• 한방안이비인후피부과학회지
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• 제36권1호
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• pp.21-39
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• 2023

Objectives : The purpose of this study was to investigate the anti-inflammatory effect of Goihwa-san water extract(GHS) in vitro & in vivo. Methods : In vitro, we evaluated the anti-inflammatory effect of GHS by comparing the Raw 264.7 cells with 10, 30, 100, 300㎍/㎖ of GHS for 1 hour before Lipopolysaccharide(LPS) to the single LPS treated group. We examined the relative cell viability by MTT assay and the relative level of LPS, Loxoribine(LOX), Peptidoglycan(PGN), Flagellin(FLA)-induced NO production by using Griess reagent and measured relative iNOS protein level and COX-2 protein level by using western blot and Image analyzing system. We measured the production of TNF-α, IL-1β, and IL-6 by each ELISA kits and then measured the relative levels of IκBα, p-IκBα in whole-cell lysate fraction and NF-κB in nuclear fraction by using western blot and Image analyzing system. In vivo, we induced the paw edema by subcutaneous injection of 100㎕/rat CA and measured the swelling volume of paw by using a plethysmometer and then measured the relative iNOS protein level by using western blot. Results : As a result, in vitro, LPS, PGN-induced NO production was significantly inhibited by pretreatment with GHS. GHS reduced LPS, PGN-induced iNOS expression, PGN-induced COX-2 expression and LPS-induced production of cytokine(TNF-α, IL-1β, IL-6). Expression of IκBα was increased by pretreatment with GHS 100㎍/㎖. And the expression of p-IκBα and NF-κB were decreased by pretreatment with GHS 100㎍/㎖. In vivo, CA-induced inflammation rat model was used for the evaluation of the anti-inflammatory effect of GHS. 0.3 or 1.0g/kg of GHS significantly reduced the increases of paw swelling and iNOS expression in paw tissues. Conclusions : These results show that GHS can decrease inflammatory response via inhibition of the NF-κB pathway in vitro. And in vivo, the anti-inflammatory effect suggest the clinical basis of GHS for the treatment of inflammatory diseases.

• Nutrition Research and Practice
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• 제16권6호
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• pp.685-699
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• 2022

BACKGROUND/OBJECTIVES: Platycodon grandiflorum (PG) has long been known as a medicinal herb effective in various diseases, including bronchitis and asthma, but is still more widely used for food. Fermentation methods are being applied to increase the pharmacological composition of PG extracts and commercialize them with high added value. This study examines the hydrolyzed and fermented PG extract (HFPGE) fermented with Lactobacillus casei in RAW 264.7 cells, and investigates the effect of amplifying the immune and the probable molecular mechanism. MATERIALS/METHODS: HFPGE's total phenolic, flavonoid, saponin, and platycodin D contents were analyzed by colorimetric analysis or high-performance liquid chromatography. Cell viability was measured by the MTT assay. Phagocytic activity was analyzed by a phagocytosis assay kit, nitric oxide (NO) production by a Griess reagent system, and cytokines by enzyme-linked immunosorbent assay kits. The mRNA expressions of inducible nitric oxide synthase (iNOS) and cytokines were analyzed by reverse transcription-polymerase chain reaction, whereas MAPK and nuclear factor (NF)-κB activation were analyzed by Western blots. RESULTS: Compared to PGE, HFPGE was determined to contain 13.76 times and 6.69 times higher contents of crude saponin and platycodin D, respectively. HFPGE promoted cell proliferation and phagocytosis in RAW 264.7 cells and regulated the NO production and iNOS expression. Treatment with HFPGE also resulted in increased production of interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, C-X-C motif chemokine ligand10, granulocyte-colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, and monocyte chemoattractant protein-1, and the mRNA expressions of these cytokines. HFPGE also resulted in significantly increasing the phosphorylation of NF-κB p65, extracellular signal-regulated kinase, and c-Jun N-terminal kinase. CONCLUSIONS: Taken together, our results imply that fermentation and hydrolysis result in the extraction of more active ingredients of PG. Furthermore, we determined that HFPGE exerts immunostimulatory activity via the MAPK and NF-κB signaling pathways.

