1.BnMTP10 regulates manganese accumulation in Brassica napus.
Yuting HE ; Zongyue LI ; Jinglin WANG ; Xingyu ZHAO ; Siying CHEN ; Sihong LIU ; Tianyu GU ; Yan GAO ; Xinke TANG ; Jiashi PENG
Chinese Journal of Biotechnology 2025;41(7):2843-2854
Stresses induced by the deficiency or excess of trace mineral elements, such as manganese (Mn), represent a common limiting factor for the production of crops like Brassica napus. To identify key genes involved in Mn allocation in B. napus and elucidate the underlying mechanisms, a member of the metal tolerance protein (MTP) family obtained in the previous screening of cDNA library of B. napus under Mn stress was selected as the research subject. Based on the sequence information and phylogenetic analysis, it was named as BnMTP10. It belongs to the Mn-cation diffusion facilitator (CDF) subfamily. Expression of BnMTP10 in yeast significantly improved the tolerance of transformants to excessive Mn and iron (Fe) and reduced the accumulation of Mn and Fe. However, the yeast transformants exhibited no significant changes in tolerance to excess cadmium, boron, aluminum, zinc, or copper. The qRT-PCR results demonstrated that the flowers of B. napus had the highest expression of BnMTP10, followed by roots and leaves. Subcellular localization studies revealed that BnMTP10 was localized in the endoplasmic reticulum (ER). Compared with wild-type plants, transgenic Arabidopsis overexpressing BnMTP10 exhibited enhanced tolerance to excessive Mn stress but showed no significant difference under Fe stress. Correspondingly, under excessive Mn stress, the Mn content in the roots of transgenic Arabidopsis increased significantly. However, under excessive Fe stress, the Fe content in transgenic Arabidopsis did not alter significantly. According to the results, we hypothesize that BnMTP10 may alleviate excessive Mn stress in plants by mediating Mn transport to the ER. This study facilitated our understanding of efficient mineral nutrients, and provided theoretical foundations and gene resources for breeding B. napus.
Brassica napus/genetics*
;
Manganese/metabolism*
;
Plants, Genetically Modified/genetics*
;
Plant Proteins/physiology*
;
Arabidopsis/metabolism*
;
Gene Expression Regulation, Plant
;
Phylogeny
;
Cation Transport Proteins/metabolism*
;
Stress, Physiological
2.BnaNRT1.5s mediates nitrate transporter to regulate nitrogen use efficiency in Brassica napus.
Shilong CHEN ; Lei YAO ; Rumeng WANG ; Jian ZENG ; Jianghe LI ; Shiyao CUI ; Xu WANG ; Haixing SONG ; Zhenhua ZHANG ; Pan GONG
Chinese Journal of Biotechnology 2025;41(7):2954-2965
Improving the nitrogen use efficiency (NUE) of Brassica napus is of significant importance for achieving the national goal of zero growth in chemical fertilizer application and ensuring the green development of the rapeseed industry. This study aims to explore the effects of the nitrate transporter gene BnaNRT1.5s on the nitrogen transport and NUE of B. napus, providing excellent genetic resources for the development of nitrogen-efficient B. napus varieties. The spatiotemporal expression of BnaA05.NRT1.5 as a key nitrogen responsive gene was profiled by qRT-PCR at different growth stages and for different tissue samples of B. napus 'Westar'. Subcellular localization was employed to examine its expression pattern in the cells. Additionally, CRISPR/Cas9 was used to create BnaNRT1.5s knockout lines, which were subjected to hydroponic experiments under high nitrogen (12.0 mmol/L) and low nitrogen (0.3 mmol/L) conditions. After the seedlings were cultivated for 21 days, root and shoot samples were collected for weighing, nitrogen content determination, xylem sap nitrate content assessment, and calculation of total nitrogen and NUE. The B. napus nitrate transporter BnaA05.NRT1.5 was localized to the cell membrane. During the seedling and early bolting stages, BnaA05.NRT1.5 was predominantly expressed in roots, while it was highly expressed in old leaves and mature silique skin during the reproductive stage. Compared with the wild type, the mutant BnaNRT1.5s showed significant increases in the dry weight and total nitrogen of seedlings under both high and low nitrogen conditions. Under low nitrogen conditions, NUE in the roots of BnaNRT1.5s significantly improved. Notably, under both high and low nitrogen conditions, the nitrate content in the shoots of BnaNRT1.5s decreased significantly, while that in the roots increased significantly, resulting in a significantly decreased shoot-to-root nitrate content ratio. BnaNRT1.5s is involved in regulating the transport of nitrate from the roots to the shoots, and its mutation enhances nitrogen absorption and utilization in B. napus seedlings, promoting seedling growth. This study not only provides references for understanding the physiological and molecular mechanisms by which BnaNRT1.5s regulates NUE but also offers valuable genetic resources for improving NUE in B. napus.
