Barley Protein LFBEP-C1 from Lactiplantibacillus plantarum dy-1 Fermented Barley Extracts by Inhibiting Lipid Accumulation in a Caenorhabditis elegans Model

Objective This study aimed to investigate the lipid-lowering activity of LFBEP-C1 in high glucose-fed Caenorhabditis elegans(C.elegans).Methods In this study,the fermented barley protein LFBEP-C1 was prepared and tested for its potential anti-obesity effects on C.elegans.The worms were fed Escherichia coli OP50(E.coli OP50),glucose,and different concentrations of LFBEP-C1.Body size,lifespan,movement,triglyceride content,and gene expression were analyzed.The results were analyzed using ANOVA and Tukey's multiple comparison test.Results Compared with the model group,the head-swing frequency of C.elegans in the group of LFBEP-C1 at 20μg/mL increased by 33.88%,and the body-bending frequency increased by 27.09%.This indicated that LFBEP-C1 improved the locomotive ability of C.elegans.The average lifespan of C.elegans reached 13.55 days,and the body length and width of the C.elegans decreased after LFBEP-C1 intake.Additionally,LFBEP-C1 reduced the content of lipid accumulation and triglyceride levels.The expression levels of sbp-1,daf-2,and mdt-15 significantly decreased,while those of daf-16,tph-1,mod-1,and ser-4 significantly increased after LFBEP-C1 intake.Changes in these genes explain the signaling pathways that regulate lipid metabolism.Conclusion LFBEP-C1 significantly reduced lipid deposition in C.elegans fed a high-glucose diet and alleviated the adverse effects of a high-glucose diet on the development,lifespan,and exercise behavior of C.elegans.In addition,LFBEP-C1 regulated lipid metabolism mainly by mediating the expression of genes in the sterol regulatory element-binding protein,insulin,and 5-hydroxytryptamine signaling pathways.

Network Pharmacology and Experimental Study of MomordicineⅠand MomordicineⅡfrom Bitter Melon Saponins in Inhibiting Fat Accumulation

Bitter melon(Momordica Charania L.),a member of the Cucurbitaceae family,is widely distributed across tropical and subtropical regions.Saponin,an important functional component of bitter melon,has been proven to exert hypoglycemic effects similarly to insulin,and also possesses lipid-lowering properties inhibiting preadipocyte differentiation and fat synthesis^([1]).

LonP1表达与线粒体稳态以及精子活力的关联性研究

目的探究线粒体lon蛋白水解酶(LonP1)对精子线粒体稳态的维持调节作用及其可能存在的调节机制。方法收集2016年6月至2017年6月于温州医科大学附属第一医院生殖中心进行精液常规检查的120名男性精液标本,根据精子活力分为正常活力精子(NS)和弱精子症精子(AS)两组,采用计算机辅助精液分析系统(CASA)检测各组精子运动参数,通过免疫荧光法和免疫蛋白印迹法检测精子中LonP1的蛋白表达水平,流式细胞术检测各组精子的线粒体膜电位强度(MMP,以绿色荧光强度/红色荧光强度比值表示,比值越小代表膜电位越高)和线粒体内活性氧(mROS)水平,比较两组各指标参数差异,并分析LonP1表达水平和精子活力及精子线粒体功能相关指标(MMP,mROS)的关联性。另外,NS组精液经密度梯度离心法处理后,沉淀精子加入培养液重悬,以LonP1功能抑制剂CDDO-ME 2.5μmoL/L和5μmol/L为终浓度进行体外培养,同时设置空白对照组(0μmol/L),比较各组精子运动参数、LonP1表达水平以及精子线粒体功能相关指标。结果 (1)人类成熟精子中存在LonP1的表达,且稳定表达于精子中段的线粒体鞘部;(2)NS组和AS组精子LonP1表达水平(0.52vs.0.22)与精子活力(69.89%vs.26.10%)、MMP(4.72%vs.12.73%)呈正相关(P<0.05),与mROS含量呈负相关(418.85vs.460.52)(P<0.05);(3)经LonP1功能抑制剂CDDO-ME处理后,与空白对照组比较,CDDO-ME药物处理组的LonP1蛋白表达水平随CDDO-ME作用浓度增加而显著降低(P<0.05),5μmol/L浓度组的MMP水平显著下降(78.81%vs.7.34%)、mROS含量增加(640.68vs.462.26),差异均有统计学意义(P<0.05);(4)CDDO-ME处理后,除精子活率(PR+NP)外,精子运动参数平均路径(VAP)和曲线速率(VSL)均随CDDO-ME作用浓度增加而下降(P<0.05),与空白对照组相比,5μmol/L浓度组的精子活力(PR)(52.08%vs.65.15%)、曲线速率(VCL)(48.28μm/s vs.78.78μm/s)、侧摆幅度(ALH)(1.28μm vs.1.93μm)、直线性(LIN)(38.83%vs.56.05%)和前向性(STR)(45.53%vs.81.25%)均显著下降,差异有统计学意义(P<0.05)。结论精子线粒体鞘部存在LonP1的稳定表达,LonP1能够通过精子线粒体稳态维持改善精子活力。

