淫羊藿总黄酮、淫羊藿苷、淫羊藿次苷Ⅱ延长健康寿限及其作用机制的研究
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摘要
研究背景：
衰老相关疾病与衰老进程本身具有同源性。干预衰老能够降低衰老相关疾病的发生发展,延长健康寿命,是实现健康老龄化的关键环节,但目前仍缺乏有效的延缓衰老手段。中医药学在养生健体、延缓衰老方面历史悠长。在前期补肾延缓衰老系列研究中,我们发现淫羊藿总黄酮(EF)为补肾抗衰老复方改善内分泌免疫衰老的主要有效成分。寿命为评价衰老干预手段的核心指标,但是EF能否延长寿命还属未知。淫羊藿苷(Icariin, ICA)是EF的主要成分,但ICA是否为EF延长寿命的有效成分需进一步研究。
延长寿命更重要的是延长有功能意义的健康寿命,因此检测衰老干预手段对老年机体健康状态的改善作用尤为重要。目前对老年机体健康状态的检测主要包括压力应激能力、衰老退行性改变以及衰老相关疾病等3个方面。ICA能否改善机体的健康状态,延长健康寿命还属未知。
现代药理研究证实,ICA经吸收后在体内被迅速代谢为淫羊藿次苷Ⅰ(Icariside Ⅰ, ICS Ⅰ)、次苷Ⅱ (Icariside Ⅱ, ICS Ⅱ)及淫羊藿素(Icaritiin, ICT)。ICA是以其原型还是代谢衍生物发挥延长健康寿命的作用值得研究。目前研究证实,Insulin/IGF-1通路(IIS)、Target of rapamycin通路(TOR)、沉默信息调节子Sirtuins家族以及AMP kinase (AMPK)等为寿命调控的核心分子机制。其中IIS通路已被证实能够调节包括低等和高等生物在内的一系列物种的寿命,其作用在进化上高度保守。寻找这些寿命调控通路的靶向药物是抗衰老研究的一个主要目标。在揭示ICA为EF发挥作用的单一成分后,ICA及其体内活性代谢衍生物是否通过上述衰老调控通路发挥作用值得探索。
模式生物秀丽隐杆线虫(以下简称线虫)是衰老研究的经典模式生物,具有实验操作简单、遗传背景清晰等优点,易于进行寿命实验和作用机制的探索,但低等生物毕竟与人类相差太远,药物的确切效果还需进一步通过哺乳动物和人类细胞的验证。因此,在本研究中我们采用了线虫、C57BL/6小鼠及人类成纤维二倍体细胞模型MRC-5等3种实验模型,结合各实验模型的优势来探索药物延缓衰老的作用及其机制。
目的：
1.研究EF及其主要成分ICA干预寿命的效果。
2.评价ICA对老年健康状态的改善作用；
3.探讨ICA延长健康寿命的体内活性成分及其分子机制。
方法：
1.EF及ICA干预寿命的研究
1.1野生型线虫N2分组为对照组、EF高(100mg/mL)、中(50mg/mL)、低剂量组(25mg/mL);对照组、ICA高(75μM)、中(45μM)、低(15μM)剂量组进行寿命实验；
1.2C57BL/6雄性小鼠分组为正常对照组、EF组(0.06%,w/w)及ICA组(0.02%,w/w),进行寿命实验,记录各组小鼠体重、饲料日耗量、饮水量、大便小便量,小鼠自然死亡后行大体解剖观察肿瘤发生及皮肤溃疡情况。
1.3人二倍体成纤维细胞MRC-5分组为对照组、EF组(3mg/mL)及ICA组(10-6mol/L),检测细胞传代次数(PD)；
2.ICA影响老年健康状态的研究
2.1压力应激能力：野生型线虫N2分组为对照组、ICA组(45μM),进行热压力应激及氧化压力应激实验,观察ICA对压力应激能力的影响；
2.2衰老退行性病变：MRC-5细胞分组为对照组、ICA组(10-6mol/L),进行细胞衰老相关β-半乳糖苷酶染色实验；野生型线虫N2分组为对照组、ICA组(45μM),进行游泳能力测试；线虫突变体PD4251分组为对照组、ICA组(45μM),进行肌肉细胞计数；C57BL/6小鼠的分组对照组、ICA组,同时取3月龄小鼠作为青年对照,进行转棒实验、骨密度检测及重要脏器病理观察,观察ICA对衰老退行性病变的影响；
2.3衰老相关疾病：线虫疾病模型突变体AM140、CL4176分组为对照组、ICA组(45μM),观察ICA对谷氨酰胺重复序列(PolyQ)及人类β-淀粉样肽(Aβ1-42)蛋白毒性诱导的神经退行性病变的作用,检测ICA对衰老相关疾病的作用。
3.ICA的体内代谢衍生物ICSⅡ延长健康寿命作用机制的研究
3.1野生型线虫N2分组为对照组及药物干预组,观察ICA及其衍生物ICSI、ICSII、ICT对寿命的影响并结合高效液相(HPLC)检测ICA和各衍生物在野生型线虫体内的浓度；
3.2野生型线虫N2分组为对照组、ICS Ⅱ组(20μM),进行压力应激实验、游泳能力测试、肌肉细胞计数,观察药物对衰老相关退化的改善作用；线虫疾病模型突变株AM140、CI4176分组为对照组、ICS Ⅱ组(20μM),观察药物对PolyQ及Aβ1-42蛋白毒性诱导的神经退行性病变的影响。
3.3不同遗传背景的线虫突变体分组为对照组、ICS Ⅱ组(20μM),进行寿命实验(Genetic Epistasis Experiment),探讨药物作用机制,并进一步用实时定量RT-PCR、Western Blot及相关基因GFP报告体检测药物对相关通路下游靶基因的表达的影响。
结果：
1.EF及其主要成分ICA能显著延长多个物种实验模型的寿命
EF及ICA呈剂量依赖性右移野生型线虫的生存曲线,提高线虫平均寿命及最大寿命；显著右移小鼠的生存曲线,显著提高小鼠平均寿命,对小鼠的最大寿命无明显影响。药物干预对小鼠自发肿瘤及皮肤溃疡发生率无明显影响。EF及ICA能显著提高人类二倍体细胞MRC-5的传代次数；