• 대한본초학회지
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• 제29권2호
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• pp.1-6
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• 2014

Objectives : Reactive oxygen species (ROS) are involved in a wide spectrum of diseases including chronic inflammation and cancer. In this study, we investigated the antioxidant activities and anti-inflammatory effects of the extracts from the herbal teas such as Lonicera japonica Thunberg (L. japonica), Chrysanthemum morifolium Ramat (C. morifolium), Mentha arvensis L. (M. arvensis), and P.rhizoma. Methods : Anti-oxidant activity was evaluated using DPPH radical scavenging assay and $Fe^{2+}$ chelating assay. And DNA cleavage assay was performed to evaluate an anti-oxidative effect. Anti-inflammatory effect was performed using NO generation assay and western blot in LPS-stimulated RAW264.7 cell line. Results : L. japonica scavenged DPPH radical by 9.8% at 12.5 ${\mu}g/ml$, 24.8% at 25 ${\mu}g/ml$, 34.3% at 50 ${\mu}g/ml$, 61.1% at 100 ${\mu}g/ml$ and 75.8% at 200 ${\mu}g/ml$, respectively. In addition, C. morifolium and M. arvensis removed DPPH radical by 15.6% and 10.4% at 12.5 ${\mu}g/ml$, 34.8% and 22.8% at 25 ${\mu}g/ml$, 66.9% and 43.3% at 50 ${\mu}g/ml$, 87.4% and 69.1% at 100 ${\mu}g/ml$, and 92.1% and 73.2% at 200 ${\mu}g/ml$, respectively. However, P. rhizoma did not affect on DPPH radical scavenging. The $Fe^{2+}$ chelating activity was highest in L. japonica, but lowest in P. rhizoma among the herbal teas. In addition, the extracts from L. japonica, C. morifolium and M. arvensis inhibited oxidative DNA damage via its anti-oxidant activity. In anti-inflammatory effect, the extracts from C. morifolium inhibited NO production. In addition, it suppressed the $NF-{\kappa}B$ signaling pathway in LPS-stimulated RAW 264.7 cells. Conclusions : Together, this study indicates that L. japonica, M. arvensis and C. morifolium possess the protective effect against the oxidative DNA damage. Furthermore, C. morifolium exerts an anti-inflammatory effect.

• 대한본초학회지
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• 제38권2호
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• pp.17-26
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• 2023

Objectives : A persistent inflammatory response can cause diseases such as fibrosis, cancer, and allergies. This study aimed to investigate the anti-inflammatory activity of Curcumae longae Rhizoma and Cinnamomi Ramulus Mixture (CCM) in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Methods : The total polyphenol and flavonoid contents of CCM were confirmed through an in vitro experiment. Also, radical scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and Hydroxyl were confirmed. Moreover, ferric reducing antioxidant power (FRAP) activity were confirmed. After, CCM (50, 100, and 200 ㎍/mL) were applied to 0.1 ㎍/mL LPS-stimulated RAW264.7 cells. The levels of nitric oxide (NO) and pro-inflammatory cytokines in the supernatant fraction were determined. Also, the expressions of mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) pathways were detected using Western blot. Results : As a result of in vitro experiments, there was an excellent antioxidant activity in CCM-treated cells. In addition, in RAW264.7 cells stimulated with LPS, the increased NO level was inhibited in a concentration-dependent manner by the treatment of CCM. In addition, inflammatory cytokines production were significantly inhibited in a concentration-dependent manner in CCM-treated group. CCM treatment significantly decreased the protein expressions of MAPKs. Moreover, the expressions of NF-κBp65 and cyclooxygenase-2 (COX-2) were significantly decreased when 200 mg/kg of CCM was applied, and phospho-inhibitor of nuclear factor kappa B-α (p-IκBα) and inducible nitric oxide synthase (iNOS) were significantly decreased at all concentrations treated with CCM. Conclusion : Our findings show that CCM exhibited excellent antioxidant activity and exhibited superior anti-inflammatory effect through the MAPKs and NF-κB pathways in LPS-stimulated RAW 264.7 macrophages.