Antitumor Activities and Apoptosis-regulated Mechanisms of Fermented Wheat Germ Extract in the Transplantation Tumor Model of Human HT-29 Cells in Nude Mice

Objective A subcutaneous transplantation tumor model of human HT-29 cells in nude mice was established to evaluate anticarcinogenic activities, and the apoptosis-regulated mechanism effect of aqueous extract of fermented wheat germ with Lactobacillus plantarum dy-1 （LFWGE）. Methods The HT-29 cells were transplanted via subcutaneous injection of 1×10^7cells into the right flank of each nude mouse. Then, nude mice were treated for 30 d with LFWGE （high-dose 2 g/kg/d; low-dose 1 g/kg/d） and for 7 d with 5-fluorouracil （5-FU, 25 mg/kg/d） by gavage and intraperitoneal injection, respectively. An inhibition of tumor growth was observed. Results Tumor volume and weights decreased significantly in both groups of nude mice treated with LFWGE. In addition, the cell apoptosis rate of the LFWGE group （2 g/kg/d, 60.2%＋4.4%; 1 g/kg/d, 58.6%＋6.9%） was significantly higher than that of the control group （11.5%＋1.6%） and 5-FU group （32.1%＋3.5%） as measured by the TUNEL assay. Moreover, the real-time fluorescent quantitative PCR and Western blot method further confirmed these enhancing apoptosis and growth inhibition effects. The involvement of LFWGE in inducing apoptosis was confirmed by the expression of Bax, Bcl-2, Caspase-3, and CyclinD1. Conclusion The results showed that LFWGE could induce subcutaneous transplantation tumor apoptosis in nude mice and could be as a natural nutrient supplements or chemopreventive agent in the treatment of human colon cancer.

Supplementation of Fermented Barley Extracts with Lactobacillus Plantarum dy-1 Inhibits Obesity via a UCP1-dependent Mechanism

Objective We aimed to explore how fermented barley extracts with Lactobacillus plantarum dy-1(LFBE) affected the browning in adipocytes and obese rats.Methods In vitro, 3T3-L1 cells were induced by LFBE, raw barley extraction(RBE) and polyphenol compounds(PC) from LFBE to evaluate the adipocyte differentiation.In vivo, obese SD rats induced by high fat diet(HFD) were randomly divided into three groups treated with oral gavage:(a) normal control diet with distilled water,(b) HFD with distilled water,(c) HFD with 800 mg LFBE/kg body weight(bw).Results In vitro, LFBE and the PC in the extraction significantly inhibited adipogenesis and potentiated browning of 3T3-L1 preadipocytes, rather than RBE.In vivo, we observed remarkable decreases in the body weight, serum lipid levels, white adipose tissue(WAT) weights and cell sizes of brown adipose tissues(BAT) in the LFBE group after 10 weeks.LFBE group could gain more mass of interscapular BAT(IBAT) and promote the dehydrogenase activity in the mitochondria.And LFBE may potentiate process of the IBAT thermogenesis and epididymis adipose tissue(EAT) browning via activating the uncoupling protein 1(UCP1)-dependent mechanism to suppress the obesity.Conclusion These results demonstrated that LFBE decreased obesity partly by increasing the BAT mass and the energy expenditure by activating BAT thermogenesis and WAT browning in a UCP1-dependent mechanism.

