• Title/Summary/Keyword: protein metabolism

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Biochemical Characteristics for the Cofactor Free Mutant of Yeast Homocysteine Catalyzing Enzyme, Cystathionine ${\beta}$-Synthase (조효소를 함유하지 않는 효모의 Homocysteine 분해효소, Cystathionine ${\beta}$-Synthase의 생화학적 특성)

  • Jhee, Kwang-Hwan;Cho, Hyun-Nam;Yang, Seun-Ah;Lee, In-Seun
    • Microbiology and Biotechnology Letters
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    • v.35 no.3
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    • pp.196-202
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    • 2007
  • Mutations in the cystathionine ${\beta}$-synthase (CBS) gene cause homocystinuria, the most frequent inherited disorder in sulfur metabolism. CBS is the unique enzyme using both heme and pyridoxal 5-phosphate (PLP) for activity. Among the reported 140 mutations, one of the most common disease-causing alterations in human CBS is G307S mutation. To investigate the pathogenic mechanism of G307S by spectroscopic methods, we engineered the full length and the truncated G247S mutation of yeast CBS that is corresponding mutation to human G307S. Yeast CBS does not contain heme and thus gives a merit to study the spectroscopic properties. The UV-visible spectra of the purified full length and the truncated G247S yeast CBSs showed the total absence of PLP in the protein. The absence of PLP in G247S mutation was also confirmed by the PLP-cyanide adduct formation experiment, which was conducted by the incubation of the purified enzyme with KCN. The adducts were detected using a circular dichroism (CD) and a spectrofluorimeter. Radio isotope activity assay of full length and truncated G247S proteins also gave no activity. Our yeast G247S mutation data suggested that G307S might make the distortion of the active site so that cofactor PLP and substrate can not fit inside the active site. Our yeast CBS study addressed the reason why the G307S mutation in human CBS makes the enzyme inactive that consequently leads to severe clinical phenotype.

Effect of Paecilomyes japonica on Lipid Metabolism In Rats Fed High Fat Diet (눈꽃동충하초가 고지방 식이를 섭취한 흰쥐의 지질대사에 미치는 영향)

  • 고진복;최미애
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.2
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    • pp.238-243
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    • 2003
  • The effects of Paecilomyes japonica on weight gains, food intakes, food efficiency ratios, serum and hepatic lipid concentrations, serum protein levels and serum enzyme activities, were studied in adult male rats. Sprague-Dawley rats,35 weeks old, were given four different types of diets for a succeeding period of five weeks: either a normal diet (5% corn oil), a control diet (high fat; 5% corn oil + 15% lard), a PF diet (control diet + 3% fruiting body of Paecilomyes japonica), or a PM diet (control diet+.3% mycelium of Paecilomyes japonica). The body weight gains, hepatic weights and food efficiency ratios of rats fed the PF or PM diets were significantly lower than those fed the control diet, but were similar to those fed the normal diet. The concentrations of hepatic total lipids, cholesterol and triglyceric, and serum triglyceride, of rats given the PF or PM diets were significantly lower than those given the control diet. Hut the concentrations of total cholesterol, HDL-cholesterol, LDL-cholesterol and phospholipid in the serum of rats fed the control, PF or PM diets were significantly higher than those fed the normal diet. In the serum of rats fed the PF diet, the HDL-cholesterol/total cholesterol ratio was significantly higher and the atherogenic index was significantly lower than those fed the control diet, while such effect was not observed in rats fed the PM diet. The alkaline phosphatase activity in the serum of rats fed the control and PM diets was more significantly decreased compared to rats fed the PF and normal diet. No differences were noted in the weights of the pancreas, kidney and heart, the serum concentrations of glucose, hemoglobin and albumin, and the activities of GOT, GPT and ${\gamma}$-GTP, among the rats on all the experimental diets. In conclusion, the rats fed the PF or PM diets maintained normal body and hepatic weights. Despite of the high intake of fats in the PF and PM diets, the concentrations of hepatic total lipids, cholesterol and triglyceride, and serum triglyceride were decreased.