Brassica napus/genetics*
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Anion Transport Proteins/metabolism*
;
Nitrogen/metabolism*
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Nitrate Transporters
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Plant Proteins/metabolism*
;
Nitrates/metabolism*
;
Gene Expression Regulation, Plant
;
Biological Transport
3.Identification and expression analysis of NHX gene family in Chinese cabbage.
Xuehua WANG ; Jia HAN ; Jizhong MA ; Xiting YANG ; Huali MAN ; Yali QIAO ; Xueqin GAO ; Linli HU
Chinese Journal of Biotechnology 2023;39(2):552-565
Na+/H+ antiporter (NHX) gene subfamily plays an important role in plant response to salt stress. In this study, we identified the NHX gene family members of Chinese cabbage and analyzed the expression patterns of BrNHXs gene in response to abiotic stresses such as high temperature, low temperature, drought and salt stress. The results showed that there were 9 members of the NHX gene family in Chinese cabbage, which were distributed on 6 chromosomes respectively. The number of amino acids was 513-1 154 aa, the relative molecular weight was 56 804.22-127 856.66 kDa, the isoelectric point was 5.35-7.68. Members of BrNHX gene family mainly existed in vacuoles, the gene structure is complete, and the number of exons is 11-22. The secondary structures of the proteins encoded by the NHX gene family in Chinese cabbage had alpha helix, beta turn and random coil, and the alpha helix occurred more frequently. Quantitative real-time PCR (qRT-PCR) analysis showed that the gene family members had different responses to high temperature, low temperature, drought and salt stress, and their expression levels differed significantly in different time periods. BrNHX02 and BrNHX09 had the most significant responses to these four stresses, and their expression levels were significantly up-regulated at 72 h after treatments, which could be used as candidate genes to further verify their functions.
Genome, Plant
;
Multigene Family
;
Stress, Physiological/genetics*
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Brassica/metabolism*
;
Gene Expression Regulation, Plant
;
Phylogeny
;
Plant Proteins/metabolism*
4.Phylogenetic and expression analysis of SEPALLATA-like gene in Brassica oleracea L. var. acephala.
Yuanping XIANG ; Yuntong HUANG ; Hongjun HE ; Qijiang XU
Chinese Journal of Biotechnology 2020;36(11):2398-2412
The E class MADS-box genes SEPALLATA (SEP)-like play critical roles in angiosperm reproductive growth, especially in floral organ differentiation. To analyze the sequence characteristics and spatio-temporal expression patterns of E-function MADS-box SEP-like genes during kale (Brassica oleracea L. var. acephala) flower development, BroaSEP1/2/3 (GenBank No. KC967957, KC967958, KC967960) homologues, three kale SEP MADS-box gene, were isolated from the kale variety 'Fourteen Line' using Rapid amplification of cDNA ends (RACE). Sequence and phylogenetic analysis indicated that these three SEP genes had a high degree of identity with SEP1, SEP2, SEP3 from Brassica oleracea var. oleracea, Brassica rapa, Raphanus sativus and Brassica napus, respectively. Alignment of the predicted amino acid sequences from these genes, along with previously published subfamily members, demonstrated that these genes comprise four regions of the typical MIKC-type MADS-box proteins: the MADS domain, intervening (I) domain and keratin-like (K) domain, and the C-terminal domain SEPⅠ and SEP Ⅱ motif. The longest open reading frame deduced from the cDNA sequences of BroaSEP1, BroaSEP2, and BroaSEP3 appeared to be 801 bp, 759 bp, 753 bp in length, respectively, which encoded proteins of 266, 252, and 250 amino acids respectively. Expression analyses using semi-quantitative RT-PCR and quantitative real-time PCR indicate that BroaSEP1/2/3 are specifically expressed in floral buds of kale during flower development process. The expression levels of the three genes are very different at different developmental stages, also in wild type, mutant flower with increased petals, and mutant flower with decreased petals. These different patterns of gene expression maybe cause the flowers to increase or decrease the petal number.
