Study On The Material Basis Of Qingyi Recipe And The Mechanism Of Action Of Interventional Acne Model Rats

Objectives1.We clarified the efficacy and pharmacological mechanisms of TCM Qing Cuo Formula(QCF)ameliorating the acne on Golden Hamster Model by observing the changes in the spot area and tissue structure of sebaceous gland,inflammatory factors and sex hormone levels in serum before and after the TCM QCF intervention.2.We established the TCM Blood Chemical Constituent's Chromatographic Fingerprints Database based on the TCM Serum Drug Chemistry Theory by using UPLC-LTQ Orbitrap MS Technology,in order to identify the chemical constituents in QCF original decoction and serum after administrated QCF.3.We tried to find the "Chemical Spectrum-Pharmaceutical Effect" Relationship of QCF by docking the relationship between the original/serum chemical constituents and pharmaceutical effect on Golden Hamster acne,so that we could determine the activecomponents and effective mechanism of QCF treating acne.MethodsSixty male Golden Hamsters were selected as damp-heat acne model for their back sebaceous gland spots.The hamsters were randomly grouped into four groups by their weight,including control group,QCF low-dose group,QCF medium-dose group,QCF high-dose group,and positive drug group.The spironolactone was chosen as positive drug.After intra-gastric administrated QCF and spironolactone for 30 days,the hamster blood was collected to test serum testosterone(T),dihydrotestosterone(DHT),estradiol(E2),interleukin-1?(IL-1?),free fatty acid(FFA),and tumor necrosis factor ?(TNF-?)level.Combing the changes in the area of sebaceous gland spots and pathological sections,the macro and micro changes of spots structure were found to evaluate the effect of QCF.Under the theory of TCM Serum Drug Chemistry,the chemical components of original decoction,normal plasma and QCF plasma were tested by using UPLC-LTQ Orbitrap MS Technology.By analyzing the fragments and distinct micro molecule to speculate the primitive structure of relative chemicals by Xcalibur 2.2 and HighChem Mass frontier 7.0 software,the active chemicals in original decoction and QCF plasma were determined.Result1.Contrast to control group,the area of sebaceous gland spots in all QCF groups and positive drug group became smaller;the micro-structure of sebaceous gland became loose and gland lobes shrink.The reduce range of spots in high-dose group were the most obvious(P<0.05).2.Contrast to control group,the level of serum T,E2,and DHT in low-dose and medium-dose group declined,which the decline of serum T and DHT in low-dose and positive-drug group was significantly different(P<0.05);the level of IL-1?,TNF-? in low-dose,medium-dose and positive drug group declined,which the decline of IL-la in low-dose,positive drug group was significantly different(P<0.05);the level of FFA in low-dose and positive drug group declined significantly(P<0.05).3.After established the QCF In Vitro Chemical Components Database,75 chemicals in decoction and 26 chemicals in plasma were identified.The chemicals in plasma including 16 original components and 10 metabolites.After comprehensively analyzing the active chemicals,QCF had effects in anti-bacterial,anti-inflammatory,and androgen-inhibiting.Conclusion1.QCF had obvious effect on the structure of sebaceous glands of Golden Hamsters,which could inhibit cell proliferation,and reduce sebum secretion.The spots faded and the area became small.These indicated that QCF could inhibit the overactive sebaceous gland function,meliorate its micro-structure,reduce excess sebum secretion,which means QCF has definite therapeutic effect on acne.2.QCF could decline the level of serum IL-1?,TNF-? in Golden Hamsters,indicating the formula has anti-inflammation effect.QCF could decrease the level of androgen,T,DHT and FFA,indicating that the formula could regulate the hormones level in order to reduce the acne caused by the excessive androgen.In this experiment,the low-dose had best effect on acne,which conformed to the thinking in TCM Constitution Regulation Theory.3.By testing the chemical components contained in original decoction and drug plasma,a variety of active chemical components were identified which had effects on antibacterial,anti-inflammatory,and androgen-inhibiting.These components provided the material basis for further studies on the potential mechanism of QCF treating damp-heat acne.