pathway of SIRT1 and Nrf2, which can inhibit irreversible damage of cardiomyocytes in myocardial ischemia/reperfusion injury and protect normal function of cardiac tissue.

9.
Article in Chinese | WPRIM | ID: wpr-345619

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the relationship of leptin gene polymorphism with obesity in ethnic minority Hui and Uygur children in China.</p><p><b>METHODS</b>Sixty-eight ethnic minority (35 Hui and 33 Uygur) children with obesity and 69 age-matched minority (36 Hui and 33 Uygur) children without obesity were recruited from six primary schools in the sub-urban areas of Urumqi. Venous blood was sampled from all subjects after fasting for 12 hours. Leptin gene C2549A polymorphism was determined by polymerase chain reaction (PCR) and restriction fragment length polymorphism methods. Blood concentrations of lipids, leptin and insulin were measured with biochemical methods and radioimmunoassys, respectively.</p><p><b>RESULTS</b>In the 137 children tested, the prevalence of AA, AC and CC genotype was 9.5%, 33.6% and 56.9%, respectively. A allele frequency was significantly different between the two ethnic (i.e. Hui and Uygur) groups (P<0.05). A allele frequency and AA+ AC genotype frequency were not significantly different between obese and non-obese children in both ethnic groups (P>0.05). Blood leptin levels were not significantly different between obese and non-obese children with an AA+AC or CC genotype in both ethnic groups (P>0.05).</p><p><b>CONCLUSIONS</b>Leptin gene polymorphisms exist in Hui and Uygur children. The C2549A polymorphism is not significantly associated with the prevalence of obesity in both Hui and Uygur children.</p>


Subject(s)
Child , China , Ethnology , Female , Genotype , Humans , Leptin , Blood , Genetics , Lipids , Blood , Male , Obesity , Blood , Genetics , Polymorphism, Genetic
10.
Article in Chinese | WPRIM | ID: wpr-294366

ABSTRACT

<p><b>OBJECTIVE</b>To observe the clinical effect of Guilu Erxian Glue Cataplasm (GEGC) on carcinoma of the large intestine patients with myelosuppression after chemotherapy, and further to confirm its efficiency and safety.</p><p><b>METHODS</b>Totally 60 patients with carcinoma of the large intestine were randomly assigned to two groups. Meanwhile, they all accepted FOLFIRI chemotherapy. Patients in the treatment group were additionally applied at Shenque (RN8), exchanging once per every other day, for 14 successive days. Patients in the control group took placebos with the same dose and dosage as the treatment group. The blood cell counts (WBC, NE, and PLT) were detected before chemotherapy, at day 7, 10, and 14. The TCM symptoms integrals, Karnofsky performance score (KPS), liver and kidney functions were observed before chemotherapy, at day 7 and day 14. Adverse skin reactions were observed each day. And the usage of hematopoietic growth factors was recorded.</p><p><b>RESULTS</b>(1) The KPS score at day 7 was more stable in the treatment group than in the control group; the WBC and NE counts in the peripheral blood at day 14 were higher in the treatment group than in the control group; and TCM symptoms integrals at day 14 was lower in the treatment group than in the control group, all with statistical difference (P < 0.05). (2) Compared with the control group, the PLT count was higher in the treatment group than in the control group, the usage of rhG-CSF and antibiotics was less in the treatment group than in the control group, all with no statistical difference (P > 0.05). (3) No obvious adverse reactions such as liver injury, renal injury, or skin allergy were observed.</p><p><b>CONCLUSIONS</b>Adjuvant treatment of GEGC could improve carcinoma of the large intestine patients with myelosuppression to some extent. No relevant adverse reactions were found.</p>


Subject(s)
Adjuvants, Immunologic , Therapeutic Uses , Adult , Aged , Bone Marrow Diseases , Drug Therapy , Colorectal Neoplasms , Drug Therapy , Drugs, Chinese Herbal , Therapeutic Uses , Female , Humans , Male , Middle Aged
11.
Article in Chinese | WPRIM | ID: wpr-251710

