L-6/GAPDH ratioIL-8/GAPDH ratio60 40 20 0 Standard DNCB 25 50 100 200 CP001 (mg/kg)(c)0 Standard DNCB 25 50 one hundred 200 CP001 (mg/kg)(d)Figure four: Effect of CP001 on the cytokine mRNA expression in mouse skin tissue. The IL-4, IL-13, IL6, and IL-8 mRNA expression were measured by real-time PCR (a), (b) and RT-PCR (c), (d) in mouse skin tissue. The columns and the error bars represent imply ?SD ( = five mice/group). 0.05.Blood IgE level0 Regular DNCB 25 50 100 CP001 (mg/kg)Figure 5: Measurement of plasma IgE level. Total IgE level was determined by ELISA. The columns plus the error bars represent mean ?SEMs ( = 4 mice/group). 0.05.the lots of unwanted side effects. Therefore, we discover a new drug, that is helpful inside the therapy of AD with no any unwanted effects. Not too long ago, we reported that topical application of KM110329 (CP001 modified drug) lowered ovalbumin- and DNCBinduced atopic dermatitis [23]. Therefore, we have been asking yourself whether or not CP001 oral administration might inhibit DNCBinduced atopic dermatitis. Mast cells degranulation may be regulated by the recruitment, trafficking, and function of inflammatory response. One example is, IL-4 and IL-13 induce cell adhesion molecules on endothelium which might be recruitment of leukocytes [27?9]. Also, the production of IL-4 cytokine in epidermal cells has been identified to be the principle element for initiation of AD [30]. In our data, we show that cytokine production and mRNA levels of IL-4, IL-13, IL-6, and IL-8 had been suppressed by CP001 in HMC-1. Also, quantitative real-time PCR of the skin lesions also showed that oral administration of CP001 diminished the mRNA degree of IL-4 and IL-13 within the AD-like skin lesions. Also, we discovered that CP001 reduces mast cell inIgE production (pg/mL)Mediators of InflammationCP001 (g/mL)-+5 +20 +50 +100 +PMA + A23187 IL-GAPDH IL-6/GAPDH ratio- two.IL-6 production (pg/mL)6000 4000 20001.five 1.0 0.5 0.+++++ PMA + A23187CP001 (g/mL)(a) (b)CP001 (g/mL)-+5 +20 +50 +100 +PMA + A23187 IL-GAPDHIL-8 production (pg/mL)5000 IL-8/GAPDH ratio-2.4000 3000 2000 1000 0 + + 5 + 20 + 50 + PMA + A231872.0 1.five 1.0 0.five 0.CP001 (g/mL)(c) (d)Figure six: Impact of CP001 on PMA plus A23187-stimulated proinflammatory cytokine expression in HMC-1. HMC-1 have been pretreated with a variety of concentrations of CP001 for 1 h and after that treated with PMA and A23187 for 24 h.1196507-58-0 Chemscene The levels of IL-6 and IL-8 in culture supernatants were measured by ELISA assay (a), (c).Phosphatidylcholines,soya web The IL-6 and IL-8 mRNA levels were measured by RT-PCR (b), (d). Information represent the imply ?SEMs of three independent experiments.CP001 (g/mL)-Mediators of InflammationCP001 (g/mL) one hundred + PMA + A23187 IL–+5 +20 +50 ++5 +20 +50 +100 +PMA + A23187 IL-GAPDHIL-13/GAPDH ratio IL-4/GAPDH ratioGAPDH 150 one hundred 50 0 Con PMA100 50 0 Con PMA(a)(b)Figure 7: Impact of CP001 on PMA plus A23187-stimulated Th2 cytokine expression in HMC-1.PMID:23614016 HMC-1 have been pretreated with a variety of concentrations of CP001 for 1 h after which treated with PMA and A23187 for 24 h. The IL-4 and IL-13 mRNA levels have been measured by RTPCR (a), (b). Data represent the mean ?SEMs of 3 independent experiments.OH140 120 100 (mAU) 80 60 40 20-HO OHOOOHCatalpolOH OHO O O OH OEllagic acidHO OH HO O O200 (mAU)OHHO OHOO OHO150 Quercitrin one hundred 50HO OOH OH(a)ten 12 (min)(b)25 30 (min)Figure eight: Typical HPLC chromatograms of CP001 for (a) catalpol, (b) ellagic acid, and quercitrin.DNCB-induced AD-like skin lesion. It seems that inhibition of infiltration of mast cell downregulates secretion of IL-4 and IL-13 cytokines and it may inhibit recr.