Myo-inositol이 돼지 난모세포의 체외성숙에 미치는 영향

조인식,한효원,이상미,박효영,정영희,문승주,강승률,강만종 충남대학교 형질전환복제돼지연구센터 2004 논문집 Vol. No.8

본 연구는 미성숙 돼지 난모세포의 체외 성숙에 있어서 myo-inositol의 영향을 알아보기 위하여 실시하였다. 미성숙 돼지 난모세포을 myo-inositol을 포함하는 또는 포함하지 않은 체외 성숙배지에서 44시간 체외 성숙을 유도하였을 때 성숙율은 myo-inositol을 첨가한 성숙배지에서 성숙시킨 실험구에 유의하게 높았다(P<0.05). Myo-inositol에 의한 성숙율 향상에 있어서 난구세포의 영향을 알아보기 위하여 난추세포의 치밀도에 따라 분류하여 미성숙 난모세포을 myo-inositol을 포함하는 체외 성숙배지에서 배양하였을 때 난구세포가 치밀한 미성숙 난모세포에서가 더 높은 성숙율을 나타내었다(P<0.05). 그러나 난구세포가 치밀하지 않은 미성숙 난모세포도 myo-inositol을 포함하는 성숙배지에서 배양하면 성숙율은 대조구에 비하여 유의하게 높았다(P<0.05). 이러한 결과는 돼지 난모세포의 체외 성숙배지에 myo-inositol의 첨가는 체외 성숙율을 향상시킬 수 있음을 나타내고 있다. This study was carried out to assess whether the addition of myo-inositol to maturation medium could improve porcine oocyte maturation in vitro. Oocytes were cultured for the first 22 h in Witten's medium containing lO1U/ml PMSG, 10 IU/ml HCG supplemented with or without myo-inositol. Subsequently, they were cultured for additional 22 h in Witten's medium without hormone supplemented with or without myo-inositol. When the porcine oocytes were cultured in maturation medium containing myo-inositol, the proportion of metaphase II oocytes 44h after culture was higher in the myo-inositol group(P<O.05). To study effects of cumulus cell on the maturation induced by myo-inositol, we examined the maturation status of cumulus-enclosed or cumulus-denuded porcine follicular oocytes. The rates of maturation were significantly higher in the cumulus-enclosed oocytes(P<O.05). However, the maturation rates of cumulus-denuded oocytes cultured in medium containing myo-inositol were higher than those of control group(P<O.05). Our results suggest that myo-inositol may affect meiotic progression of porcine follicular oocytes and supplementation of myo-inositol in maturation medium may be useful for the in vitro maturation of porcine follicular oocytes.

Myo-inositol이 돼지 난모세포의 체외성숙에 미치는 영향

조인식,한효원,이상미,박효영,정영희,문승주,강승률,강만종 한국동물생명공학회(구 한국동물번식학회) 2004 Reproductive & Developmental Biology(Supplement) Vol.28 No.2s

본 연구는 미성숙돼지 난모세포의 체외 성숙에 있어서 myo-inositol의 영향을 알아보기 위하여 실시하였다. 미성숙 돼지 난모세포을 myo-inositol을 포함하는 또는 포함하지 않은 체외 성숙배지에서 44시간 체외 성숙을 유도하였을 때 성숙율은 myo-inositol을 첨가한 성숙배지에서 성숙시킨 실험구에 유의하게 높았다(P<0.05). Myo-inositol 에 의한 성숙율 향상에 있어서 난구세포의 영향을 알아보기 위하여 난구세포의 치밀도에 따라 분류하여 미성숙 난모세포을 myo-inositol을 포함하는 체외 성숙배지에서 배양하였을 때 난구세포가 치밀한 미성숙 난모세포에서가 더 높은 성숙율을 나타내었다(P<0.05). 그러나 난구세포가 치밀하지 않은 미성숙 난모세포도 myo-inositol을 포함하는 성숙배지에서 배양하면 성숙율은 대조구에 비하여 유의하게 높았다(P<0.05). 이러한 결과는 돼지 난모세포의 체외 성숙배지에 myo-inositol의 첨가는 체외 성숙율을 향상시킬 수 있음을 나타내고 있다. This study was carried out to assess whether the addition of myo-inositol to maturation medium could improve porcine oocyte maturation in vitro. Oocytes were cultured for the first 22 h in Witten''''s medium containing 10IU/㎖ PMSG, 10 IU/㎖ HCG supplemented with or without myo-inositol. Subsequently, they were cultured for additional 22 h in Witten''''s medium without hormone supplemented with or without myo-inositol. When the porcine oocytes were cultured in maturation medium containing myo-inositol, the proportion of metaphase Ⅱ oocytes 44h after culture was higher in the myo-inositol group(P<0.05). To study effects of cumulus cell on the maturation induced by myo-inositol, we examined the maturation status of cumulus-enclosed or cumulus-denuded porcine follicular oocytes. The rates of maturation were significantly higher in the cumulus-enclosed oocytes(P<0.05). However, the maturation rates of cumulus-denuded oocytes cultured in medium containing myo-inositol were higher than those of control group(P<0.05). Our results suggest that myo-inositol may affect meiotic progression of porcine follicular oocytes and supplementation of myo-inositol in maturation medium may be useful for the in vitro maturation of porcine follicular oocytes.