Brassica/metabolism*
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Flowers/genetics*
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Gene Expression Regulation, Plant
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MADS Domain Proteins/metabolism*
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Phylogeny
;
Plant Proteins/metabolism*
5.The effects of Brassica juncea L. leaf extract on obesity and lipid profiles of rats fed a high-fat/high-cholesterol diet.
Jae Joon LEE ; Hyun A KIM ; Joomin LEE
Nutrition Research and Practice 2018;12(4):298-306
BACKGROUND/OBJECTIVES: Obesity is a global health problem of significant importance which increases mortality. In place of anti-obesity drugs, natural products are being developed as alternative therapeutic materials. In this study, we investigated the effect of Brassica juncea L. leaf extract (BLE) on fat deposition and lipid profiles in high-fat, high-cholesterol diet (HFC)-induced obese rats. MATERIALS/METHODS: Male Sprague-Dawley rats were divided into four groups (n = 8 per group) according to diet: normal diet group (ND), high-fat/high-cholesterol diet group (HFC), HFC with 3% BLE diet group (HFC-A1), and HFC with 5% BLE diet group (HFC-A2). Each group was fed for 6 weeks. Rat body and adipose tissue weights, serum biochemical parameters, and tissue lipid contents were determined. The expression levels of mRNA and proteins involved in lipid and cholesterol metabolism were determined by reverse transcription polymerase chain reaction and western blot analysis, respectively. RESULTS: The HFC-A2 group showed significantly lower body weight gain and food efficiency ratio than the HFC group. BLE supplementation caused mesenteric, epididymal, and total adipose tissue weights to decrease. The serum levels of triglyceride, total cholesterol, and low-density lipoprotein cholesterol were significantly reduced, and high-density lipoprotein cholesterol was significantly increased in rats fed BLE. These results were related to lower glucose-6-phosphate dehydrogenase, acetyl-coA carboxylase, and fatty acid synthase mRNA expression, and to higher expression of the cholesterol 7α-hydroxylase and low density lipoprotein-receptor, as well as increased protein levels of peroxisome proliferator-activated receptor α. Histological analysis of the liver revealed decreased lipid droplets in HFC rats treated with BLE. CONCLUSIONS: Supplementation of HFC with 3% or 5% BLE inhibited body fat accumulation, improved lipid profiles, and modulated lipogenesis- and cholesterol metabolism-related gene and protein expression.
Acetyl-CoA Carboxylase
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Adipose Tissue
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Animals
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Anti-Obesity Agents
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Biological Products
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Blotting, Western
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Body Weight
;
Brassica*
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Cholesterol
;
Diet*
;
Diet, High-Fat
;
Global Health
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Glucosephosphate Dehydrogenase
;
Humans
;
Lipid Droplets
;
Lipoproteins
;
Liver
;
Male
;
Metabolism
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Mortality
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Mustard Plant*
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Obesity*
;
Peroxisomes
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Polymerase Chain Reaction
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Rats*
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Rats, Sprague-Dawley
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Reverse Transcription
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RNA, Messenger
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Triglycerides
;
Weights and Measures
6.Role of jasmonic acid in improving tolerance of rapeseed (Brassica napus L.) to Cd toxicity.
