痘苗病毒致炎兔皮提取物促进小胶质细胞M1向M2极化、抑制炎性因子分泌的体外研究

doi:10.3969/j.issn.1006-9771.2020.06.007

Abstract

Abstract:

Objective To investigate the effect of analgecine (AGC) on inflammatory response in the cell model of ischemic stroke and its mechanism.Methods Sodium hydrosulfite (Na₂S₂O₂) combined with sugar-free culture-medium was used to stimulate the model of ischemic stroke in vitro. BV2 cells were divided into six groups: control group, control with 0.5 U/ml AGC group, oxygen deprivation and recovery (OGD/R) group, OGD/R with AGC (0.25 U/ml, 0.5 U/ml, 1 U/ml) groups. After oxygen and glucose deprivation for 1.5 hours, they were changed to normal medium and given different concentrations of AGC in OGD/R with AGC groups. After co-incubation for three hours, the cells were treated. The content of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the supernatant was detected. The expression of M1-type microglia marker CD₁₆+CD₃₂ and M2-type microglia marker CD₂₀₆ were detected with immunofluorescent staining. BV2 cells were divided into seven groups: control group, control with 0.5 U/ml AGC group, IL-4 group, IL-4 + lipopolysaccharide (LPS) + interferon (IFN)-γ group, IL-4 + LPS + IFN-γ with AGC (0.25 U/ml, 0.5 U/ml, 1 U/ml) groups. After 24 hours of IL-4 treatment, LPS + IFN-γ were added for 18 hours, they were changed to normal medium and given different concentrations of AGC for 24 hours, the expression of CD₁₆+CD₃₂ and CD₂₀₆ were observed by flow cytometry.Results Compared with the control group, the IL-6 and TNF-α level increased (P< 0.01), the number of CD₁₆⁺+CD₃₂⁺ increased and the number of CD₂₀₆⁺ decreased in OGD/R group. Compared with the OGD/R group, the IL-6 and TNF-α level decreased (P< 0.01), the number of CD₁₆⁺+CD₃₂⁺ decreased and the number of CD₂₀₆⁺ increased in AGC groups. Compared with the control group, the number of CD₂₀₆ tended to increase, and the number of CD₁₆+CD₃₂ tended to decrease in IL-4 group; compared with IL-4 group, the number of CD₁₆+CD₃₂ tended to increase, and the number of CD₂₀₆ tended to decrease in IL-4 + LPS + IFN-γ group; compared with IL-4 + LPS + IFN-γ group, the number of CD₁₆+CD₃₂ tended to decrease, and the number of CD₂₀₆ tended to increase in IL-4 + LPS + IFN-γ + 0.25 U/ml AGC group and IL-4 + LPS + IFN-γ + 0.5 U/ml AGC group, while the number of CD₂₀₆ increased in IL-4 + LPS + IFN-γ + 1.0 U/ml AGC group (P < 0.05). Conclusion AGC could inhibit the secretion of inflammation factors by promoting the polarization of microglia from M1 phenotype to M2 phenotype.

Key words: ischemic stroke, analgecine, microglia polarization, inflammation

CLC Number:

R743.3

GONG Shi-li,YANG Cui-cui,HU Chao-ying,WANG Ming-yang,ZHANG Lan. Promotion of Analgecine on Polarization of M1 Microglia to M2 and Inhibition on Secretion of Inflammatory Factors in Vitro[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2020, 26(6): 662-667.

Figures/Tables 5

References 26

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组别	n	TNF-α	IL-6
对照1组	3	48.70±1.637	5.093±0.5215
AGC1组	3	25.73±5.660	1.863±0.635
OGD/R组	3	342.3±22.070^a	31.13±3.003^b
L1组	3	212.3±14.560^c	7.148±1.924^c
M1组	3	138.4±14.280^c	6.127±1.966^c
H1组	3	74.85±9.630^d	3.793±0.463^d
F值		83.600	40.840
P值		<0.0001	<0.0001

组别	n	CD₁₆+CD₃₂	CD₂₀₆
对照2组	4	46.60±5.796	5.093±0.5215
AGC2组	4	26.60±8.821	1.863±0.6350
IL-4组	4	39.00±8.256	7.163±2.115
IFN-γ组	4	55.00±9.906	2.567±0.9602
L2组	4	39.65±10.98	4.437±1.865
M2组	4	39.40±11.120	7.353±2.969
H2组	4	40.97±11.180	6.037±0.3755^a
F值		0.8271	0.917
P值		0.5582	0.508

Promotion of Analgecine on Polarization of M1 Microglia to M2 and Inhibition on Secretion of Inflammatory Factors in Vitro

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