Measurement of Total Plasma Homocysteine in Patients with Chronic Renal Failure Using HPLC/FLD

Kyung-Ok Lee(이경옥),Bo-Kyung Kang(강보경),Hyung-Jin Roh(노형진),Kwang-Suk Ryoo(류광석),Jeong-Yeon Yoo(유정연),Man-Jeong Paik(백만정),Kang-Hyeob Lee(이강협) 대한의생명과학회 1997 Biomedical Science Letters Vol.3 No.1

본 연구에서는 한국인 만성신부전증 환자에서 심혈관계 질환의 조기진단을 위한 생화학적 표지자로서 homocysteine의 임상적 유용성을 검토하기 위하여, 만성신부전증 환자와 건강인에서 HPLC/FLD (high performance liquid chromatography/fluorescence detector)를 이용하여 혈장 homocysteine 농도를 측정하고 그 결과를 비교하였다. 본 실험방법의 회수율은 98.6±5.8%를 나타내었으며, 0.2 n㏖/L보다 낮은 농도까지 측정이 가능하였고, 2~50 n㏖/㎖까지 직선성이 성립하였다 (correlation coefficient =0.9997). 한국인 중 건강한 정상인 (20명)과 만성신부전증 환자 (90명)에서 혈장 homocysteine은 각각 6.81±1.54 n㏖/㎖과 27.28±14.94n ㏖/㎖이었으며, 환자군에서는 정상인군에 비하여 약 4 배 정도 높은 수치를 나타내었다 (p<0.05). 본 실험의 결과로 볼 때 HPLC/FLD를 이용한 homocysteine 측정은 예민도와 재현성이 높아 routine 실험실 방법으로 유용성이 높을 것으로 생각되며, 또한 혈장 homocysteine의 측정은 만성신부전증 환자의 주요한 사망원인이 되는 폐쇄성 동맥질환을 조기에 진단하거나, 혹은 homocysteine 농도를 저하시키는 치료를 실시한 후 치료효과를 판단하기 위한 생화학적 marker로 활용될 수 있을 것으로 기대된다. Cardiovascular disease has been the leading cause of death among patients with chronic renal failure. Many reports have been described that homocysteine is one of the independent risk factor to the occulsive vascular disease. In this study, HPLC/FLD (high performance liquid chromatography-fluorescence detector) technique was used to measure homocysteine level in patients with chronic renal failure and normal control group. The detection limit and recovery of total plasma homocysteine using HPLC/FLD were 98.6±5.8% and 0.2 n㏖/㎖, respectively. The linearity of this method was established in concentration range of 2~50 n㏖/㎖ (correlation coefficient=0.9997). The concentrations of total plasma homocysteine were 6.81 ± 1.54 n㏖/㎖ and 27.28 ± 14.94 n㏖/㎖ in normal control (n=20) and patient group (n=90), respectively (p<0.05). In this study, the HPLC/FLD method showed high sensitivity and reproducibility for a routine clinical laboratory testing. Moreover determination of homocysteine level in plasma might be useful for a biochemical marker for predicting the cardiovascular diseases and for monitoring of therapeutic effect of lowering homocysteine in patients with chronic renal failure.

Inhibition of homocysteine-induced endoplasmic reticulum stress and endothelial cell damage by <small>L</small>-serine and glycine

Sim, Woo-Cheol,Han, Inhoi,Lee, Wonseok,Choi, You-Jin,Lee, Kang-Yo,Kim, Dong Gwang,Jung, Seung-Hwan,Oh, Seon-Hee,Lee, Byung-Hoon Elsevier 2016 Toxicology in vitro Vol.34 No.-