2.ICA能显著改善老年健康状态
ICA能显著降低MRC-5细胞衰老特异性p-半乳糖苷酶的表达,提高线虫的热应激、氧化应激能力及运动能力,改善线虫衰老相关肌肉萎缩,延长小鼠转棒潜伏期时间；ICA能降低蛋白毒性介导的线虫瘫痪表型,提高老年小鼠股骨骨密度。经ICA干预过的小鼠肝组织中衰老相关病理改变明显减轻。药物长期干预对小鼠主要脏器无明显毒性影响。
3.ICA及其活性代谢物衍生物ICS Ⅱ通过ⅡS通路延长健康寿命。
在野生型线虫上对ICA及ICA代谢衍生物进行寿命实验显示20μM ICSⅡ能获得与45μM ICA同等的延长寿命的效果,HPLC结果显示在秀丽线虫中测得的化合物主要为ICA及其代谢衍生物ICSⅡ,提示ICSⅡ为ICA发挥抗衰老作用的体内活性衍生物。ICSⅡ能显著提高野生型线虫的热应激及氧化应激能力,提高老年线虫的运动能力,降低衰老相关肌肉萎缩,减低PolyQ及Aβ1-42的蛋白毒性。ICA及ICSⅡ不能影响ⅡS通路上游基因daf-2突变体、下游核心转录因子daf-16突变体及协助转录因子hsf-1突变体的寿命,提示药物作用机理与ⅡS通路相关。进一步的实时定量RT-PCR、Western Blot及SOD-3绿色荧光报告实验结果结果显示,ICS Ⅱ能增强IIS通路的核心转录因子DAF-16的转录活性,提高其下游有益靶基因的表达,从而延长健康寿命。
结论：
综上所述,我们首次发现EF、ICA及ICS Ⅱ能改善多个物种实验模型的衰老相关性退化、降低衰老相关疾病的发生发展,延长健康寿命。我们发现ICS Ⅱ为ICA发挥抗衰老作用的主要体内活性衍生物；ICA及ICS Ⅱ通过抑制ⅡS通路从而增强FOXO/DAF-16转录活性发挥抗衰老作用。本研究提示ICA及ICS Ⅱ能延缓从低等至高等多个物种实验模型的衰老,并通过进化保守的ⅡS通路发挥作用,极有可能延缓人类衰老。临床使用经验及本研究小鼠实验显示ICA可长期使用,安全性高,有望走向人类抗衰老临床使用。
Introduction
A major goal of current research on aging is to identify compounds that delay age-related diseases and extend healthspan in humans. Herba epimedii is a popular herbal tonic used in traditional Chinese medicine, with proven efficacy in treating several age-related diseases including osteoporosis, cardiovascular diseases, neurodegenerative diseases and sexual dysfunction. Our previous studies on Epimedium Flavones (EF), the raw extract of Epimedium which contains icarrin (ICA) as a major constituent, show that EF delays aging in Drosophila melanogaster and cell senescence model. Interestingly, EF also resets the age-related metabolites (fatty acids, carnosine, ergothioneine and deoxycholic acid et al) to the juvenile level in rat plasma and urine. These findings prompted us to investigate the anti-aging potential of EF and its major pharmacologically active flavonol diglycoside, icariin. Various studies indicate the anti-oxidative effect of icariin and derivatives on DNA damage, β-amyloid mediated neurotoxicity, and vein endothelial cell oxidative injury. Meanwhile, icariin and its derivatives function as signalling modulators to exert beneficial effects in a multitude of age-dependent disease states, including bone loss, cancer, cardiovascular disease, and neurodegenerative disorders. These researches further strengthen the possibility of the effects for EF, icariin and its derivatives to extend healthspan.
