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目的探讨盐酸小檗碱作用α7烟碱型乙酰胆碱受体(nAchR)与改善氓的分子机制。方法建立IR HepG2细胞模型,用盐酸小檗碱(BBR)、烟碱(NIC)干预,并设正常对照(NC)组和模型组。用现代分子生物学检测细胞培养液葡萄糖消耗量、乙酰胆碱酯酶(AchE)、乙酰胆碱(Ach)、白介素-6(Ⅱ-6)、核因子-κB(NF-κB p65)、IκB激酶βSer181(ⅡKKβSer181)、α7nAchR、磷酸肌醇3激酶(PI3K)、葡萄糖转运因子4(GluT4)等相关指标,用荧光标记方法检测细胞摄取葡萄糖的荧光强度。结果葡糖糖消耗量NC组、HepG2细胞模型组、BBR组和NIC组依次为(25.33±0.77)mmol/L、(11.95±1.94)mmol/L、(16.85±0.33)mmol/L和(16.27±1.09)mmol/L。细胞内2-NBDG(荧光标记葡萄糖)荧光强度增强,NC组、HepG2细胞模型组、BBR组和NIC组依次为(118.17±14.30)、(92.33±9.35)、(120.50±26.26)和(127.67±16.07)。BBR能抑制细胞AchE活性,抑制率为53%,NIC对AchE活性无影响。BBR和NIC能激活α7nAchR,抑制IKKβSer181和NF-κBp65的表达,降低炎性细胞因子IL-6,上调PI3K、GluT4,与HepG2细胞模型组比较,差异有统计学意义(P<0.05或P<0.01)。结论 BBR可能通过抑制AchE活性以激活α7nAchR,从而抑制IKKβSer181、NFκB p65和炎症因子IL-6的表达,促进PI3K、GluT4上调,发挥其改善IR、降低葡萄糖的作用。NIC虽不能抑制AchE活性,但它是特异性激动剂,可直接激活α7nAchR。
Objective To investigate the molecular mechanism of berberine hydrochloride on α7 nicotinic acetylcholine receptor (nAchR) and its improvement. Methods IR HepG2 cell model was established. The cells were treated with berberine hydrochloride (BBR) and nicotine (NIC), and normal control group (NC) and model group were established. The effects of AchE, Ach, IL-6 (Ⅱ-6), NF-κB p65 and IκB kinase βSer181 (ⅡKKβSer181) were detected by modern molecular biology methods. ), Α7nAchR, phosphoinositide 3 kinase (PI3K), glucose transporter 4 (GluT4) and other related indicators, fluorescence labeling method was used to detect the fluorescence intensity of glucose uptake. Results The glucose consumption in NC group, HepG2 cell model group, BBR group and NIC group were (25.33 ± 0.77) mmol / L, (11.95 ± 1.94) mmol / L, (16.85 ± 0.33) mmol / L and ± 1.09) mmol / L. Fluorescence intensities of 2-NBDG (fluorescent labeled glucose) were enhanced in NC group, HepG2 cell model group, BBR group and NIC group (118.17 ± 14.30), (92.33 ± 9.35), (120.50 ± 26.26) and 16.07). BBR can inhibit AchE activity, the inhibition rate was 53%, NIC had no effect on AchE activity. BBR and NIC could activate α7nAchR, inhibit the expression of IKKβSer181 and NF-κBp65, decrease inflammatory cytokines IL-6, upregulate PI3K and GluT4, and have statistical significance compared with HepG2 cell model group (P <0.05 or P <0.01) ). Conclusion BBR may inhibit the expression of IKKβSer181, NFκB p65 and inflammatory cytokines IL-6 by inhibiting AchE activity and activating α7nAchR. It may up-regulate PI3K and GluT4 and play an important role in improving IR and decreasing glucose. Although NIC does not inhibit AchE activity, it is a specific agonist that directly activates α7nAchR.