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1.
Diabetes Metab Syndr Obes ; 16: 565-574, 2023.
Article in English | MEDLINE | ID: mdl-36883138

ABSTRACT

Background: Gestational diabetes mellitus (GDM) is the most common pregnant disorder worldwide. In this study, we aimed to explore whether vitamin E (VE) treatment alone could protect against GDM in a mouse model. Methods: 6-week-old C57BL/6J female mice were fed on high-fat diet for two weeks and continued with high-fat diet after pregnancy to induce GDM. The pregnant mice were orally administrated with 2.5, 25 or 250 mg/kg VE twice per day during pregnancy together with high-fat diet. Oral glucose tolerance test, insulin amounts, oxidative stress and inflammation were then measured. Results: Only 250 mg/kg VE could improve glucose tolerance and insulin level in pregnant mice. VE (250 mg/kg) effectively inhibited GDM-induced hyperlipidemia, and secretion of inflammatory cytokines such as tumor necrosis factor-α and interleukin-6. VE also significantly ameliorated maternal oxidative stress at the late stage of pregnancy, and also improved reproductive outcomes, including increasing the litter size and birth weight in GDM mice. Moreover, VE also activated GDM-reduced nuclear factor-erythroid factor 2-related factor 2 (Nrf2) / heme oxygenase-1 signaling pathway in the maternal liver tissues of GDM mice. Conclusion: Our data clearly demonstrated that 250 mg/kg VE twice a day during pregnancy could significantly ameliorate the symptoms of GDM by alleviating oxidative stress, inflammation, hyperglycemia, and hyperlipidemia through Nrf2/HO-1 signaling pathway in GDM mice. Thus, additional VE supplement might be beneficial to GDM.

2.
Environ Sci Pollut Res Int ; 30(3): 5758-5773, 2023 Jan.
Article in English | MEDLINE | ID: mdl-35978248

ABSTRACT

To obtain systematic knowledge on the waterborne pollution status and ecological and human health risk of total petroleum hydrocarbons (TPHs) and metals in the southeastern Bohai Sea, seawater samples were collected in three seasons from 2014 to 2018. TPHs and mercury (Hg) levels were determined by ultraviolet spectrophotometry and cold atomic absorption spectrometry, respectively, and concentrations of copper (Cu), lead (Pb), and cadmium (Cd) were detected by anodic stripping voltammetry. Spatial distribution patterns indicated that these waterborne pollutants are mainly sourced from terrestrial inputs. Temporal variation showed that Pb contents decreased in the past five years, and summer exhibited higher concentrations of Hg, Cu, and Cd than spring and autumn. Spearman's rank correlation coefficients demonstrated that temperature correlated positively with Cu content, while dissolved oxygen, pH, and suspended particulate material correlated negatively with pollutant concentrations. While hazard quotient values were lower than 1 for TPHs, Hg, Pb, and Cd, the hazard quotient of Cu (4.88) was greater than 1, suggesting potential ecological risks of this element in seawater of the southeastern Bohai Sea. The total target hazard quotients of Hg, Cu, Pb, and Cd in seawater of the southeastern Bohai Sea were all lower than 1, which indicated that there was no noncarcinogenic risk caused by heavy metals in seawater of the southeastern Bohai Sea. However, the carcinogenic risk of Cd (1.54 × 10-5) was in the range of 10-6-10-4, which may lead to the occurrence of cancer. This study sounds an alarm for stricter control of metal emissions into this sea area.


Subject(s)
Mercury , Metals, Heavy , Petroleum , Water Pollutants, Chemical , Humans , Cadmium/analysis , Petroleum/analysis , Lead/analysis , Water Pollutants, Chemical/analysis , Environmental Monitoring/methods , Seawater/chemistry , Metals, Heavy/analysis , Mercury/analysis , China , Hydrocarbons/analysis , Risk Assessment
3.
Phytomedicine ; 99: 154022, 2022 May.
Article in English | MEDLINE | ID: mdl-35255283