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the antioxidative effects of two cysteinyl leukotriene receptors antagonists (CysLT1R and CysLT2R) montelukast and HAMI 3379 on ischemic injury of rat cortical neurons in vitro.</p><p><b>METHODS</b>Cultured rat cortical neurons were pretreated with CysLT1R antagonist montelukast and CysLT2R antagonist HAMI 3379, and then exposed to oxygen-glucose deprivation/recovery (OGD/R)or H2O2. Reactive oxygen species (ROS) mitochondrial membrane potential (MMP) depolarization, neuronal viability and lactate dehydrogenase (LDH) release were determined. Meanwhile, RNA interference was used to inhibit the expression of CysLT1R and CysLT2R,and the effects were observed.</p><p><b>RESULTS</b>ROS production in neurons was significantly increased after 1 h OGD, which reached the peak at 30 min and lasted for 1.5 h after recovery. Montelukast and HAMI 3379 at 0.01-1μmol/L moderately decreased OGD/R-induced ROS production (P<0.05). Montelukast mildly attenuated OGD/R-induced MMP depolarization (P<0.05),but HAMI 3379 had no effect. H2O2 reduced neuronal viability and increased LDH release, namely inducing neuronal injury. Montelukast and HAMI 3379 at 0.1-1μmol/L moderately attenuated H2O2-induced neuronal injury (P<0.05). However, both CysLT1R siRNA and CysLT2R shRNA did not significantly affect the responses mentioned above.</p><p><b>CONCLUSION</b>In ischemic neuronal injury, montelukast and HAMI 3379 exert a moderate antioxidative effect, and this effect may be receptor-independent.</p>


Subject(s)
Acetates , Pharmacology , Animals , Antioxidants , Pharmacology , Cell Hypoxia , Cell Survival , Cells, Cultured , Cerebral Cortex , Cell Biology , Cyclohexanecarboxylic Acids , Pharmacology , Leukotriene Antagonists , Pharmacology , Neurons , Metabolism , Phthalic Acids , Pharmacology , Quinolines , Pharmacology , Rats , Reactive Oxygen Species , Metabolism
12.
Article in Chinese | WPRIM | ID: wpr-251709

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the protective effect of histone deacetylase inhibitor NL101 on L-homocysteine (HCA)-induced toxicity in rat neurons, and the toxic effect on normal rat neurons.</p><p><b>METHODS</b>In the presence of NL101 at various concentrations, HCA (5 mmol/L)-induced changes in cell density, necrosis, and viability were determined in the mixed cultures of rat cortical cells and the primary cultures of rat neurons. The direct effect of NL101 on primary neurons was also observed in the absence of HCA. Histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA) was used as the control. After the treatments, cell viability, the density, and morphology of neurons and glial cells, and cell necrosis were determined.</p><p><b>RESULTS</b>In the mixed cultures of cortical cells, NL101 had no effect on HCA (5 mmol/L)-induced cell number reduction at 0.001-10μmol/L; however, it significantly attenuated necrosis at 1-10 μmol/L, and increased neuronal number at 1 μmol/L. NL101 had no effect on the mixed cortical cells in the absence of HCA. In the primary neurons, NL101 reduced neuronal viability and mildly increased necrosis at 1-10 μmol/L in the absence of HCA, while it significantly attenuated HCA-induced neuronal viability reduction at 0.01-10 μmol/L and reduced neuronal necrosis at 1-10 μmol/L. The effects of NL101 were apparently similar to those of SAHA.</p><p><b>CONCLUSION</b>NL101 has protective effect on HCA-induced neuronal injury but it is neurotoxic at high concentrations, which is similar to the typical histone deacetylase inhibitor SAHA.</p>


Subject(s)
Animals , Cell Survival , Cells, Cultured , Histone Deacetylase Inhibitors , Pharmacology , Neurons , Rats
13.
Article in Chinese | WPRIM | ID: wpr-251707

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the effect of water channel aquaporin 4 (AQP4) on bleomycin-induced lung fibrosis in mice.</p><p><b>METHODS</b>In wild type and AQP4 gene knockout (AQP4-/-) mice, lung fibrosis was induced by injection of bleomycin (3 mg/kg) into the trachea and saline injection was used as a control. At d3, 7, 14, 28 after bleomycin-treatment, mice were randomly sacrificed in batch and the lung coefficient was determined. Serum levels of TGF-β1 and TNF-α were measured by ELISA and hydroxyproline contents in lung tissue were determined by Alkaline hydrolysis method. H-E staining and Masson's staining were performed to examine the pathological changes of lung tissues after bleomycin-treatment.</p><p><b>RESULTS</b>On d14 after bleomycin-treatment, the lung coefficients in wild type mice and AQP4-/- mice were 1.9-fold (12.69 ± 6.05 vs 6.80 ± 0.82, q=4.204, P<0.05) and 2.3-fold (14.05 ± 5.82 vs 6.05± 0.58, q=5.172, P<0.01) of that in control, respectively, but no significant difference was found between wild type and AQP4-/- mice in the lung coefficient value (P>0.05). The hydroxyproline contents in the lung increased after bleomycin-treatment; on d28, the lung hydroxyproline contents in wild type and in AQP4-/- mice were 1.55-fold (0.85 ± 0.22 g/mg vs 0.55 ± 0.14 μg/mg, q=4.313, P<0.05) and 1.4-fold (0.84 ± 0.13 μg/mg vs 0.60 ± 0.14μg/mg, q=4.595,P<0.05) of that in control, respectively, but no significant difference was noticed between wild type and AQP4-/- mice in lung hydroxyproline contents. There was a tendency that serum TGF-β1 and TNF-α levels increased in bleomycin-treated mice, but no significant difference was found between wild type and AQP4-/- mice. AQP4-knockout showed no effects on pathological changes of lung tissues with H-E staining and Masson's staining in mice with bleomycin-induced lung fibrosis.</p><p><b>CONCLUSION</b>AQP4 might not be involved in bleomycin-induced lung fibrosis in mice.</p>


