Helicobacter pylori 감염과 관련된 철 결핍성 빈혈에서 Lactoferrin Sequestration의 역할

문광빈,강창규,최연호,한혜승,송순욱,Moon, Kwang-Bin,Kang, Chang-Kyu,Choe, Yon-Ho,Han, Hye-Seung,Song, Sun-Uk 대한소아소화기영양학회 2002 Pediatric gastroenterology, hepatology & nutrition Vol.5 No.1

배경: Lactoferrin은 위장 점막에서 H. pylori의 철분의 공급원으로 알려져 있다. 본 연구는 H. pylori에 감염된 위장 점막의 조직검체에서 lactoferrin 값을 측정하고 철 결핍성 빈혈의 유무에 따라 lactoferrin 발현의 주요 위치를 정하는 방법을 이용하여 lactoferrin과 철 결핍성 빈혈을 동반한 H. pylori감염 사이의 연관성을 밝히기 위해 시행하였다. 방법: 상부 위장관 내시경을 시행한 55명의 환아를 세 군으로 분류하였다: 정상군(NL, n=19), H. pylori 감염군(HP, n=15), H. pylori 감염이 동반된 철 결핍성 빈혈군(IDA, n=21). 조직 병리학 소견은 Updated Sydney System을 이용하여 정도를 나누었다. 위장 점막의 lactoferrin 값은 면역 측정법으로 측정하였다. lactoferrin의 발현 위치와 양을 측정하는 것은 면역조직화학적 염색방법을 이용하였다. 결과: 위 전정부에서의 lactoferrin의 값은 조직 검체의 H. pylori 밀도나 다형핵세포의 침습정도, 만성 염증에 비례하여 의미있게 증가되었다. HP와 IDA군에서는 정상군에 비하여 lactoferrin의 수치가 통계학적으로 유의하게 증가되었다(p=0.0001). IDA군은 HP군에 비하여 lactoferrin의 수치가 높은 경향이 있었다(p=0.2614). 면역조직화학적 방법에 의한 lactoferrin 발현의 주요부위는 상피세포내의 분비샘과 호중구에 있었다. Lactoferrin은 정상군에서는 약하게 염색되었고 HP와 IDA군에서는 강하게 염색되었다. 결론: IDA군에서 위장 점막내 lactoferrin 격리가 현저하였으며 이 소견은 H. pylori 감염이 철 결핍성 빈혈에 영향을 미치는 기전을 밝히는데 중요한 단서로 작용할 것이다. Purpose: It is known that lactoferrin serves as a source of iron for H. pylori in gastric mucosa. This study was undertaken to investigate the relationship between lactoferrin and H. pylori infection coexistent with iron-deficiency anemia by determining the lactoferrin levels in gastric biopsy specimens, and by locating the major sites of lactoferrin expression, according to the presence or absence of iron-deficiency anemia. Methods: Fifty-five adolescents that underwent gastroduodenoscopy were divided into three groups: NL (n=19) for normal controls, HP (n=15) for patients with H. pylori, and IDA (n=21) for patients with H. pylori gastritis and coexisting iron-deficiency anemia. Histopathologic features were graded from to marked on the basis of the Updated Sydney System. The gastric mucosal levels of lactoferrin were measured by immunoassay. Immunohistochemical technique was used to allow identification of the location and quantification of the lactoferrin expression. Results: Lactoferrin levels in the antrum increased significantly, in proportion to, H. pylori density, polymorphonuclear cell infiltration, and chronic inflammation in the histologic specimens. Patients in the HP and IDA groups showed significantly increased mucosal levels of lactoferrin compared with that observed in the normal group (p=0.0001). The lactoferrin level in IDA group tended to be higher than that in the HP group (p=0.2614). The major sites of lactoferrin expression by immunohistochemistry were in glands and neutrophils within epithelium. Lactoferrin was stained weakly in NL, and strongly in HP and IDA. Conclusion: The lactoferrin sequestration in the gastric mucosa of IDA was remarkable, and this finding seems to give a clue that leads to the clarification of the mechanism by which H. pylori infection contributes to iron-deficiency anemia.

