串叶松香草超氧化物歧化酶提取工艺优化及对D-半乳糖致衰老模型小鼠的保护作用研究

Optimization of extraction process of superoxide dismutase from Silphium perfoliatum L. and its protective effect on D-galactose induced aging model mice

中文关键词:  串叶松香草  超氧化物歧化酶  响应面法  D-半乳糖致衰小鼠  抗衰老

英文关键词:Silphium perfoliatum L.  superoxide dismutase  response surface methodology  D-galactose-induced ageing mice  anti-aging

中文摘要:

      目的 优化串叶松香草超氧化物歧化酶(superoxide dismutase, SOD)的提取工艺, 并研究串叶松香草SOD对D-半乳糖致衰老模型小鼠的保护作用。方法 采用高压脉冲电场法提取SOD, 以串叶松香草为原料, 进行最佳提取工艺的研究, 以SOD酶活力为指标, 在单因素实验基础上, 进一步通过响应面实验对提取工艺进行优化。同时采用随机法把小鼠分为空白对照组、模型组、串叶松香草SOD低、中、高剂量组、维生素C (vitamin C, VC)阳性对照组。除空白对照组外, 其余5组均皮下连续注射D-半乳糖, 采用注射D-半乳糖构建衰老小鼠模型。通过观察和测定各组小鼠的行为和体质量, 研究其衰老的变化情况; 采用分光光度计法测定小鼠血清中SOD、谷胱甘肽过氧化物酶(glutathione peroxidase, GSH-Px)、过氧化氢酶(catalase, CAT)活性和丙二醛(malondialdehyde, MDA)含量。结果 最佳提取工艺条件为: 电场强度20 kV/cm、脉冲数10个、提取时间32 min, 在此条件下, 提取的串叶松香草SOD酶活力为3878.67 U/g。串叶松香草SOD对D-半乳糖致衰老模型小鼠的抗衰老作用表明: 串叶松香草SOD能改善衰老小鼠的体征, 提高衰老小鼠的体质量, 具有较好的抗肝萎缩和脑萎缩作用, 同时还能提高衰老小鼠血清中SOD、GSH-Px、CAT的活力, 降低其MDA含量, 达到延缓衰老的作用。结论 该研究获得了串叶松香草SOD的最佳提取工艺, 得到的串叶松香草SOD能够延缓小鼠衰老。

英文摘要:

      Objective To optimize the extraction process of superoxide dismutase (SOD) from Silphium perfoliatum L., and study the protective effect of SOD from Silphium perfoliatum L. on aging model mice induced by D-galactose. Methods The method of high voltage pulsed electric field was used to extract SOD, and the optimum extraction process was studied with Silphium perfoliatum L. as the raw material. With SOD enzyme activity as the index, the extraction process was further optimized through response surface experiment on the basis of single factor experiment. At the same time, the mouse were randomly divided into blank control group, model group, low, medium and high dose groups of SOD from Silphium perfoliatum L., and vitamin C (VC) positive control group. In addition to the blank control group, the other 5 groups were continuously injected with D-galactose subcutaneously, and the aging mouse model was established by injecting D-galactose. The changes of aging were studied by observing and measuring the behavior and body mass of mice in each group. The activities of SOD, glutathione peroxidase (GSH-Px), catalase (CAT) and the content of malondialdehyde (MDA) in serum of mice were measured by spectrophotometer. Results The optimum extraction conditions were: The electric field intensity was 20 kV/cm, the number of pulses was 10, and the extraction time was 32 minutes. Under this condition, the SOD activity of the extracted from Silphium perfoliatum L. was 3878.67 U/g. The anti-aging effect of SOD from Silphium perfoliatum L. on aging model mice induced by D-galactose showed that SOD from Silphium perfoliatum L. could improve the physical signs of aging mice, improve the body mass of aging mice, have a better anti liver atrophy and brain atrophy effect, and could also increase the activities of SOD, GSH-Px, CAT in the serum of aging mice, reduce its MDA content, and achieve the effect of delaying aging. Conclusion The study obtains the best extraction process of SOD from Silphium perfoliatum L., and the obtained SOD from Silphium perfoliatum L. can delay the aging of mice.