Saccharomyces cerevisiae에서 myo-Inositor 결핍에 의한 Respiratory Capacity의 감소

정경환,이준식 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.4

Saccharomyces cerevisiad (S. cerevisiae)의 growth factor중의 하나인 myo-Inositol은 membrane-associated 효소인 phosphatidylinositol 합성효소에 의해 세포막의 구성 성분인 phosphatidylinositol로 incorporation 되는 것으로 알려져있다. 그래서 myo-inositol이 결핍되면 막구조와 기능에 좋지 않은 영향을 주는 것으로 알려져 있다. 이러한 myo-inositol의 생화학적 기능을 근거로 myo-inositol의 결핍이 호기적 포도당 대사에 미치는 영향을 조사하기 위하여 S. cerevisiae의 respiratory capacity를 대변하는 인자인 specific oxygen uptake rate (Q_O2)를 회분배양과 연속배양을 이용하여 측정하였다. 그리고, S. cerevisiae의 호기적 포도당 대상의 respiratory capacity를 myo-inositol이 결핍된 경우와 그렇지 않은 경우에 연속배양에서 포도당 pulse-addition을 한 후 관찰하였다. 그 결과 회분배양과 연속배양에서 myo-inositol이 결핍된 경우 maximum specific oxygen uptake rate (Q_O2. max)이 감소되는 현상을 관찰하였고, 포도당 pulse-addition 실험에서 얻은 결과를 정량적으로 분석하여 본 결과 myo-inositol의 결핍이 respiratory capacity의 감소를 초래하는 것이 관찰되었다. myo-Inositol, a growth factor for Saccharomyces cerevisiad (S. cerevisiae), has been know to be incorporated into phosphatidylinositol (PI), which is a kind of phospholipid in the cell membrane, by a membrane-associated PI-synthesizing enzyme. The deficiency of myo-inositol in S. cerevisiae adversely affected the membrane structure and function. On the basis of biochemical functions of myo-inositol, the effect of deficiency of myo-inositol on the aerobic glucose metabolism was investigated by measuring specific oxygen uptake rate(Q_O2) used as an indicator representing the respiratory capacity of S. cerevisiae in batch and continuous cultures. The respiratory capacity of aerobic glucose metabolism in S cerevisiae was also monitored after glucose pulse-addition in a continuous culture (D=0.2, 1/hr), in which glucose was utilized through respiratory metabolism. The deficiency of myo-inositol was found to lead to both the decrease of the maximum specific oxygen uptake rate (Q_O2. max) observed from the batch as well as in the continuous culture experiment and the decrease of the respiratory capacity of aerobic glucose metabolism of S. cerevisiae determined from the glucose pulse-addition experiment, in which the glucose flux into respiratory and fermentative metabolism was quantitatively analyzed.

