Research On Chemical Components And Physiological Functions And Resource Utilization Of Codonopsis Pilosula Leaves

The non-medicinal parts of Chinese medicinal materials usually have high value of research and utilization.How to use these resources efficiently,reasonably and economically is of great significance.Codonopsis pilosula is a bulk medicinal material used both for medicine and food.The traditional medicinal parts are only the roots.The plant’s above-ground parts including stems,leaves and flowers contain a large number of chemical substances,so they have great development potential.However,the exploration of their medicinal utilization values and ways remains to be studied.To explore the potential developing value of the aerial parts of Codonopsis pilosula deeply,the Codonopsis pilosula leaves（CL）were studied in this research.By using UPLC-MS/MS and other chemical analysis methods,the main active substances in CL were studied.Based on the dominant chemical constituents in CL,the possible important physiological activities were explored.The results showed CL had significant in vitro antioxidant activity,which inhibited free radicals effectively.Therefore,the in vivo anti-aging and anti-fatigue activities were carried out.Caenorhabditis elegans（C.elegans）aging model and Kun Ming（KM）mice fatigue model were used to explore anti-aging and anti-fatigue activity and related mechanisms.Through investigating the effects of CL on endogenous oxidative metabolites,and by selecting daf-16 gene mutation C.elegans and daf-16 and SOD-3 gene green fluorescent protein fusion C.elegans,the mechanism of anti-aging effect was revealed.By investigating the effects of CL on blood and tissue metabolism indexes of mice and the regulation of PGC-1α,PPARαand MHC-1 genes,the anti-fatigue mechanism was revealed.As a breakthrough point,based on the health promoting physiological activities including anti-oxidant,anti-aging and anti-fatigue of CL,and combining the advantages of high safety,CL were developed into Codonopsis pilosula leaf tea（CLT）.The key processing technology of CLT was explored by response surface optimization model combined with weight analysis method.The CLT with good sensory quality and high active substance content was developed.Processing the leaves into tea provided a feasible way for the comprehensive utilization of Codonopsis pilosula leaves with high-profit.The main results are as follows:（1）The results showed Codonopsis pilosula leaves had high levels of polyphenols,flavonoids,and lobetyolin,and they were materials rich in amino acids and mineral elements.Though the contents of polysaccharides and extracts in leaves were significantly lower than those in roots,the contents of polyphenols,flavonoids and other substances with important activities were significantly higher than those in the roots.By UPLC-MS/MS method,52 phenolic acids,55 flavonoids,22 sugars and alcohols,14 alkaloids,17 organic acids,and 2 terpenes Class,6 lignans and 5 coumarins were identified.Hence,the chemical material foundation for the utilization of Codonopsis pilosula leaves is sound.（2）The in vitro antioxidant test showed that CL inhibited DPPH and ABTS free radicals efficiently.The total antioxidant capacity was strong.In vivo tests showed that CL had notable anti-aging and anti-fatigue effects,and a dose-effect relationship was found.The mean lifespan of C.elegans in the groups treated with CL increased by 7.10%-18.88%,the groups treated with 0.25 mg/m L of CL performed best.For the groups treated with CL,the contents of lipofuscin in intestine reduced significantly;the exercise capacity improved significantly;after heat and oxidative stress,the survival time of C.elegans increased significantly;the levels of ROS and MDA decreased;the activities of Superoxide dismutase（SOD）increased.The regulation of daf-16 gene is an important anti-aging mechanism of CL,which induced the expression of downstream target gene of SOD-3 gene by activating the protein nuclear translocation of daf-16.Through the above regulation,CL presented notable anti-aging effect.The CL also presented a significant anti-fatigue effect.The weight-loaded swimming time of the mice treated with CL prolonged significantly.The group treated with 1 g/kg/d performed the best effect.The mechanism of anti-fatigue effect of CL was found by significantly reducing the levels of BUN,LDH,BLA in serum,the levels of MDA and glycogen consumption liver,and increasing the SOD enzyme activity in liver.The results found that the m RNA expressions of PGC-1α gene were up-regulated under the synergistic effect of PPARαand MHC-1 genes in AMPK signaling pathway.Thus,CL has strong health promoting activities,providing an important theoretical basis and breakthrough point for its utilization.（3）The safety evaluation results of CL showed that no toxicity was observed in the acute toxicity test.LD₅₀ could not be detected out in the dose range of 13.36g/kg～26.72 g/kg,there was no death or physiological abnormality.And the maximum tolerable dose was 26.72 g/kg.Therefore,CL is safe materials reaching the non-toxic level.The results of pesticide residue test showed that all indicators of CL used in this research meet the standards.Therefore,CL has relatively safe performance,providing a good safety basis for its comprehensive utilization.（4）Taking the physiological activity of CL as the starting point,and based on the advantages of high safety,CL was developed into CLT with good sensory quality and high active substance content.The processing technology of CLT was explored by using response surface optimization model and weight analysis method.The key processing technology of CLT mainly includes spreading,green fixation,rolling,and drying.The response surface optimization model combined with the weight analysis method was used to calculate the comprehensive score to explore its key processing parameters.The optimal green fixation temperature obtained by the response surface optimization model was 266℃with the time of 3 min;the amount of leaves was 51g;the drying temperature was 75℃,the baking time was 27 min with the temperature of 110℃.The development of CL into CLT provided a feasible way to promote the utilization of these resources.This research provides an important reference for the promotion and efficient utilization with added economic value of CL,and has important guiding and reference value for the comprehensive utilization of Codonopsis pilosula resources.