The highly conserved IIS pathway plays a key role in aging. It has been demonstrated in multiple species that inhibition of the IIS pathway extends lifespan. In C. elegans, the daf-2gene encodes an insulin/IGF-1receptor. Mutations in daf-2suppress the IIS which lead to the nuclear localization of FOXO/DAF-16transcription factor. The activated FOXO/DAF-16regulates a series of genes involved in lifespan control, stress tolerance and protein misfolding suppression. Nuclear localization of DAF-16in the daf-2mutants requires the heat shock transcription factor (HSF)-1, which modulates the expression of heat shock proteins and protease responsible for the stress tolerance and protein folding. Despite the well established role of a number of genes in the IIS pathway in modulating aging, pharmacological tools that inhibit IIS pathway to extend lifespan are not commonly available, which limits its translation to mammalian model systems.
Objectives
1. To investigate the delaying aging effects of EF and ICA in the experimental systems from invertebrate to mammalian.
2. To measure the effects of ICA in extending healthspan.
3. To ascertain the bioactive formation of ICA in vivo for the healthspan extension and investigate the molecular mechanisms of these effects.
Methods
In this study, we employed three experimental models including the cell senescence model MRC-5, the simple animal model C. elegans and the mammalian model C57BL/6mice to investigate the delaying aging effects of the compounds. Lifespan is the major measurement to evaluate the effect of the compounds. Stress-resistant assays, mobility assay, qualification of muscle cells, rotarod experiment, protein-misfolding disease models are used to evaluate the effects of the compounds in healthspan. HPLC was used to measure the level of ICA and its derivatives in vivo. Genetic epistasis experiments were used to study the mechanism. Three major genetic modulate pathways in longevity were tested in this study, the insulin/IGF-1pathway, the dietary restriction and the target of rapamycin (TOR) pathway. Real-time PCR, western blot were used to measure the downstream targets of the pathway to ascertain the molecular mechanism.
Results
In this study, we tested the anti-aging properties of EF, ICA and its three derivatives, icariside Ⅰ (ICS Ⅰ), icariside Ⅱ (ICS Ⅱ) and icaritin (ICT) in the cell senescence model MRC-5, the simple animal model C. elegans and the mammalian model C57BL/6mice. We found that EF has a conserved role in extending lifespan from simple animal to mammalian model. ICA is the major pharmacological element of EF to extend lifespan. Additionally, ICA promotes the stress resistance and mobility in late life of the animals. Meanwhile it ameliorates protein aggregation and protetoxicity-mediated paralysis phenotype. And one of its derivatives, ICS Ⅱ prolonged adult lifespan. Chemical profiles of ICA treated animals revealed that ICS Ⅱ was the predominant bioactive form of ICA in vivo. Furthermore, we found that ICS Ⅱ treated animals have delayed age-associated phenotypes suggesting ICS Ⅱ enhances the healthy aging significantly. Finally, our genetic analysis indicates that Icariside Ⅱ may act through the IIS pathway to affect lifespan.
Conclusion
Our findings reveal a novel role for EF, ICA, along with its bioactive form ICS II in extending healthspan via the well conserved pathway IIS. Given the extensive protective effects and safe long term use of ICA and ICS II in humans they may serve as promising anti-aging candidates in the future.
