Study Of The Anti-aging Effects Of TCMs That Could Promote Qi And Activate Blood Or Nourish Kidney And Liver On C.elegans And The Underlying Molecular Mechanisms

The 21st century is the era of aging population, aging and overgrowth of the population bring great challenges to social economy, culture,education and medical care.Therefore,it is an important ergent task to explore the mechanisms of aging and find out the effective way to delay aging.The mechanisms of aging are important contents of aging research. So far, there are more than 200 theories about aging, including damage of free radicals, disorder of neuroendocrine system, disorder of metabolism, damage of mitochondria, dietary restriction, etc. But all in all, the theories above are all regulated by gene expression. There are always some related gene expression changes corresponding to any kind of the aging theories. So, the theories could be integrated at the genetic level to investigate the aging mechanisms and the effects of medicine on it.During the long-term clinical practices of traditional Chinese medicine (TCM), many unique aging theories have been formed, a lot of TCM formulas that could retard aging effectively have been found as well with great value for research and application. In TCM, kidney and liver have close relationships with aging that kidney deficiency is the foundation of aging while liver deficiency is the aid of it. Erzhi Wan is a famous formula with founction of nourishing kidney and liver(NKL), which has been widely used in clinical practices.Since its anti-aging effects has been proved, it is necessary to explore the underlying molecular mechanisms. Qi stagnation and blood stasis(QSBS) is another important TCM theory of aging. The formulas with founctions of promoting Qi and activating Blood (PQAB)could delay aging through improving microcirculation and immunity, strengthening the ability of stess resistence. Chuanxiong is a typical TCM with functions mentioned above that deserves further research.Caenorhabditis elegans(C.elegans) has been an excellent organism for aging research with good characters of short lifespan, cheap and easy to culture, huge amounts of offspring, small numbers of cells, well-understood genetic background, ect.In the study, we chose C.elegans as a organism model to screen the medicines that could prolong lifespan of the animal from more than 30 kinds of TCMs. Among them, Erzhi Wan and Chuanxiong were the typical ones.The effects of them on reproductivity and acute heat-resisting ability of the nematodes were also investigated. In addition, we used realtime RT-PCR to explore the different underlying molecular mechanisms between these two TCMs.The method of culturing C.elegans with TCMs was established that the water extract of TCMs could be added directly to the mediums; By lifespan assay,7 of 30 TCMs were screened out for lifespan-prolonging effects on C. elegans, Erzhi Wan and Chuanxiong are the typical ones.We found that the water extract of Erzhi Wan could prolong lifespan of C.elegans effectively and the optimal concentration is 500mg/L. The mean lifespan could be extended by 10.6% (P<0.01), and the maximum lifespan was extended by 16.2% (P <0.01). Furthermore, the acute heat-resisting ability of the animal was increased to a certain extent while the reproductivity remained unchanged. By using realtime RT-PCR, we found that the anti-aging effects of Erzhi Wan pertain to the regulation of genes expression in IIS pathway,neuroendocrine and clock genes,ect.We found that the water extract of CXE could prolong lifespan of C.elegans effectively and the optimal concentration is 25mg/L. The mean lifespan could be extended by 29.9% (P<0.01), and the maximum lifespan was extended by 9.4% (P< 0.01). Furthermore, the acute heat-resisting ability of the animal was increased to a certain extent while the reproductivity remained unchanged. By using realtime RT-PCR, we found that:â‘ In the progress of aging, the expression of aging genes in IIS and dietary restriction pathway increased, while the expression of longevity genes decreased;â‘¡CXE entended the lifespan through regulating genes expression of IIS, inhibiting TOR signal, improving antioxidation, stress resisting and mitochondria activity, which contributed to affecting the physiology of C.elegans.In the study, we used a model organism-C.elegans, which was suitable for large-scale medicine screening and mechanism research, to screen medicines that could delay senility effectively. Among more than 30 kinds of TCMs,7 TCMs were screened out for medicines that could prolong lifespan effectively, Erzhi Wan (NKL) and Chuanxiong (PQAB) were the typical ones.Both Chuanxiong and Erzhi Wan could extend the lifespan and improve the heat-resisting ability of C.elegans with no reduction of reproductivity. But there were similarities and differences of molecular mechanisms between these two TCMs. The similarity was that both of them have relationships with IIS, the longevity genes of the pathway were upregulated while aging genes were downregulated.The differences were:besides IIS, Erzhi Wan affected aging mainly through regulating neuroendocrine genes and clock genes while Chuanxiong mostly affected genes of energe metabolisms, such as dietary restriction, mitochondrial activity and fat storage, etc.Consequently, different anti-aging TCM has different molecular mechanisms.We wondered whether this character could be used to study the efficacy and component compatability of TCMs proceeded with molecular targets in the future.In this study, we integrated several aging theories at the genetic level, which contributes to understand the relationship and regulating network among them more easily, helps to seek for the real function genes affecting aging. We considered the methods would bring great benefits for mechanism research and development of new medicine.