Anti-aging Effects of L-Carnitine on Human Skin (L-카르니틴의 사람피부에 대한 항노화 효과)

  • Lee Bum-Chun;Choe Tae-Boo;Sim Gwan-Sub;Lee Geun-Soo;Park Sung-Min;Lee Chun-Il;Pyo Hyeong-Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.3 s.47
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    • pp.393-397
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    • 2004
  • L-Carnitine $({\beta}-hydroxy-{\gamma}-trimethyl-ammoniumbutyric{\;}acid)$ is a small water-soluble molecule important in mammalian fat metabolism. It is essential for the normal oxidation of fatty acids by the mitochondria, and is involved in the trans-esterification and excretion of acyl-CoA esters. In this paper, to investigate the relationship between aging and L-carnitine, we investigated the effects of in vitro matrix-metalloproteinase (MMP) inhibition and activity and expression of UYA-induced MMPs in human skin fibroblasts. Also, we studied to develop as anti-aging cosmetics with L-carnitine. Fluorometric assays of the proteolytic activities of MMP-1 (collagenase) were performed using fluorescent collagen substrates. ELISA (enzyme linked immune sorbent assay), gelatin-substrate zymography, RT-PCR ELISA techniques were used for the effects of L-carnitine on MMP expression, activity, and MMP mRNA expression in UVA irradiated fibroblast $(5\;J/cm^2)$, respectively. In addition, we performed clinical study with L-carnitine cream. L-carnitine inhibited the activities of MMP-1 in a dose-dependent manner and the $IC_{50}$ values calculated from semi-log plots were 2.45 mM, and L-carnitine showed strong inhibition on MMP-2 (gelatinase) activity in UVA irradiated fibroblast by zymography. Also, UVA induced MMP-1, 2 expression was reduced $43\%,\;53\%$ by treated with L-carnitine at 1.25 mM, and MMP-1 mRNA expression was reduced dose-dependent manner. Therefore L-carnitine was able to significantly inhibit the MMP activity, and regulate MMP expression in protein and mRNA level. The results of clinical study showed that $1.0\%$ L-carnitine treated group reduced wrinkle significantly compared with placebo treated group (P<0.05). All these results suggest that L-carnitine may be useful as new anti-aging cosmetics for protection against UVA induced Mm expression and activity.

The Effects of Environment-Friendly Diets on the Growth Performance, Nutrient Digestibility, Fecal Excretion, Nitrogen Excretion and Emission Gases in Manure for Growing Pigs (환경친화적인 사료의 급여가 육성돈의 성장 능력, 영양소 소화율, 분 배설량, 분뇨내 질소배설량 및 악취 가스에 미치는 영향)

