• 대한한방부인과학회지
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• 제28권2호
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• pp.40-54
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• 2015

Objectives : In the theory of Korean medicine, Citri Reticulatae Viride Pericarpium (CRVP) can soothe the liver to break qi stagnation, eliminate mass and relieve dyspepsia. This study was carried out to investigate the effects of CRVP on the apoptotic cell death in breast cancer cells. Methods : In the present experiment, the effects of CRVP on proliferation rates, type of cell death, cell cycle distribution, and intracellular oxidative stress were investigated using MDA-MB-231 cells in vitro. In addition, the effects on expression levels of caspase 3, caspase 9, Bax and Bcl-2 were also investigated. Results : Treatment with CRVP decreased proliferation rates in a dose dependent manner. ID50 (50% inhibitory dosage) was 175.4 μg/ml. In the CRVP treated group, cell volumes showed smaller than non-treated normal. In addition, CRVP increased percentage of apoptotic and sub G1 arrested cells respectively. 200 μg/ml of CRVP treatment increased intracellular ROS level significantly. Finaly the expression level of caspase 3 and Bax/Bcl-2 ratio were elevated by treatment with CRVP respectively. Conclusions : These results suggest that CRVP can trigger intrinsic apoptotic pathway in MDA-MB-231 cells.

• 대한본초학회지
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• 제26권3호
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• pp.1-6
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• 2011

Objectives : The present study was undertaken to determine whether an ethanol extract of Citri Pericarpium (CP-E), which is the pericarp of Citrus unshiu Markovich is efficacious against collagen-induced arthritis (CIA) in mice. Methods : CIA was induced in male DBA/1J mice by intradermal injection of bovine collagen-II in Freund's incomplete adjuvant (IFA). The mice in the onset of arthritis were treated daily with oral administration of CP-E ethanol extract at different doses (50 and 100 mg/kg/bw) for 28 days. Arthritis index, histopathologic changes and the levels of TNF-${\alpha}$ as well as anti-CII IgG in blood were evaluated to confirm the anti-arthritic effect of CP-E on CIA in rats. Results : The results showed that comparing with untreated CIA mice, treated with CP-E extract significantly decreased the arthritic scores and the pathological changes of knee joint tissues, and also reduced the serum levels of TNF-${\alpha}$ and anti-CII IgG in CIA-mice. These results indicate that CP-E extract may effectively alleviate inflammatory response on CIA, and its anti-inflammation can be attributed, at least partially, to the inhibition of proinflamamtory cytokine, TNF-${\alpha}$ in CIA. Conclusions : This study suggest that CP-E has a therapeutic potential in inflammatory joint diseases such as rheumatoid arthritis.

• 대한한방부인과학회지
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• 제34권1호
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• pp.34-47
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• 2021

Objectives: This study was designed to examine anti-inflammatory effect and mechanism of Citri Reticulatae Viride Pericarpium water extract (CRE). Methods: Cell cytotoxicity was tested with RAW 264.7 cells. To investigate anti-inflammatory effect of CRE in lipopolysaccharide (LPS)-induced RAW 264.7 cell, we measured nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and interleukin-10 (IL-10). In addition, mitrogen-activated protein kinase (MAPK) and nuclear factor kappa B (NF-κB) were examined by western blotting in LPS-induced RAW 264.7 cell with treated CRE. Results: In cytotoxicity analysis, CRE does not affect cell cytotoxicity. As compared with the control group, the expression of NO, PGE2, TNF-α, IL-6 were significantly decreased, and IL-10 was significantly increased in LPS-induced RAW 264.7 cell with treated CRE. As a result of Western blotting, there was concentration-dependent inhibition of pp38, pERK in MAPK pathway and significant reduction of pp65 in the NF-κB pathway. Conclusions: CRE might have anti-inflammatory effect in LPS-induced macrophages by promoting the production of IL-10.

• 대한본초학회지
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• 제36권1호
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• pp.1-8
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• 2021

Objective : Citri Unshius Pericarpium (Citrus unshiu peel) has been used in Korean medicine to treat indigestion, vomiting, coughing and phlegm. This study investigated the hepatoprotective effect of ethanol extract of Citrus unshiu peel (CEE) in cadmium (CdCl2)-treated mouse model. Methods : CEE was dissolved in water and administered orally to mice once a day for 7 consecutive days. The mice were then exposed to a single intraperitoneal (i.p.) injection of cadmium (4 mg/kg body weight) to induce acute hepatotoxicity. At the end of the experiment, blood and liver tissue samples were collected, analyzed for alanine aminotransferase (ALT), aspartate aminotransferase (AST), and histopathological evaluation. Liver damage was assessed as the percentage of degenerative areas of the hepatic parenchyma, the number of degenerative hepatocytes, and the number of infiltrated inflammatory cells. Results : In cadmium-treated rats, pretreatment with CEE significantly reduced the serum ALT and AST levels associated with liver damage. Histopathologically, CEE prevented degenerative changes on the hepatic tissues including confluent necrosis, congestions and infiltration of inflammatory cells. CEE also reduced the elevation of oxidative stress markers (nitrotyrosine and 4-hydroxynonenal) and apoptosis markers (cleaved caspase-3 and cleaved PARP) positive cells. PARP protein expression in liver tissue was also restored by CEE. Conclusion : This study showed that CEE exerted antioxidant and anti-apoptotic effects against cadmium-induced liver injury. Thus, it can be concluded that CEE can be used to prevent liver damage caused by cadmium.

