ABSTRACT
Objective To investigate the expression of Wnt7b, β -catenin, and c-Myc in asthmatic mice and the intervention of the leukotriene receptor antagonist montelukast on airway remodeling. Methods The asthma model was established by ovalbumin (OVA) induction. HE staining was used to observe the pathological changes in lung tissue. Serum OVA-sIgE levels were determined by ELISA. The level of Wnt7b, β -catenin, and c-Myc protein and mRNA in the lung tissue of mice was analyzed by Western blotting and real-time PCR. The basement membrane perimeter (PBM), wall area of bronchial tube (WAt), wall area of bronchial smooth muscle (WAm), and the number of smooth muscle cells were measured using medical image analysis software and standardized based on the PBM. Results The amount of OVA-slgE in the asthma group was significantly higher than in the control and montelukast groups (P ＜ 0. 05). Western blotting and real-time PCR showed that the expression of Wnt7b, β -catenin, and c-Myc in the asthma group was higher than the expression in the control and montelukast groups (P ＜ 0. 05). Image analysis showed that the WAt/PBM and WAm/PBM ratios in the montelukast group were significantly lower than those in the asthma group (P ＜ 0. 05). Conclusion The Wnt/ β -catenin signaling pathway may be an important factor in the pathogenesis of asthma; montelukast may attenuate airway remodeling in asthmatic patients by decreasing the expression of Wnt7b, β -catenin, and c-Myc.
ABSTRACT
OBJECTIVE:To establish the method for simultaneous determination of the content of berberine hydrochloride, baicalin and osthole in Jinchan zhiyang capsules. METHODS:HPLC method was adopted. The determination was performed in Hypersil BDS C18 column with mobile phase consisted of methanol-acetonitrile-0.1%phosphoric acid-three triethylamine(50∶30∶19∶1, V/V/V/V) at the flow rate of 1.0 mL/min. The detection wavelengths were set at 265 nm (berberine hydrochloride),280 nm (baicalin)and 322 nm(osthole). The column temperature was set at 30 ℃,and sample size was 10 μL. RESULTS:The linear range of berberine hydrochloride,baicalin and osthole were 80-800 ng(r＝0.999 8),60-600 ng(r＝0.999 9),60-600 ng(r＝0.999 6),respectively. RSDs of precision,stability and reproducibility tests were all lower than 2.0%. The limits of quantitation were 80,60,60 ng,respectively,and the limits of detection were 24,20,20 ng,respectively. The recovery rates were 97.4%-98.3%(RSD＝0.33%,n＝6),98.4%-99.6%(RSD＝0.42%,n＝6)and 96.9%-99.0%(RSD＝0.92%,n＝6),respectively. RSDs of durability tests were all lower than 1.2%. CONCLUSIONS:The method is simple, accurate, precise, stable, reproducible and durable. It can be used for simultaneous determination of berberine hydrochloride,baicalin and osthole in Jinchan zhiyang capsules.
ABSTRACT
OBJECTIVE:To establish the method for simultaneous determination of berberine hydrochloride and baicalin in Jianpi zhixiening granules. METHODS:HPLC switching walvelength method was adopted. The determination was performed on Hypersil BDS C18 column with mobile phase consisted of methanol-0.45％ phosphoric acid solution-triethylamine(50:49:1,V/V/V) at the flow rate of 1.0 mL/min. The detection wavelength was set at 265 nm(berberine hydrochloride)and 280 nm(baicalin). The column temperature was set at 30 ℃,and sample size was 10 μL. RESULTS:The linear range of berberine hydrochloride and baicalin were 60.3-312.8 ng(r=0.9997)and 81.5-368.9 ng(r=0.9999). The limits of quantitation were 0.6668,0.7740 ng,andthe limits of detection were 0.2226,0.2580 ng,respectively. RSDs of intermediate precision,stability and repeatability tests were all lower than 1.0％. The recoveries were 96.48％-99.30％(RSD=1.06％,n=6) and 95.20％-99.39％(RSD=1.66％,n=6), respectively. RSDs of durability test were all lower than 2.0％. CONCLUSIONS:The method is simple, precise, stable, reproducible,accurate and durable. It can be used for simultaneous determination of berberine hydrochloride and baicalin in Jianpi zhixiening granules.