衰老相关疾病与衰老进程本身具有同源性。干预衰老能够降低衰老相关疾病的发生发展,延长健康寿命,是实现健康老龄化的关键环节,但目前仍缺乏有效的延缓衰老手段。中医药学在养生健体、延缓衰老方面历史悠长。在前期补肾延缓衰老系列研究中,我们发现淫羊藿总黄酮(EF)为补肾抗衰老复方改善内分泌免疫衰老的主要有效成分。寿命为评价衰老干预手段的核心指标,但是EF能否延长寿命还属未知。淫羊藿苷(Icariin, ICA)是EF的主要成分,但ICA是否为EF延长寿命的有效成分需进一步研究。
延长寿命更重要的是延长有功能意义的健康寿命,因此检测衰老干预手段对老年机体健康状态的改善作用尤为重要。目前对老年机体健康状态的检测主要包括压力应激能力、衰老退行性改变以及衰老相关疾病等3个方面。ICA能否改善机体的健康状态,延长健康寿命还属未知。
现代药理研究证实,ICA经吸收后在体内被迅速代谢为淫羊藿次苷Ⅰ(Icariside Ⅰ, ICS Ⅰ)、次苷Ⅱ (Icariside Ⅱ, ICS Ⅱ)及淫羊藿素(Icaritiin, ICT)。ICA是以其原型还是代谢衍生物发挥延长健康寿命的作用值得研究。目前研究证实,Insulin/IGF-1通路(IIS)、Target of rapamycin通路(TOR)、沉默信息调节子Sirtuins家族以及AMP kinase (AMPK)等为寿命调控的核心分子机制。其中IIS通路已被证实能够调节包括低等和高等生物在内的一系列物种的寿命,其作用在进化上高度保守。寻找这些寿命调控通路的靶向药物是抗衰老研究的一个主要目标。在揭示ICA为EF发挥作用的单一成分后,ICA及其体内活性代谢衍生物是否通过上述衰老调控通路发挥作用值得探索。
模式生物秀丽隐杆线虫(以下简称线虫)是衰老研究的经典模式生物,具有实验操作简单、遗传背景清晰等优点,易于进行寿命实验和作用机制的探索,但低等生物毕竟与人类相差太远,药物的确切效果还需进一步通过哺乳动物和人类细胞的验证。因此,在本研究中我们采用了线虫、C57BL/6小鼠及人类成纤维二倍体细胞模型MRC-5等3种实验模型,结合各实验模型的优势来探索药物延缓衰老的作用及其机制。
目的：
1.研究EF及其主要成分ICA干预寿命的效果。
2.评价ICA对老年健康状态的改善作用；
3.探讨ICA延长健康寿命的体内活性成分及其分子机制。
方法：
1.EF及ICA干预寿命的研究
1.1野生型线虫N2分组为对照组、EF高(100mg/mL)、中(50mg/mL)、低剂量组(25mg/mL);对照组、ICA高(75μM)、中(45μM)、低(15μM)剂量组进行寿命实验；
1.2C57BL/6雄性小鼠分组为正常对照组、EF组(0.06%,w/w)及ICA组(0.02%,w/w),进行寿命实验,记录各组小鼠体重、饲料日耗量、饮水量、大便小便量,小鼠自然死亡后行大体解剖观察肿瘤发生及皮肤溃疡情况。
1.3人二倍体成纤维细胞MRC-5分组为对照组、EF组(3mg/mL)及ICA组(10-6mol/L),检测细胞传代次数(PD)；
2.ICA影响老年健康状态的研究
2.1压力应激能力：野生型线虫N2分组为对照组、ICA组(45μM),进行热压力应激及氧化压力应激实验,观察ICA对压力应激能力的影响；
2.2衰老退行性病变：MRC-5细胞分组为对照组、ICA组(10-6mol/L),进行细胞衰老相关β-半乳糖苷酶染色实验；野生型线虫N2分组为对照组、ICA组(45μM),进行游泳能力测试；线虫突变体PD4251分组为对照组、ICA组(45μM),进行肌肉细胞计数；C57BL/6小鼠的分组对照组、ICA组,同时取3月龄小鼠作为青年对照,进行转棒实验、骨密度检测及重要脏器病理观察,观察ICA对衰老退行性病变的影响；
2.3衰老相关疾病：线虫疾病模型突变体AM140、CL4176分组为对照组、ICA组(45μM),观察ICA对谷氨酰胺重复序列(PolyQ)及人类β-淀粉样肽(Aβ1-42)蛋白毒性诱导的神经退行性病变的作用,检测ICA对衰老相关疾病的作用。
3.ICA的体内代谢衍生物ICSⅡ延长健康寿命作用机制的研究
3.1野生型线虫N2分组为对照组及药物干预组,观察ICA及其衍生物ICSI、ICSII、ICT对寿命的影响并结合高效液相(HPLC)检测ICA和各衍生物在野生型线虫体内的浓度；
3.2野生型线虫N2分组为对照组、ICS Ⅱ组(20μM),进行压力应激实验、游泳能力测试、肌肉细胞计数,观察药物对衰老相关退化的改善作用；线虫疾病模型突变株AM140、CI4176分组为对照组、ICS Ⅱ组(20μM),观察药物对PolyQ及Aβ1-42蛋白毒性诱导的神经退行性病变的影响。
3.3不同遗传背景的线虫突变体分组为对照组、ICS Ⅱ组(20μM),进行寿命实验(Genetic Epistasis Experiment),探讨药物作用机制,并进一步用实时定量RT-PCR、Western Blot及相关基因GFP报告体检测药物对相关通路下游靶基因的表达的影响。
结果：
1.EF及其主要成分ICA能显著延长多个物种实验模型的寿命
EF及ICA呈剂量依赖性右移野生型线虫的生存曲线,提高线虫平均寿命及最大寿命；显著右移小鼠的生存曲线,显著提高小鼠平均寿命,对小鼠的最大寿命无明显影响。药物干预对小鼠自发肿瘤及皮肤溃疡发生率无明显影响。EF及ICA能显著提高人类二倍体细胞MRC-5的传代次数；