무균성 뇌수막염 환자의 뇌척수액과 혈청에서 Nitric Oxide, Macrophage Inflammatory Protein(MIP)-1α, Lactoferrin 값의 비교

서영,심정연,심재원,김덕수,정혜림,박문수,Seo, Young,Sim, Jung Yeon,Shim, Jae Won,Kim, Deok Su,Jung, Hye Lim,Park, Moon Soo 대한소아청소년과학회 2005 Clinical and Experimental Pediatrics (CEP) Vol.48 No.1

목 적 : 중추신경계감염의 병리학적인 정보는 아직 미약하고 정확하게 규명된 바가 없지만 최근의 여러 연구들에 의하면 병원균의 직접적인 침습보다는 숙주의 염증반응이 뇌 손상의 중요한 인자가 됨을 밝힘으로써 여러 사이토카인이나 독소, 인터루킨 등이 뇌수막 염증반응을 유도한다고 밝혀져 있다. 최근 세균성 뇌수막염에서 NO, MIP-$1{\alpha}$, lactoferrin의 역할에 대한 보고들이 있었으나 아직 무균성 뇌수막염 환자를 대상으로 한 보고는 별로 없었고 특히 국내에서 보고된 바는 없었다. 이에 저자들은 무균성 뇌수막염 환아의 혈액과 뇌척수액에서 NO, MIP-$1{\alpha}$, lactoferrin의 농도를 측정하여 대조군과 비교하고, 다른 뇌수막염 관련인자들 사이의 상관관계를 비교 분석하고자 하였다. 방 법 : 2002년 6월부터 7월까지 강북삼성병원 소아과 입원환자 중 발열과 뇌막자극증상을 보인 40명의 환아들을 대상으로 뇌척수액 검사를 시행하여, 무균성 뇌수막염 소견을 보인 25명을 뇌수막염군, 정상소견을 보인 15명을 대조군으로 하였다. 입원당일 혈액과 뇌척수액을 채취하여 혈액에서 백혈구수와 CRP를 측정하고, 뇌척수액에서 백혈구수와 당, 단백농도, 뇌압을 측정하였다. 나머지 혈액 및 뇌척수액 검체는 실온에서 10분간 2,000 rpm으로 원심 분리하여 영하 70도에서 보관 후 ELISA 방법을 이용하여 각각의 검체에서 일시에 NO, MIP-$1{\alpha}$, lactoferrin의 농도를 측정하였다. 무균성 뇌수막염군에서 신경학적 후유증을 보인 경우는 없었다. 결 과 : 1) 혈액과 뇌척수액의 NO의 농도는 대조군과 무균성 뇌수막염 사이에 차이를 보이지 않았다. 2) 혈액과 뇌척수액의 MIP-$1{\alpha}$의 농도는 대조군과 무균성 뇌수막염 사이에 차이를 보이지 않았다. 3) 무균성 뇌수막염군의 뇌척수액 lactoferrin 농도는 대조군에 비해 의미 있게 증가되었으며 혈액 lactoferrin 농도는 대조군에 비해 의미 있게 감소되었다. 4) 뇌척수액 lactoferrin 농도는 뇌척수액 백혈구수($r_s=0.449$, P=0.007), 특히 중성구 수와 양의 상관관계를 보였으며($r_s=0.574$, P<0.001), 뇌척수액 단백 농도와 양의 상관관계를 보였다($r_s=0.508$, P=0.002). 결 론 : 무균성 뇌수막염 환아의 뇌척수액에서 lactoferrin은 의미 있게 증가하여 무균성 뇌수막염의 면역 반응에 중요한 역할을 할 것으로 생각되며, 이는 뇌-혈관 장막 붕괴로 인해 혈액에서 뇌척수액으로 유입된 중성구에서 분비되었을 가능성과, 뇌-혈관 장막 붕괴로 인해 혈액에서 뇌척수액으로 직접 유입되었을 가능성이 있는 것으로 생각된다. Purpose : The pathologic mechanisms of central nervous system(CNS) injuries in human meningitis are not yet completely understood. Recent studies indicate that the host inflammatory responses are as important in brain damage as the infecting organisms and toxins. There have been some reports on the relationship of nitric oxide(NO), macrophage inflammatory protein-$1{\alpha}$(MIP-$1{\alpha}$), and lactoferrin in bacterial meningitis, but few reports in aseptic meningitis. Thus, we investigated the concentrations of NO, MIP-$1{\alpha}$ and lactoferrin in cerebrospinal fluid(CSF) and serum of patients with aseptic meningitis and control subjects and evaluated their relationship with other parameters of meningitis. Methods : CSF and blood were obtained from 25 subjects with aseptic meningitis and 15 control subjects. After centrifugation, supernatants were stored at $-70^{\circ}C$ and we assayed the concentrations of NO, MIP-$1{\alpha}$ and lactoferrin with the ELISA method. There were no patients with neurologic sequelae after being recovered from aseptic meningitis. Results : Concentrations of CSF and serum NO, MIP-$1{\alpha}$ were not increased in aseptic meningitis subjects compared to control subjects. Concentration of CSF lactoferrin was significantly elevated in patients with aseptic meningitis and concentration of serum lactoferrin was significantly decreased in patients with aseptic meningitis compared with those in control subjects(P<0.05). CSF lactoferrin level was positively correlated with CSF WBC counts($r_s=0.449$, P=0.007), especially with neutrophil counts($r_s=0.574$, P<0.001) and CSF protein level($r_s=0.508$, P=0.002). Conclusion : Lactoferrin plays an important role in aseptic meningitis and may be released from neutrophils recruited from blood to the CSF through breakdown of blood-brain barrier. NO and MIP-$1{\alpha}$ may not be important factors in the pathogenesis of aseptic meningitis without neurologic sequelae.

