[關鍵詞]
[摘要]
目的 觀察毛酸漿果乙醇提取物(AEFPP)的抗炎作用,并探索其作用機制。方法 取二甲苯涂于小鼠耳廓制備耳廓腫脹急性炎癥模型,將棉花植入大鼠皮下構建棉球肉芽腫慢性炎癥模型,觀察AEFPP的在體抗炎作用;采用MTT法檢測質量濃度為0.25~20.00 mg/mL的AEFPP對RAW 264.7細胞的細胞毒性;質量濃度為0.625、1.250、2.500 mg/mL的AEFPP預處理3 h后,脂多糖(LPS)作用于RAW 264.7細胞制備炎癥模型,ELISA法檢測上清中腫瘤壞死因子(TNF-α)和白細胞介素-6(IL-6)的水平;實時定量PCR(qRT-PCR)法檢測NF-κB p65 mRNA表達;western blotting法測定NF-κB p65蛋白表達。結果 0.8 g/kg AEFPP可顯著抑制小鼠耳廓腫脹率;0.1、0.2、0.4 g/kg劑量組對大鼠棉球肉芽腫具顯著的抑制作用,且表現(xiàn)出劑量依賴性;AEFPP在0~3 mg/mL范圍內對細胞無明顯毒性;另外,0.625、1.250、2.500 mg/mL AEFPP均可顯著抑制RAW 264.7細胞上清液中的TNF-α和IL-6的水平;0.625、1.250 mg/mL AEFPP顯著抑制NF-κB p65 mRNA及其蛋白的表達。結論 AEFPP具良好的抗炎活性,其作用機制可能與下調p65蛋白的表達和抑制炎癥因子TNF-α和IL-6的釋放有關。
[Key word]
[Abstract]
Objective To investigate the anti-inflammatory effect of the alcohol extract from fruit of Physalis pubescens (AEFPP) and explore its mechanism. Methods To observe the anti-inflammatory effect of AEFPP in vivo, dimethylbenzene was daubed on mice auricle to replicate mouse ear edema acute inflammatory model, and cotton ball was sc implanted to establish rat cotton ball granuloma model. The cytotoxicity of 0.25-20.00 mg/mL AEFPP was examined with MTT method. Pretreatment with 0.625, 1.250, and 2.500 mg/mL AEFPP for 3 h, RAW 264.7 mononuclear macrophages cells were stimulated with lipopolysaccharides so as to build an inflammatory cell model. The culture supernatant of the cells was collected in order to determine tumor necrosis factor-α (TNF-α) and interleukin 6 (IL-6) by ELISA. The expression of NF-κB p65 and its anti-inflammatory activity were assayed by Western blotting and real-time quantitative PCR (qRT-PCR). Results High dose (0.8 g/kg) of AEFPP could diminish mouse ear edema, which had been induced by dimethylbenzene. Besides, all the three dosages of AEFPP (0.1, 0.2 and 0.4 g/kg) could markedly inhibit rats cotton ball granuloma. AEFPP was not toxic to the cells in the range of 0-3 mg/mL. Furthermore, the other three dosages of AEFPP (0.625, 1.250, and 2.500 mg/mL) significantly decreased the concentration of TNF-α and IL-6 in the culture supernatants. AEFPP of 0.625 and 1.25 mg/mL could markedly inhibit the expression levels of NF-κB p65 mRNA and its protein. Conclusion AEFPP has a wonderful anti-inflammatory effect. The mechanism might be related with down-regulating the expression of p65 protein and inhibiting the release of inflammatory factors, such as TNF-α and IL-6.
[中圖分類號]
[基金項目]
吉林省科技廳科技發(fā)展計劃-醫(yī)藥產業(yè)發(fā)展專項資金計劃(20130727066YY);長春市科技計劃-重大科技攻關計劃(14KG061)