Potential mechanism of Huatan Qushi decoction on improving phlegm-dampness constitution using microRNA array and RT-qPCR targeting on hsa-miR-1237-3p

摘要

Background:Metabolic diseases pose considerable burden on the healthcare system worldwide,indicating the significance of prevention and treatment.In constitution theory of traditional Chinese medicine,phlegm-dampness constitution(PDC)is the common basis of metabolic diseases.In clinical practice,Huatan Qushi(HTQS)decoction targeting on PDC can effectively improve metabolic indicators.However,its underlying biochemical mechanism still remains unclear.Methods:Eight PDC participants received HTQS decoction for three months.Their blood was collected at baseline and 1 and 3 months after intervention started.Related biomedical indicators were detected.High-throughput sequencing and RT-qPCR were used for validation.Due to the missing data,repeated measures with missing values in mixed models were used.Results:After 3-month treatment,HDL-C level increased(P<.001)and FBG,FINS,and HbA1c all showed decreasing trend at different time points(all P<.05).After miRNA high-throughput sequencing,compared with the baseline,differential miRNAs at 1 and 3 months were screened,and target gene prediction and KEGG pathway enrichment analysis were performed.The results displayed that metabolic disease-related pathways mainly included pathways in cancer,PI3K-Akt signaling pathway,etc.Further,RT-qPCR showed that hsa-miR-1237e3p differed statistically(P=.008).Then we validated the target genes of hsa-miR-1237e3p in the“Pathways in Cancer”pathway including SDF1,AC,CRK,and HGF,also known as upstream target genes of PI3K/AKT pathway.The results showed that two indicators of CRK and HGF were in statistical significance(P=.045 and P=.036,respectively).Conclusion:PDC serves as a common basis for various metabolic diseases.Through adjusting PDC,HTQS decoction can improve biomedical indicators including blood glucose,HbA1c,insulin,and HDL-C.The target pathway is“Pathways in cancer”.Specifically,HTQS decoction acts on targets of CRK and HGF by regulating hsa-miR-1237e3p,and probably exerts effects on their downstream PI3K/AKT pathway.