  • Yoo, J.S.;Cho, J.H.;Chen, Y.G.;Kim, H.J.;Wang, Q.;Hyun, Y.;Ko, T.G.;Park, C.S.;Kim, I.H.
    • Journal of Animal Science and Technology
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    • v.49 no.4
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    • pp.491-500
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    • 2007
  • Two experiments were used to determine the effects of environment-friendly diets on growth performance, fecal excretion, nitrogen excretion and emission gases in manure for growing pigs. In experiment 1, ninety six crossed pigs(Landrace×Yorkshire×Duroc) were allocated into four treatments. Treatments were AME(adequate ME diet, 3,265 kcal/kg), LME(lower ME diet, 3,100 kcal/kg), LME 0.05(lower ME diet+α- galactosidase & β-mannanase 0.05%) and LME 0.10(lower ME diet+α-galactosidase & β-mannanase 0.10%). Pigs fed AME diet had lower ADFI(Average Daily Feed Intake) than pigs fed other diets(p<0.05). DM(Dry Matter) digestibility in pigs fed AME and LME 0.10 diets had greater than pigs fed LME diet(p<0.05). Energy digestibility is higher in pigs fed AME and LME 0.10 diets than other treatments(p<0.05). In experiment 2, twenty four crossbred pigs(33.71 kg average BW) were used in a 14-d metabolism experiment. The pigs were housed in individual cages equipped with plastic bed flooring. Treatments were CP(Crude protein) 18% without Bacillus sp., CP 18% diet+Bacillus sp. 0.05%, CP 14% without Bacillus sp. and CP 14% diet+Bacillus sp. 0.05%. Nitrogen intake was higher for CP 18% diets than CP 14% diets(p<0.05). DM, N(Nitrogen) and energy digestibility were affected by probiotics(p<0.05). With the high CP in diets, Energy and N digestibility, urine N percent, urine N excretion and total N excretion were increased significantly compared to low CP in diets(p<0.05). Among the treatments, DM and N digestibilities, feces N excretion, N absorption were decreased significantly(p<0.05), however, feces excretion, feces N, urine N percent, urine N excretion and total N excretion were increased significantly(p<0.05) when pigs fed without probiotics diets compare to pigs fed with probiotics diets. DM and N digestibility, feces excretion, feces N excretion, urine N percent, urine N excretion, total N excretion, N absorption and N adsorption ratio were CP×probiotic interactions in p<0.05. Ammonia(p<0.01) and H2S(p<0.05) in manure were lower in CP 14% diets than CP 18% diets. Also, ammonia and H2S in manure were CP×probiotic interactions in p<0.05. In conclusion, low energy and reduction of CP dietary added enzyme and probiotics improved nutrient digestibility and reduced odors emission in manure for growing pigs.

Inhibition of Pancreatic Lipase Activity and Adipocyte Differentiation in 3T3-L1 Cells Treated with Purple Corn Husk and Cob Extracts (자색옥수수 포엽과 속대 추출물의 리파아제 저해활성 및 3T3-L1 지방전구세포에서의 지방분화 억제효과)

  • Lee, Ki Yeon;Hong, Soo Young;Kim, Tae Hee;Kim, Jai Eun;Park, A-Reum;Noh, Hee Sun;Kim, Si Chang;Park, Jong Yeol;Ahn, Mun Seob;Jeong, Won Jin;Kim, Hee Yeon
    • Journal of Food Hygiene and Safety
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    • v.33 no.2
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    • pp.131-139
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    • 2018
  • Our review begins with the maize hybrid for grain, called 'Seakso 1,' which was developed in 2008 by the Gangwon Agricultural Research and Extension Services in Korea, and subsequently registered in 2011. In this study, we aimed to investigate the lipid metabolic enzyme activity and inhibitory effect on the adipocyte differentiation, in 3T3-L1 cells of the identified Seakso 1 corn husk and cob extracts (EHCS). We investigated the pancreatic lipase inhibitory effect and anti-adipogenic effect of EHCS.The lipid accumulation and adipocyte differentiation were measured by the procedure of Oil Red O staining, Real-time PCR and the Western blot analysis. The pancreatic lipase inhibitory activity of EHCS was measured at higher levels than those of the positive control (orlistat) at 100, 500, and $1,000{\mu}g/mL$. In particular, EHCS was noted as being significantly inhibited and including a measured adipocyte differentiation and lipid accumulation, when treated during the adipocyte differentiation process in 3T3-L1 cells. Based on the Oil Red O staining, EHCS inhibited lipid accumulation at 19.19%, 33.30% at $1000{\mu}g/mL$, $2000{\mu}g/mL$, respectively. The real-time PCR and Western blot analysis showed that EHCS significantly decreased in the mRNA expression and protein level of obesity-related factors, such as peroxisome-proliferatorsactivated-receptor-${\gamma}$ ($PPAR{\gamma}$) and CCAAT enhancer-binding-proteins ${\alpha}$ ($C/EBP{\alpha}$). This study potentially suggests that the Saekso 1 corn husk and cob extracts may improve lipid metabolism and reduce lipid accumulation.

