Imperatorin is Transported through Blood-Brain Barrier by Carrier-Mediated Transporters

( Temdara Tun ),( Young-sook Kang ) 한국응용약물학회 2017 Biomolecules & Therapeutics(구 응용약물학회지) Vol.25 No.4

Imperatorin, a major bioactive furanocoumarin with multifunctions, can be used for treating neurodegenerative diseases. In this study, we investigated the characteristics of imperatorin transport in the brain. Experiments of the present study were designed to study imperatorin transport across the blood-brain barrier both in vivo and in vitro. In vivo study was performed in rats using single intravenous injection and in situ carotid artery perfusion technique. Conditionally immortalized rat brain capillary endothelial cells were as an in vitro model of blood-brain barrier to examine the transport mechanism of imperatorin. Brain distribution volume of imperatorin was about 6 fold greater than that of sucrose, suggesting that the transport of imperatorin was through the blood-brain barrier in physiological state. Both in vivo and in vitro imperatorin transport studies demonstrated that imperatorin could be transported in a concentration-dependent manner with high affinity. Imperatorin uptake was dependent on proton gradient in an opposite direction. It was significantly reduced by pretreatment with sodium azide. However, its uptake was not inhibited by replacing extracellular sodium with potassium or N-methylglucamine. The uptake of imperatorin was inhibited by various cationic compounds, but not inhibited by TEA, choline and organic anion substances. Transfection of plasma membrane monoamine transporter, organic cation transporter 2 and organic cation/carnitine transporter 2/1 siRNA failed to alter imperatorin transport in brain capillary endothelial cells. Especially, tramadol, clonidine and pyrilamine inhibited the uptake of [³H]imperatorin competitively. Therefore, imperatorin is actively transported from blood to brain across the blood-brain barrier by passive and carrier-mediated transporter.

Imperatorin is Transported through Blood-Brain Barrier by Carrier-Mediated Transporters

Tun, Temdara,Kang, Young-Sook The Korean Society of Applied Pharmacology 2017 Biomolecules & Therapeutics(구 응용약물학회지) Vol.25 No.4

Imperatorin, a major bioactive furanocoumarin with multifunctions, can be used for treating neurodegenerative diseases. In this study, we investigated the characteristics of imperatorin transport in the brain. Experiments of the present study were designed to study imperatorin transport across the blood-brain barrier both in vivo and in vitro. In vivo study was performed in rats using single intravenous injection and in situ carotid artery perfusion technique. Conditionally immortalized rat brain capillary endothelial cells were as an in vitro model of blood-brain barrier to examine the transport mechanism of imperatorin. Brain distribution volume of imperatorin was about 6 fold greater than that of sucrose, suggesting that the transport of imperatorin was through the blood-brain barrier in physiological state. Both in vivo and in vitro imperatorin transport studies demonstrated that imperatorin could be transported in a concentration-dependent manner with high affinity. Imperatorin uptake was dependent on proton gradient in an opposite direction. It was significantly reduced by pretreatment with sodium azide. However, its uptake was not inhibited by replacing extracellular sodium with potassium or N-methylglucamine. The uptake of imperatorin was inhibited by various cationic compounds, but not inhibited by TEA, choline and organic anion substances. Transfection of plasma membrane monoamine transporter, organic cation transporter 2 and organic cation/carnitine transporter 2/1 siRNA failed to alter imperatorin transport in brain capillary endothelial cells. Especially, tramadol, clonidine and pyrilamine inhibited the uptake of [$^3H$]imperatorin competitively. Therefore, imperatorin is actively transported from blood to brain across the blood-brain barrier by passive and carrier-mediated transporter.