Subject(s)
Animals , Aquaporin 4 , Genetics , Bleomycin , Toxicity , Male , Mice , Mice, Knockout , Pulmonary Fibrosis , Genetics
14.
Article in Chinese | WPRIM | ID: wpr-251706

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of cysteinyl leukotriene (CysLT) receptor agonist leukotriene D4 (LTD4) on proliferation and migration in lung epithelial A549 cells.</p><p><b>METHODS</b>The expression of CysLT1 receptor and CysLT2 receptor was determined by immunofluoresence staining in A549 cells. A549 cells were treated with LTD4 (0.01-100 nmol/L) for 24-72 h. Cell viability was detected by MTT reduction assay. Cell migration was determined by modified scratch and healing model.</p><p><b>RESULTS</b>In A549 cells, CysLT1 receptor and CysLT2 receptor were mainly expressed in the cytoplasm, membrane and few in the nuclei. The treatment of LTD4 (0.01-100 nmol/L) for 24-72 h caused no effect on cell viability (Ps>0.05); when A549 cells were treated with 100 nmol/L LTD4 for 24, 48 and 72 h the cell viability was (103.00±4.46)%,(107.00±9.45)% and (105.00±9.02)% of control, respectively (Ps>0.05). The migration rate of A549 cells after scratching during the first 24 h was markedly greater than that during the second and third 24 h in the same concentration groups; however, no significant difference in migration rate was noticed when the cells were treated with different concentrations of LTD4 (0.01-100 nmol/L)(Ps>0.05). The migration of A549 cells was 1.15-fold, 1.21-fold and 1.06-fold of that of control when the cells were treated with 100 nmol/L LTD4 for 24, 48 and 72 h, respectively (Ps>0.05).</p><p><b>CONCLUSION</b>The proliferation and migration of A549 cells are not changed when treated with 0.01-100 nmol LTD4 for up to 72h.</p>


Subject(s)
Cell Line , Cell Movement , Cell Proliferation , Epithelial Cells , Cell Biology , Humans , Leukotriene D4 , Pharmacology , Pulmonary Alveoli , Cell Biology
15.
Article in Chinese | WPRIM | ID: wpr-251701

ABSTRACT

<p><b>OBJECTIVE</b>To establish a method for screening nicotinamide phosphoribosyl transferase (NAMPT) inhibitors based on endogenous fluorescence determination.</p><p><b>METHODS</b>The double mutants of NAMPT, G355C/D393C, was cross-linked by using 1, 4-Bismaleimidobutane (BMB) to block the entrance of enzymatic active site of NAMPT. The binding of compounds to NAMPT was evaluated according to the change of spontaneous fluorescence of NAMPT and BMB-NAMPT with 280 nm excitation and 333 nm emmision. The in vitro enzamatic activity of NAMPT was determined by nuclear magnetic resonance. The cell viability was determined by MTT assay.</p><p><b>RESULTS</b>FK866 significantly decreased the spontaneous fluorescence of NAMPT but not of BMB-NAMPT. Rosmaric, cynarine and 1, 3-dicaffeoylquinic acid also decreased the spontaneous fluorescence of both NAMPT and BMB-NAMPT. However, the inhibition on two proteins was equivalent. FK866 significantly inhibit the catalysis of NAMPT. Rosmarinic acid, cynarine and 1, 3-dicaffeoylquinic acid failed to inhibit the catalysis of NAMPT. FK866 inhibited the viability of A549 cells, but rosmarinic acid, cynarine and 1, 3-dicaffeoylquinic acid did not.</p><p><b>CONCLUSION</b>Endogenous fluorescence spectrometry based on NAMPT and BMB-NAMPT protein can be used for screening compounds that bind with NAMPT, and distinguishing the binding site - either within the enzymatic active site or not. Rosmarinic acid, cynarine and 1, 3-dicoffeoylquinic acid can bind to NAMPT out its enzymatic active site.</p>


