Screening of Dried Plant Seed Extracts for Adiponectin Production Activity and Tumor Necrosis Factor-Alpha Inhibitory Activity on 3T3-L1 Adipocytes

To search for dried plant seeds with potent anti-diabetes activity, we conducted a large scale screening for inhibitory activity on tumor necrosis factor-alpha and facilitating activity on adiponectin production in vitro. These activities in 3T3-L1 adipocytes were screened from ethanol extracts of 20 kinds of dried plant seed marketed in Japan. komatsuna (Brassica rapa var. perviridis), common bean (Phaseolus vulgaris L.), qing geng cai (Brassica rapa var. chinensis), green soybean (Glycine max), spinach (Spinacia oleracea L.) and sugar snap pea (Pisum sativum L.) markedly enhanced adiponectin production (11.3 ~ 12.7 ng/ml) but Japanese radish (Raphanus sativus), edible burdock (Arctium lappa L.), bitter melon (Momordica charantia) and broccoli (Brassica oleracea var. italica) did not (0.9 ~ 2.7 ng/ml). All adiponectin-production-enhancing seeds except spinach (2.7 pg/ml) and okra (Abelmoschus esculentus) (6.6 pg/ml) effectively decreased tumor necrosis factor-alpha levels (0.0 pg/ml). We further examined the effects on free radical scavenging activities in the dried seed extracts. Although scavenging activity correlated well with total phenolic content of samples, no correlation was observed with adiponectin production. These results point to the potential of dried seed extracts as a means to modify the activity of tumor necrosis factor-alpha for the adiponectin production.     

六堡茶对胰岛素抵抗3T3-L1脂肪细胞糖脂代谢的影响

采用软脂酸诱导3T3-L1脂肪细胞胰岛素抵抗后,给予不同剂量的六堡茶水提物治疗,探讨六堡茶对胰岛素抵抗3T3-L1脂肪细胞的糖脂代谢的影响。结果表明,1 mmol·L^-1的软脂酸对3T3-L1脂肪细胞作用24 h后,葡萄糖摄取量减少,建立胰岛素抵抗模型。六堡茶对模型细胞干预24 h后,与模型组比较,细胞培养基的残留液中葡萄糖和游离脂肪酸的含量降低;对糖脂代谢相关关键酶（己糖激酶、6-磷酸果糖激酶-1、丙酮酸激酶、甘油-3-磷酸酰基转移酶）和葡萄糖转运子基因表达都有上调作用,并下调乙酰辅酶A羧化酶、蛋白酪氨酸磷酸酯酶1B基因表达。低剂量的六堡茶下调肉碱脂酰转移酶-I基因的表达,但是高剂量能显著上调肉碱脂酰转移酶-I基因表达。实验证明了六堡茶能够提高胰岛素抵抗3T3-L1脂肪细胞对葡萄糖的摄取能力,同时促进细胞内糖脂代谢,六堡茶有改善3T3-L1脂肪细胞胰岛素抵抗的作用。     

Ameliorating Effects of Fermented Rice Bran Extract on Oxidative Stress Induced by High Glucose and Hydrogen Peroxide in 3T3-L1 Adipocytes

In this study, we investigated whether fermented rice bran (FRB) can ameliorate the oxidative stress induced by high glucose and hydrogen peroxide (H₂O₂) in 3T3-L1 adipocytes by analyzing reactive oxygen species (ROS), oil red O staining, as well as the expression of mRNAs related to glucose homeostasis and adipogenesis. It was first confirmed that rice bran fermented by Issatchenkia orientalis MFST1 extract increased free phenolic content compared to non-fermented rice bran. The FRB extract strongly inhibited ROS generation and upregulated the expression of PPAR-γ and adiponectin. Moreover, FRB upregulated GLUT4 related to glucose transportation and insulin sensitivity. Taken together, FRB extract ameliorated oxidative stress-induced insulin resistance by neutralizing free radicals and upregulating adiponectin in adipocytes. Our results provide information toward understanding the beneficial effects of FRB on oxidative stress.     