Dietary behaviors and nutritional status according to the bone mineral density status among adult female North Korean refugees in South Korea (한국에 거주하고 있는 북한이탈주민 여성의 골밀도에 따른 식생활과 영양상태)

  • Kim, Su-Hyeon;Lee, Soo-Kyung;Kim, Sin-Gon
    • Journal of Nutrition and Health
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    • v.52 no.5
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    • pp.449-464
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    • 2019
  • Purpose: North Koreans could be at higher risk for their bone health because of previous periods of severe famine and the continuing low availability of food. This study determined the bone mineral density (BMD) status and its relationship with dietary behaviors and nutrient intake of North Korean refugees (NKR) in South Korea (SK). Methods: This cross-sectional study analyzed 110 female NKR from a NORNS cohort of a non-probability sample of adult NKR in Seoul. BMD examined by DEXA was used to divide participants into the normal group (NG) and the non-normal group (NNG) according to the WHO guideline. A self-administered questionnaire included questions on age, the socioeconomic situation in North Korea (NK) and SK, the food security in NK and SK, and the health behaviors, dietary behaviors, and food frequency questionnaire administered in SK. A one-day 24-hr recall was conducted and the results were analyzed by using CanPro. SPSS was used to analyze whether BMD and related dietary behaviors and nutrient intakes differed according to the groups. Results: NG (62.7%) was significantly younger and had a lower abdominal obesity score than NNG (p < 0.001). While 14.5% of NG reported experiencing menopause, all of NNG reported experiencing menopause. The NG more frequently consumed the dairy group of foods (9.6 times a week) than did the NNG (4.8 times a week) after the statistics were adjusted for age (p < 0.007). The NG consumed significantly more animal protein and animal calcium than did the NNG (p = 0.01, p = 0.009, respectively). Calcium intake was low with 49.3% of NG, and 78.0% of the NNG reported consuming calcium lower than the estimated average requirement. Only calcium showed an index of nutrient quality lower than one in both groups. Conclusion: These results showed that NKR women and possibly all North Korean women are at high risk for bone health and they consumed low levels of bone-related nutrients, and this should be considered for the nutrition policy for NKR and North Korea.

Effect of Sulgidduk containing pine needle juice on lipid metabolism in high fat-cholesterol diet induced dyslipidemic rats (이상지질혈증 동물 모델을 이용한 솔잎 착즙액 첨가 설기떡의 지질개선 효과)

  • Lee, Yunjung;Park, Jae-Hee;Park, Eunju
    • Journal of Nutrition and Health
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    • v.52 no.1
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    • pp.6-16
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    • 2019
  • Purpose: Dyslipidemia is a major risk factor for cardiovascular disease. Pine needles (Pinus densiflora seib et Zucc) are a traditional medicine used to treat dyslipidemia in clinical settings. This study examined the potential effects of sulgidduk, a Korean traditional rice cake containing pine needle juice to protect against dyslipidemia induced by a high-fat/sugidduk diet in a rat model. Methods: Twenty one male Sprague-Dawley rats were divided randomly into three groups: normal control (NC), Sulgidduk diet (SD), Sulgidduk diet containing pine needle juice (PSD). The blood lipid levels, production of lipid peroxide in the plasma and liver, total cholesterol and triglyceride in the liver and feces, antioxidant enzyme activities in plasma and erythrocytes were measured to assess the effects of PSD on dyslipidemia. Results: A high-fat/Sulgidduk diet induced dyslipidemia, which was characterized by significantly altered lipid profiles in the plasma and liver. The food intake was similar in the three groups, but weight gain and food efficiency ratio (FER) were reduced significantly in the PSD group compared to those in the SD group. The level of total cholesterol, LDL-cholesterol and TBARS in the plasma showed tendencies to decrease in the PSD group compared to those in the SD group. The levels of high-fat/Sulgidduk diet-induced sterol regulatory element-binding protein 2 (SREBP2) gene expression were reduced significantly in the PSD group. The supplementation of PSD reduced the hepatic triglyceride and total cholesterol levels significantly, and enhanced the fecal excretion of triglyceride and hepatic antioxidant enzyme activities compared to the SD group. Conclusion: These results suggest that the addition of 0.4% pine needle juice to Sulgidduk may be an alternative snack to control dyslipidemia.