Subject(s)
Apoptosis , Cell Line, Tumor , Drug Evaluation, Preclinical , Methods , Fluorescence , Humans , Nicotinamide Phosphoribosyltransferase
16.
Article in Chinese | WPRIM | ID: wpr-251698

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the application of locomotor activity test in functional injury after global cerebral ischemia (GCI) in C57BL/6 mice.</p><p><b>METHODS</b>GCI was induced by bilateral carotid arteries occlusion for 30 min in C57BL/6 mice. Mice were divided into sham group, GCI group and minocycline group. Saline or minocycline (45 mg/kg) was i.p. injected once daily for 6 d after ischemia. At Day 6 after ischemia, locomotor activity was recorded for 1 h in open field test. Total distance, central distance, central distance ratio, periphery distance, periphery distance ratio, central time and periphery time were used to evaluate the behavior characteristics of locomotor activity in C57BL/6 mice after ischemia. The survival neuron density was detected by Nissl staining in hippocampus, cortex and striatum.</p><p><b>RESULTS</b>Compared with sham group, total distance, central distance and central time increased and periphery time decreased in C57BL/6 mice after GCI (Ps<0.05). However, minocycline significantly reduced the central distance and central time and increased the periphery time (Ps<0.05). Neurons were damaged in hippocampus, cortex and striatum after GCI, which manifested by decreased neurons and the most serious damage in hippocampal CA1 region. Minocycline significantly improved the neuron appearance and increased the neuron number in hippocampus and striatum (P<0.001 or P<0.05).</p><p><b>CONCLUSION</b>Locomotor activity in open field test can objectively evaluate the behavior injury after GCI in mice. Central distance and central time can be used as indexes of quantitative assessment.</p>


Subject(s)
Animals , Apoptosis , Brain Ischemia , Disease Models, Animal , Mice , Mice, Inbred C57BL , Motor Activity , Physiology , Neurons , Pathology , Reperfusion Injury
17.
Article in Chinese | WPRIM | ID: wpr-251697

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the efficacy of novel object recognition (NOR) test in assessment of learning and memory ability in ICR mice in different experimental conditions.</p><p><b>METHODS</b>One hundred and thirty male ICR mice were randomly divided into 10 groups: 4 groups for different inter-trial intervals (ITI: 10 min, 90 min, 4 h, 24 h), 4 groups for different object materials (wood-wood, plastic-plastic, plastic-wood, wood-plastic) and 2 groups for repeated test (measured once a day or every 3 days, totally three times in each group). The locomotor tracks in the open field were recorded. The amount of time spent exploring the novel and familiar objects, the discrimination ratio (DR) and the discrimination index (DI) were analyzed.</p><p><b>RESULTS</b>Compared with familiar object, DR and DI of novel object were both increased at ITI of 10 min and 90 min (P<0.01). Exploring time, DR and DI were greatly influenced by different object materials. DR and DI remained stable by using identical object material. NOR test could be done repeatedly in the same batch of mice.</p><p><b>CONCLUSION</b>NOR test can be used to assess the learning and memory ability in mice at shorter ITI and with identical material. It can be done repeatedly.</p>


Subject(s)
Animals , Learning , Male , Memory , Mice , Mice, Inbred ICR , Time Factors
18.
Article in Chinese | WPRIM | ID: wpr-344754

ABSTRACT

<p><b>OBJECTIVE</b>To discuss the feasibility of vascular bundle implantation combined with allogeneic bone marrow stromal cells (BMSCs) transplantation in treating rabbit femoral head osteonecrosis and bone defect, in order to explore a new method for the treatment of femoral head necrosis.</p><p><b>METHODS</b>Thirty-six New Zealand rabbits were randomly divided into three groups,with 12 rabbits in each group. Bilateral femoral heads of the rabbits were studied in the experiment. The models were made by liquid nitrogen frozen, and the femoral heads were drilled to cause bone defect. Group A was the control group,group B was stem cells transplantaion group of allograft marrow stromal,and group C was stem cells transplantation group of allograft marrow stromal combined with vascular bundle implantation. Three rabbits of each group were sacrificed respectively at 2, 4, 8, 12 weeks after operation. All specimens of the femoral heads were sliced for HE staining. Furthermore ,vascular density and the percentage of new bone trabecula of femoral head coronary section in defect area were measured and analyzed statistically.</p><p><b>RESULTS</b>In group C,new bone trabecula and original micrangium formed at the 2nd week after operation; new bone trabecula was lamellar and interlaced with abundant micrangium at the 8th week;at the 12th week,the broadened,coarsened bone trabecula lined up regularly,and the mature bone trabecula and new marrow were visible. At the 2nd week after operation,there was no statistical significance in the percentage of new bone trabecula of femoral head coronary section in defect area between group B and C. While at 4, 8, 12 week after operation, vascular density and the percentage of new bone trabecula of femoral head coronary section in defect area of group C was higher than that of group B.</p><p><b>CONCLUSION</b>Allogeneic bone marrow stromal cells cultured in vivo can form new bone trabecula, and can be applied to allotransplant. Vascular bundle implanted into the bone defect area of femoral head necrosis could improve blood supply, and promote the formation of bone trabecula.</p>


