Heme oxygenase-1 (HO-1)/carbon monoxide (CO) axis suppresses RANKL-induced osteoclastic differentiation by inhibiting redox-sensitive NF-κB activation

( Sun-uk Bak ),( Suji Kim ),( Hae-jun Hwang ),( Jung-a Yun ),( Wan-sung Kim ),( Moo-ho Won ),( Ji-yoon Kim ),( Kwon-soo Ha ),( Young-guen Kwon ),( Young-myeong Kim ) 생화학분자생물학회(구 한국생화학분자생물학회) 2017 BMB Reports Vol.50 No.2

Heme oxygenase (HO-1) catalyzes heme to carbon monoxide (CO), biliverdin/bilirubin, and iron and is known to prevent the pathogenesis of several human diseases. We assessed the beneficial effect of heme degradation products on osteoclastogenesis induced by receptor activator of NF-κB ligand (RANKL). Treatment of RAW264.7 cells with CORM-2 (a CO donor) and bilirubin, but not with iron, decreased RANKLinduced osteoclastogenesis, with CORM-2 having a more potent anti-osteogenic effect. CORM-2 also inhibited RANKLinduced osteoclastogenesis and osteoclastic resorption activity in marrow-derived macrophages. Treatment with hemin, a HO-1 inducer, strongly inhibited RANKL-induced osteoclastogenesis in wild-type macrophages, but was ineffective in HO-1^+/-cells. CORM-2 reduced RANKL-induced NFATc1 expression by inhibiting IKK-dependent NF-κB activation and reactive oxygen species production. These results suggest that CO potently inhibits RANKL-induced osteoclastogenesis by inhibiting redox-sensitive NF-κB-mediated NFATc1 expression. Our findings indicate that HO-1/CO can act as an antiresorption agent and reduce bone loss by blocking osteoclast differentiation. [BMB Reports 2017; 50(2): 103-108]

Carbon monoxide prevents TNF-α-induced eNOS downregulation by inhibiting NF-κB-responsive miR-155-5p biogenesis

Choi, Seunghwan,Kim, Joohwan,Kim, Ji-Hee,Lee, Dong-Keon,Park, Wonjin,Park, Minsik,Kim, Suji,Hwang, Jong Yun,Won, Moo-Ho,Choi, Yoon Kyung,Ryoo, Sungwoo,Ha, Kwon-Soo,Kwon, Young-Guen,Kim, Young-Myeong Nature Publishing Group 2017 Experimental and molecular medicine Vol.49 No.11

<P>Heme oxygenase-1-derived carbon monoxide prevents inflammatory vascular disorders. To date, there is no clear evidence that HO-1/CO prevents endothelial dysfunction associated with the downregulation of endothelial NO synthesis in human endothelial cells stimulated with TNF-α. Here, we found that the CO-releasing compound CORM-2 prevented TNF-α-mediated decreases in eNOS expression and NO/cGMP production, without affecting eNOS promoter activity, by maintaining the functional activity of the <I>eNOS</I> mRNA 3′-untranslated region. By contrast, CORM-2 inhibited MIR155HG expression and miR-155-5p biogenesis in TNF-α-stimulated endothelial cells, resulting in recovery of the 3′-UTR activity of <I>eNOS</I> mRNA, a target of miR-155-5p. The beneficial effect of CORM-2 was blocked by an NF-κB inhibitor, a miR-155-5p mimic, a HO-1 inhibitor and siRNA against HO-1, indicating that CO rescues TNF-α-induced eNOS downregulation through NF-κB-responsive miR-155-5p expression via HO-1 induction; similar protective effects of ectopic HO-1 expression and bilirubin were observed in endothelial cells treated with TNF-α. Moreover, heme degradation products, except iron and <I>N</I>-acetylcysteine prevented H<SUB>2</SUB>O<SUB>2</SUB>-mediated miR-155-5p biogenesis and eNOS downregulation. These data demonstrate that CO prevents TNF-α-mediated eNOS downregulation by inhibiting redox-sensitive miR-155-5p biogenesis through a positive forward circuit between CO and HO-1 induction. This circuit may play an important preventive role in inflammatory endothelial dysfunction associated with human vascular diseases.</P>

Ethanol Extract of Ganoderma lucidum Augments Cellular Anti-oxidant Defense through Activation of Nrf2/HO-1