Preventive effects of imperatorin on perfluorohexanesulfonate-induced neuronal apoptosis via inhibition of intracellular calcium-mediated ERK pathway

Lee, Eunkyung,Choi, So-Young,Yang, Jae-Ho,Lee, Youn Ju The Korean Society of Pharmacology 2016 The Korean Journal of Physiology & Pharmacology Vol.20 No.4

Early life neuronal exposure to environmental toxicants has been suggested to be an important etiology of neurodegenerative disease development. Perfluorohexanesulfonate (PFHxS), one of the major perfluoroalkyl compounds, is widely distributed environmental contaminants. We have reported that PFHxS induces neuronal apoptosis via ERK-mediated pathway. Imperatorin is a furanocoumarin found in various edible plants and has a wide range of pharmacological effects including neuroprotection. In this study, the effects of imperatorin on PFHxS-induced neuronal apoptosis and the underlying mechanisms are examined using cerebellar granule cells (CGC). CGC were isolated from seven-day old rats and were grown in culture for seven days. Caspase-3 activity and TUNEL staining were used to determine neuronal apoptosis. PFHxS-induced apoptosis of CGC was significantly reduced by imperatorin and PD98059, an ERK pathway inhibitor. PFHxS induced a persistent increase in intracellular calcium, which was significantly blocked by imperatorin, NMDA receptor antagonist, MK801 and the L-type voltage-dependent calcium channel blockers, diltiazem and nifedipine. The activation of caspase-3 by PFHxS was also inhibited by MK801, diltiazem and nifedipine. PFHxS-increased ERK activation was inhibited by imperatorin, MK801, diltiazem and nifedipine. Taken together, imperatorin protects CGC against PFHxS-induced apoptosis via inhibition of NMDA receptor/intracellular calcium-mediated ERK pathway.

Preventive effects of imperatorin on perfluorohexanesulfonateinduced neuronal apoptosis via inhibition of intracellular calcium-mediated ERK pathway

이은경,최소영,양재호,이윤주 대한약리학회 2016 The Korean Journal of Physiology & Pharmacology Vol.20 No.4

Early life neuronal exposure to environmental toxicants has been suggested to be an important etiology of neurodegenerative disease development. Perfluorohexanesulfonate (PFHxS), one of the major perfluoroalkyl compounds, is widely distributed environmental contaminants. We have reported that PFHxS induces neuronal apoptosis via ERK-mediated pathway. Imperatorin is a furanocoumarin found in various edible plants and has a wide range of pharmacological effects including neuroprotection. In this study, the effects of imperatorin on PFHxS-induced neuronal apoptosis and the underlying mechanisms are examined using cerebellar granule cells (CGC). CGC were isolated from sevenday old rats and were grown in culture for seven days. Caspase-3 activity and TUNEL staining were used to determine neuronal apoptosis. PFHxS-induced apoptosis of CGC was significantly reduced by imperatorin and PD98059, an ERK pathway inhibitor. PFHxS induced a persistent increase in intracellular calcium, which was significantly blocked by imperatorin, NMDA receptor antagonist, MK801 and the L-type voltage-dependent calcium channel blockers, diltiazem and nifedipine. The activation of caspase-3 by PFHxS was also inhibited by MK801, diltiazem and nifedipine. PFHxS-increased ERK activation was inhibited by imperatorin, MK801, diltiazem and nifedipine. Taken together, imperatorin protects CGC against PFHxSinduced apoptosis via inhibition of NMDA receptor/intracellular calcium-mediated ERK pathway.

Effect of Imperatorin on Adenosine Triphosphate-stimulated Mucin Secretion from Airway Epithelial Cells

( Ho Jin Heo ),( Cheol Su KIM ),( Hyun Jae Lee ),( Seung Won Shin ),( Young Sik Kim ),( Sam Sik Kang ),( Yang Chun Park ),( Yun Hee Kim ),( Un Kyo Seo ),( Jeong Ho Seok ),( Choong Jae Lee ) 한국응용약물학회 2006 Biomolecules & Therapeutics(구 응용약물학회지) Vol.14 No.4