Relationship between porcine miR-20a and its putative target low-density lipoprotein receptor based on dual luciferase reporter gene assays

  • Ding, Yueyun;Zhu, Shujiao;Wu, Chaodong;Qian, Li;Li, DengTao;Wang, Li;Wan, Yuanlang;Zhang, Wei;Yang, Min;Ding, Jian;Wu, Xudong;Zhang, Xiaodong;Gao, Yafei;Yin, Zongjun
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.7
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    • pp.922-929
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    • 2019
  • Objective: Mutations in low-density lipoprotein receptor (LDLR), which encodes a critical protein for cholesterol homeostasis and lipid metabolism in mammals, are involved in cardiometabolic diseases, such as familial hypercholesterolemia in pigs. Whereas microRNAs (miRNAs) can control LDLR regulation, their involvement in circulating cholesterol and lipid levels with respect to cardiometabolic diseases in pigs is unclear. We aimed to identify and analyze LDLR as a potential target gene of SSC-miR-20a. Methods: Bioinformatic analysis predicted that porcine LDLR is a target of SSC-miR-20a. Wild-type and mutant LDLR 3'-untranslated region (UTR) fragments were generated by polymerase chain reaction (PCR) and cloned into the pGL3-Control vector to construct pGL3 Control LDLR wild-3'-UTR and pGL3 Control LDLR mutant-3'-UTR recombinant plasmids, respectively. An miR-20a expression plasmid was constructed by inserting the porcine premiR-20a-coding sequence between the HindIII and BamHI sites in pMR-mCherry, and constructs were confirmed by sequencing. HEK293T cells were co-transfected with the miR-20a expression or pMR-mCherry control plasmids and constructs harboring the corresponding 3'-UTR, and relative luciferase activity was determined. The relative expression levels of miR-20a and LDLR mRNA and their correlation in terms of expression levels in porcine liver tissue were analyzed using reverse-transcription quantitative PCR. Results: Gel electrophoresis and sequencing showed that target gene fragments were successfully cloned, and the three recombinant vectors were successfully constructed. Compared to pMR-mCherry, the miR-20a expression vector significantly inhibited wild-type LDLR3'-UTR-driven (p<0.01), but not mutant LDLR-3'-UTR-driven (p>0.05), luciferase reporter activity. Further, miR-20a and LDLR were expressed at relatively high levels in porcine liver tissues. Pearson correlation analysis revealed that porcine liver miR-20a and LDLR levels were significantly negatively correlated (r = -0.656, p<0.05). Conclusion: LDLR is a potential target of miR-20a, which might directly bind the LDLR 3'-UTR to post-transcriptionally inhibit expression. These results have implications in understanding the pathogenesis and progression of porcine cardiovascular diseases.

Comparative Transcriptome Analysis of the Response of Two Lines of Rapeseed (Brassica napus L.) to Cold Stress (유채 두 계통에서 저온 스트레스에 반응하는 전사체 발현 비교 분석)

