玉竹多糖干预后衰老模型鼠抗氧化系统及免疫功能的变化

Effect of polygonatum odoratum polysaccharide on anti-oxidation system and immunological function of senile mice

目的:观察具有扶正固本,养阴润燥和滋补强壮功效的百合科黄精属植物药玉竹的提取物玉竹多糖对D-半乳糖致亚急性衰老模型小鼠血清中超氧化物歧化酶活性和丙二醛含量的抗氧化系统作用及免疫功能的影响,探讨玉竹多糖抗衰老的可能作用途径。方法:实验于2004-07/2005-05锦州医学院免疫教研室完成。①取昆明小鼠50只,随机分为5组:正常对照组、衰老模型组和低、中、高剂量玉竹多糖治疗组。除正常对照组外,其余各组每只颈背部皮下注射50g/LD-半乳糖0.5mL,连续6周。同时低、中、高剂量玉竹多糖治疗组分别腹腔注射0.5,1,2g/kg剂量的玉竹多糖(玉竹为百合科黄精属植物药,药用根茎,由锦州医学院免疫教研室采摘,经中国医科大学免疫教研室潘兴瑜教授乙醇回流脱脂、水提纯、浓缩、醇沉、除蛋白、离心、真空干燥等程序,得到玉竹多糖;玉竹产自锦州凌海市温滴楼乡岩井寺工区)溶液0.5mL,正常对照组和衰老模型组小鼠给予腹腔注射生理盐水0.5mL。②给药6周后,麻醉下将小鼠摘除眼球取血,常规分离血清,在4℃下3000r/min离心5min,取上清待测。用分光光度计检测小鼠血清中超氧化物歧化酶活性和丙二醛水平,并将小鼠称质量并剖取小鼠胸腺和脾脏,用电子天平称质量,以脏器湿质量(mg/g)表示小鼠脾指数和胸腺指数,用MTT法测定脾淋巴细胞转化指数,用免疫荧光法测定脾T淋巴细胞亚群,观察玉竹多糖对免疫功能的影响。③多组间样本均数比较采用方差分析。结果:昆明种小鼠50只均进入结果分析。①衰老模型组小鼠血清中超氧化物歧化酶活性明显低于正常对照组,丙二醛水平明显高于正常对照组(P<0.05)。高剂量玉竹多糖治疗组小鼠血清中超氧化物歧化酶活性明显高于衰老模型组,丙二醛水平明显低于衰老模型组(P<0.01,0.05);中、低剂量玉竹多糖治疗组与衰老模型组血清中超氧化物歧化酶活性和丙二醛水平比较,差异不明显(P>0.05)。②衰老模型组小鼠胸腺指数和脾指数明显低于正常对照组(P<0.01,0.05);低、中、高剂量玉竹多糖治疗组的胸腺指数和脾指数均明显高于衰老模型组(P<0.05～0.01),以中剂量玉竹多糖治疗组作用最明显(P<0.01);3个剂量玉竹多糖治疗组之间胸腺指数和脾指数差异不明显。③衰老模型组小鼠脾T和B淋巴细胞转化刺激指数均明显低于正常对照组(P<0.05,0.01);高剂量玉竹多糖治疗组小鼠脾T淋巴细胞转化刺激指数和各剂量玉竹多糖治疗组小鼠脾B淋巴细胞转化刺激指数明显高于衰老模型组(P<0.05)。④各组小鼠脾脏中CD4+细胞数差异不明显(P>0.05);正常对照组和各剂量玉竹多糖治疗组小鼠脾脏中CD8+细胞数明显多于衰老模型组(P<0.05～0.01),CD4+/CD8+比值则明显低于衰老模型组(P<0.01)。结论:①玉竹多糖可能通过提高衰老小鼠血清超氧化物歧化酶活性,增强对自由基的清除能力,抑制脂质过氧化,降低丙二醛含量,从而减轻衰老小鼠机体组织损伤程度,以延缓衰老。②玉竹多糖对亚急性衰老小鼠免疫器官的功能具有一定的调节作用,可改善机体的免疫失衡状态,从而增强机体的细胞及体液免疫功能,延缓衰老。

