Comparative Study Of The Inhibition Effect On Preadipocyte Differentiation And Anti-inflammatory Effect Of Paecilomyces Hepiali And Hirsutella Sinensis

Cordyceps sinensis is a precious traditional Chinese medicine with a variety of pharmacological activities.However,the resource is scarce and expensive which limit its use.With the isolation of fungal anamorphs from the natural Cordyceps sinensis,the artificial mycelia gradually become the Cordyceps's substitutes.Among them,Paecilomyces hepiali is comprehensive in the activity evaluation and widely used in Chinese traditional patent medicines as one well-studied substitute of cordyceps sinensis.More and more studies are showing that Hirsutella sinensis is the true anamorph of Cordyceps sinensis,but the high cost of fermentation and the lack of bioavailability research limit the research and utilization of the Hirsutella sinensis resources.Paecilomyces hepiali has hypolipidemic and anti-inflammatory effects,but the study of the Hirsutella sinensis on fat metabolism has been rarely reported,meanwhile the anti-inflammatory mechanism of Hirsutella sinensis is unclear enough.Therefore,this paper used the adipocyte and macrophage cell model to study the action of Hirsutella sinensis in the regulation of fat metabolism and anti-inflammatory effects and was compared with the activities and mechanisms of Paecilomyces hepiali.·First,this study measured the main components of the mycelia of Paecilomyces hepiali and Hirsutella sinensis.The results showed that the contents of adenosine,total sugar,reduced sugar,total amino acids and mannitol were 0.038%,3.188%,0.252%,41.198%and 6.1%in the mycelia of Paecilomyces hepiali,and 0.013%,3.846%,0.310%,40.112%and 6.0%in the mycelia of Hirsutella sinensis.Among them,the mycelia of Paecilomyces hepiali was high in adenosine,the total amino acids and mannitol,while Hirsutella sinensis was high in the total sugar and the reduced sugar.Then,the mycelia of Paecilomyces hepiali and Hirsutella sinensis were extracted with methanol and then with water to get four extracts.The contents of adenosine in the methanol extract of Paecilomyces hepiali(PHME)and Hirsutella sinensis(HSME)were 0.296%and 0.112%,and mannitol 17.62%and 34.61%.The contents of total sugar in the water extract of Paecilomyces hepiali(PHWE)and Hirsutella sinensis(HS WE)were 8.591%and 18.398%,and reduced sugar 2.670%and 3.252%.Next,we compared the effects of the extracts on the differentiation of 3T3-L1 preadipocytes into adipocytes by Oil Red O staining experiments.Both the methanol and water extracts of Hirsutella sinensis performed well in inhibiting 3T3-L1 preadipocyte differentiation,but only the water extract of Paecilomyces hepiali performed well in it.We explored the possible mechanism of the PHWE,HSME and HSWE on the differentiation of 3T3-L1 preadipocytes and the results showed that the three extracts could decreased the adipogenic-related gene expressions,such as PPARy,C/EBPa and SREBPI-c.Further studies showed that both the extracts of Hirsutella sinensis might inhibit the preadipocyte differentiation by activating the ERK signal pathways,but the water extract of Paecilomyces hepiali also suppressed the JNK and p3 8 signaling pathways while activating the ERK signal patthways.We used the lipopolysaccharide induced RAW264.7 macrophages to compare the anti-inflammation activity of Paecilomyces hepiali and Hirsutella sinensis in vitro-The results suggested that HSME suppressed the expression of inflammatory factors and exhibited the anti-inflammatory activity.However,PHME,PHWE and HSWE did not exhibit anti-inflammatory activity.Further study of the anti-inflammatory mechanism showed that HSWE maybe down regulate the expression of TNF-?,IL-6,IL-1? and iNOS via the suppression of ERK,p38 and NF-?B activation in LPS-induced RAW264.7 cells.In conclusion,Hirsutella sinensis performs well in the regulation of fat metabolism and anti-inflammatory effects and is superior to Paecilomyces hepiali with the good promising applications.This paper may enrich the efficacy research and provide theoretical basis for the efficacy exploitation of the Hirsutella sinensis and has the practical significance for promoting the development and use of Hirsutella sinensis.