Subject(s)
Animals , Blood Vessels , Transplantation , Combined Modality Therapy , Female , Femur Head Necrosis , Pathology , General Surgery , Male , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells , Rabbits , Transplantation, Homologous
19.
Article in Chinese | WPRIM | ID: wpr-252636

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of CysLT receptor agonist leukotriene D4(LTD4) and antagonists on activation of microglia BV2 cells.</p><p><b>METHODS</b>The expression of CysLT1 and CysLT2 protein was determined by Western blotting and immunostaining in microglia BV2 cells. BV2 cells were pretreated with or without CysLT1 receptor selective antagonist montelukast, CysLT2 receptor selective antagonist HAMI 3379, or CysLT1/CysLT2 receptor dual antagonist BAY u9773 for 30 min, then the cells were treated with LTD4 for 24 h. Cell viability was detected by MTT reduction assay. Phagocytosis and mRNA expression of IL-6 were determined by fluorescent bead tracking and RT-PCR, respectively.</p><p><b>RESULTS</b>In BV2 cells, LTD4 did not affect proliferation but significantly enhanced phagocytosis and increased IL-6 mRNA expression in a concentration-dependent manner. LTD4 at 100 nmol/L induced a 1.4-fold increase of phagocytic index and a 2-fold up-regulation of IL-6 mRNA expression (P<0.01). HAMI 3379 and BAY u9773 (100 nmol/L) further increased LTD4-induced phagocytosis; BAY u9773 and montelukast decreased LTD4-induced IL-6 mRNA expression, while HAMI 3379 had no effect on that.</p><p><b>CONCLUSION</b>LTD4 activates BV2 cells in vitro and enhances IL-6 mRNA expression mediated by CysLT1 receptor, LTD4 induces phagocytosis which might be negatively regulated by CysLT2 receptor in BV2 cells.</p>


Subject(s)
Acetates , Pharmacology , Cell Line , Cell Proliferation , Cyclohexanecarboxylic Acids , Pharmacology , Humans , Interleukin-6 , Metabolism , Leukotriene Antagonists , Pharmacology , Leukotriene D4 , Pharmacology , Microglia , Cell Biology , Metabolism , Phagocytosis , Phthalic Acids , Pharmacology , Quinolines , Pharmacology , Receptors, Leukotriene , Metabolism , SRS-A , Pharmacology
20.
Article in Chinese | WPRIM | ID: wpr-733221

ABSTRACT

Objective To build the rat animal model of neurogenic bladder induced by spinal cord injury and investigate the changes of urodynamics and P2X1 receptor expression in bladder detrusor after the spinal shock stage.Methods A total of 40 female Wistar rats were randomly allocated into control group,suprasacral injury group and sacral injury group.Complete spinal cord transection was established by cutting out of dura at T10-L2 in suprasacral injury group.The sacral injury group was done by cutting out of dura at S2-4.Urodynamic monitor was performed on rats after the spinal shock stage(6 weeks after operation).The P2X1 receptor expression in bladder detrusor among the 3 groups was checked by using immunohistochemical method.Results The suprasacral injury group exhibited detrusor hyperreflexia during bladder filling,elevated leak point pressure and compliance.The sacral injury group exhibited detrusor lower reflection,reduced leak point pressure and elevated compliance.The level of P2X1 receptor expression in rats' detrusor of suprasacral injury group was increased,and the level of sacral injury group was reduced.Conclusions Neurogenic bladder induced by spinal cord injury reveals significant changes in urodynamic after the spinal shock stage.The enhancement of the P2X1 receptor expression in detrusor may be one of the mechanisms of detrusor hyperreflexia.

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