• Nutrition Research and Practice
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• 제14권4호
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• pp.334-351
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• 2020

BACKGROUND/OBJECTIVES: This study was designed to investigate the improvement effect of white ginseng extract (GS-KG9) on D-galactosamine (Ga1N)-induced oxidative stress and liver injury. SUBJECTS/METHODS: Sixty Sprague-Dawley rats were divided into 6 groups. Rats were orally administrated with GS-KG9 (300, 500, or 700 mg/kg) or silymarin (25 mg/kg) for 2 weeks. The rats of the GS-KG9- and silymarin-treated groups and a control group were then intraperitoneally injected Ga1N at a concentration of 650 mg/kg for 4 days. To investigate the protective effect of GS-KG9 against GalN-induced liver injury, blood liver function indicators, anti-oxidative stress indicators, and histopathological features were analyzed. RESULTS: Serum biochemical analysis indicated that GS-KG9 ameliorated the elevation of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) in GalN-treated rats. The hepatoprotective effects of GS-KG9 involved enhancing components of the hepatic antioxidant defense system, including glutathione, glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT). In addition, GS-KG9 treatment inhibited reactive oxygen species (ROS) production induced by GalN treatment in hepatocytes and significantly increased the expression levels of nuclear factor erythroid-2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) proteins, which are antioxidant proteins. In particular, by histological analyses bases on hematoxylin and eosin, Masson's trichrome, α-smooth muscle actin, and transforming growth factor-β1 staining, we determined that the administration of 500 mg/kg GS-KG9 inhibited hepatic inflammation and fibrosis due to the excessive accumulation of collagen. CONCLUSIONS: These findings demonstrate that GS-KG9 improves GalN-induced liver inflammation, necrosis, and fibrosis by attenuating oxidative stress. Therefore, GS-KG9 may be considered a useful candidate in the development of a natural preventive agent against liver injury.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 추계학술대회
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• pp.103-103
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• 2018

Oenothera biennis, commonly known as evening primrose, a potential source of natural bioactive substances: flavonoids, steroids, tannins, fatty acids and terpenoids responsible for a diverse range of pharmacological functions. However, whether extract prepared from aerial part of O. biennis (APOB) protects skin against oxidative stress remains unknown. To investigate the protective effects of APOB against oxidative stress-induced cellular damage and elucidated the underlying mechanisms in the HaCaT human skin keratinocytes. Our results revealed that treatment with APOB prior to hydrogen peroxide ($H_2O_2$) exposure significantly increased viability, and the highest DPPH radical-scavenging activities and reducing power of HaCaT cells. APOB also effectively attenuated H2O2-induced comet tail formation and inhibited the $H_2O_2$-induced phosphorylation levels of the histone ${\gamma}H2AX$, as well as the number of apoptotic bodies and Annexin V-positive cells. In addition, APOB exhibited scavenging activity against intracellular reactive oxygen species (ROS) accumulation and restored the mitochondrial membrane potential loss by $H_2O_2$. Moreover, $H_2O_2$ enhanced the cleavage of caspase-3 and degradation of poly (ADP-ribose)-polymerase (PARP), a typical substrate protein of activated caspase-3, as well as DNA fragmentation; however, these events were almost totally reversed by pretreatment with APOB. Furthermore, APOB increased the levels of heme oxygenase-1 (HO-1), which is a potent antioxidant enzyme, associated with the induction of nuclear factor-erythroid 2-related factor 2 (Nrf2). According to our data, APOB is able to protect HaCaT cells from $H_2O_2$-induced DNA damage and cell death through blocking cellular damage related to oxidative stress through a mechanism that would affect ROS elimination and activating the Nri2/HO-1 signaling pathway.

• 대한한의학방제학회지
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• 제25권1호
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• pp.51-68
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• 2017

