The aim of this study was to explore the molecular network regulation mechanism of Coptidis Rhizoma and Scutellariae Radix (SRCR) on treatment of Helicobacter pylori (Hp) associated peptic ulcer (PU) based on network pharmacology and bioinformatics methods,and to screen out the core genes and key microRNAs.Firstly,the main active ingredients and possible targets of SRCR were retrieved through the Chinese Medicine System Pharmacology Database and Analysis Platform (TCMSP).GSE70394 gene expression profile data was downloaded from the GEO database.The microarray data of differentially expressed genes in Hp infected human gastric epithelial cells were homogenized by R software,and the volcano map and cluster heat map of differentially expressed genes were drawn.Secondly,STRING database was used to construct the protein interaction network between SRCR component target and Hp- infected human GES cells.The functions of GO and the enrichment of KEGG pathways of the above-mentioned characteristic genes were analyzed by DAVID database.Then,the core genes involved in the treatment of Hp related PU by SRCR were screened out by cytohubba plug-in unit.The key microRNAs regulating the core genes were predicted by using the TargetScan and miRBase databases.Finally,41 active components and 216 potential targets of SRCR were retrieved.The quality of GEO chip data was good.With P< 0.05 and︱logFC︱≥ 2 as the screening conditions,128 differentially expressed genes were obtained,including 77 up-regulated genes and 51 down-regulated genes.127 characteristic gene targets were further obtained for the treatment of HpPU by SRCR.Go analysis of these targets showed that the biological process involved 55 items such as inflammatory response,immune response,etc.Cell components included 11 items such as extracellular space,exosomes,nuclei,etc.Molecular functions included 15 items such as growth factor activity,epidermal growth factor receptor binding,etc.KEGG results showed that 33 signaling pathways,including cytokine receptor interaction pathway,nod like receptor signaling pathway,NF-κB signaling pathway,etc.were closely related to the treatment of Hp related PU by SRCR.Ten core genes regulated by SRCR,such as CXCL8,IL10,IL1B,etc.were screened out by cytohubba.Ten of the most functional microRNAs,such as mir-93-5p,mir-374b-5p,mir-3937,etc.which may be regulated by SRCR,were further screened out.The above results indicate that the treatment of HpPU with SRCR has the characteristics of multi-component,multi-target,and multi-channel synergy.SRCR can exert its therapeutic effect on HpPU by regulating the coding RNA (gene) and non-coding RNA (microRNA).

Aimed to provid a new approach for the exploitation of natural product oroxylin A,this reasearch focused on obtaining oroxylin A from baicalin by biotransformation applying endophytic fungus-Penicillium sp.R3 from Scutellaria baicalensis Georgi,identifying its biotransformation pathway and optimizing the fermentation conditions.The culture and biotransformation reaction were carried out in potato dextrose broth(PDB).Cell growth was measured by dry weight and the maximum biomass of Penicillium sp.R3 was gained on the day 4,then it entered the stationary phase.The biotransformation pathway was analyzed by the method of adding different substrates and the products were measured and identified by HPLC.After fermentation baicalin was converted to oroxylin A accompanied with baicalein which was identified as intermediate metabolite.Single factor analysis and orthogonal test were applied to optimized fermentation conditions.The optimal fermentation conditions were as follows:the liquid medium volume was 10% of the bottle;the concentration of inoculated spores was 1×10⁶-2×10⁶/mL and the temperature and shaker speed were 30 ℃,150 rpm respectively.The substrate baicalin was added on the day 3 with proportion of 0.08%,then the biotransformation process sustained 6 days.Under these optimal conditions the molar conversion rate and concentration of oroxylin A reached 42.44% and 177 mg/L respectively.The growth regularity and the biotransformation pathway of Penicillium sp.R3 was clarified,the stable and optimal conditions were obtained with a satisfactory yeild.All these findings lay a foundation for the further development and utilization of oroxylin A.