ABSTRACT

BACKGROUND: Silibinin, a major component of milk thistle extract silymarin, promotes hypoglycemia by activating estrogen receptor (ER) α and ß-mediated pathways in pancreatic ß-cells. Glucagon-like peptide-1 (GLP-1) is the enteroendocrine peptide produced in L-cells, and it controls glucose homeostasis through multiple pathways. The effect of silibinin on L-cell mass and function is still unknown. PURPOSE: The protective effect of silibinin on palmitate (PA)-treated intestinal L-cell line GLUTag cells and the SHRSP•Z-Leprfa/Izm-Dmcr (SP•ZF) diabetic rat model was investigated in current study. METHODS: After pre-incubation with 50 µM silibinin for 4 h, GLUTag cells were treated with 0.125 mM PA. MTT, Annexin V/PI apoptosis, Hoechst 33342 staining, western blot, DCFH-DA, GLP-1 ELISA, qRT-PCR and immunofluorescence analyses were undertaken to determine ER-dependent protection of silibinin against PA-induced cellular damage. The differential protein expression of GLUTag cells under different treatments was examined by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). The SP•ZF diabetic rat model was chosen for in vivo study. After 4 weeks of gastric gavage with 100 or 300 mg kg-1 of silibinin, the physiological indexes of the rats were measured. Cells expressing GLP-1, 8­hydroxy-2'-deoxyguanosine (8-OHdG), ERα, and/or ERß in duodenum tissues were detected by immunofluorescence. RESULTS: The current study showed that the GLUTag cells preincubated with silibinin activated the transcription factor nuclear erythroid-2 like factor-2 (Nrf2)-antioxidant pathway, reduced reactive oxygen species (ROS) generation, and improved cell survival and GLP-1 content, while the antioxidative effect of silibinin was blocked by the selective ERα antagonist MPP or ERß antagonist PHTPP in GLUTag cells. Our proteomics data further revealed that ERα or ß inactivation reduced glutathione peroxide and proteins associated with endocytosis and reproduction, thus at least partially reversing the protective effect of silibinin. SP•ZF rats received silibinin treatment showed increased serum GLP-1 content and improved glucose homeostasis. Furthermore, silibinin upregulated ERα and ß levels and reduced the level of 8-OHdG in GLP-1-positive cells. CONCLUSIONS: Our study showed that silibinin improved L-cell mass and function through an ER-mediated antioxidant pathway, and the proteomics analysis revealed for the first time the differential regulation of proteins by PA and silibinin in GLUTag cells.

4.
J Proteomics ; 260: 104574, 2022 05 30.
Article in English | MEDLINE | ID: mdl-35354085

ABSTRACT

Protein phosphorylation plays an important role in animal reproduction. However, its role in the onset of puberty in goats remains largely unexplored. Accordingly, in the present study, the molecular changes controlling the onset of puberty in goats were investigated by identifying the differentially phosphorylated proteins (DPPs) and phosphorylation sites (DPSs) in the hypothalami of prepubertal and pubertal female goats using LC-MS/MS and tandem mass tag labelling. A total of 3265 phosphopeptides corresponding to 1628 phosphoproteins were identified, including 234 upregulated and 342 downregulated phosphopeptides. The DPSs HTT, MAP1B, CAMKK1, MAP2, DNAJC5, and GAP43 were identified. These DPPs are enriched in the endocytosis, cAMP signaling, Rap1 signaling, melanogenesis, and insulin secretion pathways. These pathways are related to gonadotropin-releasing hormone and puberty. In particular, glucose-6-phosphate isomerase, fructose-bisphosphate aldolase C, and fructose-bisphosphate aldolase A occupy important locations in the protein-protein interaction network. These data provide evidence for a complex interaction network in goat hypothalamus proteins that affects puberty. Furthermore, they may help identify new puberty-regulating candidates and/or serve as an important resource for exploring the physiological mechanism of puberty onset in mammals. SIGNIFICANCE: This study provides evidence for a complex interaction network in goat hypothalamus proteins that affects puberty. Furthermore, our data may help identify new puberty-regulating candidates and/or serve as an important resource for exploring the physiological mechanism of puberty onset in mammals.