Lee, Yoo-hwan,Kim, Jung-hee,Song, Choon-ho,Jang, Kyung-jeon,kim, Cheol-hong,Kang, Ji-Sook,Choi, Yung-hyun,Yoon, Hyun-Min KOREAN PHARMACOPUNCTURE INSTITUTE 2016 Journal of pharmacopuncture Vol.19 No.1

Objectives: The mushroom Ganoderma lucidum has been widely used as a traditional herbal medicine for many years. Although several studies have focused on the anti-oxidative activity of this mushroom, the molecular mechanisms underlying its activity have not yet been clearly established. The present study investigated the cytoprotective effect of ethanol extract of Ganoderma lucidum (EGL) against oxidative stress (hydrogen peroxide, $H_2O_2$) and elucidated the underlying mechanisms in a C2C12 myoblast cell line. Methods: Oxidative stress markers were determined by using the comet assay to measure reactive oxygen species (ROS) generation and deoxyribonucleic acid (DNA) damage. Cell viability and Western blotting analyses were employed to evaluate the cellular response to EGL and $H_2O_2$ in C2C12 cells. Transfection with nuclear factor erythroid 2-related factor 2 (Nrf2)-specific small interfering ribonucleic acid (siRNA) was conducted to understand the relationship between Nrf2 expression and $H_2O_2$-induced growth inhibition. Results: The results showed that EGL effectively inhibited $H_2O_2$-induced growth and the generation of ROS. EGL markedly suppressed $H_2O_2$-induced comet-like DNA formation and phosphorylation of histone H2AX at serine 139 ($p-{\gamma}H2AX$), a widely used marker of DNA damage, suggesting that EGL prevented $H_2O_2$-induced DNA damage. Furthermore, the EGL treatment effectively induced the expression of Nrf2, as well as heme oxygenase-1 (HO-1), with parallel phosphorylation and nuclear translocation of Nrf2 in the C2C12 myoblasts. However, zinc protoporphyrin IX, a HO-1 inhibitor, significantly abolished the protective effects of EGL against $H_2O_2$-induced accumulation of ROS and reduced cell growth. Notably, transient transfection with Nrf2-specific siRNA attenuated the cytoprotective effects and HO-1 induction by EGL, indicating that EGL induced the expression of HO-1 in an Nrf2-dependent manner. Conclusion: Collectively, these results demonstrate that EGL augments the cellular anti-oxidant defense capacity through activation of Nrf2/HO-1, thereby protecting C2C12 myoblasts from $H_2O_2$-induced oxidative cytotoxicity.

ORiginal Article : Gastroprotective Effects of PMK-S005 against Ethanol-Induced Acute Gastric Damage in Rats

( Yoon Jeong Choi ),( Nayoung Kim ),( Ju Yup Lee ),( Ryoung Hee Nam ),( Ji Hyung Seo ),( Seonmin Lee ),( Hee Jin Kim ),( Yoon Jin Choi ),( Hye Seung Lee ),( Dong Ho Lee ) 대한간학회 2016 Gut and Liver Vol.10 No.3

Background/Aims: This study aimed to examine the gastroprotective effects of PMK-S005, which is a synthetic S-allyl-Lcysteine (SAC; a sulfur-containing amino acid), against acute ethanol-induced gastric damage in rats. Methods: Sprague- Dawley rats were divided into six groups, including a nonethanol group, groups treated with absolute ethanol 1 hour after pretreatment with various doses of PMK-S005 (1, 5, and 10 mg/kg) or rebamipide (50 mg/kg), and an absolute ethanolonly group. Ethanol-induced gross ulcer and mucus levels were measured. Myeloperoxidase, tumor necrosis factor a, interleukin 1β, PGE2, LTB4, cPLA2, COX-1, and COX-2 levels were estimated by enzyme-linked immunosorbent assay or Western blot analysis. Furthermore, the protein expression levels of antioxidant enzymes, including heme oxygenase-1 (HO-1), NAD(P)H:quinine oxidoreductase 1 (NQO-1), GCLC, and GCLM, were assessed. Results: PMK-S005 significantly attenuated the ethanol-induced gastric damage; it reduced mucosal inflammatory cytokine production and increased mucus levels. The expression levels of cPLA2, COX-1, and COX-2 were decreased by PMK-S005. PMK-S005 did not affect PGE2 synthesis, but LTB4 production was significantly suppressed. In addition, long-term administration of PMKS005 significantly increased the expression of HO-1, NQO-1, GCLC, and GCLM. Conclusions: These results strongly suggest that PMK-S005 prevents gastric mucosal damage and that these gastroprotective activities are due to anti-inflammatory effects and enhancement of the gastric defense system, including antioxidant enzymes. (Gut Liver 2016;10:348- 355)