Lactococcus lactis subsp. cremoris FC에 대한 Lactoferrin과 Transferrin의 생장촉진효과

김완섭,Kim, Woan-Sub 한국유가공학회 2017 Journal of Dairy Science and Biotechnology (JMSB) Vol.35 No.3

Lactococcus lactis subsp. cremoris FC는 카스피해 요구르트의 명칭으로 강한 점성을 가지는 것을 특징으로 하는 코카커스 지방 유래의 발효유로부터 분리된 균주로 있다. 본 연구는 Lc. lactis subsp. cremoris FC의 생장에 대한 lactoferrin group(native-lactoferrin, apo-lactoferrin, 그리고 holo-lactoferrin)과 transferrin (apo-transferrin과 holo-transferrin)의 첨가가 생장에 미치는 영향을 알아보기 위하여 수행하였다. Lc. lactis subsp. cremoris FC의 생장에 lactoferrin과 transferrin의 첨가효과는 0.5 또는 1 mg/mL의 농도로 첨가하는 것이 생장 촉진에 좋은 효과를 나타내는 것으로 조사되었다. Recent studies have reported that certain lactic acid bacteria and their metabolites, such as exopolysaccharides (EPSs), have immunological effects and can modulate the immune system following oral administration. Lactococcus lactis subsp. cremoris FC is a lactic acid bacteria isolated from fermented milk from Caucasians and has been shown to produce EPSs. In this study, the effects of lactoferrins (apo-lactoferrin, holo-lactoferrin, and native-lactoferrin) and transferrins (apo-transferrin and holo-lactoferrin) on the growth of L. lactis subsp. cremoris FC were examined. The addition of lactoferrins and transferrins to L. lactis subsp. cremoris FC cultures was found to be effective at concentrations of 0.5 or 1 mg/mL.