Subject(s)
Goats , Phosphopeptides , Animals , Chromatography, Liquid , Female , Fructose-Bisphosphate Aldolase/metabolism , Goats/metabolism , Hypothalamus/metabolism , Phosphopeptides/metabolism , Phosphorylation , Tandem Mass Spectrometry
5.
J Cell Mol Med ; 25(24): 11278-11289, 2021 12.
Article in English | MEDLINE | ID: mdl-34786818

ABSTRACT

Oxidative stress (OS)-induced mitochondrial damage and the subsequent osteoblast dysfunction contributes to the initiation and progression of osteoporosis. Notoginsenoside R1 (NGR1), isolated from Panax notoginseng, has potent antioxidant effects and has been widely used in traditional Chinese medicine. This study aimed to investigate the protective property and mechanism of NGR1 on oxidative-damaged osteoblast. Osteoblastic MC3T3-E1 cells were pretreated with NGR1 24 h before hydrogen peroxide administration simulating OS attack. Cell viability, apoptosis rate, osteogenic activity and markers of mitochondrial function were examined. The role of C-Jun N-terminal kinase (JNK) signalling pathway on oxidative injured osteoblast and mitochondrial function was also detected. Our data indicate that NGR1 (25 µM) could reduce apoptosis as well as restore osteoblast viability and osteogenic differentiation. NGR1 also reduced OS-induced mitochondrial ROS and restored mitochondrial membrane potential, adenosine triphosphate production and mitochondrial DNA copy number. NGR1 could block JNK pathway and antagonize the destructive effects of OS. JNK inhibitor (SP600125) mimicked the protective effects of NGR1while JNK agonist (Anisomycin) abolished it. These data indicated that NGR1 could significantly attenuate OS-induced mitochondrial damage and restore osteogenic differentiation of osteoblast via suppressing JNK signalling pathway activation, thus becoming a promising agent in treating osteoporosis.


Subject(s)
Ginsenosides/pharmacology , MAP Kinase Signaling System/drug effects , Osteoblasts/drug effects , Osteoblasts/metabolism , Oxidative Stress/drug effects , Adenosine Triphosphate/metabolism , Animals , Apoptosis/drug effects , Biomarkers , Cell Line , Cell Survival/drug effects , Mice , Mitochondria/drug effects , Mitochondria/metabolism , Superoxides/metabolism
6.
BMC Complement Altern Med ; 19(1): 111, 2019 May 30.
Article in English | MEDLINE | ID: mdl-31146723

ABSTRACT

BACKGROUND: Atherosclerosis is a condition with the vascular accumulation of lipid plaques, and its main major contributing factor is endothelial injury induced by oxidized low-density lipoprotein (ox-LDL). Salidroside (SAL) is the primary active ingredient of Rhodiola rosea, and exhibits antioxidant properties on endothelial cells and alleviates atherosclerosis. However, the effect of SAL on autophagy in ox-LDL-induced vascular endothelial injury remains unclear. Here, we investigated the effect and underlying mechanisms of SAL on autophagy in human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs were incubated with ox-LDL to induce in vitro atherosclerosis model. The cell viability and injury were evaluated by cell counting kit-8 (CCK-8) assay and lactate dehydrogenase (LDH) release assay. The oxidative stress was evaluated by NADPH oxidase, malondialdehyde (MDA) and superoxide dismutase (SOD) activities. Immunofluorescence was performed to detect autophagy using LC3ß antibody. Quantitative real-time PCR (qRT-PCR) and western blot were performed to measure the mRNA expressions of SIRT1 and Forkhead box O1 (FOXO1). Nicotinamide (NAM) and AS1842856 were used to inhibit activities of SIRT1 and FOXO1, respectively. RESULTS: Exposure of HUVECs to ox-LDL (100 µg/mL) reduced cell viability, increased cellular MDA, and reduced SOD in a concentration-dependent manner. The pretreatment with SAL (20, 50 and 100 µM) significantly enhanced the cell viability and decreased LDH release in HUVECs exposed to ox-LDL (100 µg/mL). ox-LDL induced autophagy in HUVECs, which was further enhanced by pretreatment with SAL. However, SAL attenuated increase in oxidative stress in HUVECs induced by ox-LDL. ox-LDL reduced mRNA and protein expressions of SIRT1 and FOXO1, which could be reversed by SAL. The protective, anti-oxidative and pro-autophagic effects of SAL could be obviously abolished by cotreatment with SIRT1 inhibitor or FOXO1 inhibitor. CONCLUSION: Salidroside shows protective effect on endothelial cell induced by ox-LDL, and the mechanisms might be related to autophagy induction via increasing SIRT1 and FoxO1 expressions.


