1.Study on the change of migration ability of dendritic cells induced by intermittent hypoxia and the mechanism of its pathway
Ke HU ; Guangwei WANG ; Yu YANG ; Guoyao XU
Chongqing Medicine 2016;45(30):4200-4202,4206
Objective By simulating the intermittent hypoxia(IH ) environment of human obstructive sleep apnea syndrome (OSAS) ,to reveal the effect of IH on migratory ability of human peripheral blood derived dendritic cells(DCs) ,and through the in‐tervention of RelB ,p38 expression in order to explore the possible mechanism of the change of DCs migration ability induced by IH . Methods DCs were divided into RelB ,p38 siRNA interfering and non interfering plasmid group before cultivation .The environment of hypoxia was created by a intermittent hypoxia cabin ,among them ,oxygen concentration was 0 .5% ,1 .5% ,5 .0% ,10 .0% ,hypox‐ia/reoxygenation time ratio was set as 1∶1 ,1∶3 ,1∶5 and 1∶9 ,while sustained oxygen was supplied to the contrast at a normal concentration of 21 .0% .The content of RelB and p38 was tested by Western blotting after culture in vitro ,migration ability of DCs was detected by invasion chamber .Results Compared with normoxia ,DCs under IH tended to have declined migratory ability , which was confirmed to be correlated with the average oxygen partial pressure level under IH .IH could promote the expression of RelB and p38 in DCs ,while the migratory ability of DCs was not reversed after intervening the expression of RelB and p38 .Conclu‐sion IH in vitro could cause a decline in migratory ability of DCs ,which may not be induced by activation of RelB or p38 in DCs .
2.Effect of Kv channel-interacting protein 1 over-expression on excitability of primary cultured hippocampal neurons
Meng LI ; Linming ZHANG ; Guoyao XU ; Yonghong YI ; Tao SU
Chinese Journal of Neuromedicine 2014;13(10):990-993
Objective To investigate the effect of Kv channel-interacting protein 1 (KCNIP1) over-expression on K+ currents and neuronal excitability in primary cultured hippocampal neurons.Methods Enhanced green fluorescent protein plasmids carried KCNIP1 (pEGFP-KCNIP1) were established; empty pEGFP vectors were used as controls; primary cultured hippocampal neurons were transfected with pEGFP-KCNIP1 and control vectors.Whole-cell patch clamp technique was used for electrophysiological recording.Results The cultured neurons transfected with pEGFP-KCNIP1 led to KCNIP1 over-expression.The amplitudes of A-type K+ currents in the KCNIP1-overexpress neurons were significantly higher than that in the control group ([0.96±0.17] nA vs.[0.72±0.09] nA,P<0.05),while no significant difference was found between the component of steady-state outwards K+ currents and controls.Current clamp analysis revealed significantly decreased frequency of evoked discharges and subthreshold membrane potential oscillations,and statistically increased membrane resistance of the hippocampal neurons in the group of KCNIP1 over-expression as compared with those in the controls (P<0.05).Conclusion Over-expression of KCNIP1 could inhibit neuronal discharges possibly via its potentiation on A-type K+ currents.
3.Screening of active fractions of Jiegu ointment for promoting fracture healing in New Zealand rabbits
Guoyao ZHENG ; Lei PENG ; Yuancui XU ; Fei WU ; Bo WU
China Pharmacy 2023;34(11):1349-1353
OBJECTIVE To screen the active site of Jiegu ointment in promoting fracture healing in New Zealand rabbits. METHODS The ethanol extract of Jiegu ointment, as well as the ethyl acetate and n-butanol parts, were prepared and mixed with honey to form a plaster with appropriate viscosity. The radial fracture model of left forelimb in New Zealand rabbit was established and divided into model control group, ethanol extract group, ethyl acetate fraction group and n-butanol fraction group, with 6 rabbits in each group. Except for model control group, rabbits of all other groups were treated with corresponding polar part of Jiegu ointment for external application, for 4 weeks. The radial fracture healing of rabbits was studied by X-ray examination. Enzyme-linked immunosorbent assay was used to detect the serum levels of interleukin-6 (IL-6), tumor necrosis factor α (TNF-α), osteocalcin (OC), vascular endothelial growth factor A (VEGFA), basic fibroblast growth factor 2 (bFGF2) and alkaline phosphatase (ALP). HE staining was adopted to observe the changes of pathological morphology of rabbit fracture site, and immunohistochemical method was used to detect the protein expression of bFGF2 in fracture site of rabbits. RESULTS The healing speed of the fracture site in the n-butanol fraction group was the fastest, followed by ethanol extract group, and the ethyl acetate fraction group was the slowest; the serum levels of TNF-α and IL-6 in n-butanol fraction group decreased the fastest, while the levels of ALP, bFGF2, OC and VEGFA increased the fastest [significant increase compared with ethanol extract group (P<0.01)]; the chondrocytes at the fracture fraction completely disappeared, forming a large number of bone marrow cavities, and the bone trabeculae in the bone marrow cavity were officially formed. The expression level of bFGF2 was also higher than ethanol extract group. CONCLUSIONS The effect of n-butanol fraction on promoting fracture healing is more significant than ethyl acetate fraction and ethanol extract, and n-butanol fraction is the active fraction of Jiegu ointment to promote fracture healing.