Objectives : Oxidative stress due to excessive accumulation of reactive oxygen species (ROS) is one of the risk factors for the development of several chronic diseases, including neurodegenerative diseases. Methods : In the present study, we investigated the protective effects of cheongnoemyeongsin-hwan (CNMSH) against oxidative stress‑induced cellular damage and elucidated the underlying mechanisms in neuronal-derived SH-SY5Y cells. Results : Our results revealed that treatment with CNMSH prior to hydrogen peroxide (H2O2) exposure significantly increased the SH-SY5Y cell viability, indicating that the exposure of the SH-SY5Y cells to CNMSH conferred a protective effect against oxidative stress. CNMSH also effectively attenuated H2O2‑induced comet tail formation, and decreased the phosphorylation levels of the histone ${\gamma}H2AX$, as well as the number of apoptotic bodies and Annexin V‑positive cells. In addition, CNMSH exhibited scavenging activity against intracellular ROS generation and restored the mitochondria membrane potential (MMP) loss that were induced by H2O2, suggesting that CNMSH prevents H2O2‑induced DNA damage and cell apoptosis. Moreover, H2O2 enhanced the cleavage of caspase-3 and degradation of poly (ADP-ribose)-polymerase, a typical substrate protein of activated caspase-3, as well as DNA fragmentation; however, these events were almost totally reversed by pretreatment with CNMSH. Furthermore, CNMSH increased the levels of heme oxygenase-1 (HO-1), which is a potent antioxidant enzyme, associated with the induction of nuclear factor-erythroid 2-related factor 2 (Nrf2). According to our data, CNMSH is able to protect SH-SY5Y cells from H2O2-induced apoptosis throughout blocking cellular damage related to oxidative stress through a mechanism that would affect ROS elimination and activating Nrf2/HO-1 signaling pathway. Conclusions : Therefore, we believed that CNMSH may potentially serve as an agent for the treatment and prevention of neurodegenerative diseases caused by oxidative stress.

• 생명과학회지
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• 제27권7호
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• pp.783-789
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• 2017

Malus melliana (Hand.-Mazz.) Rehder (M. melliana)는 장미과에 속하는 중국 자생 식물 중 하나로 현재까지 보고된 생리활성은 전무하다. 본 연구에서는 M. melliana 에탄올 추출물(MMEE)의 항산화 및 항염증 생리활성을 DPPH 라디칼 소거능, ROS 소거능, NO 생성 저해능 및 Western blot hybridization을 통한 연관 단백질 발현분석을 통해 평가하였다. MMEE의 항산화능을 DPPH 라디칼 소거능을 통해 분석한 결과 양성 대조군으로 사용한 대표적인 항산화제인 아스코르빈산과 유사한 정도의 높은 소거활성을 보여 MMEE가 매우 강한 항산화능을 보유함을 확인하였다. 또한 RAW 264.7 세포주에서 $H_2O_2$에 의해 유도된 ROS에 대한 MMEE의 소거능을 분석한 결과, 농도의존적인 강한 ROS 소거능을 보였다. 뿐만 아니라 대표적인 항산화 효소인 HO-1 및 그 전사 인자인 Nrf2의 단백질 발현에 미치는 영향을 분석한 결과 MMEE에 의해 HO-1 및 Nrf2의 발현이 증가됨을 보였다. 한편 MMEE가 LPS에 의해 유도된 NO 생성에 미치는 영향을 분석한 결과 농도의존적인 NO 생성 저해능을 보였으며 이는 NO 생성 단백질인 iNOS의 발현 저해에서 기인함을 확인하였다. 이러한 결과를 통해 MMEE의 높은 항산화능과 항염증 활성을 확인하였으며 향후 잠재적인 기능성 소재로서 유용하게 활용될 수 있을 것으로 판단된다. 추후 계속적인 연구를 통해 활성 물질의 규명이 필요할 것으로 판단된다.

• 한방재활의학과학회지
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• 제29권4호
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• pp.29-45
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• 2019

Objectives This study is to investigate anti-obesity effects of Banggihwanggi-tang-hap-yeonggyechulgam-tang (BY), an herbal formula, in high fat diet induced obese mice model. Methods Fourty five male C57Bl/6J mice were randomly assigned to normal group fed with normal research diet (Nor, n=9), high fat diet control group treated with water (Veh, n=9), high fat diet group treated with orlistat (Oris; n=9, Orlistat 40 mg/kg), high fat diet group treated with low concentraion BY (BYL; n=6, BY 0.87 g/kg) and high fat diet group treated with high concentration BY (BYH; n=6, BY 1.74 g/kg). Results Seven weeks later, antioxidative capacity, body weight, epididymal fat pad and liver weight, reactive oxygen species (ROS), peroxynitrite ($ONOO^-$), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol, triglyceride, high density lipoprotein (HDL), low density lipoprotein (LDL), superoxide dismutase (SOD), catalase, glutathione peroxidase (Gpx), heme oxygenase (HO)-1 and histology of liver were evaluated. In the BYH group, 1,1-diphenyl-2-picrylhydrazyl and 2,2'-azinobis (3 ethybenzothiazoline-6-sulfonic acid) radical scavenging activity were more than L-ascorbic acid. Body weight gain were significantly less than Veh group. Epididymal fat pad and liver weight gain were significantly less than Veh group. ROS and $ONOO^-$ were significantly less than with Veh group. ALT and AST were significantly less than with Veh group. Total cholesterol, triglyceride and LDL were significantly less, HDL were significantly more than Veh group. SOD, catalase, Gpx, HO-1 significantly increased compared with Veh group. Injury on liver was lesser than Veh group. Conclusions It can be suggested that BY has anti-obesity effects in high fat diet induced obese mice model.