Subject(s)
Autophagy/drug effects , Endothelial Cells/drug effects , Glucosides/pharmacology , Phenols/pharmacology , Plant Extracts/pharmacology , Rhodiola , Atherosclerosis/prevention & control , Drug Evaluation, Preclinical , Endothelial Cells/metabolism , Forkhead Box Protein O1/metabolism , Glucosides/therapeutic use , Human Umbilical Vein Endothelial Cells , Humans , Lipoproteins, LDL , Oxidative Stress/drug effects , Phenols/therapeutic use , Phytotherapy , Plant Extracts/therapeutic use , Sirtuin 1/metabolism
7.
Reprod Biol Endocrinol ; 15(1): 81, 2017 Oct 06.
Article in English | MEDLINE | ID: mdl-28985764

ABSTRACT

BACKGROUND: There are many variables affecting the onset of puberty in animals, including genetic, nutritional, and environmental factors. Recent studies suggest that epigenetic regulation, especially DNA methylation, plays a majorrole in the regulation of puberty. However, there have been no reports on DNA methylation of the pubertal genome. METHODS: We investigated DNA methylation in the female rat hypothalamus at prepuberty and puberty using reduced representation bisulfite sequencing technology. The identified genes and signaling pathways exhibiting changes to DNA methylation in pubertal rats were determined by Gene Ontogeny and Kyoto Encyclopedia of Genes and Genomes analysis. RESULTS: The distribution of the three types of methylated C bases in promoter and CpG island (CGI) regions in the hypothalamus was as follows: 87.79% CG, 3.05% CHG, 9.16% CHH for promoters, and 88.35% CG, 3.21% CHG, 88.35% CHH for CGI in prepubertal rats; and 90.78% CG, 2.13% CHG, 7.09% CHH for promoters, and 88.59% CG, 88.59% CHG, 8.35% CHH for CGI in pubertal animals. CG showed the highest percentage of methylation, and was the highest methylation state in CGI. Compared to prepubertal hyoyhalamus samples, we identified ten genes with altered methylation in promoter regions in the pubertal hypothalamus samples, and 43 genes with altered methylation in the CGI. Changes in DNA methylation were found in gonadotropin-releasing hormone signaling pathways, and the oocyte meiosis pathway. CONCLUSION: Our results demonstrate changes in DNA methylation occur in female rats from prepuberty to puberty suggestng DNA methylation may play a crucial role in the regulation of puberty onset. This study provides essential information for future studies on the role of epigenetics in the regulation of puberty.


Subject(s)
DNA Methylation , Epigenesis, Genetic , Hypothalamus/metabolism , Promoter Regions, Genetic , Sexual Maturation/genetics , Animals , CpG Islands , Female , Gonadotropin-Releasing Hormone/genetics , Rats , Sequence Analysis, DNA/methods , Sulfites/chemistry
8.
Anim Sci J ; 88(9): 1298-1310, 2017 Sep.
Article in English | MEDLINE | ID: mdl-28349625

