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Up-regulation on cytochromes P450 in rat mediated by total alkaloid extract from Corydalis yanhusuo.

Yan J, He X, Feng S, Zhai Y, Ma Y, Liang S, Jin C - BMC Complement Altern Med (2014)

Bottom Line: The effects of TAE on five CYPs activity and mRNA levels were quantitated by cocktail probe drugs using a rapid chromatography/tandem mass spectrometry (LC-MS/MS) method and reverse transcription-polymerase chain reaction (RT-PCR), respectively.Moreover, the mRNA levels of CYP2E1 and CYP3A1 in the rat liver, lung, and intestine were significantly up-regulated with TAE from 6 and 30 mg/kg, respectively.These results suggest that TAE-induced CYPs activity in the rat liver results from the elevated mRNA levels of CYPs.

View Article: PubMed Central - PubMed

Affiliation: School of Chinese Materia Medica, Tianjin University of Traditional Chinese Medicine, 312 Anshanxi Road, Tianjin 300193, Nankai District, China. hexintn@163.com.

ABSTRACT

Background: Yanhusuo (Corydalis yanhusuo W.T. Wang; YHS), is a well-known traditional Chinese herbal medicine, has been used in China for treating pain including chest pain, epigastric pain, and dysmenorrhea. Its alkaloid ingredients including tetrahydropalmatine are reported to inhibit cytochromes P450 (CYPs) activity in vitro. The present study is aimed to assess the potential of total alkaloid extract (TAE) from YHS to effect the activity and mRNA levels of five cytochromes P450 (CYPs) in rat.

Methods: Rats were administered TAE from YHS (0, 6, 30, and 150 mg/kg, daily) for 14 days, alanine aminotransferase (ALT) levels in serum were assayed, and hematoxylin and eosin-stained sections of the liver were prepared for light microscopy. The effects of TAE on five CYPs activity and mRNA levels were quantitated by cocktail probe drugs using a rapid chromatography/tandem mass spectrometry (LC-MS/MS) method and reverse transcription-polymerase chain reaction (RT-PCR), respectively.

Results: In general, serum ALT levels showed no significant changes, and the histopathology appeared largely normal compared with that in the control rats. At 30 and 150 mg/kg TAE dosages, an increase in liver CYP2E1 and CYP3A1 enzyme activity were observed. Moreover, the mRNA levels of CYP2E1 and CYP3A1 in the rat liver, lung, and intestine were significantly up-regulated with TAE from 6 and 30 mg/kg, respectively. Furthermore, treatment with TAE (150 mg/kg) enhanced the activities and the mRNA levels of CYP1A2 and CYP2C11 in rats. However, the activity or mRNA level of CYP2D1 remained unchanged.

Conclusions: These results suggest that TAE-induced CYPs activity in the rat liver results from the elevated mRNA levels of CYPs. Co-administration of prescriptions containing YHS should consider a potential herb (drug)-drug interaction mediated by the induction of CYP2E1 and CYP3A1 enzymes.

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HPLC-fingerprint chromatograms of total alkaloids of YHS. (A) Chromatogram of the total alkaloid extract (TAE); (B) chromatogram of reference standards; and (C) chromatogram of the blank solvent; 1, protopine; 2, allocryptopin; 3, dehydrocorydaline; 4, tetrahydropalmatine; 5, corydaline; 6, tetrahydroberberine;7, glaucine.
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Fig1: HPLC-fingerprint chromatograms of total alkaloids of YHS. (A) Chromatogram of the total alkaloid extract (TAE); (B) chromatogram of reference standards; and (C) chromatogram of the blank solvent; 1, protopine; 2, allocryptopin; 3, dehydrocorydaline; 4, tetrahydropalmatine; 5, corydaline; 6, tetrahydroberberine;7, glaucine.

Mentions: The dried and pulverized tubers of yanhusuo (2 Kg) were refluxed with 70% alcohol for three times. The pooled extract was concentrated under vacuum drying at 50°C. The dry powder was then dissolved in 200 mL water. The extracted solution was purified using 732 strong acid cation exchange resin, and the following elution program was used: six times water, and six times 70% ethanol:water containing 5% ammonia. A solution of 70% ethanol:water containing 5% ammonia was obtained, and the solvent was evaporated under reduced pressure. Preliminary analysis of the extracts was performed using an Agilent 1200 series high performance liquid chromatography (HPLC) instrument (Agilent Technologies, USA). Chromatographic separation was performed using an Agilent Zorbax HC-C18 (4.6 mm × 250 mm × 5 μm) column (Agilent, USA). The mobile phase were (A) 0.5% phosphate buffer (pH 5.0) and (B) methanol and the elution system was as follows: 0 - 25 min, from 85% A to 70% A; 25 - 75 min, from 70% A to 65% A; 75 -103 min, from 65% A to 25% A; 103 - 105 min, from 25% A to 20% A; 105 - 115 min, from 20% A to 85% A;115 - 125 min, 85% A. The flow rate was 1.0 mL/min, and the UV wavelength was 280 nm. The major alkaloids in YHS extract were quantified, and HPLC (Figure 1) showed that the contents (w/w) of protopine, allocryptopin, dehydrocorydaline, tetrahydropalmatine, corydaline, tetrahydroberberine, and glaucine were 2.52%, 1.67%, 0.34%, 3.58%, 3.0%, 0.22%, and 0.66%, respectively. Figure 2 presents the structures of the alkaloid components. The total alkaloid content (>45% in the extract) was determined by UV spectroscopy.Figure 1


Up-regulation on cytochromes P450 in rat mediated by total alkaloid extract from Corydalis yanhusuo.

