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BACKGROUND:Previous studies showed that extracts of Sambucus adnata Wall.have the ability to promote the proliferation and differentiation of osteoblasts,fracture healing,anti-inflammatory and antioxidant effects,which can effectively alleviate the development of osteoarthritis.Vascular endothelial growth factor,on the other hand,is a biomarker for the evaluation of osteoarthritis severity. OBJECTIVE:To investigate the effect and mechanism of two extracts of Sambucus adnata Wall.(methanol extract SAW-ME and dichloromethane extract SAW-DCE)on angiogenesis in osteoarthritis. METHODS:(1)Rat models of osteoarthritis were established using anterior cruciate ligament transection and given SAW-ME and SAW-DCE.A sham group was set as a control.Immunohistochemistry and immunofluorescence were used to detect the changes of articular vascular endothelial growth factor A in joint tissue and vascular endothelial growth factor and"H"type blood vessels in serum of osteoarthritis rats.(2)Vascular endothelial cells EA.hy926 were used as the research object and intervened with SAW-ME and SAW-DCE.Cell proliferation was then detected by MTT assay.Vascular endothelial growth factor was used to induce EA.hy926 cells,and the model of angiogenesis was replicated.Cell scratch assay and tube formation assay were performed to study the role and mechanism.(3)EA.hy926 cells were used for transcriptome sequencing to analyze the characteristic changes of cell differential genes and related functions after SAW-DCE intervention. RESULTS AND CONCLUSION:(1)SAW-ME and SAW-DCE downregulated the expression of vascular endothelial growth factor A in the rat knee cartilage and reduced the formation of"H"type vessels in osteoarthritis rats.SAW-ME could significantly decrease the level of vascular endothelial growth factor in serum of osteoarthritis rats(P<0.05).SAW-DCE could also decrease the level of vascular endothelial growth factor in serum of osteoarthritis rats,but there was no significant change.(2)Both SAW-ME and SAW-DCE significantly inhibited vascular endothelial cell migration and tube formation,and downregulated the expression of Ang1 and Tie2 proteins.(3)Transcriptome sequencing analysis found that abnormal angiogenesis in osteoarthritis was related to the PI3K/AKT signaling pathway.(4)To conclude,SAW-ME and SAW-DCE can inhibit angiogenesis in the rat model of osteoarthritis,and the mechanism may be related to the Ang1/Tie2 and PI3K/AKT signaling pathways.
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Objective:To explore whether different phospholipid peptides could induce different pathological features of EAE in Lewis rats.Methods:Lewis rats were immunized with myelin basic protein 82-99 (MBP82-99),MBP68-86,and myelin oligodendroglia glycoprotein 35-55 (MOG35-55),respectively,and evaluated everyday for neurological scores.The cerebrum,brain stem,cerebellum and spinal cord of every rat were removed and pathological features observed.Results: Rats immunized with the two MBP peptides exhibited neurological signs of EAE wherein the central nervous system had extensive inflammation infiltration.The number of infiltrates in spinal cords in the two MBP peptides groups was higher than that in the cerebrums,brain stems and cerebellums.In spinal cords, there was no statistical difference of infiltrates between MBP82-99 and MBP68-86 groups;however,the both groups had more infiltrates than MOG35-55 group.Inflammatory cells were observed only in spinal cords of animals immunized with MOG35-55.Conclusion:This study provides certain evidence for understanding the diversity of pathological manifestations of EAE in Lewis rats.
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Objective To investigate the expression of connective tissue growth factor( CTGF) and the effects of azithromycin in airway remodeling in asthmatic mice.Methods BALB/c female mice were randomly divided into 3 groups;control group(A),asthma group(B) ,and azithromycin treated group(C),with 10 mice in each group.Mice were sensitized and challenged continually with ovalbumin( OVA).The distribution and types of collagens were detected by cirius red staining.Lung hydroxyproline content was determined by acid hydrolysis.CTCF expression in mice pulmonary tissues was detected by SABC immunohistochemistry.Results After continual challenging with OVA,collagen hyperplasia was found in the airway wall and CTCF positive expression in airway epithelium.CTCF expression was closely associated with lung hydroxyproline content(r = 0.65,P <0.01).Conclusion Asthma continually challenged with OVA could result sub-epithelial fibrosis and remodeling in airway,which could be associated with the upregulation of CTCF over-expression.Azithromycin inhibited airway remodeling in asthmatic mice,possibly by inhibiting the expression of CTGF.