• Nutrition Research and Practice
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• 제17권5호
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• pp.899-916
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• 2023

BACKGROUND/OBJECTIVES: Oxidative stress is a fundamental neurodegenerative disease trigger that damages and decimates nerve cells. Neurodegenerative diseases are chronic central nervous system disorders that progress and result from neuronal degradation and loss. Recent studies have extensively focused on neurodegenerative disease treatment and prevention using dietary compounds. Heseperetin is an aglycone hesperidin form with various physiological activities, such as anti-inflammation, antioxidant, and antitumor. However, few studies have considered hesperetin's neuroprotective effects and mechanisms; thus, our study investigated this in hydrogen peroxide (H₂O₂)-treated SH-SY5Y cells. MATERIALS/METHODS: SH-SY5Y cells were treated with H₂O₂ (400 µM) in hesperetin absence or presence (10-40 µM) for 24 h. Three-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assays detected cell viability, and 4',6-diamidino-2-phenylindole staining allowed us to observe nuclear morphology changes such as chromatin condensation and apoptotic nuclei. Reactive oxygen species (ROS) detection assays measured intracellular ROS production; Griess reaction assays assessed nitric oxide (NO) production. Western blotting and quantitative polymerase chain reactions quantified corresponding mRNA and proteins. RESULTS: Subsequent experiments utilized various non-toxic hesperetin concentrations, establishing that hesperetin notably decreased intracellular ROS and NO production in H₂O₂-treated SH-SY5Y cells (P < 0.05). Furthermore, hesperetin inhibited H₂O₂-induced inflammation-related gene expression, including interluekin-6, tumor necrosis factor-α, and nuclear factor kappa B (NF-κB) p65 activation. In addition, hesperetin inhibited NF-κB translocation into H₂O₂-treated SH-SY5Y cell nuclei and suppressed mitogen-activated protein kinase protein expression, an essential apoptotic cell death regulator. Various apoptosis hallmarks, including shrinkage and nuclear condensation in H₂O₂-treated cells, were suppressed dose-dependently. Additionally, hesperetin treatment down-regulated Bax/Bcl-2 expression ratios and activated AMP-activated protein kinase-mammalian target of rapamycin autophagy pathways. CONCLUSION: These results substantiate that hesperetin activates autophagy and inhibits apoptosis and inflammation. Hesperetin is a potentially potent dietary agent that reduces neurodegenerative disease onset, progression, and prevention.

• Korean Journal of Radiology
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• 제25권3호
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• pp.267-276
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• 2024

Objective: To evaluate the diagnostic performance of susceptibility map-weighted imaging (SMwI) taken in different acquisition planes for discriminating patients with neurodegenerative parkinsonism from those without. Materials and Methods: This retrospective, observational, single-institution study enrolled consecutive patients who visited movement disorder clinics and underwent brain MRI and ¹⁸F-FP-CIT PET between September 2021 and December 2021. SMwI images were acquired in both the oblique (perpendicular to the midbrain) and the anterior commissure-posterior commissure (AC-PC) planes. Hyperintensity in the substantia nigra was determined by two neuroradiologists. ¹⁸F-FP-CIT PET was used as the reference standard. Inter-rater agreement was assessed using Cohen;s kappa coefficient. The diagnostic performance of SMwI in the two planes was analyzed separately for the right and left substantia nigra. Multivariable logistic regression analysis with generalized estimating equations was applied to compare the diagnostic performance of the two planes. Results: In total, 194 patients were included, of whom 105 and 103 had positive results on ¹⁸F-FP-CIT PET in the left and right substantia nigra, respectively. Good inter-rater agreement in the oblique (κ = 0.772/0.658 for left/right) and AC-PC planes (0.730/0.741 for left/right) was confirmed. The pooled sensitivities for two readers were 86.4% (178/206, left) and 83.3% (175/210, right) in the oblique plane and 87.4% (180/206, left) and 87.6% (184/210, right) in the AC-PC plane. The pooled specificities for two readers were 83.5% (152/182, left) and 82.0% (146/178, right) in the oblique plane, and 83.5% (152/182, left) and 86.0% (153/178, right) in the AC-PC plane. There were no significant differences in the diagnostic performance between the two planes (P > 0.05). Conclusion: There are no significant difference in the diagnostic performance of SMwI performed in the oblique and AC-PC plane in discriminating patients with parkinsonism from those without. This finding affirms that each institution may choose the imaging plane for SMwI according to their clinical settings.