  • Lee, Ji-Eun;Kim, Kwang-Soo;Cha, Young-Lok;An, Da-Hee;Byun, Jong-Won;Kang, Yong-Ku
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.66 no.1
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    • pp.37-71
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    • 2021
  • Rapeseed is a typical winter crop, and its freezing stress tolerance is a major feature for winter survival. Therefore, it is important to comprehend clearly the physical and molecular mechanisms of rapeseed under freezing stress conditions. This study investigates the physical and transcriptome changes of two rapeseed lines, 'J8634-B-30' and 'EMS26', under cold acclimation and freezing temperature treatments. The proline content of 'J8634-B-30' at 5 ℃ increased 8.7-fold compared to that before treatment, and there was no significant change in that of 'EMS26' RNA-sequencing analysis revealed 5,083 differentially expressed genes (DEGs) of 'J8634-B-30' under cold acclimation condition. Among the genes, 2,784 (54.8%) were up-regulated and 2,299 (45.2%) were down-regulated. The DEGs of 'EMS26' under cold acclimation condition were 5,831 genes, and contained 2,199 up-regulated genes (37.7%) and 3,632 down-regulated genes (62.3%). Among them, only DEGs annotated in the cold response-related signaling pathways were selected, and their expression in the two rapeseed lines was compared. Comparative DEGs analysis indicated that cold response related signaling pathways are proline metabolism and ABA (Abscisic acid) signaling. And ICE (Inducer of CBF expression) - CBF (C-repeat-binding factor) - COR (Cold-regulated) signaling were the significantly differentially expressed transcripts in the two rapeseed lines. The major induced transcripts of 'J8634-B-30' induced P5CS (Δ'-pyrroline-5-carboxylate synthetase), which is related to proline biosynthesis, PYL (pyrabactin resistance-like protein, ABA receptor) and COR413 (cold-regulated 413 plasma membrane 1). In conclusion, these result provide a foundation for understanding the mechanisms of freezing stress tolerance in rapeseeds. Further functional studies should be performed on the freezing stress-related genes identified in this study, which can contribute to the transgenic and molecular breeding for freezing stress tolerance in rapeseed.

Anti-obesogenic Effect of Brassica juncea Extract on Bisphenol-A Induced Adipogenesis of 3T3-L1 Cells (비스페놀 A (Bisphenol-A)로 유도된 지방세포 분화에 미치는 갓 추출물의 항오비소겐 효과)

  • Lee, Se-jeong;Na, Uoon-Joo;Choi, Sun-Il;Han, Xionggao;Men, Xiao;Lee, Youn Hwan;Kim, Hyun Duk;Kim, Yoon Jung;Lee, Ok-Hwan
    • Journal of Food Hygiene and Safety
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    • v.36 no.6
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    • pp.528-536
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    • 2021
  • The purpose of the study was to investigate the content of sinigrin, an index component, in Brassica juncea extract and to evaluate the differentiation of lipocytes, inhibition of production of reactive oxygen species (ROS) and reduction of protein production by lipogenic factors (PPARγ, C/EBPα, aP2) in the processing of Brassica juncea extract and sinigrin in 3T3-L1 preadipocytes which induces Bisphenol A (BPA), an endocrine disrupting environmental hormone. From the investigation, the content of sinigrin in Brassica juncea extract, measured by HPLC, is found to be 21.27±0.2 mg/g. The XTT assay result on BPA-derived 3T3-L1 adipocytes shows there is no cytotoxicity found from 180 µM of sinigrin and 300 ㎍/mL of Brassica juncea extract. Moreover, both intracellular lipid accumulation and ROS production during differentiation of lipocyte are significantly reduced in cells processed with Brassica juncea extract and sinigrin. Lastly, it was also found that the production of transcription factors of lipocyte differentiation, PPARγ, C/EBPα and aP2, were found to be suppressed by the application of Brassica juncea extract and sinigrin. Such results reveals that Brassica juncea is effective in not only suppressing lipid accumulation in the environmental hormone bisphenol A-derived lipocyte, but also in reducing the ROS. The sinigrin-containing Brassica juncea is highly expected to be used in natural functional supplements that prevents the lipid metabolism disorders caused by BPA. There are necessities for additional clinical research and follow-up studies on the in vivo model to verify the relevant mechanisms.