Involvement of a Novel Organic Cation Transporter in Paeonol Transport Across the Blood-Brain Barrier

Asmita Gyawali,Sokhoeurn Krol,강영숙 한국응용약물학회 2019 Biomolecules & Therapeutics(구 응용약물학회지) Vol.27 No.3

Paeonol has neuroprotective function, which could be useful for improving central nervous system disorder. The purpose of this study was to characterize the functional mechanism involved in brain transport of paeonol through blood-brain barrier (BBB). Brain transport of paeonol was characterized by internal carotid artery perfusion (ICAP), carotid artery single injection technique (brain uptake index, BUI) and intravenous (IV) injection technique in vivo. The transport mechanism of paeonol was examined using conditionally immortalized rat brain capillary endothelial cell line (TR-BBB) as an in vitro model of BBB. Brain volume of distribution (VD) of [3H]paeonol in rat brain was about 6-fold higher than that of [14C]sucrose, the vascular space marker of BBB. The uptake of [3H]paeonol was concentration-dependent. Brain volume of distribution of paeonol and BUI as in vivo and inhibition of analog as in vitro studies presented significant reduction effect in the presence of unlabeled lipophilic compounds such as paeonol, imperatorin, diphenhydramine, pyrilamine, tramadol and ALC during the uptake of [3H]paeonol. In addition, the uptake significantly decreased and increased at the acidic and alkaline pH in both extracellular and intracellular study, respectively. In the presence of metabolic inhibitor, the uptake reduced significantly but not affected by sodium free or membrane potential disruption. Similarly, paeonol uptake was not affected on OCTN2 or rPMAT siRNA transfection BBB cells. Interestingly. Paeonol is actively transported from the blood to brain across the BBB by a carrier mediated transporter system.

Paeonol Exerts Anti-angiogenic and Anti-metastatic Activities through Downmodulation of Akt Activation and Inactivation of Matrix Metalloproteinases

Kim, Seung-Ae,Lee, Hyo-Jeong,Ahn, Kwang Seok,Lee, Hyo-Jung,Lee, Eun-Ok,Ahn, Kyoo-Seok,Choi, Seung-Hoon,Jung, Soo-Jin,Kim, Ji Young,Baek, Namin,Kim, Sung-Hoon Pharmaceutical Society of Japan 2009 Biological & pharmaceutical bulletin Vol.32 No.7

<P>Paeonol (2′-hydroxy-4′-methoxyacetophenone) is known to possess anti-inflammatory and anti-proliferative activities. Recently there is evidence that anti-inflammatory agents may be useful in the setting of angiogenesis-related diseases. Thus in the present study the anti-angiogenic activity of paeonol and its mechanism were investigated <I>in vitro</I> and <I>in vivo</I>. Paeonol significantly inhibited proliferation of basic fibroblast growth factor (bFGF)-stimulated human umbilical vein endothelial cells (HUVECs). Paeonol also significantly inhibited migration and tube formation of bFGF-stimulated HUVECs <I>in vitro</I>. In addition, paeonol significantly suppressed neovessel formation on bFGF-treated chick chorioallantoic membrane (CAM) and disrupted bFGF-induced neovascularization in Matrigel plug assay <I>in vivo</I>. Furthermore, paeonol downregluated Akt phosphorylation in bFGF-stimulated HUVECs and reduced expression of matrix metalloproteinases-2 and -9 in HT1080 human fibrosarcoma cells. The Akt inhibitor LY294002 synergistically potentiated paeonol-induced inactivation of Akt and vascular endothelial growth factor in bFGF-treated HUVECs. Taken together, these findings suggest that paeonol can be a potent suppressor of angiogenesis and metastasis partially through inhibition of Akt signaling pathway and matrix metalloproteinase activity.</P>

Paeonol에 의한 표피줄기세포 활성화

김도형 ( Do Hyung Kim ),김효진 ( Hyo Jin Kim ),여혜린 ( Hyerin Yeo ),이천구 ( Cheon Goo Lee ),이상화 ( Sang Hwa Lee ) 대한화장품학회 2016 대한화장품학회지 Vol.42 No.2