• 대한본초학회지
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• 제35권3호
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• pp.1-8
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• 2020

Objectives : In this study, we examined the estrogenic activities and anti-osteo clastogenesis effects of PCE17, a mixture of PE (an extract of Puerariae Flos), and CE (an extract of Citri Unshius Pericarpium). Methods : The estrogenic effect of PCE17, PE and CE were examined by ER-β/ERE reporter gene assay and proliferation assay in 293 T and MCF-7 cells. The expression of estrogen-responsive gene and protein were checked by Real Time-PCR (RT-PCR) and Western blotting in MCF-7 cells. Inhibitory effect of PCE17, PE and CE on RANKL-induced osteoclast differentiation were evaluated by TRAP staining and RT-PCR in primary osteoclast precursors from rat bone marrow cells. Results : PCE17 and PE bind to ERs (estrogen receptors) and show estrogenic activities in 293T cells. They also stimulated the proliferation of MCF-7 cells and increased the expression of ER response gene, pS2. Tectorigenin, an active ingredient of PE, shows similar estrogenic activities in MCF-7 cells. PCE17 and CE inhibited RANKL-induced osteoclastogenesis in rat primary osteoclast precursor cells and down-regulated the osteoclast-specific genes of Nfatc1, Ctsk, and Acp5. Conclusions : In conclusion, PCE17 may have therapeutic potential in cases of menopause and osteoporosis.

• 대한한의학회지
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• 제43권4호
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• pp.52-73
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• 2022

Objectives: The anti-inflammatory and anti-oxidative activities of LCVC (Lonicerae Flos, Citri Pericarpium and Violae Herba Complex) have not been fully elucidated. The purpose of this study was to investigate the mechanisms underlying these effects in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. Methods: The evaluation of the anti-oxidative activity of LCVC was completed via DPPH and ABTS radical scavenging capacity, FRAP assay, measurement of polyphenol and flavonoid, assessment of ROS and NO levels in LPS-induced RAW 264.7 cells. The anti-inflammatory activity was defined by measuring the production of biomarkers (PGE2, IL-1B, IL-6 and TNF-𝛼), proteins (ERK, JNK, P38, Nrf2, Keap1, HO-1 and NQO1) and expressions of genes (iNOS, COX-2, IL-1𝛽, IL-6, TNF-𝛼, Nrf2, Keap1, HO-1 and NQO1) in LPS-induced RAW 264.7 cells. Results: LCVC have polyphenol and flavonoid contents. The results of DPPH and ABTS free radical scavenging capacity and FRAP assay showed that the anti-oxidative activity was increased. Production of ROS, NO, IL-6, TNF-𝛼, mRNA expressions of IL-1𝛽, IL-6, TNF-𝛼, Keap1, iNOS and COX-2 were decreased, and NQO1, Nrf2, and HO-1 were increased. In protein expression, JNK and Keap1 were decreased, NQO1, Nrf2 and HO-1 were increased, and no relationships were observed with the ERK and P38 by LCVC. Conclusions: These results suggest that LCVC may offer protective effects against LPS-induced inflammatory and oxidative responses through attenuating Nrf2/HO-1 pathway and MAPKs pathway. Therefore, we propose that LCVC has anti-inflammatory and anti-oxidative activities that have therapeutic potential in the treatment of inflammatory and oxidative disorders caused by the over-activation of macrophages.

• 대한한의학방제학회지
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• 제31권4호
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• pp.265-281
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• 2023

Objectives : Ukgan-san plus Citri Pericarpium and Pinelliae Rhizoma (UCP) is used as a traditional herbal formula in Korea and Japan for treatment of fever, fever-induced convulsions, and liver dysfunction and so on. In this study, we investigated the cytoprotective effect and underlying mechanism of UCP against oxidative stress induced by cotreatment of arachidonic acid (AA) and iron. Methods : To evaluate the hepatoprotective effects of UCP against AA + iron-induced oxidative stress in HepG2 cell, cell viability and changes on apoptosis-related proteins were assessed by MTT and immunoblot analyses. The changes in intracellular reactive oxygen species (ROS), glutathione (GSH), and mitochondrial membrane permeability (MMP) were investigated against to the oxidative stress. Furthermore, to verify underlying molecular mechanism, NF-E2-related factor 2 (Nrf2) and its downstream target genes were examined by immunoblot analysis. Results : Treatment of UCP increased the cell viability and altered the expression levels of apoptosis-related proteins such as PARP, caspase-9, caspase-3, Bcl-2. UCP also inhibited the GSH depletion, excessive ROS production and mitochondrial dysfunction induced by AA + iron. In addition, the Nrf2 and the Nrf2 target genes activation were increased by UCP. Conclusions : These results indicated that UCP has the ability to protect against oxidative stress-induced hepatocyte damage, which may be mediated with Nrf2 pathway.