<P><B>Abstract</B></P> <P>Hyperhomocysteinemia is an independent risk factor for several cardiovascular diseases. The use of vitamins to modulate homocysteine metabolism substantially lowers the risk by reducing plasma homocysteine levels. In this study, we evaluated the effects of <SMALL>L</SMALL>-serine and related amino acids on homocysteine-induced endoplasmic reticulum (ER) stress and endothelial cell damage using EA.hy926 human endothelial cells. Homocysteine treatment decreased cell viability and increased apoptosis, which were reversed by cotreatment with <SMALL>L</SMALL>-serine. <SMALL>L</SMALL>-Serine inhibited homocysteine-induced ER stress as verified by decreased glucose-regulated protein 78kDa (GRP78) and C/EBP homologous protein (CHOP) expression as well as <I>X-box binding protein 1</I> (<I>xbp1</I>) mRNA splicing. The effects of <SMALL>L</SMALL>-serine on homocysteine-induced ER stress are not attributed to intracellular homocysteine metabolism, but instead to decreased homocysteine uptake. Glycine exerted effects on homocysteine-induced ER stress, apoptosis, and cell viability that were comparable to those of <SMALL>L</SMALL>-serine. Although glycine did not affect homocysteine uptake or export, coincubation of homocysteine with glycine for 24h reduced the intracellular concentration of homocysteine. Taken together, <SMALL>L</SMALL>-serine and glycine cause homocysteine-induced endothelial cell damage by reducing the level of intracellular homocysteine. <SMALL>L</SMALL>-Serine acts by competitively inhibiting homocysteine uptake in the cells. However, the mechanism(s) by which glycine lowers homocysteine levels are unclear.</P> <P><B>Highlights</B></P> <P> <UL> <LI> <SMALL>L</SMALL>-Serine and glycine ameliorated cell viability and apoptosis induced by homocysteine. </LI> <LI> <SMALL>L</SMALL>-Serine reduced homocysteine-induced ER stress by inhibiting homocysteine uptake. </LI> <LI> Glycine decreased intracellular homocysteine level and attenuated homocysteine-induced ER stress. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

서울중앙병원에서 IMx를 이용한 혈장내 총Homocysteine의 측정

한태진 ( Tae Jin Han ),신연수 ( Yeun Soo Shin ),유승용 ( Seung Yong Yu ),장진연 ( Jin Yeon Jang ),조병철 ( Byoung Cheol Cho ),전사일 ( Sa Il Chun ),민원기 ( Won Ki Min ) 대한임상검사과학회 2000 대한임상검사과학회지(KJCLS) Vol.32 No.3

Background : Elevated plasma homocysteine (HCY) levels are considered to be an independent risk factor in coronary 따tery disease, cerebrovascular disease, and peripheral vascular disease. Currently HPLC fluoremetric detection has been most widely used for the quantitation of homocysteine. Recently IMx homocysteine assay by use of a fluorescence polarization immlUloassay(FPIA) has been introduced that enables rapid quantitation of plasma homocysteine levels. We evaluated the performance of the IMx@ Homocysteine assay and compared it to routinely used HPLC method. Methods Within-run precision, between-run precision, degree of correlation, linearity, and reportable ranges were analyzed, according to National Committee for Clinical Laboratory Standarili잉 guidelines as much as possible. To establish a reference interval 148 apparently healthy adults were investigated (81 males, 67 females). For the methods comparison We selected 20 patients admitted to the Asan Medical Center. The total plasma homocysteine concentraton was measure by FPIA (lMx""J Homocysteine assay, Abbotte Laboratories, IL, USA) and by HPLC (HPll00, Hewlett Packard com., Germany) using Bio-rad@ Homocysteine kit (Bio-Rad Laboratories, CA, USA). The caIculations were performed with computer software, EP Suit(MarChem Associates, Concord, USA). Result : The within run coefficients of variations (CVs) of the low, medium, and of the high control were 1.8%, 1.5% and 1.5%. The between run CVs were 2.8%, 1.4% and 1.0%, respectively. Method was linear and passed lack of fit. Reportable range was found to be 0.5-50.0 μ molfL. The reference interval from hea1thy adu1ts was 10.3 i: 5.54(mean i: SD) μ molfL. Total plasma homocysteine levels according to sex were 11.9 i: 6. 89(mean i: SD) μ molfL for male, 8.39i: 2.03(meani: SD) μ mol/L for female, respectively. The comparison between FPIA (y) and HPLC(x) resu1ted in a linear regression analysis described by the equation : y=0.9463x-0.2364 with a correlation coefficient r=0.9961 Conclusions Linearity, pricision, reportable ranges of the IMx homocysteine were satisfactory. The IMx homocysteine also correlated well with HPLC method. The normal range, determined by testing apparent1y hea1thy adu1ts, is in agreement with the literature. πle IMx homocysteine assay is fast and easy to perform and very useful for routine determinations of plasma homocysteine levels.