메밀에서 비생물학적 스트레스에 의한 D-chiro-inositol의 함량 증가

박훤범 수원대학교 기능성생명소재연구소 2007 자연과학연구논문집 Vol.6 No.1

D-chiro-inositol which is the isomer of myo-inositol is well known drug for the type Ⅱ diabetes. The methylated form of D-chiro-inositol, pinitol and D-chiro-inositol are synthesized when the plants are exposed to the abiotic stresses such as drought, salinity and low temperature as osmoprotectants. In soybean, myo-inositol is converted to ononitol by O-methyltransferase and ononitol is converted to pinitol by ononitol epimerase and finally converted to D-chiro-inositol by demethylase. However there have been some reports that in buckwheat, myo-inositol tan be converted to D-chiro-inositol directly Thus we checked the cyclitol contents after abiotic stresses in buckwheat, and confirmed that myo-inositol is directly converted to D-chiro-inositol by myo-inositol epimerase.

납이 HL 60 세포 내 Myo-inositol 수송 체계에 미치는 영향

정명규(Myung Kiu Chung) 한국환경보건학회 1999 한국환경보건학회지 Vol.25 No.2

Lead is the most ubiquitous toxic metal and released to the environment from natural and man-made sources. Presence of lead in the environment has led to an increasing awareness and concern of deterimental effect on ecosystem and human health. But there are several limits for toxicity assessment and protection human from toxicity of lead because the toxic effects of lead are detectable in practice only at high exposure concentraion. Therfore, it is very important to search sensitive biomakers at low exposure concentration which could not be observed any practical symptoms such as anemia, encephalopathy, and neuropathy in human. In the present study, to investigate myo-inositol concentration in cell could be a biomaker for lead cytotoxicity, the toxic effects of lead on myo-inositol uptake system were studied in the HL 60 cell line from three groups : normal control, lOppm-lead treated group, 80 ppm-lead treated group. Lead significantly reduced specific myo-inositol uptake in lead-treated groups, but did not affected nonspecific uptake. The magnitude of reduction in myo-inositol uptake was directly propotional to exposure dose. But myo-inositol uptakes in lead treated groups were significantly normalized by addition sodium ion to culture medium. These results suggest that myo-inositol concentration in the cell could be a useful biomaker in toxicity test with cell line and the toxic mechanism of lead on HL 60 cell might be a reduction in myo-inositol uptake, which induced by the perturbation of the sodiumion metabolism.

Urine myo-inositol as a novel prognostic biomarker for diabetic kidney disease: a targeted metabolomics study using nuclear magnetic resonance

( Soie Kwon ),( Jin Seong Hyeon ),( Youngae Jung ),( Lilin Li ),( Jung Nam An ),( Yong Chul Kim ),( Seung Hee Yang ),( Tammy Kim ),( Dong Ki Kim ),( Chun Soo Lim ),( Geum-sook Hwang ),( Jung Pyo Lee ) 대한신장학회 2023 Kidney Research and Clinical Practice Vol.42 No.4

Background: As a leading cause of chronic kidney disease, clinical demand for noninvasive biomarkers of diabetic kidney disease (DKD) beyond proteinuria is increasing. Metabolomics is a popular method to identify mechanisms and biomarkers. We investigated urinary targeted metabolomics in DKD patients. Methods: We conducted a targeted metabolomics study of 26 urinary metabolites in consecutive patients with DKD stage 1 to 5 (n = 208) and healthy controls (n = 26). The relationships between estimated glomerular filtration rate (eGFR) or urine protein-creatinine ratio (UPCR) and metabolites were evaluated. Multivariate Cox analysis was used to estimate relationships between urinary metabolites and the target outcome, end-stage renal disease (ESRD). C statistics and time-dependent receiver operating characteristics (ROC) were used to assess diagnostic validity. Results: During a median 4.5 years of follow-up, 103 patients (44.0%) progressed to ESRD and 65 (27.8%) died. The median fold changes of nine metabolites belonged to monosaccharide and tricarboxylic acid (TCA) cycle metabolites tended to increase with DKD stage. Myo-inositol, choline, and citrates were correlated with eGFR and choline, while mannose and myo-inositol were correlated with UPCR. Elevated urinary monosaccharide and TCA cycle metabolites showed associations with increased morality and ESRD progression. The predictive power of ESRD progression was high, in the order of choline, myo-inositol, and citrate. Although urinary metabolites alone were less predictive than serum creatinine or UPCR, myo-inositol had additive effect with serum creatinine and UPCR. In time-dependent ROC, myo-inositol was more predictive than UPCR of 1-year ESRD progression prediction. Conclusion: Myo-inositol can be used as an additive biomarker of ESRD progression in DKD.