Characterization of Protein Disulfide Isomerase during Lactoferrin Polypeptide Structural Maturation in the Endoplasmic Reticulum

Lee, Dong-Hee,Kang, Seung-Ha,Choi, Yun-Jaie Korean Society for Biochemistry and Molecular Biol 2001 Journal of biochemistry and molecular biology Vol.34 No.2

A time-dependent folding process was used to determine whether or not protein disulfide isomerase (PDI) plays an important role in the maturation of nascent lactoferrin polypeptides. Interaction between lactoferrin and PDI was analyzed according to the co-immunoprecipitation of the two proteins. The results indicate that lactoferrin folding requires a significant interaction with PDI and its binding is relatively brief compared to other nascent polypeptides. The amount of lactoferrin interacting with PDI increases up to half a minute and sharply decreases beyond this time point. During the refolding process that follows reduction by DTT, lactoferrin polypeptides heavily interact with PDI and the interaction period was extended compared to the normal folding process. In terms of the temperature effect on PDI-lactoferrin interaction, PDI binds to lactoferrin polypeptides longer at a lower temperature (here, $25^{\circ}C$) than $37^{\circ}C$. The lactoferrin-PDI interaction was also studied in vitro. According to the in vitro experiment data, PDI was still functional in cell lysates assisting lactoferrin folding into the mature form. PDI interacts with lactoferrin polypeptides for an extended period during the folding in vitro. During the refolding process in vitro, intermolecular aggregates and refolding oligomers matured into a functional form after PDI binds to the lactoferrin. These results suggest that PDI provides a prolonged chaperoning activity in the refolding processes and that there appears to be a greater requirement for PDI chaperone activity in the refolding of lactoferrin polypeptides.

PMA로 자극되어진 세포에서 염증 Cytokine 발현에 미치는 Bovine Lactoferrin의 생물활성 영향

정승희,강호범,김재화,윤성식,남명수,Chung, Sung-Hee,Kang, Ho-Bum,Kim, Jae-Wha,Yoon, Sung-Sik,Nam, Myoung-Soo 한국축산식품학회 2012 한국축산식품학회지 Vol.32 No.3

유청단백질의 일종인 lactoferrin은 이미 많은 보고에 의해서 여러 가지 생리활성기능이 있는 것으로 밝혀지고 있는데, U937, NK92, 수지상세포의 분화된 상태인 mutz-3와 muDC를 이용하여 과민반응과 천식, 면역 관련 유전자의 발현에 미치는 영향을 조사 한 결과 의미 있는 결과를 도출하였다. Lactoferrin 단독 또는 면역증강물질인 PMA와 혼합하여 처리 한 경우 상승효과 작용으로 과민반응과 천식, 면역 관련 유전자의 발현을 강하게 유도하였다. 이는 유청단백질의 주요 성분 중 하나인 lactoferrin이 면역기전에 중요한 역할을 하고 있다는 결과로 사료된다. Bovine lactoferrin is well known as biological activator in defense mechanism related some cells. In this study, we was investigated about the immune modulator as a role of lactoferrin through the transcriptional regulation of genes associated with hypersensitivity such as allergy, athma and inflammatory disease. Effects of inflammatory reaction of bovine lactoferrin was carried out by RT-PCR analysis from isolated total RNA treated with lactoferrin 0, 10, 50, 100, 500 ${\mu}g/mL$ and PMA 100 ng/mL. The expression of the TYROBP, PITPNA, IL-10, SLP1, DC-stamp and ICAM-1 mRNA were increased by synergy effect of bovine lactoferrin and PMA. The results of RT-PCR showed that bovine lactoferrin and PMA had an effect of immune modulator by enhancement of TYROBP, PITPNA, SLP1, DC-stamp, IL-10 and ICAM-1 gene transcription in U937, Mutz-3 and NK92 cells, respectively. Bovine lactoferrin showed a potential of biological function which could be used for industrial applications as a material of food and pharmaceutical.