• 동의생리병리학회지
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• 제28권3호
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• pp.303-309
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• 2014

The fruit of Gleditsia sinensis has been extensively used as a key ingredient of an herbal remedy for the treatment of various inflammatory diseases in traditional Korean Medicine. However, the reason of using the fruit of G. sinensis for the remedy is unclear. Since Nuclear factor (erythroid-derived 2)-like 2 (Nrf2) is a key anti-inflammatory transcription factor, which is activated by the fruit of G. sinesis, we examined whether other plant parts of G. sinensis are also capable of suppressing inflammatory responses by activating Nrf2. Water extracts of various parts of G. sinensis were prepared and tested for Nrf2 activation by reporter assay and western blot analysis. Our results show that the hull of G. sinensis is the most potent in activating Nrf2. Sequential organic solvent extraction of the hull show that all the fractions had a higher potency in activating Nrf2 than the water extract, albeit differential degrees. The hull originated from Korea in general activated Nrf2 strongly compared to that of China. Chloroform fraction of the hull was further examined, showing that the fraction induced nuclear localization of Nrf2, indicative of activated Nrf2, and Nrf2-dependent gene expression including NAD(P)H dehydrogenase quinone 1 (NQO-1), glutamate-cysteine ligase catalytic subunit (GCLC), and heme oxygenase - 1 (HO-1). Therefore, our results show that, among other plant parts examined in this study, the hull of G. sinensis is the most potent, providing the experimental basis for the use of the hull of G. sinensis as an active ingredient for an anti-inflammatory remedy.

• Journal of Acupuncture Research
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• 제31권4호
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• pp.33-43
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• 2014

This study was performed to investigate the protective effects of acupuncture on the liver in the oxidative stress caused by cadmium accumulation. Sprague-Dawley male($150{\pm}30g$) rats were stabilized for a week and divided into 5 groups which is normal, control, $LR_3$ acupuncture, $BL_{23}$ acupuncture and sham acupuncture group. For three days experimental groups were received oral doses of cadmium 2 mg/kg twice a day. Acupuncture was given bilaterally at each point 10 times for two weeks. The depth of stimulation was 1 mm at right angles and torsion of acupuncture was produced 2 times per second for 1 minutes. The liver was shipped off and taken weight at the last day of two weeks, and hepatic functions was confirmed through alanine transaminase(ALT) and aspartate amino-transferase(AST). We measured reactive oxygen species of serum, liver and kidney, and compared expression levels of superoxide dismutase(SOD), catalase, glutathione peroxidase(Gpx), nuclear factor erythroid derived 2-related factor 2(Nrf-2), heme oxygenase-1(HO-1), nuclear factor-${\kappa}B$ (NF-${\kappa}B$), cyclooxygenase-2(COX-2), inducible nitric oxide synthase(iNOS), Bax and Cytochrome c. $BL_{23}$ acupuncture group significantly increased liver weight and decreased ALT compared to control group. For the oxidative stress, $LR_3$ acupuncture group significantly reduced reactive oxygen species, and $BL_{23}$ acupuncture group significantly reduced reactive oxygen species and inflammation-related protein compared to control group. But $LR_3$ acupuncture group and $BL_{23}$ acupuncture group didn't significantly reduce apoptosis-related protein. Therefore $LR_3$ and $BL_{23}$ acupuncture showed the effects of antioxidant and anti-inflammatory, especially $BL_{23}$ acupuncture was more effective than $LR_3$ acupuncture on the protection of liver in the oxidative stress.

• 대한한의학회지
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• 제25권3호
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• pp.123-136
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• 2004

