桦褐孔菌有效破壁方法及其对免疫功能的影响

The effective wall breaking method of Inonotus obliquus and its effect on immunefunction

目的探讨超声粉碎仪在何种工作条件下粉碎桦褐孔菌的效果最好,探索桦褐孔菌破壁水溶液对免疫系统的影响。方法取桦褐孔菌共120g分为4组,置于超声粉碎仪中,分别在不同的工作条件下,粉碎制成溶液,通过蛋白浓度的测定选择超声粉碎仪的最佳工作条件;分别将桦褐孔菌破壁水溶液、4倍稀释的桦褐孔菌水溶液、PBS液置于淋巴细胞培养基中,观察并比较体外淋巴细胞增殖情况,分别将桦褐孔菌破壁水溶液、生理盐水喂服结肠癌模型鼠,比较结肠癌模型鼠体内细胞因子TNF-α、IL-2、IL-17、TGF-β浓度和自然杀伤细胞(N K细胞)活性。结果在设置功率55W,工作时间15min时测定的蛋白浓度最高(8.15±0.88)mg/ml(P<0.05);桦褐孔菌破壁水溶液组、4倍稀释后桦褐孔菌破壁水溶液组、PBS液组的淋巴细胞增值指数PI分别为(158.82±2.77)%、(146.08±6.93)%、(100±0)%,差异有统计学意义;桦褐孔菌组TNF-α、IL-2浓度分别是(168.47±11.02)、(1.96±0.28)pg/ml均高于生理盐水组,差异有统计学意义;而IL-17、TGF-β浓度分别是(42.52±3.18)、(28.44±1.01)pg/ml,均低于生理盐水组,差异无统计学意义;桦褐孔菌组NK细胞活性为(17.18±0.17)%,高于对照组(15.84±0.58)%(P<0.05)。结论超声粉碎仪在功率55W,工作时间15min时测得蛋白浓度最高,粉碎桦褐孔菌的效果最好;桦褐孔菌破壁水溶液能促进淋巴细胞增殖、提高细胞因子TNF-α、IL-2浓度、增强NK细胞杀伤活性。

Objective To select the optimum condition of ultrasonic cell crusher to break the cytodcrm of Inonotus obliquus and explore effect of Inonotus obliquus on the immune system.Methods A total of 120g of inonotus obliquus was divided equally into 4 groups.Then,they were crushed by ultrasonic cell crusher in different conditions.The protein concentration were measured respectively,to acquire the optimum condition of ultrasonic cell crusher.The lymphocytes were cμ ltured in mediums respectively containing extract of inonotus obliquus,4-fold diluted extract and PBS.The proliferation of them was compared.The model mice with colon cancer were fed with the extract of inonotus obliquus and saline,respectively.The concentration of TNF-α,IL-2,IL-17 and TGF-β and the activity of natural killer cells(NK cells)were determined and compared in the colon cancer model mice.Results The highest protein concentration was 8.15±0.88mg/ml(jP<0.05)when the condition of ultrasonic cell crusher was set on power 55 watts and the working-time 15 minutes.Compared with that of control group,the lymphocyte proliferation index of extract group and 4-fold diluted extract group was significantly higher,respectively was 158.82±2.77%,146.08±6.93%.Compared with that of saline group,the concentration of TNF-α,IL-2,in the inonotus obliquus group was significantly increased,and the concentrations of IL-17 and TGF-β in the inonotus obliquus group was decreased,with no statistical significance.NK cell activity in the inonotus obliquus group was 17.18±0.17%,which was higher than that in the control group(P<0.05).Conclusion The ultrasonic cell crusher has a better effect on crushing inonotus obliquus when the condition is set on power 55 watts,working-time 15minutes.The inonotus obliquuscan promotes the proliferation of lymphocytes,increases the concentration of TNF-α and IL-2 in tumor-bearing mice,and enhances the killing activity of NK cells.