The purpose of the study was to investigate the effect of different proportions of Scutellariae Radix(SR) and Fritillariae Cirrhosae Bulbus (FCB) on the content of main chemical components in SR and the anti-pneumonia effect of equal proportions.SR and FCB was mixed in the ratios of 3∶0,3∶1,3∶2,3∶3,2∶3,1∶3,0∶3,and then extracted with water and determined by HPLC to determine baicalin,wogonoside,baicalein and wogonin.Thirty-six healthy ICR mice were randomly divided into six groups:control group,LPS group,treatment group,SR group,FCB group,and SRFCB group,six in each group.The drug was administered for 6 consecutive days.On the 7th day,the experimental group was injected intraperitoneally with 10 mg/kg of LPS,and the control group was injected with an equal volume of normal saline.LPS was used for 6 hours to determine the levels of IL-1β,IL-6 and TNF-α in the serum of the mice.The pathological changes of lung tissue of mice in each group were observed under light microscope.The results showed that the total content of baicalin,wogonoside,baicallein,wogonin was less than 6% when the compatibility ratio were 3∶1,3∶2,3∶3 and 2∶3.When the compatibility ratio reached 1∶3,the total amount of the four chemical components decreased by 23.7%.After equal proportions compatibility of SR and FCB,the levels of IL-1β,IL-6,TNF-α in the serum of mice were significantly reduced,and the pathological morphology of lung tissue was improved,and the therapeutic effect was better than that of the single drug group.It is advisable to control the proportions compatibility of SR and FCB to 3∶1-2∶3,and they have a synergistic effect in anti-pneumonia.

In this study,the inhibitory effect and mechanism of baicalein on tumor growth and lung metastasis of human breast cancer cell line MDA-MB-231 in nude mice was investigated.Twenty female BALB/C nude mice were randomly divided into normal group,model group,paclitaxel group and baicalein group.There were 5 nude mice in each group.The nude mice model of human breast cancer xenograft was established.Baicalein group was injected with 50 mg/kg baicalein abdominal cavity,paclitaxel group was injected with 10 mg/kg paclitaxel,normal group and model group were injected with normal saline for 8 weeks.Experimental results indicated that the tumor weight and the number of lung metastasis nodule and metastasis focus in paclitaxel group and baicalein group were less than those in the model group.Baicalein alone or in combination with ROCK inhibitor Y-27632 could inhibit the migration and cytoskeleton protein expression of MDA-MB-231 cells,reduce the formation of protein bundles,and inhibit the protein levels of ROCK1,Rac1,RhoA and p-MLC.

In order to investigate the effect of wogonin on atherosclerosis and arterial inflammation in ApoE^-/- mice fed with high fat diet,ApoE^-/- mice were divided into normal diet group (ND),high-fat diet group (HFD),high-fat diet + wogonin (10 mg/kg) group (HFD + wogonin 10 mg/kg) and high-fat diet + wogonin (20 mg/kg) group (HFD + wogonin 20 mg/kg).Mice were given high-fat diet and wogonin for 12 weeks and then the arterial tissues were collected.RT-qPCR was used to detect the mRNA expression of IL-6,TNF-α,ICAM-1 and VCAM-1.Immunofluorescence staining was used to detect the expression of Moma-2 and VCAM-1 in the aortic root.Oil red O staining and HE staining were used to detect the changes of the lipid deposition and pathological changes in the aortic root.Biochemical kits were used to detect the serum lipid levels.Western blot was used to detect the phosphorylation of NF-κB and the expression of IκB-α.The results showed that compared with HFD group,the area of atherosclerotic plaque in HFD+wogonin group was decreased (P<0.05),but the serum lipid levels do not change; moreover,monocyte/monocyte macrophage infiltration,the protein expression of VCAM-1,the mRNA expression level of TNF-α,IL-6,ICAM-1 and VCAM-1 and the activation of NF-κB signal pathway in HFD+wogonin group were significantly reduced (P<0.05).These findings indicated that wogonin could effectively alleviate arterial atherosclerosis and inhibit NF-κB-mediated arterial inflammation in ApoE^-/- mice induced by high-fat diet.