2.ICA能显著改善老年健康状态
ICA能显著降低MRC-5细胞衰老特异性p-半乳糖苷酶的表达,提高线虫的热应激、氧化应激能力及运动能力,改善线虫衰老相关肌肉萎缩,延长小鼠转棒潜伏期时间；ICA能降低蛋白毒性介导的线虫瘫痪表型,提高老年小鼠股骨骨密度。经ICA干预过的小鼠肝组织中衰老相关病理改变明显减轻。药物长期干预对小鼠主要脏器无明显毒性影响。
3.ICA及其活性代谢物衍生物ICS Ⅱ通过ⅡS通路延长健康寿命。
在野生型线虫上对ICA及ICA代谢衍生物进行寿命实验显示20μM ICSⅡ能获得与45μM ICA同等的延长寿命的效果,HPLC结果显示在秀丽线虫中测得的化合物主要为ICA及其代谢衍生物ICSⅡ,提示ICSⅡ为ICA发挥抗衰老作用的体内活性衍生物。ICSⅡ能显著提高野生型线虫的热应激及氧化应激能力,提高老年线虫的运动能力,降低衰老相关肌肉萎缩,减低PolyQ及Aβ1-42的蛋白毒性。ICA及ICSⅡ不能影响ⅡS通路上游基因daf-2突变体、下游核心转录因子daf-16突变体及协助转录因子hsf-1突变体的寿命,提示药物作用机理与ⅡS通路相关。进一步的实时定量RT-PCR、Western Blot及SOD-3绿色荧光报告实验结果结果显示,ICS Ⅱ能增强IIS通路的核心转录因子DAF-16的转录活性,提高其下游有益靶基因的表达,从而延长健康寿命。
结论：
综上所述,我们首次发现EF、ICA及ICS Ⅱ能改善多个物种实验模型的衰老相关性退化、降低衰老相关疾病的发生发展,延长健康寿命。我们发现ICS Ⅱ为ICA发挥抗衰老作用的主要体内活性衍生物；ICA及ICS Ⅱ通过抑制ⅡS通路从而增强FOXO/DAF-16转录活性发挥抗衰老作用。本研究提示ICA及ICS Ⅱ能延缓从低等至高等多个物种实验模型的衰老,并通过进化保守的ⅡS通路发挥作用,极有可能延缓人类衰老。临床使用经验及本研究小鼠实验显示ICA可长期使用,安全性高,有望走向人类抗衰老临床使用。
Introduction
A major goal of current research on aging is to identify compounds that delay age-related diseases and extend healthspan in humans. Herba epimedii is a popular herbal tonic used in traditional Chinese medicine, with proven efficacy in treating several age-related diseases including osteoporosis, cardiovascular diseases, neurodegenerative diseases and sexual dysfunction. Our previous studies on Epimedium Flavones (EF), the raw extract of Epimedium which contains icarrin (ICA) as a major constituent, show that EF delays aging in Drosophila melanogaster and cell senescence model. Interestingly, EF also resets the age-related metabolites (fatty acids, carnosine, ergothioneine and deoxycholic acid et al) to the juvenile level in rat plasma and urine. These findings prompted us to investigate the anti-aging potential of EF and its major pharmacologically active flavonol diglycoside, icariin. Various studies indicate the anti-oxidative effect of icariin and derivatives on DNA damage, β-amyloid mediated neurotoxicity, and vein endothelial cell oxidative injury. Meanwhile, icariin and its derivatives function as signalling modulators to exert beneficial effects in a multitude of age-dependent disease states, including bone loss, cancer, cardiovascular disease, and neurodegenerative disorders. These researches further strengthen the possibility of the effects for EF, icariin and its derivatives to extend healthspan.