피부의 가장자리에 위치한 표피는 개체의 생존에 절대적으로 중요하며, 기저막에 위치한 표피줄기세포에 의해 평생 끊임없이 재생되고 있다. 표피줄기세포는 한정된 세포분열을 진행하고 각질세포로 분화하는 transit amplifying (TA)세포를 만들어 냄으로써 오랜 기간 재생에 필요한 많은 각질세포를 만들어 낼 수 있다. 본 연구에서는 표피줄기세포 세포분열 활성화 화합물로 목단으로부터 추출한 페오놀(paeonol)을 350여 개의 화합물들로부터 검색해 냈다. 이러한 세포분열 활성화 효능은 표피줄기세포 특이적으로 나타나며, 표피줄기세포의 지표로 알려진 p63 단백질의 발현 경향은 페오놀을 처리한 표피줄기세포에서 변화하지 않음을 유세포분석법으로 확인하였다. 콜로니형성 분석에서는 페오놀을 처리한 표피줄기세포가 1.3배 이상 더 나은 콜로니 형성능을 보여주었다. 그리고 PCR array 분석을 통해서 페오놀의 효능은 여러 신호전달에 의해 나타남을 알 수 있었다. 이러한 결과들로부터 페오놀이 줄기세포성에 영향을 주지 않고 표피줄기세포의 재생능력을 향상시키므로 표피줄기 세포 활성화 물질로서 화장품 소재로 적용될 수 있을 것으로 판단된다. Epidermis is continuously regenerated by keratinocyte stem cells (KSCs) residing in basement membrane, which is critical to the survival of an organism. KSCs are believed to persist during the whole lifetime and generate an enormous number of keratinocytes, required for the maintenance of epidermis, through transit amplifying cells dividing definite times until they become differentiated. In this report, we have developed a phenolic compound, paeonol, purified from Moutan Cortex, as a KSC proliferation activator, by screening about 350 herbal compounds. The cell proliferation activation by paeonol is specific for KSC not for keratinocyte, and no significant difference in the expression of p63 protein, a KSC marker, in KSCs treated with paeonol was observed in FACS analysis with anti-p63 antibody. In the colony forming assay, paeonol-treated KSC showed improved colony forming activity more than 1.3 fold. In addition, the result of PCR array shows that the activity of paeonol is through several signal pathways involving stem cell functions. These results suggest that paeonol could enhance KSC proliferation activity without reduction in stemness and could be applied to cosmetics as a KSC activating ingredient.

모란 추출액에서 paeoniflorin과 paeonol 동시 정량 분석 및 화장품 원료의 품질관리 기준 설정

윤기훈(Ki Hun Yun),지용하(Yong Ha Chi),이동규(Dong Kyu Lee),백수희(Soo Heui Paik) 한국유화학회 2018 한국응용과학기술학회지 Vol.35 No.1