• Korean Journal of Radiology
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• 제24권10호
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• pp.983-995
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• 2023

Objective: To compare the diagnostic performance and interobserver agreement between contrast-enhanced computed tomography (CECT) and contrast-enhanced magnetic resonance imaging (CE-MRI) with magnetic resonance cholangiopancreatography (MRCP) for evaluating the resectability in patients with extrahepatic cholangiocarcinoma (eCCA). Materials and Methods: This retrospective study included treatment-naïve patients with pathologically confirmed eCCA, who underwent both CECT and CE-MRI with MRCP using extracellular contrast media between January 2015 and December 2020. Among the 214 patients (146 males; mean age ± standard deviation, 68 ± 9 years) included, 121 (56.5%) had perihilar cholangiocarcinoma. R0 resection was achieved in 108 of the 153 (70.6%) patients who underwent curative-intent surgery. Four fellowship-trained radiologists independently reviewed the findings of both CECT and CE-MRI with MRCP to assess the local tumor extent and distant metastasis for determining resectability. The pooled area under the receiver operating characteristic curve (AUC), sensitivity, and specificity of CECT and CE-MRI with MRCP were compared using clinical, surgical, and pathological findings as reference standards. The interobserver agreement of resectability was evaluated using Fleiss kappa (κ). Results: No significant differences were observed between CECT and CE-MRI with MRCP in the pooled AUC (0.753 vs. 0.767), sensitivity (84.7% [366/432] vs. 90.3% [390/432]), and specificity (52.6% [223/424] vs. 51.4% [218/424]) (P > 0.05 for all). The AUC for determining resectability was higher when CECT and CE-MRI with MRCP were reviewed together than when CECT was reviewed alone in patients with discrepancies between the imaging modalities or with indeterminate resectability (0.798 [0.754-0.841] vs. 0.753 [0.697-0.808], P = 0.014). The interobserver agreement for overall resectability was fair for both CECT (κ = 0.323) and CE-MRI with MRCP (κ = 0.320), without a significant difference (P = 0.884). Conclusion: CECT and CE-MRI with MRCP showed no significant differences in the diagnostic performance and interobserver agreement in determining the resectability in patients with eCCA.

• 대한한의학방제학회지
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• 제32권2호
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• pp.155-179
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• 2024

Objective : Danggwisu-san (DGSS) is an herbal formula that has been mainly used in the East Asia for the treatment of bruise, sprain and external injury. The cause of this pain is that Qi and blood become tangled and do not circulate well. DGSS can improve the tangled situation and make it well-circulated. The present study evaluated the anti-inflammatory effects of DGSS on Raw 264.7 cells and in rats with paw edema. Methods : Cell viability was measured using the MTT assay. The amount of nitric oxide (NO) production was measured the amount of nitrite content in the cultured medium using Griess reagent. The amount of tumor necrosis factor-α, monocyte chemoattractant protein 1, interleukin (IL)-1βand IL-6 in the cultured supernatant were measured by ELISA kit. Proteins expression were detected by Western blot. Furthermore, the effect of DGSS on acute inflammation was observed in rat paw edema model. Results : The DGSS ameliorates the lipopolysaccharide-activated changes in NO production, iNOS expression and pro-inflammatory cytokines. Additionally, DGSS significantly suppressed expression of p-JNK, p-ERK and nuclear NF-κB. As expected, in rat paw edema study, 1.0 g/kg of DGSS significantly reduced the carrageenan-induced paw edema and iNOS expression for 1-4 h. Moreover, administration of 1.0 g/kg (4 days) of DGSS used in this study did not show any significant change on ALT and AST. Conclusion : These results demonstrate that DGSS has anti-inflammatory effects in vitro and in vivo. Therefore, this present study can put scientific evidences up for the anti-inflammatory effect of DGSS.