Angelicae Koreanae Radix has been used for controlling inflammatory respiratory diseases in folk medicine and their components, imperatorin, marmesinin and oxypeucedanin were reported to have diverse biological effects. In this study, we investigated whether imperatorin, marmesinin and oxypeucedanin affect adenosine triphosphate (ATP)-induced mucin secretion from cultured airway epithelial cells. Confluent primary hamster tracheal surface epithelial (HTSE) cells were metabolically radiolabeled using 3H-glucosamine for 24 h and chased for 30 min in the presence of varying concentrations of each agent to assess the effects on 3H-mucin secretion. The results were as follows: 1) imperatorin significantly inhibited ATP-induced mucin secretion; 2) However, marmesinin and oxypeucedanin did not affect ATP-induced mucin secretion, significantly. We conclude that imperatorin might inhibit ATP-induced mucin secretion by directly acting on airway mucin-secreting cells. Therefore, imperatorin should further be investigated for the possible use as mucoregulators in the treatment of inflammatory airway diseases.

Anti-inflammatory and antioxidant effects of coumarins isolated from <i>Foeniculum vulgare</i> in lipopolysaccharide-stimulated macrophages and 12-<i>O</i>-tetradecanoylphorbol-13-acetate-stimulated mice

Yang, In Jun,Lee, Dong Ung,Shin, Heung Mook Taylor & Francis 2015 IMMUNOPHARMACOLOGY AND IMMUNOTOXICOLOGY Vol.37 No.3

<P><I>Context</I>: <I>Foeniculum vulgare</I> (<I>F. vulgare</I>) is traditionally used to treat inflammatory diseases. Recently, anti-inflammatory and antioxidant activities of methanol extract of the fruits of <I>F. vulgare</I> were reported. To identify biologically active compounds responsible for the anti-inflammatory activity, we isolated four coumarins, scopoletin, 8-methoxypsoralen, bergapten and imperatorin from the fruits of <I>F. vulgare.</I></P><P><I>Objective</I>: This study assessed the anti-inflammatory and antioxidant effects of coumarins isolated from <I>F. vulgare</I> in lipopolysaccharide (LPS)-stimulated macrophages and 12-<I>O</I>-tetradecanoylphorbol-13-acetate (TPA)-stimulated mice.</P><P><I>Materials and methods</I>: RAW 264.7 cells were treated with the coumarins (30 µM) and then stimulated with LPS (100 ng/ml). Ears of ICR mice were treated with TPA (1 µg/ear) once a day. Ten microliters each of the four coumarins (200 &mgr;g/ml) were topically applied to the ears for 3 days. Antioxidant activities were examined using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis 3-ethylbenzthiazoline-6-sulfonic acid (ABTS) scavenging assays.</P><P><I>Results</I>: All the tested coumarins showed excellent antioxidant activities in DPPH and ABTS radical scavenging assays. Among the coumarins, imperatorin had the greatest anti-inflammatory activities as measured by inhibition of the pro-inflammatory cytokines production including interleukin (IL)-6 and tumor necrosis factor (TNF)-&agr; in LPS-stimulated RAW 264.7 cells through blockade of the I&kgr;B kinase (IKK)/inhibitor of kappa B (I&kgr;B)/nuclear factor-&kgr;B (NF-&kgr;B) pathway. <I>In vivo</I> experiments showed that imperatorin reduced TPA-induced ear thickness/weight, cutaneous cytokines expression and improved histopathological features.</P><P><I>Conclusion</I>: Although four coumarins isolated from the fruits of <I>F. vulgare</I> provide effective anti-inflammatory and antioxidant activities, imperatorin is most potent.</P>

구릿대 뿌리의 부위별 성분 비교

윤의중,유재국,진전성,나민균,민병선,정현주,서은경,배기환 한국생약학회 2010 생약학회지 Vol.41 No.3

In this study, we analyzed and quantified the amounts of bioactive phenolic constituents, xanthotoxin (1), oxypeucedanin (2), and imperatorin (3) in each part of the root of Angelica dahurica by HPLC, which validated by ICH guide lines comparing the linearity, intra day precision, inter-day precision. As a result, the amount of imperatorin 2.96% in the rootlet was two fold higher than that of the main root 1.32%. On the other hand, the amounts of xanthotoxin and oxypeucedanin in the rootlet showed similar to those of main root. In addition, the cortex of root is more plentiful of three consitituents (0.66%,0.53%, and 1.85%) than those of xylem (0.29%, 0.05%, and 0.07%). These results show that the rootlet and cortex contain a large amount of bioactive phenolic constituent including xanthotoxin, oxypeucedanin, and imperatorin than other parts of the root.

