中药蜣螂酶解提取工艺及抗肿瘤药效研究

Studies on the Enzymolysis Extraction Technology and Anti-Tumor Efficacy of Chinese Medicine Catharsius

目的研究用四种不同蛋白酶对蜣螂粉进行酶解并分别进行抗肿瘤实验,从而优选出最佳用酶及酶解液抗肿瘤方面的药效。方法将蜣螂粉分别用胰蛋白酶,胰凝乳蛋白酶,胃蛋白酶,弹性蛋白酶进行酶解,以蜣螂水提物作为对照组,采用茚三酮法测定蛋白质的水解度并利用MTT法研究水提物对肺腺癌细胞A549的抑制效果,评价4种蛋白酶对蜣螂粉的水解效果,研究蜣螂粉酶解液抗肿瘤方面的药效。结果胰蛋白酶、弹性蛋白酶和胰凝乳蛋白酶的水解度分别为26.62%、24.99%、21.76%,在较低药物浓度下,酶解蜣螂粉对肺腺癌A549无明显抑制作用,而在较高浓度下,酶解蜣螂粉对肺腺癌A549的抑制率大小中,胰凝乳蛋白酶>弹性蛋白酶>胰蛋白酶。胃蛋白酶的水解度为7.64%,其酶解液对肺腺癌A549细胞几乎没有抑制作用。4种酶解液对肺腺癌A549的抑制率均小于酶解前的水提液。结论在蜣螂酶解工艺4种蛋白酶中,无最佳用酶。蜣螂经酶解后会使其抗肿瘤药理活性降低,进而说明中药蜣螂在抗肿瘤方面不宜口服入药。

Objective： Studies on the Catharsius powder was enzymolysised by four different kinds of enzymes and respectively being Anti-tumor experiment,In order to selecting the best enzyme and enzymatic hydrolysate antitumor effects. Method： The Catharsius powder was respectively hydrolyzed by trypsin,chymotrypsin,pepsin,elastase. Using indene three ketone colorimetric method measured the DH of protrin,meanwhile using MTT method measured inhibition rate on A549 cell line compared the enzymatic hydrolysate with the control group of water extracts of the Catharsius powders. Studies on the effect of Catharsius powder hydrolysate of anti-tumor. Results： Trypsin,elastase and chymotrypsin hydrolysis degree was 26. 62%,24. 99% and 21. 76% respectively. At low concentration,Catharsius powder was enzymolysised had no significant inhibitory effect on lung adenocarcinoma A549,but at higher concentrations,Catharsius was enzymolysised on lung adenocarcinoma A549 inhibition rate,chymotrypsin trypsin elastase. The pepsin hydrolysis degree is 7. 64%,the enzymatic hydrolysate on lung adenocarcinoma A549 cells had no inhibitory effect. The inhibition rate of 4 kinds of enzymatic hydrolysate on lung adenocarcinoma A549 was less than that of water extract before enzymatic hydrolysis. Conclusion： In enzymolysis technology of Catharsius powders has no best enzyme. Catharsius by enzymolysis will make its antitumor activity is reduced,and the Chinese medicine catharsius should not be oral medicine in anti-tumor aspect.