The highly conserved IIS pathway plays a key role in aging. It has been demonstrated in multiple species that inhibition of the IIS pathway extends lifespan. In C. elegans, the daf-2gene encodes an insulin/IGF-1receptor. Mutations in daf-2suppress the IIS which lead to the nuclear localization of FOXO/DAF-16transcription factor. The activated FOXO/DAF-16regulates a series of genes involved in lifespan control, stress tolerance and protein misfolding suppression. Nuclear localization of DAF-16in the daf-2mutants requires the heat shock transcription factor (HSF)-1, which modulates the expression of heat shock proteins and protease responsible for the stress tolerance and protein folding. Despite the well established role of a number of genes in the IIS pathway in modulating aging, pharmacological tools that inhibit IIS pathway to extend lifespan are not commonly available, which limits its translation to mammalian model systems.
Objectives
1. To investigate the delaying aging effects of EF and ICA in the experimental systems from invertebrate to mammalian.
2. To measure the effects of ICA in extending healthspan.
3. To ascertain the bioactive formation of ICA in vivo for the healthspan extension and investigate the molecular mechanisms of these effects.
Methods
In this study, we employed three experimental models including the cell senescence model MRC-5, the simple animal model C. elegans and the mammalian model C57BL/6mice to investigate the delaying aging effects of the compounds. Lifespan is the major measurement to evaluate the effect of the compounds. Stress-resistant assays, mobility assay, qualification of muscle cells, rotarod experiment, protein-misfolding disease models are used to evaluate the effects of the compounds in healthspan. HPLC was used to measure the level of ICA and its derivatives in vivo. Genetic epistasis experiments were used to study the mechanism. Three major genetic modulate pathways in longevity were tested in this study, the insulin/IGF-1pathway, the dietary restriction and the target of rapamycin (TOR) pathway. Real-time PCR, western blot were used to measure the downstream targets of the pathway to ascertain the molecular mechanism.
Results
In this study, we tested the anti-aging properties of EF, ICA and its three derivatives, icariside Ⅰ (ICS Ⅰ), icariside Ⅱ (ICS Ⅱ) and icaritin (ICT) in the cell senescence model MRC-5, the simple animal model C. elegans and the mammalian model C57BL/6mice. We found that EF has a conserved role in extending lifespan from simple animal to mammalian model. ICA is the major pharmacological element of EF to extend lifespan. Additionally, ICA promotes the stress resistance and mobility in late life of the animals. Meanwhile it ameliorates protein aggregation and protetoxicity-mediated paralysis phenotype. And one of its derivatives, ICS Ⅱ prolonged adult lifespan. Chemical profiles of ICA treated animals revealed that ICS Ⅱ was the predominant bioactive form of ICA in vivo. Furthermore, we found that ICS Ⅱ treated animals have delayed age-associated phenotypes suggesting ICS Ⅱ enhances the healthy aging significantly. Finally, our genetic analysis indicates that Icariside Ⅱ may act through the IIS pathway to affect lifespan.
Conclusion
Our findings reveal a novel role for EF, ICA, along with its bioactive form ICS II in extending healthspan via the well conserved pathway IIS. Given the extensive protective effects and safe long term use of ICA and ICS II in humans they may serve as promising anti-aging candidates in the future.
引文
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