ABSTRACT

Melatonin (MLT) is an endogenous hormone with roles in animal germ cell development. However, the effect of MLT on porcine oocyte maturation and its underlying mechanisms remain largely unknown. Here, we investigated the effects of exogenous MLT on oocyte maturation, histone acetylation, autophagy and subsequent embryonic development. We found that 1 nmol/L MLT supplemented in maturation medium was the optimal concentration to promote porcine oocyte maturation and subsequent developmental competence and quality of parthenogenetic embryos. Interestingly, the beneficial effects of 1 nmol/L MLT treatment on porcine oocyte maturation and embryo development were mainly attributed to the first half period of in vitro maturation. Simultaneously, MLT treatment could also improve maturation of small follicle-derived oocytes, morphologically poor (cumulus cell layer ≤1) and even artificially denuded oocytes and their subsequent embryo development. Furthermore, MLT treatment not only could decrease the levels of H3K27ac and H4K16ac in metaphase II (MII) oocytes, but also could increase the expression abundances of genes associated with cumulus cell expansion, meiotic maturation, histone acetylation and autophagy in cumulus cells or MII oocytes. These results indicate that MLT treatment can facilitate porcine oocyte maturation and subsequent embryonic development probably, through improvements in histone acetylation and autophagy in oocytes.


Subject(s)
Acetylation/drug effects , Autophagy/drug effects , Embryonic Development/drug effects , Histones/metabolism , Melatonin/pharmacology , Oocytes/growth & development , Oocytes/physiology , Animals , Cells, Cultured , Female , In Vitro Oocyte Maturation Techniques , Oocytes/metabolism , Stimulation, Chemical , Swine
9.
PLoS One ; 11(10): e0165327, 2016.
Article in English | MEDLINE | ID: mdl-27788248

ABSTRACT

Female pubertal development is tightly controlled by complex mechanisms, including neuroendocrine and epigenetic regulatory pathways. Specific gene expression patterns can be influenced by DNA methylation changes in the hypothalamus, which can in turn regulate timing of puberty onset. In order to understand the relationship between DNA methylation changes and gene expression patterns in the hypothalamus of pubertal goats, whole-genome bisulfite sequencing and RNA-sequencing analyses were carried out. There was a decline in DNA methylation levels in the hypothalamus during puberty and 268 differentially methylated regions (DMR) in the genome, with differential patterns in different gene regions. There were 1049 genes identified with distinct expression patterns. High levels of DNA methylation were detected in promoters, introns and 3'-untranslated regions (UTRs). Levels of methylation decreased gradually from promoters to 5'-UTRs and increased from 5'-UTRs to introns. Methylation density analysis demonstrated that methylation level variation was consistent with the density in the promoter, exon, intron, 5'-UTRs and 3'-UTRs. Analyses of CpG island (CGI) sites showed that the enriched gene contents were gene bodies, intergenic regions and introns, and these CGI sites were hypermethylated. Our study demonstrated that DNA methylation changes may influence gene expression profiles in the hypothalamus of goats during the onset of puberty, which may provide new insights into the mechanisms involved in pubertal onset.


Subject(s)
DNA Methylation , Goats/genetics , Hypothalamus/metabolism , Animals , CpG Islands/genetics , Female , Gene Expression Regulation, Developmental , Goats/growth & development , Hypothalamus/growth & development
10.
Sci Rep ; 6: 32877, 2016 09 07.
Article in English | MEDLINE | ID: mdl-27599613

ABSTRACT

Nesfatin-1 is an important molecule in the regulation of reproduction. However, its role in the reproductive axis in male animals remains to be understood. Here, we found that nesfatin-1 was mainly distributed in the arcuate nucleus (ARC), paraventricular nucleus (PVN), periventricular nucleus (PeN), and lateral hypothalamic area (LHA) of the hypothalamus; adenohypophysis and Leydig cells in male rats. Moreover, the concentrations of serum nesfatin-1 and its mRNA in hypothalamo-pituitary-gonadal axis (HPGA) vary with the age of the male rat. After intracerebroventricular injection of nesfatin-1, the hypothalamic genes for gonadotrophin releasing hormone (GnRH), kisspeptin (Kiss-1), pituitary genes for follicle-stimulate hormone ß(FSHß), luteinizing hormone ß(LHß), and genes for testicular steroidogenic acute regulatory (StAR) expression levels were decreased significantly. Nesfatin-1 significantly increased the expression of genes for 3ß-hydroxysteroid dehydrogenase (3ß-HSD), 17ß-hydroxysteroid dehydrogenase (17ß-HSD), and cytochrome P450 cleavage (P450scc) in the testis of pubertal rats, but their levels decreased in adult rats (P < 0.05), along with the serum FSH, LH, and testosterone (T) concentrations. After nesfatin-1 addition in vitro, T concentrations of the supernatant were significantly higher than that in the control group. These results were suggestive of the role of nesfatin-1 in the regulation of the reproductive axis in male rats.