Effect of Germinated Soybean Protein Hydrolysates on Adipogenesis and Adipolysis in 3T3-L1 Cells

Germination of soybeans increases the bioavailability of some nutrients. An evaluation was done to determine if germination increased the anti-adipogenic and lipolytic effects of soybean. Soybeans were germinated for 0 to 6 days and protein concentrates extracted from beans germinated at each period. Soy protein concentrates can retain notable amounts of phytochemicals with anti-adipogenic activity. For this reason, it was evaluated the effect of protein hydrolysates with and without phytochemicals in the adipocyte-like cells after 3T3-L1 (murine fibroblasts) cell line differentiation. Cell viability decreased with exposure to the germinated soybean protein hydrolysates during the differentiation stage, but not during the fibroblast or mature adipocyte stages. Adipogenesis and triglycerides accumulation were strongly inhibited by the hydrolysate from soybeans germinated for 2 days (with ethanol-soluble phytochemicals), when compared to ungerminated soybean. Adipolysis increased with exposure to hydrolysates from beans germinated for 2 days (with phytochemicals) and 5 days (without phytochemicals). Germinated soy protein hydrolysates had an effect on inhibition of lipid storage in adypocites and increasing lipolysis, which was improved by changes of the protein and increased phytochemical content due to germination.     

Lipid Inhibitory Effect of (−)-loliolide Isolated from Sargassum horneri in 3T3-L1 Adipocytes: Inhibitory Mechanism of Adipose-Specific Proteins

Sargassum horneri (S. horneri) is a well-known brown seaweed widely distributed worldwide. Several biological activities of S. horneri have been reported. However, its effects on lipid metabolism and the underlying mechanisms remain elusive. In the present study, we examined the inhibitory effect of the active compound “(−)-loliolide ((6S,7aR)-6-hydroxy-4,4,7a-trimethyl-5,6,7,7a-tetrahydro-1-benzofuran-2(4H)-one (HTT))” from S. horneri extract on lipid accumulation in differentiated adipocytes. MTT assays demonstrated that (−)-loliolide is not toxic to 3T3-L1 adipocytes in a range of concentrations. (−)-loliolide significantly reduced intracellular lipid accumulation in the differentiated phase of 3T3-L1 adipocytes as shown by Oil Red O staining. Western blot analysis revealed that (−)-loliolide increased the expression of lipolytic protein phospho-hormone-sensitive lipase (p-HSL) and thermogenic protein peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1). Additionally, (−)-loliolide decreased expression of adipogenic and lipogenic proteins, including sterol regulatory element-binding protein-1 (SREBP-1), peroxisome proliferator-activated receptor-γ (PPAR-γ), CCAAT/enhancer-binding protein-α (C/EBP-α), and fatty acid-binding protein 4 (FABP4) in 3T3-L1 adipocytes. These results indicate that (−)-loliolide from S. horneri could suppress lipid accumulation via regulation of antiadipogenic and prolipolytic mechanisms in 3T3-L1 cells. Considering the multifunctional effect of (−)-loliolide, it can be useful as a lipid-lowering agent in the management of patients who suffer from obesity.     

EGCG通过PI3K-AKT信号通路调控FOXO1磷酸化抑制3T3-L1前脂肪细胞分化

为探究EGCG对3T3-L1前脂肪细胞分化的影响及其分子机制,以3T3-L1前脂肪细胞为材料,分别添加不同浓度EGCG(0μM、5μM、10μM、50μM、100μM),通过油红O染色检测EGCG对3T3-L1脂肪细胞分化过程中脂质积累的影响,同时采用RT-PCR和免疫印迹检测不同浓度EGCG对PPARγ、FAS、PI3K、FOXO1的mRNA及蛋白表达的影响.油红O染色结果表明,EGCG可抑制3T3-L1前脂肪细胞分化,阻碍脂质积累,并具有浓度效应,EGCG浓度越高抑制作用越明显;EGCG能够直接抑制PPARγ和FAS的表达,同时抑制P-AKT、P-PI3K以及P-FOXO1的表达.表明EGCG通过介导PI3K/AKT-FOXO1级联信号通路,下调具有促进前脂肪细胞分化功能的PPARγ和FAS的表达,从而抑制前脂肪细胞分化,阻碍脂肪生成.     