모란은 항염증작용, 항알레르기작용, 항균작용, 중추억제, 위액분비억제, 진경작용 등의 약리활 성이 있을 뿐 아니라 항산화작용, 미백작용 등이 보고되면서 화장품 원료, 미용식품으로 다양하게 활용 가능성이 탐색되고 있다. 모란의 뿌리, 잎 및 꽃에서 추출용매와 온도, 시간에 따른 추출액 중 성분의 함량 변화를 비교하였다. Paeoniflorin과 paeonol을 지표성분으로 하여 두 성분의 함량을 동시에 정량분석하기 위한 HPLC 분석법을 설정하고 분석 배치마다 검량선과 QC용 검체를 사용하여 시스템적합성을 확인하면서 시료 분석을 실시하였다. Paeonol은 뿌리에서만 검출되었고 잎과 꽃에서는 검출되지 않았으며, paeoniflorin은 잎, 꽃에서의 농도가 뿌리에서보다 높았다. 추출용매와 섞을 때 뿌리의 절도는 구입 당시의 조각을 그대로 사용할 때보다 적당한 크기로 분쇄한 후 사용할 때 두 성분 모두 높은 농도로 추출되었다. 추출용매로는 30% 1,3-butylene glycol을 사용할 때 농도가 가장 높았다. 추출온도와 시간은 고온, 장시간 일수록 농도가 높아지는 경향을 보였으나 75℃, 4 h 이상에서는 농도 상승이 완만했으며, paeonol은 8 h까지 지속적으로 농도가 상승하는 경향을 보였다. 뿌리, 잎, 꽃의 배합 비율은 2+2+1g/0.5kg로 할 때 두 지표성분의 함량 기준에 도달 가능하다고 판단하였다. 최종적으로 원재료를 2+2+1g/0.5kg로 배합하고 30% 1,3-butylene glycol을 추출용매로 사용하여 75℃에서 4 h 동안 추출하는 방법이 두 지표성분의 활성 농도 기준과 원재료 및 제조공정상의 비용을 함께 고려할 때 적절한 조건이라고 판단하였고, 이 조건에서 10 kg 규모까지 추출액 제조 단위를 스케일업 하였다. Paeony has pharmacological activities such as anti-inflammatory, anti-allergic, anti-bacterial, central inhibitory, gastric secretion inhibition, and antispasmodic activities. In addition, its antioxidant activity and whitening effect being reported, thus it is being explored as raw materials for cosmetics. We compared the changes in the contents of paeoniflorin and paeonol in Peony extracts, depending on the changes of extracting solvents, temperature and time. The HPLC method was set up for simultaneous analysis, the system suitabilities were confirmed by using the calibration curves and the QC samples for each assay batch. Paeonol was detected only in roots, and paeoniflorin was higher in leaf and flower than root. Higher concentrations of both ingredients were extracted when the root was used after grinding to a suitable size, and when 30% 1,3-butylene glycol was used as the extraction solvent. Also the concentrations tended to increase at higher temperature and longer time, but the increase was gradual at over 75℃ and 4 hours. The ratio of root, leaf and flower was determined to be 2+2+1g/0.5kg of batch, reaching the contents criteria of paeoniflorin and paeonol. Finally, we selected as the best extraction condition when the raw materials are mixed with 2+2+1g/0.5kg and extracted with 30% 1,3-butylene glycol as an extraction solvent at 75℃ for 4 hours, considering both the concentrations of two components and the cost of raw materials and manufacturing process, The extraction units were scaled up to 10 kg under this condition.

Inhibitory Effects of Paeonol on Morphine-Induced Locomotor Sensitization and Conditioned Place Preference in Mice

Eun, Jae-Soon,Bae, Ki-Hwan,Yun, Yeo-Pyo,Hong, Jin-Tae,Kwon, Han-Na,Oh, Ki-Wan The Pharmaceutical Society of Korea 2006 Archives of Pharmacal Research Vol.29 No.10

The inhibitory effects of paeonol, a major compound of Paeoniae radix, on the development of locomotor sensitization, conditioned place preference (CPP) and dopamine receptor supersensitivity induced by the repeated administration of morphine were investigated through behavioral experiments. A single administration of morphine produces hyperlocomotion. Repeated administration of morphine develops sensitization (reverse tolerance), a progressive enhancement of locomotion, which is used as a model for studying the drug-induced drug-seeking behaviors, and CPP, which is used as a model for studying drug reinforcement. Paeonol inhibited morphine-induced hyperlocomotion, sensitization and CPP. In addition, paeonol inhibited the development of postsynaptic dopamine receptors supersensitivity, which may be an underlying common mechanism that mediates the morphine-induced dopaminergic behaviors such as sensitization and CPP. Apomorphine (a dopamine agonist)-induced climbing behaviors also were inhibited by a single direct administration of paeonol. These results provide evidence that paeonol exerts anti-dopaminergic activity, and it is suggested that paeonol may be useful for the prevention and therapy of these adverse actions of morphine.