Subject(s)
Calcium-Binding Proteins/physiology , DNA-Binding Proteins/physiology , Hypothalamus/metabolism , Leydig Cells/metabolism , Nerve Tissue Proteins/physiology , Pituitary Gland, Anterior/metabolism , 17-Hydroxysteroid Dehydrogenases/metabolism , Animals , Arcuate Nucleus of Hypothalamus/metabolism , Follicle Stimulating Hormone/metabolism , Follicle Stimulating Hormone, beta Subunit/metabolism , Hypothalamus, Posterior/metabolism , Luteinizing Hormone/metabolism , Male , Nucleobindins , Paraventricular Hypothalamic Nucleus/metabolism , Pituitary Gland/metabolism , Rats , Testis/metabolism , Testosterone/metabolism
11.
Foodborne Pathog Dis ; 13(4): 196-204, 2016 Apr.
Article in English | MEDLINE | ID: mdl-26919471

ABSTRACT

Cronobacter sakazakii is an opportunistic pathogen transmitted by food that affects mainly newborns, infants, and immune-compromised adults. In this study, the antibacterial activity of ferulic acid was tested against C. sakazakii strains. Minimum inhibitory concentration of ferulic acid against C. sakazakii strains was determined using the agar dilution method. Changes in intracellular pH, membrane potential and intracellular ATP concentration were measured to elucidate the possible antibacterial mechanism. Moreover, SYTO 9 nucleic acid staining was used to assess the effect of ferulic acid on bacterial membrane integrity. Cell morphology changes were observed under a field emission scanning electron microscope. The minimum inhibitory concentrations of ferulic acid against C. sakazakii strains ranged from 2.5 to 5.0 mg/mL. Addition of ferulic acid exerted an immediate and sustained inhibition of C. sakazakii proliferation. Ferulic acid affected the membrane integrity of C. sakazakii, as evidenced by intracellular ATP concentration decrease. Moreover, reduction of intracellular pH and cell membrane hyperpolarization were detected in C. sakazakii after exposure to ferulic acid. Reduction of green fluorescence indicated the injury of cell membrane. Electronic microscopy confirmed that cell membrane of C. sakazakii was damaged by ferulic acid. Our results demonstrate that ferulic acid has moderate antimicrobial activity against C. sakazakii. It exerts its antimicrobial action partly through causing cell membrane dysfunction and changes in cellular morphology. Considering its antimicrobial properties, together with its well-known nutritional functions, ferulic acid has potential to be developed as a supplement in infant formula or other foods to control C. sakazakii.


Subject(s)
Anti-Bacterial Agents/metabolism , Coumaric Acids/metabolism , Cronobacter sakazakii/metabolism , Dietary Supplements , Food Preservatives/metabolism , Adenosine Triphosphate/metabolism , Anti-Bacterial Agents/pharmacology , Cell Membrane/drug effects , Cell Membrane/metabolism , Cell Membrane/ultrastructure , Cell Proliferation/drug effects , China , Colony Count, Microbial , Coumaric Acids/pharmacology , Cronobacter sakazakii/drug effects , Cronobacter sakazakii/growth & development , Cronobacter sakazakii/ultrastructure , Drug Resistance, Bacterial , Food Preservatives/pharmacology , Humans , Hydrogen-Ion Concentration , Infant , Infant Food/microbiology , Infant Formula/microbiology , Intracellular Fluid/drug effects , Intracellular Fluid/metabolism , Membrane Potentials/drug effects , Microbial Sensitivity Tests , Microbial Viability/drug effects , Microscopy, Electron, Scanning , Species Specificity
12.
Theriogenology ; 84(9): 1556-64, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26365586