Decreased Insulin Resistance by Myo-Inositol Is Associated with Suppressed Interleukin 6/Phospho-STAT3 Signaling in a Rat Polycystic Ovary Syndrome Model

Yulong Zhang,Changzhong Li,Wenhui Zhang,Xiangqin Zheng,Xiujuan Chen 한국식품영양과학회 2020 Journal of medicinal food Vol.23 No.4

Myo-inositol supplementation may reduce insulin resistance (IR) with few serious side effects in patients with polycystic ovary syndrome (PCOS). To explore the mechanism of this action in an animal model, a PCOS-IR rat model was generated. Enzyme-linked immunosorbent assay was used to assess changes in ovulation function during treatment with a myo-inositol supplement, and Western blotting, real-time polymerase chain reaction, and immunohistochemistry were performed to investigate the underlying molecular mechanisms. The results showed that the myo-inositol supplement decreased the homeostatic model assessment of insulin resistance (HOMA-IR) index and significantly decreased the serum levels of luteinizing hormone (LH), LH/follicle-stimulating hormone ratio, and testosterone, while increasing the serum level of estradiol. Upregulation of interleukin 6 (IL-6), phospho-STAT3 (p-STAT3), Mir-21, and Mir-155 and significant downregulation of PPAR-γ and GLUT4 were detected in the untreated PCOS-IR rat model. However, downregulation of IL-6, p-STAT3, miR-21, and miR-155 and significant upregulation of PPAR-γ and GLUT4 were detected with myo-inositol supplementation. Thus, myo-inositol supplementation may reduce Mir-21 and Mir-155 levels by downregulating IL-6 and p-STAT3 and, subsequently, reverse the expression of PPAR-γ and GLUT4, leading to a decreased HOMA-IR index. In conclusion, the identification of an IL-6/p-STAT3/Mir-155/Mir-21/PPAR-γ/GLUT4 system in the PCOS-IR rat model provides insight into the pathogenesis of PCOS and may indicate a possible therapeutic strategy. Amelioration of the basal serum glucose levels and of the HOMA/HOMA-IR index may be achieved by the reversal of the expression of PPAR-γ and GLUT4 through the downregulation of IL-6, p-STAT3, miR-21, and miR-155 with myo-inositol supplementation.

