| Objective: By exploring the immunomodulating effects of yiqijiedu formula on RAW264.7 cells, the pharmacological mechanism of resisting NPC and pharmacodynamic material basis can be revealed. This project will be helpful for illuminating compatible principle,optimizing the preparation process and formulating quality standard.Method:(1)Appropriate chemical and physical methods was used to extract preparation of yiqjiedu formula each target composition, the composition of yiqijiedu formula seven herbs their effective components and pharmacological activities were analyzed, and the total polysaccharides and total saponins, flavonoids and total alkaloid composition of four categories as a target, at the same time, the visible- ultraviolet spectrophotometric method for content determination of the target component.(2)Objectives is determined by MTT method to detect yiqijiedu formula component of RAW264.7 cell proliferation activity, the influence of filter can significantly enhance RAW264.7 cell activity substance.(3) ELISA method to detect yiqijiedu formula aqueous extract and total polysaccharides of TNF-? RAW264.7 cells, the influence of the level of secretion of IL- 12 and NO.(4) Western blot method to detect yiqijiedu formula aqueous extract and total polysaccharides of RAW264.7 cells NF-? Bp65 the influence of the amount of protein expression.Result:(1)Extract 20 vice or 1260 g yiqi Jiedu formula side medicinal materials available water extracting total of 128.63 g(extract rate 10.21%), alcohol extract yield(13.33%)of the total extract of 168 g, total polysaccharide samples 14.49 g(extract rate 1.15%),total saponins extract yield(1.11%), 13.99 g sample flavonoids extract yield(1.63%),20.6 g sample sample 14.97 g of total alkaloid extract yield(1.19%); Total polysaccharide samples total polysaccharide content was 74.8%, total saponin content in the samples of total saponins was 67.4%, total content of flavonoids in the total flavonoids samples was 60.39%, total alkaloid content in the samples of total alkaloids was 61.32%.(2)Determined by MTT experiments show that yiqijiedu formula aqueous extract five concentration group(10,20,40,80,160,320 mg. L- 1) and total polysaccharides yiqijiedu formula five concentration group(5,10,25,50,100,200 mg ? L- 1) are huge bite cell in mice activity of RAW264.7 cells have different degrees of enhancement effect; And yiqijiedu formula alcohol extract, yiqijiedu formula total saponins,flavonoids, yiqijiedu formula total alkaloid concentration of their five group, some promote RAW264.7 cells huge bite cell proliferation in mice, or suppressed.(3) ELISA experiments show that yiqijiedu formula aqueous extract five concentration group(10,20,40,80,160,320 mg ? L- 1) and yiqijiedu formula total polysaccharide concentrations of five groups(5,10,25,50,100,200 mg ? L- 1) huge bite cell line can stimulate mice RAW264.7 cell secreted TNF-?, IL- 12 and NO.(4) Western blot results show that the aqueous extract yiqijiedu formula five concentration group(10,20,40,80,160,320 mg. L- 1) and total polysaccharides yiqi jiedu formula five concentration group(5,10,25,50,100,200 mg ? L- 1) respectively deal with huge bite cell in mice RAW264.7 cells after 24 hours, in the sample internal GADPH almost the same amount of protein expression, can obviously raise the NF-?Bp65 protein expression level.(5)Yiqijiedu formula aqueous extract and yiqijiedu formula total polysaccharides can regulate the immune activity of RAW264.7 cells. The results suggest that yiqi jiedu formula aqueous extract and total polysaccharides may be by activating the nf-kappa B signaling pathways, activate the corresponding TNF alpha, IL- 12 cytokines gene expression, NO so as to promote the TNF alpha, IL- 12, NO the secretion of cytokines, such as enhancing the immune function of macrophages. This may be yiqijiedu formula side play against one of the pharmacological action mechanisms of nasopharyngeal carcinoma. |
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