Simultaneous determination of imperatorin and its metabolite, xanthotoxol, in rat plasma and urine by LC-MS/MS: Applications to pharmacokinetic studies

( Ngo Thi Lien ) 한국공업화학회 2018 한국공업화학회 연구논문 초록집 Vol.2018 No.1

Imperatorin Sustained-release Tablets: In Vitro and Pharmacokinetic Studies

Jingjing Pan,Wen Lu,Changhui Li,Sicen Wang,Langchong He 대한약학회 2010 Archives of Pharmacal Research Vol.33 No.8

We prepared and evaluated imperatorin (IMP) sustained-release tablets. IMP is an active compound in Angelica dahuricae, a Chinese herbal medicine. We used different polymers, such as hydroxypropyl methylcellulose (HPMC K4M, K15M, and K100M), carbopol 934P, sodium carboxymethyl cellulose (CMC-Na), and their combinations to prepare the matrix tablets and achieve the desired sustained release profile. The in vitro release profiles of these formulations were examined and fit to various kinetic release models. We also tested the effects of polymer combination ratios on the in vitro release rate. In vivo studies were performed for the optimized formulation in six beagle dogs, and pharmacokinetic parameters were compared with plain IMP tablets. IMP sustained-release tablets exhibited a more sustained plasma concentration than the plain tablets, with a relative bioavailability of 127.25%. The in vitro releases rates and in vivo absorption correlated for the initial 8 hours. These results demonstrate that the sustained-release tablet system can effectively control the release of IMP.

Chemosensitizing effect and mechanism of imperatorin on the anti-tumor activity of doxorubicin in tumor cells and transplantation tumor model

Xin-li Liang,Miao-miao Ji,Zheng-gen Liao,Guo-wei Zhao,Xi-lan Tang,Wei Dong 대한약리학회 2022 The Korean Journal of Physiology & Pharmacology Vol.26 No.3

Multidrug resistance of tumors has been a severe obstacle to the success of cancer chemotherapy. The study wants to investigate the reversal effects of imperatorin (IMP) on doxorubicin (DOX) resistance in K562/DOX leukemia cells, A2780/Taxol cells and in NOD/SCID mice, to explore the possible molecular mechanisms. K562/ DOX and A2780/Taxol cells were treated with various concentrations of DOX and Taol with or without different concentrations of IMP, respectively. K562/DOX xenograft model was used to assess anti-tumor effect of IMP combined with DOX. MTT assay, Rhodamine 123 efflux assay, RT-PCR, and Western blot analysis were determined in vivo and in vitro. Results showed that IMP significantly enhanced the cytotoxicity of DOX and Taxol toward corresponding resistance cells. In vivo results illustrated both the tumor volume and tumor weight were significantly decreased after 2-week treatment with IMP combined with DOX compared to the DOX alone group. Western blotting and RT-PCR analyses indicated that IMP downregulated the expression of P-gp in K562/DOX xenograft tumors in NOD/SCID mice. We also evaluated glycolysis and glutamine metabolism in K562/DOX cells by measuring glucose consumption and lactate production. The results revealed that IMP could significantly reduce the glucose consumption and lactate production of K562/DOX cells. Furthermore, IMP could also remarkably repress the glutamine consumption, α-KG and ATP production of K562/DOX cells. Thus, IMP may sensitize K562/DOX cells to DOX and enhance the antitumor effect of DOX in K562/DOX xenograft tumors in NOD/SCID mice. IMP may be an adjuvant therapy to mitigate the multidrug resistance in leukemia chemotherapy.