관상동맥질환 환자에서 혈중 Homocystene농도와 Folic Acid, Vitamin B_(12) 농도와의 관계

김철현,최태형,장성근,홍세용 대한순환기학회 1998 Korean Circulation Journal Vol.28 No.4

Measurement of Total Plasma Homocysteine in Patients with chronic Renal Failure Using HPLC/FLD

Lee,Kyung-Ok,Yoo,Jeong-Yeon,Roh,Hyung-Jin,Paik,Man-Jeong,Lee,Kang-Hyeob,Kang,Bo-Kyung,Ryoo,Kwang-Suk THE KOREAN SOCIETY FOR BIOMEDICAL LABORATORY SCIEN 1997 Journal of biomedical laboratory sciences Vol.3 No.1

본 연구에서는 한국인 만성신부전증 환자에서 심혈관계 질환의 조기진단을 위한 생화학적 표지자로서 homocysteine의 임상적 유용성을 검토하기 위하여, 만성신부전증 환자와 건강인에서 HPLC/FLD (high performance liquid chromatography/fluorescence detector)을 이용하여 혈장 homocysteine 농도를 측정하고 그 결과를 비교하였다. 본 실험방법의 회수율은 98.6±5.8%를 나타내었으며, 0.2 nmol/L보다 낮은 농도까지 측정이 가능하였고, 2∼50 nmol/ml까지 직선성이 성립하였다 (correlation coefficient=0.9997). 한국인 중 건강한 정상인 (20명)과 만성신부전증 환자 (90명)에서 혈장 homocysteine은 각각 6.81±1.54 nmol/ml과 27.28±14.94 nmol/ml이었으며, 환자군에서는 정상인군에 비하여 약 4배 정도 높은 수치를 나타내었다 (p<0.05). 본 실험의 결과로 볼 때 HPLC/FLD를 이용한 homocysteine 측정은 예민도와 재현성이 높아 routine 실험실 방법으로 유용성이 높을 것으로 생각되며, 또한 혈장 homocysteine의 측정은 만성신부전증 환자의 주요한 사망원인이 되는 폐쇄성 동맥질환을 조기에 진단하거나, 혹은 homocysteine농도를 저하시키는 치료를 실시한 후 치료효과를 판단하기 위한 생화학적 marker로 활용될 수 있을 것으로 기대된다. Cardiovascular disease has been the leading cause of death among patients with chronic renal failure. Many reports have been described that homocysteine is one of the independent risk factor to the occulsive vascular disease. In this study, HPLC/FLD (high performance liquid chromatography-fluorescence detector) technique was used to measure homocysteine level in patients with chronic renal failure and normal control group. The detection limit and recovery of total plasma homocysteine using HPLC/FLD were 98.6±5.8% and 0.2 nmol/ml, respectively. The linearity of this method was established in concentration range of 2∼50 nmol/ml, (correlation coefficient=0.9997). The concentrations of total plasma homocysteine were 6.81±1.54 nmol/ml and 27.28±14.94 nmol/ml in normal control (n=20) and patient group (n=90), respectively (p<0.05). In this study, the HPLC/FLD method showed high sensitivity and reproducibility for a routine clinical laboratory testing. Moreover determination of homocysteine level in plasma might be useful for a biochemical marker for predicting the cardiovascular diseases and for monitoring of therapeutic effect of lowering homocysteine in patients with chronic renal failure.

말초동맥질환에 있어서 혈장 호모시스테인 농도의 의미

최동현,이상목,고석환,홍성화,오수명,윤충,박호철 대한혈관외과학회 2004 Vascular Specialist International Vol.20 No.1

경도인지장애와 알츠하이머형 치매 환자에서 혈중 Superoxide Dismutase 활성도와 Nitric oxide 및 Homocysteine 농도

김지은(Ji Eun Kim),정성수(Sung Soo Jung),이정구(Jung Goo Lee),신배섭(Bae Seob Shin),유영선(Young Sun You),윤성욱(Sung Wook Yoon),전동욱(Dong Wook Jeon),백준형(Jun Hyung Baek),이선정(Sun Jung Lee),박성우(Sung Woo Park),김영훈(Young Ho 대한생물치료정신의학회 2007 생물치료정신의학 Vol.13 No.2