AIM： To observe the effect of polygonatum odoratum polysaccharide, which has effects of strengthening the body resistance to consolidate the constitution, nourishing yin to moisten dryness and strengthening by means of tonics, on the anti-oxidation system and immunological function of superoxide dismutae （SOD） activity and malondialdehyde （MDA） content in mice with subacute senile induced by D-galactose, and explore the possible anti-aging action＇of polygonatum odoratum polysaccharide. METHODS： The experiment was completed in the Department of Immunology of Jinzhou Medical University between July 2004 and May 2005.①Fifty Kunming mice were randomly divided into five groups： control group, senile model group and 3 treatment groups with low, medium and high dose of polygonatum odoratum polysaccharide. The 50 g/L 0.5 mL Dgalactose were injected subcutaneously into the naps of mice except control group for six successive weeks; Meanwhile the mice of 3 treatment groups were injected intraperitoneally with 0.5 mL polygonatum odoratum polysaccharide solutions of 0.5 g/kg, 1 g/kg, 2 g/kg respectively （Polygonatum odoratum were from Linghai of Jinzhou city, and provided by the Department of Immunology, Jinzhou Medical University. Polygonatum odoratum polysaccharide were obtained by Professor Pan Xing-yu of Department of Immunology, China＇Medical University in the sequences of ethanol defathng, purification, concentration, protein-removal, centrifugation and vacuum dehydration）. The mice of control group and senile model group were injected intraperitoneally with 0.5 mL saline. ②Six weeks later, the eyeballs of mice were taken out at anesthesia stage; Serum was separated routinely and centrifuged at 3 000 r/minute for 5 minutes at 4 ℃. The upper serum was obtained for measurement. The SOD activity and MDA content in .the serum of mice were detected with spectrophotometer. Meanwhile, mice were weighted, and thymus and spleen were taken out to weight with electronic balance. Wet mass of internal organs （mg/g） was used to express the indexes of thymus and spleen. Spleen lymphocyte transforming index was measured with MTT method, and subgroups of spleen T lymphocyte were detected by immunofluorescence.③The sample averages were com- pared by analysis of variance, RESULTS： All 50 mice of Kunming species were entered into result analysis.①Compared with the control group, the activity of SOD decreased and contents of MDA increased significantly in the senile model group （P 〈 0.05）. Compared with the senile model group, the activity of SOD decreased and contents of MDA incresed significantly in the treatment group with high dose of polygonatum odoratum polysaccharide （P 〈 0.01,0.05）. There was no significant difference in the SOD activity and MDA content between the other two treatment groups and senile model group （P 〉 0.05）. ②The indexes of thymus and spleen in mice were significantly lower in the senile model group than in the control group （P 〈 0.01,0.05）, and also lower in the treatment groups than in the senile model group （P 〈 0.05- 0.01）, especially in the treatment group with medium dose of polygonatum odoratum polysaccharide （P 〈 0.01）.There was no significant difference in the indexes of thymus and spleen in three treatment groups. ③The transforming indexes of spleen T and B lymphocyte were significantly lower in senile model group than in the control group （P 〈 0.05,0.01）; The spleen T lymphocyte transforming index in the treatment group with high dose of polygonatum odoratum polysaccharide and spleen B lymphocyte transforming index in three treatment groups were remarkably higher than those of senile model group （P 〈 0.05）. ④There was no significant difference in the spleen CD4^＋ cell numbers of 5 groups （P 〉 0.05）. The number of CD8^＋ T lymphocyte of rats was remarkably higher in the control group and treatment groups than in the senile model group （P 〈 0.05-0.01）, and the ratio of CD4^＋/CD8^＋ were obviously lower than that of the senile model group （P 〈 0.01）. CONCLUSION： ①Polygonatum Odoratum polysaccharide may improve the SOD activity of senile mice, reinforce the cleaning activity of free radical, inhibit lipid peroxidation and reduce the MDA conient so as to relieve the damaged level of tissues in the senile mice and play the anti-aging effect.②Polygonatum odoratum polysaccharide also has certain effect on regulating the functions of immunological organs in the subacute senile mice and ameliorating the immunological disequilibria, thus enhance the cell immunity and humoral immunity, and postperte the senility.