Antitumor Activities and Apoptosis-regulated Mechanisms of Fermented Barley Extract in the Transplantation Tumor Model of Human HT-29 Cells in Nude Mice

Objective A subcutaneous transplantation tumor model of human HT-29 cells was established in nude mice to study the anticarcinogenic activities and apoptosis-regulatory mechanistic effect of aqueous extract of fermented barley with Lactobacillus plantarum dy-1 （LFBE）. Methods HT-29 cells were transplanted via subcutaneous injection of 1 × 107cells into the right flank of each nude mouse. Then, nude mice were treated for 30 days with LFBE （high-dose 2 g·kg-1·d-1; low-dose 1 g·kg-1·d-1） and for 7 days with 5-fluorouracil （5-FU, 25 g·kg-1·d-1） by gavage and intraperitoneal injection, respectively. Results Tumor volume and weight decreased significantly in both groups of nude mice treated with LFBE. In addition, the cell apoptosis rate of the LFBE group was significantly higher than that of the control group and 5-FU groups as measured by the TUNEL assay. Moreover, the real-time fluorescent quantitative PCR and Western blot methods further confirmed these apoptosis-enhancing and growth-inhibiting effects. The involvement of LFBE in inducing apoptosis was confirmed by the expression of Bax, Bcl-2, caspase-3, and cyclin D1. Conclusion The results showed that LFBE could induce subcutaneous transplantation tumor apoptosis in nude mice and could be used as a natural nutrient supplement or chemopreventive agent in the treatment of human colon cancer.

Fermented Barley Extracts with Lactobacillus plantarum dy-1 Rich in Vanillic Acid Modulate Glucose Consumption in Human HepG2 Cells

Objective To investigate the effect of fermented barley extracts with Lactobacillus plantarum dy-1（LFBE） for modulating glucose consumption in HepG2 cells via miR-212 regulation. Methods Hepatocellular carcinoma（HepG2） cells were treated with palmitate. After 12 h, palmitate-induced HepG2 cells were treated with LFBE and its main components. Changes in glucose consumption, proinflammatory cytokine secretion, and miRNA-212 expression in HepG2 cells was observed. Results Treatment with LFBE rich in vanillic acid（VA） increased glucose consumption and reduced proinflammatory cytokine secretion in HepG2 cells. LFBE and VA normalized the upregulation of miR-212, which led to the upregulation of dual-specificity phosphatase-9（DUSP9）, a direct target of miR-212, at both protein and mR NA levels. Downregulation of miR-212 markedly increased glucose consumption and reduced proinflammatory cytokine secretion by enhancing DUSP9 expression. Conclusion The results showed the benefit of LFBE and miR-212 downregulation in modulating glucose consumption and reducing proinflammatory cytokine secretion by targeting DUSP9. VA in LFBE was a strong regulator of palmitate-induced abnormal glucose consumption in HepG2 cells and can be a primary mediator.

Anti-obesity Action of Fermented Barley Extracts with Lactobacillus plantarum dy-1 and Associated Micro RNA Expression in High-fat Diet-induced Obese Rats

Objective To further explore associated effects of Lactobacillus plantarum dy-1(LFBE) on obesity and lipid metabolism at the gene expression level, the expression of micro RNAs(mi RNAs) was investigated in the liver of high-fat diet(HFD) induced obese rats.Methods Three groups of animal models were established. Changes in mi RNA expression in the liver of each group were analyzed by microarray and RT-q PCR, complemented by bioinformatics. Palmitateinduced hepatocellular carcinoma(Hep G2) cells were used as a model to validate the test.Results LFBE treatment groups and HFD groups were observed to be distinctly different with respect to rates of increase in body weight and body fat percentage and triglyceride(TG) and total cholesterol(TC) levels in serum and liver. In addition, the LFBE group showed upregulation of ten mi RNAs and downregulation of five mi RNAs in the liver. Downregulation of mi R-34 a and mi R-212 was observed in the livers of the LFBE group. Gene ontology and kyoto encyelopedia of geues and genomes(KEGG)pathway analysis showed that possible target genes of the deregulated mi RNAs were significantly enriched in the adrenergic and HIF-1 signaling pathways.Conclusion These results demonstrate that LFBE might regulate the expression of mi RNAs in order to inhibit obesity and fatty liver.