Shrimp Oil Extracted from Shrimp Processing By-Product Is a Rich Source of Omega-3 Fatty Acids and Astaxanthin-Esters,and Reveals Potential Anti-Adipogenic Effects in 3T3-L1 Adipocytes

The province of Newfoundland and Labrador, Canada, generates tons of shrimp processing by-product every year. Shrimp contains omega (n)-3 polyunsaturated fatty acids (PUFA) and astaxanthin (Astx), a potent antioxidant that exists in either free or esterified form (Astx-E). In this study, shrimp oil (SO) was extracted from the shrimp processing by-product using the Soxhlet method (hexane:acetone 2:3). The extracted SO was rich in phospholipids, n-3 PUFA, and Astx-E. The 3T3-L1 preadipocytes were differentiated to mature adipocytes in the presence or absence of various treatments for 8 days. The effects of SO were then investigated on fat accumulation, and the mRNA expression of genes involved in adipogenesis and lipogenesis in 3T3-L1 cells. The effects of fish oil (FO), in combination with Astx-E, on fat accumulation, and the mRNA expression of genes involved in adipogenesis and lipogenesis were also investigated. The SO decreased fat accumulation, compared to untreated cells, which coincided with lower mRNA expression of adipogenic and lipogenic genes. However, FO and FO + Astx-E increased fat accumulation, along with increased mRNA expression of adipogenic and lipogenic genes, and glucose transporter type 4 (Glut-4), compared to untreated cells. These findings have demonstrated that the SO is a rich source of n-3 PUFA and Astx-E, and has the potential to elicit anti-adipogenic effects. Moreover, the SO and FO appear to regulate adipogenesis and lipogenesis via independent pathways in 3T3-L1 cells.     

Effects of Ethanol Extracts from Grateloupia elliptica,a Red Seaweed,and Its Chlorophyll Derivative on 3T3-L1 Adipocytes: Suppression of Lipid Accumulation through Downregulation of Adipogenic Protein Expression

Grateloupia elliptica (G. elliptica) is a red seaweed with antioxidant, antidiabetic, anticancer, anti-inflammatory, and anticoagulant activities. However, the anti-obesity activity of G. elliptica has not been fully investigated. Therefore, the effect of G. elliptica ethanol extract on the suppression of intracellular lipid accumulation in 3T3-L1 cells by Oil Red O staining (ORO) was evaluated. Among the eight red seaweeds tested, G. elliptica 60% ethanol extract (GEE) exhibited the highest inhibition of lipid accumulation. GEE was the only extract to successfully suppress lipid accumulation among ethanol extracts from eight red seaweeds. In this study, we successfully isolated chlorophyll derivative (CD) from the ethyl acetate fraction (EA) of GEE by high-performance liquid chromatography and evaluated their inhibitory effect on intracellular lipid accumulation in 3T3-L1 adipocytes. CD significantly suppressed intracellular lipid accumulation. In addition, CD suppressed adipogenic protein expression such as sterol regulatory element-binding protein-1 (SREBP-1), peroxisome proliferator-activated receptor-γ (PPAR-γ), CCAAT/enhancer-binding protein-α (C/EBP-α), and fatty acid binding protein 4 (FABP4). Taken together, our results indicate that CD from GEE inhibits lipid accumulation by suppressing adipogenesis via the downregulation of adipogenic protein expressions in the differentiated adipocytes. Therefore, chlorophyll from G. elliptica has a beneficial effect on lipid metabolism and it could be utilized as a potential therapeutic agent for preventing obesity.     

Effects of Low Root Temperature on Dry Matter Production and Root Water Uptake in Rice Plants

Abstract

Chilling is a major constraint in rice production in cool climates. In rice (Oryza sativa L.) plants, both the air temperature and the water (soil) temperature affect various growth processes independently, and low root zone temperature (thus, root temperature) can inhibit rice growth and yield. In this study, we investigated the effect of low root temperature on rice growth in relation to dry matter production and root water uptake. Plants were grown in hydroponic solutions at two temperatures, one equivalent to air temperature and the other 14ºC for 15 d starting 11 d after germination. Low temperature of the solution (low root temperature) inhibited dry matter production of rice plants by decreasing leaf area rather than photosynthetic rate. The response of leaf area was affected by changes in plant water status, that is relative water content (RWC) of stem was decreased by low root temperature resulting in reduced leaf area. The decrease in RWC caused by low root temperature was related to that in root hydraulic conductance (K_r). The responses of transpiration (E) and K_r to the low root temperature depended more on root surface area than on changes in hydraulic conductance per unit root surface area (Lp_r). These results suggest that dry matter production under the low root temperature condition is controlled mainly by quantitative growth parameters such as leaf area and root surface area.     