Yan J, He X, Feng S, Zhai Y, Ma Y, Liang S, Jin C - BMC Complement Altern Med (2014)

HPLC-fingerprint chromatograms of total alkaloids of YHS. (A) Chromatogram of the total alkaloid extract (TAE); (B) chromatogram of reference standards; and (C) chromatogram of the blank solvent; 1, protopine; 2, allocryptopin; 3, dehydrocorydaline; 4, tetrahydropalmatine; 5, corydaline; 6, tetrahydroberberine;7, glaucine.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4150957&req=5

Fig1: HPLC-fingerprint chromatograms of total alkaloids of YHS. (A) Chromatogram of the total alkaloid extract (TAE); (B) chromatogram of reference standards; and (C) chromatogram of the blank solvent; 1, protopine; 2, allocryptopin; 3, dehydrocorydaline; 4, tetrahydropalmatine; 5, corydaline; 6, tetrahydroberberine;7, glaucine.
Mentions: The dried and pulverized tubers of yanhusuo (2 Kg) were refluxed with 70% alcohol for three times. The pooled extract was concentrated under vacuum drying at 50°C. The dry powder was then dissolved in 200 mL water. The extracted solution was purified using 732 strong acid cation exchange resin, and the following elution program was used: six times water, and six times 70% ethanol:water containing 5% ammonia. A solution of 70% ethanol:water containing 5% ammonia was obtained, and the solvent was evaporated under reduced pressure. Preliminary analysis of the extracts was performed using an Agilent 1200 series high performance liquid chromatography (HPLC) instrument (Agilent Technologies, USA). Chromatographic separation was performed using an Agilent Zorbax HC-C18 (4.6 mm × 250 mm × 5 μm) column (Agilent, USA). The mobile phase were (A) 0.5% phosphate buffer (pH 5.0) and (B) methanol and the elution system was as follows: 0 - 25 min, from 85% A to 70% A; 25 - 75 min, from 70% A to 65% A; 75 -103 min, from 65% A to 25% A; 103 - 105 min, from 25% A to 20% A; 105 - 115 min, from 20% A to 85% A;115 - 125 min, 85% A. The flow rate was 1.0 mL/min, and the UV wavelength was 280 nm. The major alkaloids in YHS extract were quantified, and HPLC (Figure 1) showed that the contents (w/w) of protopine, allocryptopin, dehydrocorydaline, tetrahydropalmatine, corydaline, tetrahydroberberine, and glaucine were 2.52%, 1.67%, 0.34%, 3.58%, 3.0%, 0.22%, and 0.66%, respectively. Figure 2 presents the structures of the alkaloid components. The total alkaloid content (>45% in the extract) was determined by UV spectroscopy.Figure 1

Bottom Line: The effects of TAE on five CYPs activity and mRNA levels were quantitated by cocktail probe drugs using a rapid chromatography/tandem mass spectrometry (LC-MS/MS) method and reverse transcription-polymerase chain reaction (RT-PCR), respectively.Moreover, the mRNA levels of CYP2E1 and CYP3A1 in the rat liver, lung, and intestine were significantly up-regulated with TAE from 6 and 30 mg/kg, respectively.These results suggest that TAE-induced CYPs activity in the rat liver results from the elevated mRNA levels of CYPs.

View Article: PubMed Central - PubMed

Affiliation: School of Chinese Materia Medica, Tianjin University of Traditional Chinese Medicine, 312 Anshanxi Road, Tianjin 300193, Nankai District, China. hexintn@163.com.

ABSTRACT

Background: Yanhusuo (Corydalis yanhusuo W.T. Wang; YHS), is a well-known traditional Chinese herbal medicine, has been used in China for treating pain including chest pain, epigastric pain, and dysmenorrhea. Its alkaloid ingredients including tetrahydropalmatine are reported to inhibit cytochromes P450 (CYPs) activity in vitro. The present study is aimed to assess the potential of total alkaloid extract (TAE) from YHS to effect the activity and mRNA levels of five cytochromes P450 (CYPs) in rat.

Methods: Rats were administered TAE from YHS (0, 6, 30, and 150 mg/kg, daily) for 14 days, alanine aminotransferase (ALT) levels in serum were assayed, and hematoxylin and eosin-stained sections of the liver were prepared for light microscopy. The effects of TAE on five CYPs activity and mRNA levels were quantitated by cocktail probe drugs using a rapid chromatography/tandem mass spectrometry (LC-MS/MS) method and reverse transcription-polymerase chain reaction (RT-PCR), respectively.

Results: In general, serum ALT levels showed no significant changes, and the histopathology appeared largely normal compared with that in the control rats. At 30 and 150 mg/kg TAE dosages, an increase in liver CYP2E1 and CYP3A1 enzyme activity were observed. Moreover, the mRNA levels of CYP2E1 and CYP3A1 in the rat liver, lung, and intestine were significantly up-regulated with TAE from 6 and 30 mg/kg, respectively. Furthermore, treatment with TAE (150 mg/kg) enhanced the activities and the mRNA levels of CYP1A2 and CYP2C11 in rats. However, the activity or mRNA level of CYP2D1 remained unchanged.

Conclusions: These results suggest that TAE-induced CYPs activity in the rat liver results from the elevated mRNA levels of CYPs. Co-administration of prescriptions containing YHS should consider a potential herb (drug)-drug interaction mediated by the induction of CYP2E1 and CYP3A1 enzymes.

Show MeSH
Related in: MedlinePlus