Korean Red ginseng prevents endothelial senescence by downregulating the HO-1/NF-κB/miRNA-155-5p/eNOS pathway

Tae-Hoon Kim,Ji-Yoon Kim,Jieun Bae,Young-Mi Kim,Moo-Ho Won,Kwon-Soo Ha,Young-Guen Kwon,Young-Myeong Kim 고려인삼학회 2021 Journal of Ginseng Research Vol.45 No.2

Background: Korean Red ginseng extract (KRGE) has beneficial effects on the cardiovascular system by improving endothelial cell function. However, its pharmacological effect on endothelial cell senescence has not been clearly elucidated. Therefore, we examined the effect and molecular mechanism of KRGE on the senescence of human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were grown in normal or KRGE-supplemented medium. Furthermore, they were transfected with heme oxygenase-1 (HO-1) gene or treated with its inhibitor, a NF-κB inhibitor, and a miR-155-5p mimic or inhibitor. Senescence-associated characteristics of endothelial cells were determined by biochemical and immunohistochemical analyses. Results: Treatment of HUVECs with KRGE resulted in delayed onset and progression of senescence-associated characteristics, such as increased lysosomal acidic β-galactosidase and decreased telomerase activity, angiogenic dysfunction, and abnormal cell morphology. KRGE preserved the levels of anti-senescent factors, such as eNOS-derived NO, MnSOD, and cyclins D and A: however, it decreased the levels of senescence-promoting factors, such as ROS, activated NF-κB, endothelial cell inflammation, and p21 expression. The beneficial effects of KRGE were due to the induction of HO-1 and the inhibition of NF-κB-dependent biogenesis of miR-155-5p that led to the downregulation of eNOS. Moreover, treatment with inhibitors of HO-1, NF-κB, and miR-155-5p abolished the anti-senescence effects of KRGE. Conclusion: KRGE delayed or prevented HUVEC senescence through a signaling cascade involving the induction of HO-1, the inhibition of NF-κB-dependent miR-155-5p biogenesis, and the maintenance of the eNOS/NO axis activity, suggesting that it may protect against vascular diseases associated with endothelial senescence.

나프타분해플랜트의 부탄추출공정에서 부탄증기의 연속누출에 의한 화재사고의 영향평가

윤대건,이헌창,함병호,조지훈,김태옥 韓國火災 ·消防學會 1998 한국화재소방학회논문지 Vol.10 No.4

나프타분해플랜트의 부탄추출공정에서 부탄증기의 연속 누출에 의한 제트화재와 플래쉬화재의 사고결과에 대한 영향을 평가하고 사고결과에 미치는 공정변수들의 영향을 해석하였다. 877 kPa과 346.75K의 공정조건에서 부탄증기가 87.8 kg/s의 속도로 8m의 높이에서 연속누출되는 경우에 표준조건으로 설정한 대기 및 환경조건에서 사고결과를 해석한 결과, 제트화재에서는 화염의 형태와 크기를 예측할 수 있었으며 누출지점으로부터 200m에서 API모델로 산출한 목사열은 약 5kW/m^2이었고 플래쉬화재가 일어날 수 있는 연소범위는 11.2~120.2m이었다. 그리고 사고결과에 미치는 조업압력의 영향은 조업온도보다 크며 압력이 증가할수록 사고결과 값이 증가하였다. 이때 플래쉬화재에서 X_UFL에 미치는 공정압력의 영향은 X_LFL에서보다 약 1/10이하를 나타내었다. The consequence analysis for jet and flash fire accidents by the continuous release of butane vapor was performed and effects of process variables on consequences were analyzed in standard conditions. For the continuous release(87.8 kg/s) of butane vapor at 8 m elevated height in the debutanizing process of the naphtha cracking plant operating at 877 kPa, 346.75 K, we found that for the jet fire accident, shape and size of the flame could be predicted and thermal radiation estimated by API model at 200 m distance from release point was 1.5 kW/m^2, and that for the flash fire accident, effect range was 11.2~120.2m. Also, simulation results showed that effects of operating pressures on consequences were large than those of operating temperatures and results of accidents were increased with increasing operating pressures. At this time, effects of operating pressures on X_UFL. were smaller