• 생명과학회지
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• 제23권8호
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• pp.1057-1063
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• 2013

진피(Citri Pericarpium)의 항암작용 기전 해석을 위하여 U937 백혈병 세포의 apoptosis 유발에 미치는 메탄올 추출물(EMCP)의 영향을 조사하였으며, apoptosis 조절에 중요한 몇 가지 유전자들의 발현 및 활성 변화, ROS의 생성 변화를 조사하였다. EMCP 처리에 의한 U937 세포의 증식 억제는 apoptosis 유도와 연관성이 있음을 DAPI 염색을 통한 apoptotic body 출현의 증가 및 Flow cytometry 분석에 의한 Sub-G1기 세포 빈도의 증가로 확인하였다. EMCP 처리에 의한 apoptosis 유도에서 Bax 발현 증가, caspases의 활성 및 PARP의 단편화 등이 동반되었으며, ROS 생성의 증가와 연관성이 있었다. 산화적 손상에 대해 세포나 조직을 보호하는 역할을 하는 것으로 알려진 세포 내 항산화 효소인 HO-1의 발현이 EMCP의 처리에 의해 증가되었으나 ROS 생성 억제제인 NAC의 전처리에 의해 감소된 HO-1의 발현은 전사인자인 Nrf2의 핵으로의 이동 억제와 관련되어 있었다. 이상의 결과에서 EMCP 처리에 의한 U937 세포의 apoptosis 유발에는 ROS 생성의 증가와 pNrf2에 의해 조절되는 HO-1의 발현 증가가 중요한 기전으로 작용한다는 것을 알 수 있었다. 이러한 자료는 진피의 항암기전 해석을 이해하는데 중요한 기초자료로서 활용될 수 있을 것으로 생각된다.

• 대한한방내과학회지
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• 제13권2호
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• pp.1-15
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• 1992

This study has been carried out to investigate the effects of Banhaonpaetang extract and its constituent herbs on the contractile force of Isolated Guinea pig trachea smooth muscle and to elucidate its mechanism. The results were as follows; 1. Banhaonpaetang significanly inhibitd the contractile force of isolated guinea pig trachea smooth muscle pretreated by histamine. 2. Banhaonpaetang significantly inhibited the contractile force of isolated guinea pig trachea smooth muscle pretreated by acetylcholine. 3. Radix Asari(細辛), Flos Inulae(旋覆花), Pericarpium Citri Nobilis(陳皮) and Radix Ginseng(人蔘) extract significantly inhibited the contractile force of isolated guinea pig trachea smooth muscle pretreated by histamine. 4. Radix Asari(細辛), Cortex Cinnamomi(桂心), Flos Inulae(旋覆花), Pericarpium Citri Nobilis(陳皮) and Radix Ginseng(人蔘) extract significantly inhibited the contractile force of isolated guinea pig trachea smooth muscle pretreated by acetylcholine. 5. The effects of Banhaonpaetang extract on the changes of plasma ACTH in rats were shown to be insignificant. 6. The effects of Banhaonpaetang extract on the changes of serum electrolytes were shown to be significant in $K^+$ and $Ca^{2+}$.

• 대한한의학방제학회지
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• 제8권1호
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• pp.329-342
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• 2000

Citri Reticulatae Viride Pericarpium(CRVP) is being used to regulate the flow of Q(氣) However, the mechanism of it's pharmacological actions is not well understood. The purpose of this research was to investigate effects of CRVP contractil response of isolated on abdominal and femoral artery in rabbits and renal artery in pigs. 1. Abdominal artery was relaxed by CRVP in a dose-dependent manner. 2. Femoral artery was relaxed by CRVP in a dose-dependent manner 3. Pretreatment with methylene blue and indomethacin did not inhibited CRVP induced relax in abdominal artery and femoral artery. 4. Renal artery was relaxed by $H_{2}O$ fraction in a dose-dependent manner, 5. Pretreatment with regitine inhibited $H_{2}O$ fraction(CRVP) induced relax in renal artery in a dose-dependent manner. 6. Renal artery was not relaxed by hexane fraction(CRVP) in a dose-dependent manner. 7. Pretreatment with regitine$(10^{-7}M)$ was relaxed by methylene chloride(MC)(CRVP) and $H_{2}O$ fraction in a $0.5{\mu}g/m{\ell}$.