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Objective To compare the activation of microglia in vitro induced by oligomeric and fibrillar Aβ,and research the effects of Piper Kadsura Ohwi extracts on the microglial activation.Methods Microglia were divided randomly into 4 test groups,intervened by fibrillar Aβ25-35,fibrillar Aβ25-35 + Piper Kadsura Ohwi extracts,oligomeric Aβ25-35 and oligomeric Aβ25-35 + Piper Kadsura Ohwi extracts respectively.Also a blank contrast group was set without any intervention.48 hours later,activation of microglia was tested by immunocytochemistry,using the CD68 molecule as a specific marker of microglial activation and the incidences of active microglia in different groups were compared.Results Each of the 4 test groups had a higher positive incidence of CD68 expression than that of the blank contrast (5.1％ ) (P ＜ 0.05 ) ; positive incidence of fibrillar Aβ + Piper Kadsura Ohwi extracts group (52.1％) was lower than that of the fibrillar Aβ group (60.8％) (P＜0.05) ; positive incidence of oligomeric Aβ + Piper Kadsura Ohwi extracts group (67.0％) was not significantly different with that of the oligomeric Aβ group (71.2％),P=0.101.Conclusion Both fibrillar and oligomeric Aβ has the ability to activate the silent microglia.Piper Kadsura Ohwi extracts could inhibit the activation of microglia induced by fibrillar Aβ25-35,but didn't show significant effects on the activation induced by oligomeric Aβ25-35.
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Objectives To explore the impact of Aβ oligomer(Aβo) and fibrillar Aβ( Aβf) on the ethology of model rat and to explore whether or not piper kadsura ohwi(PKO) can ameliorate the ethological changes.Methods AD animal model was made by continuous intracerebroventricular infusion of Aβ through mini-osmotic pump.After surgery,rats of Aβo + PKO group and Aβf + PKO group received intraperitoneal injection of 10％PKO everyday,rats from Aβo + DMSO group and Aβf + DMSO group received intraperitoneal injection of 2.5％ DMSO.The treatment lasted 5 weeks.Morris water maze experiment began on the 31st day after surgery.Results ( 1 ) Acquisition trials:the escape latency was longer in Aβo group ( ( 89.40 ± 7.20 ) s ) than that in Aβf group ( (65.00 ± 10.89 ) s ) ; escape latency was shorter in Aβf + PKO group ( ( 34.00 ± 11.26 ) s) than that in Aβf group and Aβf + DMSO group( (60.6 ± 6.95 ) s) ; escape latency was shorter in Aβ3o + PKO group ( ( 65.33 ±8.89) s) than that in Aβo group and Aβo + DMSO group ( ( 85.60 ± 6.02) s).( 2 ) Robe trial:the times ( 3.00 ±0.71 ) and the proportion of time (0.23 ± 0.02 ) and path(0.23 ± 0.04) spent in the target quadrant in Aβo group was less than that in Aβf group(6.00 ± 1.58,0.25 ± 0.01,0.26 ± 0.03 ) ; the three parameters in Aβf + PKOgroup were more than those in Aβ3f group and Aβf+ DMSO group; the three parameters in Aβo + PKO group were more than those in Aβo group and Aβo + DMSO group.Conclusions Aβ oligomer had a more severe impact on the ethology of AD model rats than fibrillar Aβ did; piper kadsura ohwi may ameliorate the ethological changes of AD model rats.
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Objective To investigate the effects of azithromycin on airway remodeling and the expression of transforming growth factorβ1 (TGF-β1) in asthmatic mice.Methods BALB/c male mice were random divided into 3 groups:control group(A),asthma group (B),and azithromycin treated group (C),with 10 mice in each group.Bronchoalveolar lavage fluid was collected to count the number of white blood cells and eosinophilic granulocytes EOS.The morphological parameters of the bronchi were measured by computer image analysis and the pathologic changes of the bronchi and lung tissue were observed by HE staining.The expressions of TGF-β1 were detected by immunohistochemistry.Results The number of EOS in B group was significantly higher than that in control group(8.12 ±0.54 vs 0.70 ±0.40;8.12 ±0.54 vs 0.87 ±0.25,P ＜0.01).WAt and WAi and WAm in group C was significantly lower than that in group B (10.15 ±0.95 vs 15.36 ±0.85,4.16 ±0.32 vs 10.64 ± 1.03,3.77 ±0.15 vs 7.97 ±0.17,P ＜0.01)but higher than that in group A.The expression of TGF-β1 in group C was significantly lower than that in group B but higher than that in group A.TGF-β1 expression of lung tissue in C group was significantly correlated with EOS,(r =0.840,P ＜0.01) and WAt(r =0.735,P ＜0.01) and WAm (r =0.870,P ＜0.01).Conclusion Azithromycin inhibited airway remodeling in asthmatic mice,which might possibly be achieved through inhibiting the expression of TGF-β1.
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Objective To investigate the correlation between heat-shock protein 70-2 (HSP70-2) gene+1267A/G polymorphism and coronary heart disease (CHD) in Han Chinese population. Methods Using the method of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) , the polymorphism and genotype and allele distribution of HSP70-2 gene + 1267A/G in 185 CHD patients and 149 controls were analyzed. Results The HSP70-2 gene + 1267A/G polymorphism was found in this study population.The distribution of HSP70-2 genotypes was in Hardy-Weinberg equilibrium. The frequency of G allele in CHD group was significantly higher than that in control (61.89% vs51.68%, P ＜ 0. 01). After multiple logistic regression analysis, HSP70-2 gene (GG + AG) genotype was an independent risk factor of coronary heart disease. Conclusion HSP70-2 gene + 1267A/G polymorphism was associated with CHD risk of Han Chinese population, the G allele might serve as a genetic risk factor of coronary heart disease.