生防菌BRF-1和BRF-2对大豆根腐病和产量的影响

BRF-1和BRF-2是中国科学院东北地理与农业生态农田分子生态学科组从大豆根际土壤筛选得到的2株革兰氏阳性生防细菌,为了检测它们对田间大豆根腐病的防治效果,2010年在沈阳军区龙镇农副业基地对BRF-1和BRF-2进行田间试验。结果表明:2株生防细菌均有效地抑制了大豆根腐病的发生,发病程度分别降低44.4%和8.8%,BRF-1菌株效果较好,在降低大豆根腐病病情指数的同时,能增加大豆植株花期、始粒期地上部和地下部干物重,提高大豆产量5.9%,是一株很有潜力的生防菌。     

ABA和AgNO3对小麦幼胚愈伤组织诱导和分化的影响

为解决河南小麦生产上三个主导品种（豫麦18号、豫麦49号、兰考906）生物技术改良的技术瓶颈问题。研究了不同浓度的ABA和AgNO3处理对上述三个小麦品种幼胚愈伤组织诱导和分化特性的影响。结果表明。添加0．1mg／LABA的诱导培养基以及在添加0．1mg／l ABA的基础上再添加2．5mg／L或5．0mg／LAgNO3的诱导培养基对幼胚愈伤组织形成和分化最为有利,但不同基因型在相同的处理下幼胚愈伤组织诱导率和分化率仍存在较大的差异。     

生物炭对玉米根系特性及产量的影响

采用大田试验研究生物炭对郑单958产量和不同生育时期根系生长、形态特征及生理特性的影响,明确生物炭对玉米根系与产量的效应,探明生物炭在玉米生产上应用的潜力与价值。结果表明,土壤中施入生物炭能增加玉米根系的总根长、根体积和根干重,提高玉米根系总吸收面积和活跃吸收面积。在玉米生育后期,生物炭在一定程度上延缓了根系衰老,成熟期施生物炭处理根系活力分别比对照高48.12%和42.71%;同时维持了较为适宜的根冠比,根系生理功能增强;添加生物炭后郑单958最高产量达12 076.35 kg/hm2,比对照增产10.23%,具体表现为穗长、穗行数、行粒数和百粒重的提高。     

增氧对水稻根系生长与氮代谢的影响

 以国稻6号(杂交籼稻)和秀水09(粳稻)为材料,在水培条件下研究通气增氧对水稻苗期根系生长和氮代谢的影响。结果表明,增氧处理水稻根系干物质积累量、根长、根体积、根系活力以及吸收面积较对照均有不同程度增加。水稻根系硝态氮（NO3 N）含量、游离氨基酸和可溶性糖含量均增加。谷氨酸合成酶（GS）、谷草转氨酶（GOT）和谷丙转氨酶（GPT）活性均增强。在本试验条件下,增氧能显著提高粳稻秀水09苗期根系氮代谢相关酶生理活性;但增氧对杂交籼稻国稻6号根系氮代谢相关酶生理活性影响不显著,说明通气增氧对水稻根系氮代谢生理活性的影响可能与水稻基因型有关。     

in vitro Anti-Proliferative Effect of Adiponectin on Human Endometriotic Stromal Cells through AdipoR1 and AdipoR2 Gene Receptor Expression

Background:

Endometriosis is a complex disorder in reproductive age women which consist of stromal and epithelial cells implantation outside the uterine cavity. Adiponectin is a member of cytokine family with various metabolic roles and proliferation inhibition of many cancer cells. The aim of the present research was to determine adiponectin effect on human endometriotic stromal cells (ESCs) proliferation and their expression of adiponectin receptors.

Methods:

In this experimental study, endometrial biopsies (n=7) were taken. ESCs isolation was done by enzymatic digestion and cell filtrations. ESCs of each biopsy were divided into four groups: 0 (control), 10, 100, and 200 ng/ml adiponectin concentrations in three different times (24, 48, or 72 h). The effect of adiponectin on ESC viability and expression of mRNA Adipo receptor1 (R1) and Adipo receptor2 (R2) was determined by Trypan blue staining and semi-quantitative RT-PCR, respectively. Data were analyzed by one-way ANOVA and unpaired student’s t-test, and P<0.05 was considered statistically significant.