Enterobacter sp. YB-46의 myo-Inositol dehydrogenase 유전자 클로닝과 특성분석

박찬영,김광규,윤기홍 한국미생물·생명공학회 2018 한국미생물·생명공학회지 Vol.46 No.2

A bacterial strain capable of metabolizing myo-inositol (MI) and converting to other substances was isolated from soil of orchard. The isolate, named YB-46, was grown on minimal medium supplemented with MI as the sole carbon source and was presumed to belonging to genus Enterobacter according to the 16S rDNA sequence. Escherichia coli transformant converting MI into unknown metabolites was selected from a metagenomic library prepared with fosmid pCC1FOS vector. Plasmid was isolated from the transformant, and the inserted gene was partially sequenced. From the nucleotide sequence, an iolG gene was identified to encode myo-inositol dehydrogenase (IolG) consisting of 336 amino residues. The IolG showed amino acid sequence similarity of about 50% with IolG of Enterobacter aerogenes and Bacillus subtilis. The His-tagged IolG (HtIolG) fused with hexahistidine at C-terminus was produced and purified from cell extract of recombinant E. coli. The purified HtIolG showed maximal activity at 45℃ and pH 10.5 with the highest activity for MI and D-glucose, and more than 90% of maximal activity for D-chiro-inositol, D-mannitol and D-xylose. Km and Vmax values of the HtIolG for MI were 1.83 mM and 0.724 μmol/min/mg under the optimal reaction condition, respectively. The activity of HtIolG was increased 1.7 folds by Zn2+, but was significantly inhibited by Co2+ and SDS. myo-Inositol (MI)을 대사하여 다른 물질로 전환하는 미생물을 과수원 토양으로부터 분리하였다. 분리균 YB-46은 유일한 탄소원으로 MI이 첨가된 배지에서 성장하였고 16S rDNA 염기서열에 따라 Enterobacter 속의 균주로 추정되었다. Fosmid pCC1FOS 벡터를 사용하여 제조된 거대 유전체은행으로부터 MI을 미지의 대사 물질로 전환하는 Escherichia coli 형질전환주를 선발하였다. 이로부터 플라스미드를 분리하고 삽입된 유전자의 일부 염기서열을 결정한 결과 336 아미노 잔기로 구성된 myo-inositol dehytrogenase (IolG)를암호화하는 iolG 유전자가 발견되었다. 분리균 YB-46의 IolG 는 E. aerogenes와 Bacillus subtilis의 IolG와 약 50% 수준의 상동성을 보였다. 카르복실 말단에 hexahistidine이 연결되도록 제조한 His-tagged IoG (HtIolG)의 유전자를 재조합대장균에서 발현하여 균체 파쇄액으로부터 HtIolG를 정제하였다. 정제된 HtIolG는 45℃와 pH 10.5에서 최대 활성을보였고 MI과 D-glucose에 대한 활성이 가장 높았으며 Dchiro- inositol, D-mannitol 및 D-xylose에도 90% 이상의 활성을 보였다. 최적 반응조건에서 MI을 기질로 하여 반응 동력학적 계수를 측정한 결과 Km과 Vmax가 1.83 mM과 0.724 μmol/min/mg로 확인되었다. HtIolG의 활성은 Zn2+에 의해1.7배 증가하였으며, Co2+와 SDS에 의해서는 크게 감소하였다.

대두에서 ononitol epimerase를 coding 하는 유전자의 분리와 E. coli에서 발현 및 기질 특이성 확인

박훤범 수원대학교 기능성생명소재연구소 2004 자연과학연구논문집 Vol.3 No.2

Myo-inositol plays a key role in plant development and growth. Conversion of glucose-6-phosphate to myo-inositol-1-phosphate is the first committed step in myo-inositol biosynthesis. Methylation and isomerization of myo-inositol forms 0-methyl inositols (ononitol, pinitol, etc.) which participate in abiotic stress related reponses, storage of seed products, and production of inositol-glycosides. Upon salinity stress, an 0-methyl transferase methylates C4 of myo-inositol then produces ononitol. The next step is epimerization of Cl of ononitol by ononitol epimerase then produces pinitol. However, there has been no report about the ononitol epimerase in plant yet. We have searched salinity inducible ESTs which were homologous with the NAD-dependent epimerase from the soybean EST data base. We have chosen one putative ononitol epimerase EST and sequenced the full-length cDNA. The putative ononitol epimerase cDNA was cloned into E. coli expression vector pET15b and we have checked the substrate of this putative ononitol epimerase, and found out that the substrate was ononitol by GC-FID. The transcripts level of the ononitol epimerase gene under the salinity condition was also checked.

Efficient Expression of Myo-inositol Oxygenase in Escherichia coli and Application for Conversion of Myo-inositol to Glucuronic Acid

Shuxiang Zheng,Peilian Wei,Lei Huang,Jin Cai,Zhi-nan Xu 한국식품과학회 2014 Food Science and Biotechnology Vol.23 No.2

Glucuronic acid is an important biochemicalwith wide applications in the food and medical industries. The myo-inositol oxygenase (MIOX) gene was synthesizedand expressed in Escherichia coli BL 21(DE3). Afteroptimization of induction conditions and the culturetemperature, the highest MIOX activity (45.46 kU/L) wasachieved when 0.1 mM isopropyl-thio-β-D-galactoside(IPTG) was added to cell cultures with an OD600 value 1.0followed by induction at 26oC for 8 hours. The purifiedMIOX enzyme exerted characteristics similar to the nativeform. Conversion of myo-inositol to glucuronic acid wasperformed using whole cells in a pH 8.5 buffer system. Whole cells harboring myo-inositol oxygenase were usedas a biocatalyst to produce 2.13 g/L of glucuronic acid witha conversion rate of 99%. A promising novel process forglucuronic acid production from abundant agriculturalbyproducts is presented.