목적 : 경도인지장애와 알츠하이머형 치매 환자에서 정상 대조군에 비해 혈중 Superoxide dismutase(SOD) 활성도와 nitric oxide(NO) 및 homocysteine 농도가 유의하게 차이가 나는지와 함께 전체 대상자에 대해 인지 기능 저하 정도와 각각의 상관관계에 대해 알아보고자 하였다. 방법 : 본 연구는 52명의 알츠하이머 치매 환자, 67명의 경도인지장애, 49명의 정상대조군을 대상으로 하였고 알츠하이머병의 진단은 DSM-IV와 NINCDS-ADRDA의 진단 기준을 따랐다. 인구학적 조사와 함께 임상적 평가로 Mini-Mental Status Examination-Korean version(MMSE-KC), Clinical Dementia Rating Scale (CDRS), Geriatric Depression Scale(GDS-K)을 시행하였고, 혈장 SOD 활성도와 혈청 NO 농도 및 혈장 homocysteine 농도를 측정하여 분석하였다. 결과 : 산화 스트레스인 혈중 SOD 활성도와 NO 및 homocysteine의 평균 농도는 알츠하이머형 치매군와 경도 인지장애군에서 정상 대조군과 비교했을 때 통계적으로 유의한 차이를 보이지 않았다. 그리고 전체 군을 대상으로 했을 때 MMSE-KC 점수와 혈중 homocysteine 농도 사이에는 약한 음의 상관 관계를 보였다(r=-0.18, p<0.05). 결론 : 본 연구에서는 경증의 알츠하이머형 치매에서 혈중 SOD 활성도와 NO 농도보다는 homocysteine 농도가 인지능력저하를 추정할 수 있는 더 유용한 생물학적 지표가 될 수 있을 것으로 추정하였다. Objectives:The purpose of this study was to investigate the differences in the blood superoxide dismutase(SOD) activity, concentrations of nitric oxide(NO) and homocysteine between the normal controls, mild cognitive impairment (MCI) and Alzheimer’s dementia. And we also evaluate the correlation between blood SOD activity, concentrations of NO and homocysteine and the cognitive decline level among all subjects. Methods:The study was carried out in 52 patients with Alzheimer’s dementia, 67 subjects with MCI and 49 normal controls. The subject received a diagnosis of Alzheimer’s dementia based on DSM-Ⅳ and NINCDS-ADRDA criteria. The Subjects were evaluated the demographic data and clinical data, including the Mini-Mental Status Examination-Korean version(MMSE-KC), clinical dementia rating scale(CDRS) and Geriatric Depression Scale (GDS-K). Plasma SOD activity, serum NO concentration and plasma homocysteine concentration were measured. Results:There was no significant correlation between the oxidative stress, such as blood SOD activity, concentrations of NO and homocysteine in the subjects with MCI and Alzheimer’s dementia, when compared with the normal controls. There was a weak negative correlation between the total score of MMSE-KC and plasma homocysteine level (r=-0.18;p<0.05). Conclusion:This study concluded that the plasma homocysteine level in mild Alzheimer’s dementia could be a more useful biological marker to estimate the cognitive decline compared to blood SOD activity and NO concentration.

Combined effect of folate and adiposity on homocysteine in children at three years of age

Su Jin Cho,Hye Ah Lee,Bo Hyun Park,Eun Hee Ha,Young Ju Kim,Eun Ae Park,Hyesook Park 한국영양학회 2016 Nutrition Research and Practice Vol.10 No.1