• Pediatric Infection and Vaccine
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• 제15권1호
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• pp.52-58
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• 2008

목 적 : 잠복 결핵 감염 진단 시 사용되는 고전적인 방법인 투베르쿨린 피부 반응 검사(TST) 외 전혈에서 인터페론 감마의 분비능을 측정하는 검사가 개발된 이후 많은 연구들이 그 유용성을 입증하였으나, 소아에서는 그러한 평가가 제한되어 있다. 본 연구는 결핵에 노출된 소아의 잠복 결핵 감염 진단에 있어 QuantiFERON-TB Gold 검사의 유용성을 알아보고자 하였다. 방 법 : 2006년 1월부터 2007년 12월까지 분당 서울 대학교 병원에 결핵 환자와 접촉하여 내원한 소아를 대상으로 흉부 방사선 사진, TST 및 QuantiFERON-TB Gold 검사를 시행하였다. 초기 TST가 음성인 경우, 3개월 뒤 추적 검사를 시행하였다. TST 결과에 따라 초기 TST가 양성인 경우 감염 여부는 확실치 않으나 잠복 감염의 가능성이 있는 군, 초기 TST는 음성이었으나 3개월 뒤 검사에서 양성 전환된 경우는 잠복 결핵 감염군, 처음과 3개월 뒤 TST가 모두 음성인 경우는 감염되지 않은 군으로 정의하였다. 초기 TST가 음성이었으나 추적검사를 하지 않은 경우는 분석에서 제외시켰다. 결 과 : 대상군은 총 49명(남자 22명, 여자 27명) 이었으며, 중앙 연령은 75세(범위; 3개월-14.7세)이었다. QuantiFERON-TB Gold 검사 양성을 보인 소아는 총 5명으로, 감염 여부는 확실치 않으나 잠복 감염의 가능성이 있는 13명 중 3례(23.1%), 잠복 결핵 감염으로 진단된 8명 중 1례(13%), 그리고 감염되지 않은 21명 중 1례(5%)가 포함되었다. QuantiFERON-TB Gold와 TST의 일치도는 0.72이었다(${\kappa}=0.101$). TST와 비교하였을 때 QuantiFERON-TB Gold의 민감도는 12.5%(1/8), 특이도는 95.2%(20/ 21)으로 나타났다. 결 론 : 결핵에 노출된 소아의 잠복결핵감염의 진단에 있어서 QuantiFERON-TB Gold 검사는 TST에 비하여 민감도가 매우 낮아서 임상적 유용성이 불확실하며 더 많은 연구가 필요하다.

• 대한소아치과학회지
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• 제30권2호
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• pp.254-262
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• 2003

개량형 Dentocult-SM 키트의 치아우식활성검사로서의 활용도와 임상적 지수와의 상관관계 그리고 민감도 및 특이도를 조사하기 위하여 만 5-6세 남녀아동 128명을 대상으로 구강검사와 screening strip 검사와 site strip 검사를 시행한 후 일정 부위의 치면세균막을 채취하여 뮤탄스 연쇄상구균 배양검사를 실시하여 다음과 같은 결론을 얻었다. Screening strip 및 site strip 검사 모두에서 우식활성도가 0에서부터 3까지로 증가할수록 아동들이 보유하고 있는 dft index와 dfs index는 각각 통계적으로 유의하게 증가하였고 아동들의 치면세균막 내의 뮤탄스 연쇄상구균 집락수 역시 우식활성도가 0에서 3으로 높아질수록 통계적으로 유의하게 수적인 증가를 나타내었다. Screening strip과 site strip 검사결과의 일치정도는 Spearman의 상관계수가 0.775 그리고 kappa 지수는 0.496으로 두 검사법의 일치도가 높았다. 또한 screening strip 검사의 민감도는 0.74-1.00, 특이도는 0.11-0.67로 나타났고, site strip 검사의 민감도와 특이도는 각각 0.48-1.00 및 0.11-0.66으로 계산되었다. 따라서 치과임상에서도 종래의 우식활성검사법들 보다 시간과 비용이 그리 많이 들지 않으며, 또한 각종 미생물학적, 생화학적인 기술이나 장비가 많이 필요치 않도록 간편하게 개량된 Dentocult-SM 검사법을 활용하면 치아우식증을 예측하는 진단학적인 측면이나 환자들을 대상으로 한 구강보건교육적인 측면에서 많은 도움이 될 수 있을 것으로 사료되었다.