중국 목단재배종의 채취시기 및 부위별 Paeonol과 Paeoniflorin의 함량변화

최경,조비,이옥화,최준원,이학주,권영한,박광우 韓國藥用作物學會 2005 한국약용작물학회지 Vol.13 No.1

중국에서 재배되는 목단의 품종 중 화형의 연화정도가 다른 대표적인 6개 품종의 뿌리에 대해 채취시기에 따른 paeonol과 paeoniflorin의 함량을 조사하였다. 6개 품종의 뿌리에서 paeonol의 양은 계절에 관계없이 거의 일정하여 변화가 없었다. 반면 paeoniflorin은 개화기인 5월에 가장 높은 값을 보이다가 6월에 감소, 7월에는 증가, 8월에 다시 감소하다가 9월에는 증가하는 경향을 보였다. 비교한 6개의 품종중 화형의 형태가 단순한 종류가 복잡한 종류에 비해 약리성분을 많이 함유하고 있었다. 단판형화관인 Dan Feng과 만첩형 화관의 Wu Long Feng Sheng의 채취시기에 따른 부위별 약리성분의 분포정도를 비교하였다. Paeonol은 두 품종 모두 뿌리를 제외한 잎과 줄기에 미량으로 존재하며 채취시기가 달라져도 변화가 없었다. Paeoniflorin은 뿌리와 잎 두 부분에 다량으로 존재하여 뿌리에서의 분포양이 계절적으로 증감을 반복하는 반면, 잎은 성장이 가장 왕성한 5월에 가장 높게 분포하고 시기가 지날수록 감소하였다. We analyzed six cultivars of Chinese Moutan, Paeonia suffruticosa Andrews, using HPLC for the investigation of appropriate root harvesting time and distributions of paeonol and paeoniflorin according to the seasonal changes. The contents of paeonol remained nearly constant at different harvesting times. However, the paeoniflorin contents have changed significantly during the harvesting time. These showed the increasing tendency in May, July and September. We compared the distribution of two compounds between two cultivars with different flower forms, Dan Feng and Wu Long Feng Sheng. The contents of paeonol were very low in the leaf and annual shoot. This tendency was not changed during five harvesting times. But the paeoniflorin existed abundantly in the leaf and root. The contents of paeoniflorin in the leaf were highest in May, and gradually decreased.

Antitumor and Apoptosis Induction Effects of Paeonol on Mice Bearing EMT6 Breast Carcinoma

( Yetao Ou ),( Qingwang Li ),( Jianjie Wang ),( Kun Li ),( Shaobo Zhou ) 한국응용약물학회 2014 Biomolecules & Therapeutics(구 응용약물학회지) Vol.22 No.4

Paeonol is a major phenolic micromolecular component of Moutan cortex Radicis, a traditional Chinese Medicine. It has shown antitumor effects in previous studies; however, the underlying mechanisms remain unknown. This study investigated the mechanism by giving treatments of placebo, cyclophosphamide, paeonol of 150 and 300 mg/kg to 4 groups of mice bearing EMT6 breast cancer. Apoptosis in tumor cells were confi rmed by morphology analysis, including hematoxylin, eosin staining and TUNEL staining. The results showed that the weight of EMT6 breast tumor was signifi cantly reduced in the groups treated with both 150 and 300 mg/kg of paeonol. Immunohistochemical and Western blot results showed that the expression of Bcl-2 was down-regulated while the expression of Bax, caspase 8 and caspase 3 was up-regulated respectively. These results suggest that paeonol exhibits antitumor effects and the mechanism of the inhibition is via induction of apoptosis, regulation of Bcl-2 and Bax expression, and activation of caspase 8 and caspase 3.