BACKGROUND/OBJECTIVES: Cardiovascular diseases is a major cause of death and is responsible for 23.8% of deaths in Korea. Clinical symptoms manifest in adulthood, but susceptibility begins in utero. Elevated homocysteine levels and adiposity might be linked to a greater risk in children as well as adults. We hypothesized that those who have simultaneous risk for folate and adiposity would be affected with elevated homocysteine levels at 3 years of age. SUBJECTS/METHODS: From the ongoing birth cohort at Ewha Womans University Mok-Dong Hospital, we compared adiposity parameters, serum homocysteine, and folate levels in 238 children (118 boys and 120 girls) at three years of age. The relationship between birth outcome, current weight and body mass index (BMI), postnatal growth, and homocysteine level were assessed using correlation and general linear model. Additionally, we assessed the combined effect between blood folate status and adiposity on current homocysteine levels. RESULTS: Birth characteristics were not correlated with homocysteine. Current weight, BMI, upper-arm circumference, skinfold thickness, waist circumference, and hip circumference were positively correlated with homocysteine at three years of age (P< 0.05). Folate level was negatively correlated with homocysteine at three years of age (P < 0.0001). A relative high anthropometric measure which is compatible with adiposity and low folate level was associated with high homocysteine levels. CONCLUSION: We found a combined effect of adiposity and folate levels with homocysteine levels at three years of age. This implicates the beneficial role of folate supplementation in the high-risk population at an early age.

Association of Homocysteine Levels With Blood Lead Levels and Micronutrients in the US General Population

이유미,이미경,배상근,이선화,김선영,이덕희 대한예방의학회 2012 예방의학회지 Vol.45 No.6

Objectives: Even though several epidemiological studies have observed positive associations between blood lead levels and homocysteine,no study has examined whether this association differs by the levels of micronutrients, such as folate, vitamin B6, and vitamin B12, which are involved in the metabolism of homocysteine. In this study, we examined the interactions between micronutrients and blood lead on homocysteine levels. Methods: This study was performed with 4089 adults aged ≥20 years old in the US general population using the National Health and Nutrition Examination Survey 2003-2004. Results: There were significant or marginally significant interactions between micronutrients and blood lead levels on mean homocysteine levels. Positive associations between blood lead and homocysteine were clearly observed among subjects with low levels of folate or low vitamin B6 (p-trend <0.01, respectively). However, in the case of vitamin B12, there was a stronger positive association between blood lead and homocysteine among subjects with high levels of vitamin B12, compared to those with low levels of vitamin B12. In fact, the levels of homocysteine were already high among subjects low in vitamin B12, irrespective of blood lead levels. When we used hyperhomocysteinemia (homocysteine>15 μmol/L) as the outcome, there were similar patterns of interaction, though pvalues for each interaction failed to reach statistical significance. Conclusions: In the current study, the association between blood lead and homocysteine differed based on the levels of folate, vitamin B6, or vitamin B12 present in the blood. It may be important to keep sufficient levels of these micronutrients to prevent the possible harmful effects of lead exposure on homocysteine levels.

Association of Homocysteine Levels With Blood Lead Levels and Micronutrients in the US General Population

Lee, Yu-Mi,Lee, Mi-Kyung,Bae, Sang-Geun,Lee, Seon-Hwa,Kim, Sun-Young,Lee, Duk-Hee The Korean Society for Preventive Medicine 2012 Journal of Preventive Medicine and Public Health Vol.45 No.6

Objectives: Even though several epidemiological studies have observed positive associations between blood lead levels and homocysteine, no study has examined whether this association differs by the levels of micronutrients, such as folate, vitamin B6, and vitamin B12, which are involved in the metabolism of homocysteine. In this study, we examined the interactions between micronutrients and blood lead on homocysteine levels. Methods: This study was performed with 4089 adults aged ${\geq}20$ years old in the US general population using the National Health and Nutrition Examination Survey 2003-2004. Results: There were significant or marginally significant interactions between micronutrients and blood lead levels on mean homocysteine levels. Positive associations between blood lead and homocysteine were clearly observed among subjects with low levels of folate or low vitamin B6 (p-trend <0.01, respectively). However, in the case of vitamin B12, there was a stronger positive association between blood lead and homocysteine among subjects with high levels of vitamin B12, compared to those with low levels of vitamin B12. In fact, the levels of homocysteine were already high among subjects low in vitamin B12, irrespective of blood lead levels. When we used hyperhomocysteinemia (homocysteine>15 ${\mu}mol/L$) as the outcome, there were similar patterns of interaction, though p-values for each interaction failed to reach statistical significance. Conclusions: In the current study, the association between blood lead and homocysteine differed based on the levels of folate, vitamin B6, or vitamin B12 present in the blood. It may be important to keep sufficient levels of these micronutrients to prevent the possible harmful effects of lead exposure on homocysteine levels.