青龙衣多糖对荷瘤小鼠红细胞膜流动性及封闭度的影响

目的:探讨青龙衣多糖对红细胞膜流动性及封闭度的影响。方法:首先建立肿瘤模型,并对小鼠进行分组和给药,再给药第8天处死小鼠,眼球取血,代用。在封闭度的测定中,取血离心,3000转／分,生理盐水洗涤3次,蒸馏水破膜,12000转／分离心,制成半封闭红细胞影泡,2000转／分离心40分钟,37℃温浴1小时,制成封闭影泡,按Zamudio法进行NADH-细胞色素C氧化还原酶活性的测定,并按公式计算出核瘤小鼠红细胞的封闭度。以1,6-Dipheyl-1,3,5-hexatriene(DPH)为探针剂,用荧光分光光度计以发射光波长432nm,激发光波长362nm,用荧光偏振法测定荧光偏振度(P)值和微粘度(η)研究细胞膜脂的流动性。两者的膜蛋白定量均采用考马斯亮兰法。结果:针对S180核瘤小鼠生理盐水组的红细胞膜封闭度为,流动性为4.3645±0．1173,黄芪多糖两数值分别为54．81±3．73和8．8806±0．5586,青龙衣多糖1低剂量组为48．90±1．45和6．6177±0. 2905,中剂量组为54．59±1．21和8．0146±0．1339,高剂量组为61.59±1．34和9．1415±0．2893,多糖2低剂量组为46. 71±0．84,6．8156±0．1781,中剂量组为52．61±0．74和7．9889±0．0479,高剂量组为62．69±6．78和9．1890±0．1336针对H22核瘤小鼠,生理盐水组为38．43±4．63和4．6022±0．0398,黄芪多糖组为54．81±3．73和9．6234±0．9633,多糖1低剂量组为44．89±2．13和6．7871±0．3303,中剂量组为56．39±1．46和8．0988±0．2352,高剂量组为62．32±2．29和9．1415±0．2893,多糖2低剂量组为45．69±2．18和6．6796±0．0367,中剂量组分别为57．89±1．33、0．4854±0．0059,高剂量组分别为63．44±2．48、9．4473±0．1226,各组数据与空白对照生理盐水组相比较,均有显著差异,且P<0．05。结论:青龙多糖的抑瘤作用可能通过提高红细胞膜流动性及封闭度,增强维持红细胞膜的稳定性的能力并升高红细胞膜的重新封闭的能力起积极作用。

The Effects of the Two Qinglongyi Polysaccharide on RBC Cell Membrane Fluidity and Blocking Degree of Tumor Mice

Objective: To find the effects of the two Qinglongyi polysaccharide on RBC cell membrane fluidity and blocking degree. Methods: Fist constitute the tumor former, grouping and medicining point to rice, than at the 8th day execute these rice, get the blood from eyes, substitutional. In the examination of blocking degree, we get the blood and use the centrifugal machine, 3000round/minite, than use isotonic Na chloride to cleanse it and use DW distilled water to break the cell membrane up, 12000round/minite in the centrifugal machine, than half-blocked ghost-corpuscle executed. Use the centrifugal machine at 2000round/minite for 40 minutes, temperature it at 37℃for 1 hour, we get the blocking ghost-corpuscle. Set the NADH-cytochrome C-reductase and cytochrome C oxidase out according to Zamudio method, and calculate the RBC blocking degree of tumor mice. Probed 1,6-Dipheyl-1,3,5-hexatriene(DPH), use SPF at 432nm emission spectro-wavelength, 362nm excitation wavelength to get the fluorescence polarization and microviscosimete to explore RBC cell membrane fluidity. These two membrane proteins' fixed quantity ate both according to coomassie blue method. Result: The group of isotonic Na chloride's RBC membrane blocking degree of S180 tumor rice is 42.24±0.99, fluidity is 4.3645±0.1173; Astragalan's is 54.81±3.73 and 8. 8806±0.5586; Qinglongyi polysaccharidel low dose group's is 48.90±1.45 and 6.6177±0.2905￡(?)middle dose group's is 54.59±1. 21 and 8.0146±0.1339; high dose group's is 61.59±1.34 and 9.1415±0.2893; Qinglongyi polysaccharide2 low dose group's is 46. 71±0.84 and 6.8156±0.1781; middle dose group's is 52.61±0.74 and 7.9889±0.0479,; high dose group's is 62.69±6.78 and 9. 1890±0.1336. The group of isotonic Na chloride's RBC membrane blocking degree of H22 tumor rice is 38.43±4.63, fluidity is 4. 6022±0.0398; Astragalan's is54.81±3.73 and 9.6234±0.9633; Qinglongyi polysaccharidel low dose group's is 44.89±2.13 and 6. 7871±0.3303; middle dose group's is 56.39±1.46 and 8.0988±0.2352; high dose group's is 62.32±2.29 and 9.1415±0.2893; Qinglongyi polysaccharide2 low dose group's is 45.69±2.18 and 6.6796±0.0367; middle dose group's is 57.89±1.33 and 0.4854±0.0059; high dose group's is 63.44±2.48 and 9.4473±0.1226. Compared with isotonic Na chloride group, every group all has visible difference (P<0.05). Conclusion: The inhibit effects to untitumor of Qinglongyi polysaccharide can be through improving the membrane fluidity and blocking degree of RBC, the actions to RBC can be active