수완진동 증후군에서 신경장해의 조기진단을 위한 객관화된 방법

윤재국,이헌,최나리,김석환,박형욱,이지호,유철인 大韓産業醫學會 2009 대한직업환경의학회지 Vol.21 No.2

목적: 통각과 진동감각 역치검사 및 수부와 수지의 악력, 그리고 수지의 운동기능검사(태평) 등과 근로자들이 호소했던 증상에 대한 HAVS의 스톡홀름 워크샵 분류와의 관련성을 통해 좀 더 객관화된 신경학적 조기검사 방법을 찾고 타당성을 검증하고자 하였다. 방법: 2000년 1월부터 2007년 12월까지 수부진동에 폭로되고 수부의 불편함을 호소하여 진동 장해에 대한 검사를 시행한 497명의 근로자를 대상으로 하였다. 일차적으로 문진을 통해 직업력에 대해 알아보고 과거력을 조사했다. 대상자들의 스톡홀름 워크샵 분류에 따른 소견을 확인하고 수부의 말초 순환기능 검사(손톱압박, 수지피부 온도), 신경기능 검사(통각과 진동각 역치), 운동기능 검사(악력, 태평) 등을 시행하였다. 다른 질환의 감별을 위한 몇 가지 이학적 검사를 실시하고 스톡홀름 워크샵 분류에 따른 검사 결과들을 분석하였다. 결과: 사용 도구는 그라인더 단독 사용이 265명 (67.3%)으로 가장 많았으며 평균 노출 연수는 14.8년이었다. 통각과 진동감각에서 스톡홀름 워크샵 분류 단계에 따른 역치가 증가하는 경향을 보였으나 통각은 양측 모두 통계적인 유의성이 없었고, 진동감각은 125와 250 Hz에서 양측 모두 유의했다(p=0.006-0.038). 수부와 수지의 악력은 단계가 올라갈수록 감소하는 경향을 보였지만 수부와 수지 모두 우측에서만 통계적으로 유의했다 (p=0.041, p<0.01, p=0.034). 태평 첫수는 단계가 올라 가면서 전반적으로 태핑 첫수가 감소하였으며 좌측에서만 통계적으로 유의했다(p=0.002-0.019). 결론: 본 연구는 수부 진동 노출자들의 수부 불편함에 대한 신경계통을 초기에 정량적으로 객관화하여 진단할 수 있는 단일 방법은 아직 없으며 일상생활의 불편함과 스톡홀름 워크샵 분류 등을 통한 주관적인 증상에 더하여 수지 통각과 진동감각 역치검사, 수부와 수지 악력검사, 운동기능검사(태핑) 등을 통합하면 초기에 객관적으로 진단하는 것이 가능할 것으로 판단된다. Objectives: In order to determine find out the best methods for a more objective detection of neurologic abnormality in early hand ann vibration syndrome(HAVS), early with analyzing the validity of each of the detection methods was analyzed. We evaluated the relationships between the sensorineural stage of Stockholm-revised vibration syndrome classification and the results of several tests. Methods: 497 workers were investigated for symptom, exposure duration, the types of tools used, and medical history from January 2000 to December 2007. Pain sense threshold, vibrotactile threshold, hand grasp force, finger grasp force, and a finger tapping frequency test were performed by the workers. Results: The grinder(67.3%) was the most commonly used tool and the mean exposure duration was 14.8 years. Although the pain sense and vibrotactile threshold level tended to increase according to sensorineural stage of the Stockholm classification, there was statistically significant difference in the vibrotactile threshold of 125, 250 Hz (P=0.006~0.038) but not in the pain sense threshold. Hand and finger grasp force tended to decrease according to the sensorineural stage of Stockholm classification and there was statistically significant difference(P=0.041, <0.001, 0.034) only on the right hand side. The tapping frequency also generally decreased according to the sensorineural stage of the Stockholm classification and there was statistically significant difference (P=0.002~0.019) only on the left hand side. Conclusions: Although there is no single standardized method that can objectively diagnose the Sensorineural component of early HAVS early, the combination of subjective symptoms, the sensorineural stage of Stockholm classification, the pain and vibrotactile threshold test, the hand and finger grasp force, and the finger agility (tapping) test can objectively detect sensorineural component of HAVS early.