Essa ALI ; Nazim HUSSAIN ; Imran Haider SHAMSI ; Zahra JABEEN ; Muzammil Hussain SIDDIQUI ; Li-Xi JIANG
Journal of Zhejiang University. Science. B 2018;19(2):130-146
The well-known detrimental effects of cadmium (Cd) on plants are chloroplast destruction, photosynthetic pigment inhibition, imbalance of essential plant nutrients, and membrane damage. Jasmonic acid (JA) is an alleviator against different stresses such as salinity and drought. However, the functional attributes of JA in plants such as the interactive effects of JA application and Cd on rapeseed in response to heavy metal stress remain unclear. JA at 50 µmol/L was observed in literature to have senescence effects in plants. In the present study, 25 µmol/L JA is observed to be a "stress ameliorating molecule" by improving the tolerance of rapeseed plants to Cd toxicity. JA reduces the Cd uptake in the leaves, thereby reducing membrane damage and malondialdehyde content and increasing the essential nutrient uptake. Furthermore, JA shields the chloroplast against the damaging effects of Cd, thereby increasing gas exchange and photosynthetic pigments. Moreover, JA modulates the antioxidant enzyme activity to strengthen the internal defense system. Our results demonstrate the function of JA in alleviating Cd toxicity and its underlying mechanism. Moreover, JA attenuates the damage of Cd to plants. This study enriches our knowledge regarding the use of and protection provided by JA in Cd stress.
Brassica napus/metabolism*
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Cadmium/toxicity*
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Catalase/metabolism*
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Cyclopentanes/pharmacology*
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Oxylipins/pharmacology*
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Photosynthesis
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Plant Leaves/metabolism*
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Superoxide Dismutase/metabolism*
7.Metabolic Pathways Associated with Kimchi, a Traditional Korean Food, Based on In Silico Modeling of Published Data.
Ga Hee SHIN ; Byeong Chul KANG ; Dai Ja JANG
Genomics & Informatics 2016;14(4):222-229
Kimchi is a traditional Korean food prepared by fermenting vegetables, such as Chinese cabbage and radishes, which are seasoned with various ingredients, including red pepper powder, garlic, ginger, green onion, fermented seafood (Jeotgal), and salt. The various unique microorganisms and bioactive components in kimchi show antioxidant activity and have been associated with an enhanced immune response, as well as anti-cancer and anti-diabetic effects. Red pepper inhibits decay due to microorganisms and prevents food from spoiling. The vast amount of biological information generated by academic and industrial research groups is reflected in a rapidly growing body of scientific literature and expanding data resources. However, the genome, biological pathway, and related disease data are insufficient to explain the health benefits of kimchi because of the varied and heterogeneous data types. Therefore, we have constructed an appropriate semantic data model based on an integrated food knowledge database and analyzed the functional and biological processes associated with kimchi in silico. This complex semantic network of several entities and connections was generalized to answer complex questions, and we demonstrated how specific disease pathways are related to kimchi consumption.
Asian Continental Ancestry Group
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Biological Processes
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Brassica
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Capsicum
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Computer Simulation*
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Garlic
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Genome
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Ginger
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Humans
;
Insurance Benefits
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Metabolic Networks and Pathways*
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Metabolism
;
Onions
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Raphanus
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Seafood
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Seasons
;
Semantics
;
Vegetables
8.The Effects of Broccoli Sprout Extract Containing Sulforaphane on Lipid Peroxidation and Helicobacter pylori Infection in the Gastric Mucosa.
Young Woon CHANG ; Jae Young JANG ; Yong Ho KIM ; Jung Wook KIM ; Jae Jun SHIM
Gut and Liver 2015;9(4):486-493
BACKGROUND/AIMS: The aims of this study were to investigate whether a broccoli sprout extract containing sulforaphane (BSES) inhibited the Helicobacter pylori infection density and exerted an antioxidative effect on gastric mucosal damage. METHODS: The enrolled subjects were randomized in a double-blinded manner into three groups. Finally, 33 H. pylori (+) BSES treatment subjects (group A), 28 H. pylori (+) placebo subjects (group B), and 28 H. pylori (-) BSES treatment subjects (group C) were studied. H. pylori infection density was indirectly quantified by a 13C-urea breath test (UBT), and the ammonia concentration in gastric juice aspirates was measured through gastroscopic examination. Malondialdehyde (MDA), an oxidative damage biomarker, and reduced glutathione (GSH), an antioxidant biomarker, were measured in the gastric mucosa by an enzyme-linked immunosorbent assay. RESULTS: BSES treatment did not significantly affect the UBT values or ammonia concentration in group A (p=0.634 and p=0.505, respectively). BSES treatment did significantly reduce mucosal MDA concentrations in group A (p<0.05) and group C (p<0.001), whereas the gastric mucosal GSH concentrations did not differ before and after treatment in any of the groups. CONCLUSIONS: BSES did not inhibit the H. pylori infection density. However, BSES prevented lipid peroxidation in the gastric mucosa and may play a cytoprotective role in H. pylori-induced gastritis.