Objectives : The purpose of this investigation is to evaluate the effects of Woohwangcheongsim-won (WC) on the in vitro neuronal development and alteration in gene expression in a hypoxia model using cultured rat cortical cells. Methods : E/sub 18/ rat cortical cells were grown in a neurobasal medium containing B27 supplement and various concentration of WC. Initial development of growth cone was investigated by phase-contrast microscopy, while dendritic spine formation and synaptogenesis were investigated by immunocytochemistry with SynGAPα(a postsynaptic marker) and synaptophysin (presynaptic marker) antibodies. Alteration in gene expression was analyses by microarray using rat 5K-TwinChips. Results : WC suppressed the development of growth cones and WC increased the number of dendritic spines at 20 and 50㎍/mL concentration but there was no statistical significance. Instead, it significantly decreased the number at 100㎍/mL. The expression of anti-apoptosis gene Bcl2-like 1 (Bcl211) increased (Global M=0.46), while Akt1 decreased. Proapoptosis genes Bad and PDCD2 increased. The expression of hemoglobin alpha 1 (probably neuroglobin) increased (Global M=0.93). The expression of antioxidants such as catalase, heme oxygenase (HO), and PRKAG2 gene increased. The expression PKC gene increased. The expression of retinoic acid receptor alpha (RARα) increased significantly (Global M=1.0). Conclusions : These data suggest that WC trends to suppress cellular activity slightly in normoxia and increases the expression of apoptosis-, antioxidation-, oxygen capture-related genes in hypoxia, but increases Bcl111 that anti-apoptosis gene, on the other hand increases Bad, PDCD2 that pro-apoptosis genes, too..

• 생명과학회지
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• 제21권12호
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• pp.1689-1697
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• 2011

본 연구는 고마리 추출물이 가지는 항염증 활성을 알아보기 위하여 쥐의 대식세포(RAW264.7 cell)에 Lipopolysaccharide (LPS)를 처리하여 염증반응을 유도하고 이때 발생되는 Nitric oxide (NO)의 생성 억제를 확인하였다. 또한 염증에서 중요하게 알려져 있는 Inducible nitric oxide synthase (iNOS), Cyclooxygenase-2 (COX-2), Nuclear factor-kappa B (NF-${\kappa}B$) 단백질들의 발현을 비교하였고, 추가적으로 NF-${\kappa}B$ 단백질의 핵 내부로의 이전 및 활성을 확인하였다. 메탄올 추출물은 NO 생성 및 iNOS, COX-2, NF-${\kappa}B$ 단백질의 발현을 억제하고, 세포를 보호하는 효과를 가지는 Heme oxygenase-1 (HO-1) 단백질의 발현을 증가시켰다. 위 결과를 바탕으로 하여 n-butanol, hexane, ethyl acetate 용매를 이용한 추가적인 분획을 실시하였다. 이들 분획 중 고마리의 ethyl acetate 추출물은 Prostaglandin $E_2$ ($PGE_2$), NO 생성을 억제 하였으며, iNOS, COX-2 단백질들의 발현을 감소, NF-${\kappa}B$의 핵 내부로의 이동을 억제하는 효과가 높다는 것을 확인하였다. 이러한 연구결과는 고마리 식물이 좋은 항염증 활성을 가지고 있음을 나타내며, 지속적인 분획으로 고마리 식물이 가지는 항염증 활성 물질을 선별하여 그 작용기작을 규명하는 연구가 필요하다.

• 한방재활의학과학회지
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• 제31권1호
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• pp.17-32
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• 2021

Objectives The present study was designed to find out the therapeutic effects and possible underlying mechanism of Buja-tang, a herbal complex formula on experimental monosodium iodoacetate (MIA)-induced osteoarthritis. Methods Osteoarthritis models were created via intra-joint injection of MIA (50 μL with 80 mg/mL) in rats. Rats were divided into five groups and each group consisted of seven. Normal group was not injected MIA and did a normal diet. Control group injected MIA and received distilled water. Indo injected MIA and oral administration of 5 mg/kg of indomethacin. BJTL injected MIA and oral administration of 100 mg/kg of Buja-tang. BJTH injected MIA and oral administration of 200 mg/kg of Buja-tang. We analyzed weight-bearing ability of hind paws, oxidative stress related factor, antioxidant protein, inflammatory protein, inflammatory messenger and cytokine in joint tissue. Pathological observation of knee cartilage tissue structures was also performed with hematoxylin & eosin and safranin-O chromosomes. Results Weight-bearing ability of hind paws showed a tendency to reduce pain. The incidence of nicotinamide adenine dinucleotide phosphate oxidase and p22phox in articular tissue was significantly reduced, and the incidence of nuclear factor-erythroid 2-related factor 2 and heme oxygenase-1 and superoxide dismutases was significantly increased. The incidence of phosphorylated inhibitor of κBα, nuclear factor-kappa B p65, inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor alpha, interleukin (IL)-6, and IL-1β decreased significantly. In pathological observation, cartilage tissue damaged by MIAs in biopsy has significantly recovered from Buja-tang administration. Conclusions Buja-tang has anti-inflammation, antioxidation and pain relief effects. So this is thought to inhibit the progress of osteoarthritis in rat caused by the MIA.