还少丹对老龄大鼠自由基损伤及端粒长度影响的研究
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摘要
目的：本研究从理论上探讨了古方“还少丹”延缓衰老的机理,并以老龄大鼠为研究对象,观察了“还少丹”对自由基体系相关指标以及对端粒长度的影响,初步探讨了“还少丹”抗自由基损伤的作用,从基因水平探讨“还少丹”延缓衰老的分子作用机理。
方法：采用TBA法(硫代巴比妥酸法)测定大鼠血清和肝组织中丙二醛(MDA)含量；采用DTNB法(5,5’-双硫代对硝基苯甲酸法)测定大鼠血清及肝组织中谷胱甘肽过氧化物酶(GSH-Px)活性；用酚／氯仿法提取老龄鼠肝组织中DNA,用试剂盒分析平均端粒长度。
结果：①与老龄对照组比较,“还少丹”能明显降低大鼠血清和肝组织中MDA含量(P<0.01),升高GSH-Px活性(P<0.01),表明其具有良好的抗自由基损伤作用；②Vi tE组与“还少丹”组均可降低大鼠血清及肝组织中MDA含量并提高GSH-Px活性,但“还少丹”优于Vi tE(P<0.05);③与老龄对照组比较,“还少丹”组端粒长度明显长于对照组(P<0.01),表明“还少丹”能明显减缓老龄大鼠端粒长度的缩短,对DNA具有明显保护作用。
结论：①“还少丹”能升高GSH-Px活性、降低MDA含量,具有明显的抗自由基损伤作用；②“还少丹”能延缓端粒长度的缩短,从基因水平方面说明其能保护DNA,延缓衰老。
Objective:This study explored the anti-aging mechanism of the ancient prescription of Huan Shao Dan in theories, then used aged rats as the object of study, observed the effects of Huan Shao Dan on free radical and telomere length, also discussed the role of Huan Shao Dan of anti-free radical damage and the anti-aging mechanism of the molecule from genetic level.
Methods:Using TBA method (thiobarbituric acid method) to determinate the MDA content of serum and liver tissue; Using DTNB method(5, 5'-dithiobis-2-nitrobenzoic acid method) to investigate the GSH-Px activities of serum and liver tissue; Using phenol/chloroform method to extracte the DNA of liver tissue of aged rats, then analysed the average telomere length with the kit.
Results:①Compared with the aged control group, Huan Shao Dan could decrease significantly the malondialdehyde (MDA) content of serum and liver tissue(P<0.01), and increase the GSH-Px activity(P<0.01), indicated that it had good role of anti-free radical damage;②Both VitE and Huan Shao Dan could reduce the MDA content of serum and liver tissue and improve GSH-Px activity, Huan Shao Dan was better than VitE(P<0.05);③Compared with the aged control group, the telomere length of Huan Shao Dan group was longer than the length of the control group(P<0.01), It indicated that Huan Shao Dan could significantly slow down the telomere length of aged rats to get short, and had a clear protective effect for DNA.
Conclusion:①Huan Shao Dan could increase GSH-Px activity and reduce the MDA content, so it had an obvious effect of anti-free radical injury;②Huan Shao Dan could delay the shorten of telomere length, it showed that Huan Shao Dan could protect the DNA and anti-aging from the aspects of genetic level.
方法：采用TBA法(硫代巴比妥酸法)测定大鼠血清和肝组织中丙二醛(MDA)含量；采用DTNB法(5,5’-双硫代对硝基苯甲酸法)测定大鼠血清及肝组织中谷胱甘肽过氧化物酶(GSH-Px)活性；用酚／氯仿法提取老龄鼠肝组织中DNA,用试剂盒分析平均端粒长度。
结果：①与老龄对照组比较,“还少丹”能明显降低大鼠血清和肝组织中MDA含量(P<0.01),升高GSH-Px活性(P<0.01),表明其具有良好的抗自由基损伤作用；②Vi tE组与“还少丹”组均可降低大鼠血清及肝组织中MDA含量并提高GSH-Px活性,但“还少丹”优于Vi tE(P<0.05);③与老龄对照组比较,“还少丹”组端粒长度明显长于对照组(P<0.01),表明“还少丹”能明显减缓老龄大鼠端粒长度的缩短,对DNA具有明显保护作用。
结论：①“还少丹”能升高GSH-Px活性、降低MDA含量,具有明显的抗自由基损伤作用；②“还少丹”能延缓端粒长度的缩短,从基因水平方面说明其能保护DNA,延缓衰老。
Objective:This study explored the anti-aging mechanism of the ancient prescription of Huan Shao Dan in theories, then used aged rats as the object of study, observed the effects of Huan Shao Dan on free radical and telomere length, also discussed the role of Huan Shao Dan of anti-free radical damage and the anti-aging mechanism of the molecule from genetic level.
Methods:Using TBA method (thiobarbituric acid method) to determinate the MDA content of serum and liver tissue; Using DTNB method(5, 5'-dithiobis-2-nitrobenzoic acid method) to investigate the GSH-Px activities of serum and liver tissue; Using phenol/chloroform method to extracte the DNA of liver tissue of aged rats, then analysed the average telomere length with the kit.
Results:①Compared with the aged control group, Huan Shao Dan could decrease significantly the malondialdehyde (MDA) content of serum and liver tissue(P<0.01), and increase the GSH-Px activity(P<0.01), indicated that it had good role of anti-free radical damage;②Both VitE and Huan Shao Dan could reduce the MDA content of serum and liver tissue and improve GSH-Px activity, Huan Shao Dan was better than VitE(P<0.05);③Compared with the aged control group, the telomere length of Huan Shao Dan group was longer than the length of the control group(P<0.01), It indicated that Huan Shao Dan could significantly slow down the telomere length of aged rats to get short, and had a clear protective effect for DNA.
Conclusion:①Huan Shao Dan could increase GSH-Px activity and reduce the MDA content, so it had an obvious effect of anti-free radical injury;②Huan Shao Dan could delay the shorten of telomere length, it showed that Huan Shao Dan could protect the DNA and anti-aging from the aspects of genetic level.
引文
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[4]Harley C B. Telomere loss:mitotic clock or genetic time bomb [J]. Mutat Res,1991; 256 (2):271-282
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[11]陈红,杜家经.茯苓生脉饮对离体大鼠海马神经元SOD、MDA、LF的影响[J].中国中医急症,2003,12(1)：62-63
[12]陈丽艳,刘君星,施晓光.中药四君子汤抗衰老的实验研究[J].黑龙江医药,2006,19(8)：363-364
[13]苗艳波,师海波,孙英莲,等.高山红景天总甙的抗衰老作用[J].中药药理与临床,2004,20(5)：20-21
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[16]邹俊华,梁红业,闵凌峰,等.枸杞子的抗衰老功效及增强 DNA 修复能力的作用[J].中国临床康复,2005,9(11)：132-133
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[18]同征,陆宇，包晨颖等.西红花酸对自由基发生系统诱导 DNA 损伤的保护作用[J].中国药科大学学报,2002,33(6)：506-509
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[20]张小如,吴晓平,兰风华.抗衰1号对老年小鼠肝线粒体DNA缺失的影响[J].中国老年学杂志,2005,8(25)：929-931
[21]俞仲毅,赵伟康.益气活血方药对衰老大鼠肝脏细胞增殖和凋亡的影响[J].中西医结合学报,2004,2(3)：196-198
[22]裴媛,李德新,孙松辉.独活及其醇提物对自然衰老小鼠脑组织细胞凋亡的影响[J].中国老年学杂志,2005,25(8)：959
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