Results:

Adiponectin inhibited human endometriotic stromal cell proliferation in time- and dose-dependent manners significantly (P=0.001). Expression of AdipoR1 and AdipoR2 gene receptors was increased in human ESCs significantly (P<0.05).

Conclusions:

Adiponectin can suppress endometriosis by inhibiting ESC proliferation and increased AdipoR1 and AdipoR2 expression.Key Words: Adiponectin, Adiponectin receptors, Endometriosis, Stromal cells     

The Effect of Human Platelet-Rich Plasma on Adipose-Derived Stem Cell Proliferation and Osteogenic Differentiation

Background: The cultured mesenchymal stem cells (MSC) have been used in many clinical trials; however, there are still some concerns about the cultural conditions. One concern is related to the use of FBS as a widely used xenogeneic supplement in the culture system. Human platelet-rich plasma (hPRP) is a candidate replacement for FBS. In this study, the effect of hPRP on MSC proliferation and osteogenic differentiation has been evaluated. Methods: Human adipose-derived stem cells (hADSC) were expanded. Cells from the third passage were characterized by flow cytometric analysis and used for in vitro experiments. Resazurin and alizarin red stains were used for cell proliferation and osteogenic differentiation assays, respectively. Results: Treatment with hPRP resulted in a statistically significant increase in cell proliferation compare to the negative control group (P<0.001). Cell proliferation in the 15% hPRP group was also significantly higher than that in the 10% hPRP group (P<0.05). Additionally, it caused less osteogenic differentiation of the hADSC compared to the FBS (P<0.001), but in comparison to negative control, it caused acceptable mineralization (P<0.001). Conclusion: These findings indicate that hPRP not only improves the proliferation but also it can be a suitable substitution in osteogenic differentiation for clinical purposes. However, the clinical application value of hPRP still needs more investigation. Key Words: Platelet-Rich Plasma, Adipose tissue, Stem Cells, Cell differentiation, Cell proliferation     

宿根栽培年限对桂西南香蕉生产的影响及管理建议

分析目前广西桂西南香蕉主产区不同宿根栽培年限蕉园在香蕉生长、蕉园管理难度、蕉园成本收益上的差异,提出限制延长蕉园宿根栽培年限的主要因素是种植密度的显著下降：宿根栽培2年的蕉园经济收益最高,随着宿根栽培年限的延长,蕉园经济收益逐年下降。就此提出合理延长蕉园宿根栽培年限的管理措施。     

根系局部供氮对水稻根系形态的影响及其机理

 通过砂培试验，研究了水稻根系和地上部分（苗）对局部根系供应氮素的适应性变化。局部根系供应氮素对水稻根系形态，生物量及碳、氮分配有明显的影响。无论供氮量高低，供氮一侧均能诱导根系生长发育，表现为供氮一侧侧根长、根表面积、根系体积、根重等明显高于无氮侧，但是根系直径明显变细。供氮侧根系碳水化合物含量高于无氮侧；随供氮量的增加，碳水化合物含量相对减少。根据试验结果，认为局部供应氮素诱导根系生长发育的可能机理是无氮一侧根系的饥饿信号诱导地上部碳水化合物定向运输和分配到供氮侧的根系，从而促进该侧根系的生长发育。     

施肥方式对玉米根系分布及产量的影响

采用土柱模拟法研究施肥方式对玉米根系分布及产量的影响,结果表明,垄沟追肥促使玉米根系向纵深发展,与传统的垄面撒施覆土相比,垄沟施肥3cm深处理的中层根和深层根干重分别增加8.9%～9.4%和26.6%～45.8%,总根重增加6.0%～8.5%,而百粒重和产量增加不显著.垄沟施肥8cm的中层根和深层根干重分别增加19.6%～22.6%和59.3%～91.7%,总根重增加12.3%～13.8%,百粒重增加13.9%～16.3%,产量增加10.6%～11.8%.表明玉米追肥的最佳方式为垄沟深追肥。