Adult
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Ammonia/metabolism
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Antioxidants/*pharmacology
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Biomarkers/analysis
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Brassica/*chemistry
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Breath Tests
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Double-Blind Method
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Enzyme-Linked Immunosorbent Assay
;
Female
;
Gastric Juice/enzymology
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Gastric Mucosa/*drug effects/metabolism
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Glutathione/analysis
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Helicobacter Infections/*drug therapy
;
*Helicobacter pylori
;
Humans
;
Isothiocyanates/*pharmacology
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Lipid Peroxidation/*drug effects
;
Male
;
Malondialdehyde/analysis
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Middle Aged
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Plant Extracts/chemistry/*pharmacology
;
Urea
9.Enhancement of beta-Glucan Content in the Cultivation of Cauliflower Mushroom (Sparassis latifolia) by Elicitation.
Hyun PARK ; Kang Hyeon KA ; Sung Ryul RYU
Mycobiology 2014;42(1):41-45
The effectiveness of three kinds of enzymes (chitinase, beta-glucuronidase, and lysing enzyme complex), employed as elicitors to enhance the beta-glucan content in the sawdust-based cultivation of cauliflower mushroom (Sparassis latifolia), was examined. The elicitors were applied to the cauliflower mushroom after primordium formation, by spraying the enzyme solutions at three different levels on the sawdust-based medium. Mycelial growth was fully accomplished by the treatments, but the metabolic process during the growth of fruiting bodies was affected. The application of a lysing enzyme resulted in an increase in the beta-glucan concentration by up to 31% compared to that of the control. However, the treatment resulted in a decrease in mushroom yield, which necessitated the need to evaluate its economic efficiency. Although we still need to develop a more efficient way for using elicitors to enhance functional metabolites in mushroom cultivation, the results indicate that the elicitation technique can be applied in the cultivation of medicinal/edible mushrooms.
Agaricales*
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Brassica*
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Fruit
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Glucuronidase
;
Metabolism
10.Evaluation of hypoglycemic and hypolipidemic activity of methanol extract of Brassica oleracea.
Tahira ASSAD ; Rafeeq A KHAN ; Zeeshan FEROZ
Chinese Journal of Natural Medicines (English Ed.) 2014;12(9):648-653
AIM:
The hypoglycemic and hypolipidemic effects of the methanol extract of Brassica oleracea var. capitata (MEB) was evaluated in alloxan-induced diabetic rabbits.
METHOD:
The study was conducted on twenty-eight healthy white rabbits of either sex. All animals were equally divided into four groups. After confirmation of hyperglycemia, the animals of the treated and standard groups were administered MEB (500 mg·kg(-1)) and glibenclamide (10 mg·kg(-1)), respectively for 15 and 30 days. The animals of the normal and diabetic controls received normal saline 1 mL/day equivalent to the volume of doses given to the test and standard animals. Biochemical tests were performed at the end of dosing, i.e. the 16(th) and 31(st) days.
RESULTS:
The MEB revealed a decrease of 106.6 mg·dL(-1) in fasting blood glucose as compared to diabetic control, which was almost comparable to glibenclamide; both of these changes were highly significant. The decrease in total cholesterol and low density lipoprotein was 94.3 and 96.5 mg·dL(-1), respectively, whereas the high-density lipoprotein was increased by 26.7 mg·dL(-1), as compared to diabetic control. All of the changes in lipid profile were statistically significant.
CONCLUSION
These results suggest the potential of MEB as a hypoglycemic and hypolipidemic agent.
Animals
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Blood Glucose
;
metabolism
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Brassica
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Cholesterol
;
blood
;
Diabetes Mellitus, Experimental
;
blood
;
drug therapy
;
Female
;
Hypoglycemic Agents
;
pharmacology
;
therapeutic use
;
Hypolipidemic Agents
;
pharmacology
;
therapeutic use
;
Lipoproteins
;
blood
;
Male
;
Phytotherapy
;
Plant Extracts
;
pharmacology
;
therapeutic use
;
Rabbits
;
Triglycerides
;
blood

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