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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.
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Objective:To observe the effects of Analgecine (AGC) on middle cerebral artery ischemia-reperfusion injury in rats and its mechanism. Methods:A total of 61 Sprague-Dawley rats were divided into sham group (n = 11), sham-AGC group (n = 11), model group (n = 20) and model-AGC group (n = 19). The model group and the model-AGC group were occluded the middle cerebral arteries for 1.5 hours and reperfused (2 rats in each group unsuccessful). The sham-AGC group and the model-AGC group were injected AGC 20 U/kg through tail-vein, while the sham group and the model group were injected saline of same volume. Four rats in each group were tested heat shock proteins 70 (HSP70), Bcl-2 and Bax in brain with Western blotting 48 hours after injection. The other rats were assessed with Prehensile Traction Test seven days after injection, and then, four of each group were detected ionized calcium binding adapter molecule 1 (Iba1) and glial fibrillary acidic protein (GFAP) expression with immunohistochemistry. Results:The prehensile time increased in the model-AGC group compared with that of the model group (P < 0.01), with the increase of HSP70 and Bcl-2 (P < 0.01) and decrease of Iba1 and GFAP expression (P < 0.05). Conclusion:AGC may promote the recovery of motor function in rats with cerebral ischemia-reperfusion injury, which may associate with inhibiting cell apoptosis and neruoinflammatory response.
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Objective To investigate the effect of extracts from rabbit skins inflamed by Viccinia virus vaccine (analgecine) on the proliferation of human cancer cells and on the cytokine secretion in mouse spleen lymphocytes in vitro. Methods Five human tumor cell lines, HepG2, LM3, H460, A549 and HeLa were used and the effect of analgecine (1.63, 0.815, 0.326, 0.163 and0.0815 U/ml) on the cell proliferation was evaluated by the CCK-8 assay. The mouse spleen was isolated aseptically, and the spleen lymphocyte suspension was prepared and cultured with PRMI-1640 medium containing 10％ fetal bovine serum (FBS). For detection of the cytokine IL-2, IFN-γ, IL-4 and IL-12 level, the stimulant concanavalin A (ConA) or lipoplysaccharide (LPS) was added into the lymphocyte suspension, and the lymphocytes were cultured under the presence of analgecine at the final concentration of 0.815, 0.163 and 0.0815 U/ml for 24 hours. Then, the level of the cytokines in the supernatant was detected by the ELISA kit. On the other hand, the effect of supernatant of the spleen lymphocyte cultures under the presence of analgecine at 0.815 U/ml on the proliferation HepG2 cells was also evaluated by the CCK-8 assay. The CCK-8 assay was performed after cultivation of the HepG2 cells in the whole supernatant or in its dilution with fresh medium for 24 hours. Results Analgecine showed a dose-dependent inhibitory effect on the five tested cancer cell lines, with the inhibition rate of 58.95％, 55.08％, 57.28％, 45.80％ and 48.18％ at the 1.63 U/ml on the HepG2, LM3, H460, A549 and HeLa cells, respectively. Compared with the control group, the secretion of IL-2, IFN-γ and IL-4 was significantly increased in the 0.163 and 0.815 U/ml analgecine groups (P＜0.01), while the secretion level of IL-12 was increased in the 0.0815, 0.163 and 0.815 U/ml analgecine groups (P＜0.01). The supernatant of the mouse spleen lymphocyte cultures under the presence of0.163 U/ml analgecine could inhibit the HepG2 cell proliferation in a dose-dependent manner, and the inhibitory effect of the whole supernatant was significantly stronger than the effect of the same concentration analgecine 0.163 U/ml (P＜0.01). Conclusion Analgecine could inhibit the cell proliferation of the tested five human cancer cell lines, increased the secretion of IL-2, IFN-γ, IL-4 and IL-12 cytokines in mouse spleen lymphocytes, all in vitro, and its effect on the cytokine secretion may be related to the inhibitory effect on the human cancer cell proliferation.
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Objective To compare the effect of perioperative intravenous injection of Analgecine on the analgesic efficacy and complications of patient-controlled intravenous analgesia ( PCIA ) of different doses of fentsnyl in postoperative lumhotomy patients.Methods 200 patients underweat hmbotomy in general anesthesia were randomly divided into four groups with fifty cases each.Fentanyl 1.0mg in group A,fentanyl 0.5mg in group B,fentanyl 1.0mg in group C,fentanyl 0.5mg in group D.The drugs in each group were diluted to 100ml and infused by pumps.Besides,the patients in group C and D were injected with analgecine 3.6u and 7.2u at the night before the operation,preoperation and postoperation respectively.The visual analog scale(VAS),times of PCA and incidence of side effects were recorded during the period of postoperative 24 hours.Results The VAS of group B at 2h after operation was (5.2 ± 1.9 ) points,which was significandy higher than that of group A,C and D( P ＜ 0.05 ),VAS became similar 4h later( P ＞0.05).The demanding times for supplemental bolus in group B were also significantly higher than that of A,C and D( P ＜ 0.05 ).The incidence of nausea,vomiting,itching,somnolence in group B and D were significantly less than those in group A and C( P ＜0.05 ).No respiratory depression or abnormal bleeding occurred in the four groups.Conclusion Perioperative intravenous injection of analgecine had a better effect on PCIA of fentanyl and could reduce fentanyl requirement and its side effects in lumbotomy patients.