개의 아토피 피부염에 대한 Leflunomide의 임상적 적용 1례

홍은지,윤기영,서경원,최호정,정성목,송근호,조종기,박성준 忠南大學校 獸醫科大學 附設 動物醫科學硏究所 2012 動物醫科學硏究誌 Vol.19 No.1

A Shih-tzu(8-year-old, castrated male) was referred to the Veterinary Medical Teaching Hospital of Chungnam National University with the history of relapsing pruritus, recurrent dermatitis. On the physical examination, generalized erythema, otitis externa, alopecia and lichenification were observed. Using Favrot's criteria for diagnishing atopic dermatitis, 7 criteria were satisfied out of 8 criteria. After treatment for infection, intradermal skin test was performed(positive allergen : Df, Dp). And then immunotherapy was performed, but clinical signs were not getting better. Next treatment of glucocorticoid shown little improvement of clinical signs, and adverse effect(diarrhea) was shown after cyclosporine application. So application of leflunomide which is used for human atopic dermatitis treatment in some case was planned. After 6 months of application of leflunomide, clinical signs were decreased gradually.

枇杷葉의 抗糖尿病藥理活性과 IL-1β誘發糖尿病 마우스의 膵臟 燐酸化酵素에 미치는 效果에 關한 硏究

윤철호,정지천,김철호 동국대학교 한의학연구소 1998 東國韓醫學硏究所論文集 Vol.7 No.1

淸凉生津, 解渴上嘔의 효능이 있으며, 한국등에서 항당뇨병성 한약으로서 사용되어 온 비파엽의 열수출물의 당뇨병치료효과를 검증하기 위하여 alloxan으로 유발된 당뇨병흰쥐의 혈장포도당농도 및 인슈린농도를 측정하였다. 비파엽처리군의 혈장 포도당농도는 현저히 증가하였으며 혈장인슈린농도는 크게 감소하지 않았다. 한편, 당뇨병유발의 내재적인 요소로 알려진 사이토카안인 IL-1β로 당뇨병을 유발한 뒤 그 효과를 검정했다. 즉, 실험동물을 interleukin-1β로 처리하여 실험적으로 당뇨병을 유발한 후 glucose의 燐酸化에 관여하는 glutokinase와 hexokinase의 활성을 살펴보았다. 그 결과, 비파엽엑기스는 interleukin-1β 과잉투여 당뇨병으로 인하여 증가된 혈액내 glucose농도를 감소시키고 insulin 과잉분비를 정상으로 회복시키는데 효과적이었다. 동시에 lL-1β에 의해 유도된 당뇨마우스에 대한 枇杷葉엑기스 投與가 췌장 glucokinase와 hexokinase의 활성을 촉진하는 것으로 나타나 lL-1β유발 당뇨병에 효과가 있는 것으로 사료되었다. Studies were conducted on anti-dibetic effect of the water extract from leaves of Eriobotryae folium which had been used in Korea as a remedy for dibetes. The extract was found to inhibit the Increase in the plasma level of sugar bu the not the decrease in the plasma level of insulin in alloxan-induced dibetic rata. Also, we investigated the in vivo effect of an aqueous extract(referred to as EF) from Eriobotryae folium on glucokinase and hexokinase activities of diabetes mellilus induced by Interteukin-1β (IL-1β). After 1 week of IL-1β injection, the levels of serum glucose concentration and insulin secretion were dramatically increased. However, the insulin secretion was decreased with administration of EF. The level of glucose concentration was decreased by EF administration. Furthermore, it was observed that EF was effective in recovering the levels of insulin secretion. Enzyme activities of the glucokinase and hexokinase, which are key enzymes of glucose phosphorylastion, were decreased by IL-1β. EF administration to the mice allowed proportional increasing by stimulation of induction of enzyme activities as high as normal group. These results suggested that EF is highly effective in treatment of diabetes mellitus induce by IL-1β.

鹿茸 藥針 製劑가 힌쥐 腎臟 組織의 抗酸化 作用에 미치는 影響

윤철호,정지천,신억섭 한국한의학연구원 1996 한국한의학연구원논문집 Vol.2 No.1

Cervus elaphus for herb-acupuncture solution (CEHAS) was tested for the effects of free radical generating enzyme and lipid peroxidation in rat's kidney. In vitro, levels of lipid peroxide in tissues of kidney were proportionally decreased to concentration of extracts prepared from CEHAS. They were much more decreased, when lipid perocidation was inducesd with ferrous iron(Fe II). Also, enzyme activities of xanthine oxidase were decreased. The ratio of type conversion of zanthine oxidase was lowered, too. But, it was not seen changes on enzyme activities of aldehyde oxidase. These results suggest that CEHAS decrease the activities of free radical generating enzymes such as xanthine oxidase which form lipid peroxide.