Effects of Heat-treated Bovine Lactoferrin on the Growth of Lactococcus lactis subsp. cremoris JCM 20076

Kim, Woan-Sub Korean Society of Dairy Science and Biotechnology 2019 Journal of Dairy Science and Biotechnology (JMSB) Vol.37 No.2

This study investigated the effects of heat-treated and non-heat-treated bovine lactoferrin on the growth of Lactococcus lactis subsp. cremoris JCM 20076. The addition of heat-treated and non-heat-treated bovine lactoferrin in adjusted MRS medium stimulated the growth of Lc. cremoris JCM 20076. Heat-treated bovine lactoferrin had a greater impact on the growth of Lc. cremoris JCM 20076 compared to that with non-heat-treated bovine lactoferrin. Bovine lactoferrin heated at $65^{\circ}C$ for 30 min stimulated the growth of the bacteria more than that heated at $80^{\circ}C$ for 5 min. Furthermore, the growth of Lc. cremoris JCM 20076 increased substantially with heat-treated bovine lactoferrin at a concentration of 1 mg/mL.

Effects of Heat-treated Bovine Lactoferrin on the Growth of Lactococcus lactis subsp. cremoris JCM 20076

( Woan-sub Kim ) 한국유가공학회(구 한국유가공기술과학회) 2019 Journal of Dairy Science and Biotechnology (JMSB) Vol.37 No.2

This study investigated the effects of heat-treated and non-heat-treated bovine lactoferrin on the growth of Lactococcus lactis subsp. cremoris JCM 20076. The addition of heattreated and non-heat-treated bovine lactoferrin in adjusted MRS medium stimulated the growth of Lc. cremoris JCM 20076. Heat-treated bovine lactoferrin had a greater impact on the growth of Lc. cremoris JCM 20076 compared to that with non-heat-treated bovine lactoferrin. Bovine lactoferrin heated at 65°C for 30 min stimulated the growth of the bacteria more than that heated at 80°C for 5 min. Furthermore, the growth of Lc. cremoris JCM 20076 increased substantially with heat-treated bovine lactoferrin at a concentration of 1 mg/mL.

연구단보(Research Note) : Phospholipid Liposomes과 Bovine Lactoferrin의 상호작용

김미영(Mi-young Kim),강신원(Shin-won Kang),박장수(Jang-su Park),남명수(Myoung Soo Nam) 충남대학교 농업과학연구소 2010 농업과학연구 Vol.37 No.3

Apo, holo, native-type of lactoferrin showed a similar pattern of circular dichroism(CD) spectra. Lactoferrin appeared to interact more easily with negatively charged liposome than neutral liposome. Holo-type of lactoferrin showed the highest degree of leakage, whereas apo-type of lactoferrin showed the lowest level of leakage. Holo-type lactoferrin could more easily interact with phospholipid liposomes than apo-type lactoferrin, when used as an artificial membrane.

Effects of Heat-treated Bovine Lactoferrin on the Growth of Lactococcus lactis subsp. cremoris JCM 20076

김완섭 한국낙농식품응용생물학회 2019 한국유가공기술과학회지 Vol.37 No.2

This study investigated the effects of heat-treated and non-heat-treated bovine lactoferrin on the growth of Lactococcus lactis subsp. cremoris JCM 20076. The addition of heattreated and non-heat-treated bovine lactoferrin in adjusted MRS medium stimulated the growth of Lc. cremoris JCM 20076. Heat-treated bovine lactoferrin had a greater impact on the growth of Lc. cremoris JCM 20076 compared to that with non-heat-treated bovine lactoferrin. Bovine lactoferrin heated at 65°C for 30 min stimulated the growth of the bacteria more than that heated at 80°C for 5 min. Furthermore, the growth of Lc. cremoris JCM 20076 increased substantially with heat-treated bovine lactoferrin at a concentration of 1 mg/mL.

한우 초유로부터 Lactoferrin의 분리.정제

양희진,하월규,양동훈,박기문,이수원 한국축산식품학회 2000 한국축산식품학회지 Vol.20 No.2

Lactoferrin was isolated from the colostrum of Korean native cow by using several purification steps such as batch extraction, ion exchange chromatography, gel filtration chromatography and affinity chromatography. Other whey protein components that having similar molecular weight and affinity to lactoferrin were gradually removed from crude Korean native cow's lactoferrin during the purification steps. The amount of lactoferrin collected from a liter of Korean native cow's colostrum was 65mg and the recovery rate was 29.4%. The molecular weight of the purified Korean native cow's lactoferrin was estimated approximately 81,000dalton.