Paeonol accelerates skin wound healing by regulating macrophage polarization and inflammation in diabetic rats

Zuyang Zhang,Tianhua Chen,Wei Liu,Jiepeng Xiong,Liangdong Jiang,Mingjiang Liu 대한약리학회 2023 The Korean Journal of Physiology & Pharmacology Vol.27 No.5

Diabetic ulcer is usually seen in people with uncontrolled blood sugar. Reportedly, many factors such as impaired glucose metabolism, and macrovascular and microvascular diseases caused angiogenesis disorders and delayed the healing of diabetic ulcers, thus affecting the body's metabolism, nutrition, and immune function. This study aimed to explore the effect of paeonol on skin wound healing in diabetic rats and the related mechanism. A rat model of diabetic ulcer was established. High glucose-treated mouse skin fibroblasts were co-cultured with M1 or M2- polarized macrophages treated with or without paeonol. H&E and Masson staining were used to reveal inflammatory cell infiltration and collagen deposition, respectively. Immunohistochemistry visualized the expression of Ki67, CD31, and vascular endothelial growth factor (VEGF). Western blot was used to detect interleukin (IL)-1β, tumor necrosis factor (TNF)-α, IL-4, IL-10, CD31, VEGFA, and collagen I/III. The expression of iNOS and arginase 1 was revealed by immunofluorescence staining. Paeonol treatment augmented collagen deposition and the expression of Ki67, CD31, VEGF, and macrophage M2 polarization markers (IL-4 and IL-10) and reduced wound area, inflammatory cell infiltration, and macrophage M1 polarization markers (IL-1β and TNF-α) in the ulcerated area. In vitro, paeonol treatment promoted M2-polarization and repressed M1-polarization in macrophages, thereby improving the repair of cell damage induced by high glucose. Paeonol accelerates the healing of diabetic ulcers by promoting M2 macrophage polarization and inhibiting M1 macrophage polarization.

Inhibitory Effects of Paeonol on Morphine-Induced Locomotor Sensitization and Conditioned Place Preference in Mice

은재순,배기환,윤여표,홍진태,Han-Na Kwon,오기완 대한약학회 2006 Archives of Pharmacal Research Vol.29 No.10

The inhibitory effects of paeonol, a major compound of Paeoniae radix, on the development of locomotor sensitization, conditioned place preference (CPP) and dopamine receptor supersensitivity induced by the repeated administration of morphine were investigated through behavioral experiments. A single administration of morphine produces hyperlocomotion. Repeated administration of morphine develops sensitization (reverse tolerance), a progressive enhancement of locomotion, which is used as a model for studying the drug-induced drug-seeking behaviors, and CPP, which is used as a model for studying drug reinforcement. Paeonol inhibited morphine-induced hyperlocomotion, sensitization and CPP. In addition, paeonol inhibited the development of postsynaptic dopamine receptors supersensitivity, which may be an underlying common mechanism that mediates the morphine-induced dopaminergic behaviors such as sensitization and CPP. Apomorphine (a dopamine agonist)-induced climbing behaviors also were inhibited by a single direct administration of paeonol. These results provide evidence that paeonol exerts anti-dopaminergic activity, and it is suggested that paeonol may be useful for the prevention and therapy of these adverse actions of morphine.

Antiallergic Effect of the Root of Paeonia lactiflora and Its Constituents Paeoniflorin and Paeonol

Lee, Bo-Mi,Shin, Yong-Wook,Bae, Eun-Ah,Han, San-Jun,Kim, Joo-Sun,Kang, Sam-Sik,Kim, Dong-Hyun 대한약학회 2008 Archives of Pharmacal Research Vol.31 No.4

The root of Paeonia lactiflora PALL (PL, Family Paeoniaceae) has been used frequently as an antipyretic and anti-inflammatory agent in traditional medicines of Korea, China and Japan. To evaluate antiallergic effect of PL, we isolated its main constituents, paeoniflorin and paeonol, and evaluated in vivo their inhibitory effects against passive cutaenous anaphylaxis (PCA) reaction induced by IgE-antigen complex and scratching behaviors induced by compound 48/80. PL, paeoniflorin and paeonol potently inhibited PCA reaction and scratching behaviors in mice. Paeoniflorin exhibited the most potent inhibition against scratching behaviors and the acetic acid-induced writhing syndrome in mice. Paeonol most potently inhibited PCA reaction and mast cells degranulation. These findings suggest that PL can improve IgE-induced anaphylaxis and scratching behaviors, and may be due to the effect of its constituents, paeoniflorin and paeonol.