ABSTRACT

The aim of this study was to assess whether changes in kisspeptin and GnRH levels could be attributed to sex steroids at puberty onset. We used the ovariectomy (OVX) model in rats treated with 17ß-estradiol (E2; OVX + E2), or oil (OVX + oil), and in intact rats treated with E2 (intact + E2) or oil only (intact + oil) to determine gene expression changes of Kiss1 and Gnrh1 in the hypothalamus and protein expression of kisspeptin and GnRH in the different areas of the hypothalamus. In the intact + E2 and OVX + E2 rats on the day of the onset of puberty, GnRH-immunoreactive (ir) cell numbers decreased (P < 0.05) in the arcuate nucleus but were increased in the preoptic area; Kisspeptin-ir cells increased (P < 0.05) in the arcuate nucleus, periventricular nucleus, and preoptic area; no difference (P > 0.05) was found in the paraventricularis nucleus for GnRH-ir or kisspeptin-ir cells. Additionally, levels of Kiss1 and Gnrh1 messenger RNA in the hypothalamus were significantly higher (P < 0.05) in the OVX + E2 or intact + E2 rats than in the OVX + oil or intact + oil animals, respectively. In the OVX + oil rats, OVX significantly increased (P < 0.05) levels of Gnrh1 and Kiss1 messenger RNA and the expression of GnRH and kisspeptin in the hypothalamus compared to intact + oil animals. These results suggest that kisspeptin and GnRH play major roles in modulating the activity of estrogen circuits at the onset of puberty.


Subject(s)
Estradiol/pharmacology , Estrogens/pharmacology , Gonadotropin-Releasing Hormone/metabolism , Hypothalamus/metabolism , Kisspeptins/metabolism , Sexual Maturation/physiology , Animals , Estradiol/administration & dosage , Estradiol/blood , Female , Gene Expression Regulation/physiology , Gonadotropin-Releasing Hormone/genetics , Kisspeptins/genetics , Male , Ovariectomy , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , Real-Time Polymerase Chain Reaction
13.
Theriogenology ; 81(4): 545-55, 2014 Mar 01.
Article in English | MEDLINE | ID: mdl-24368149

ABSTRACT

The genetic manipulation of spermatogonial stem cells (SSCs) can be used for the production of transgenic animals in a wide range of species. However, this technology is limited by the absence of an ideal culture system in which SSCs can be maintained and proliferated, especially in domestic animals like the goat. The aim of this study therefore was to investigate whether the addition of vitamin C (Vc) in cell culture influences the growth of caprine SSCs. Various concentrations of Vc (0, 5, 10, 25, 40, and 50 µg/mL(-1)) were added to SSC culture media, and their effect on morphology and alkaline phosphatase activity was studied. The number of caprine SSC colonies and area covered by them were measured at 10 days of culture. The expression of various germ cell and somatic cell markers such as VASA, integrins, Oct-4, GATA-4, α-SMA, vimentin, and Thy-1 was studied to identify the proliferated cells using immunostaining analyses. Further, the intracellular reactive oxygen species (ROS) level was measured at the 3rd, 6th, and 9th day after culture, and expression of Bax, Bcl-2, and P53, factors involved in the regulation of apoptosis, were analyzed on the 7th day after culture using reverse transcription polymerase chain reaction and quantitative real-time polymerase chain reaction. The results showed that the SSCs formed compact colonies and had unclear borders in the different Vc-supplemented groups at 10 days, and there were no major morphologic differences between the groups. The number and area of colonies were both the highest in the 40 µg/mL(-1) Vc group. Differential expression of markers for germ cells, undifferentiated spermatogonia, and testis somatic cells was observed. Cultured germ cell clumps were found to have alkaline phosphatase activity regardless of the Vc dose. The number of Thy-1- and Oct-4-positive cells was the most in the 40 µg/mL(-1) Vc group. Moreover, the level of ROS was dependent on the Vc dose and culture time. The Vc dose 40 µg/mL(-1) was found to be optimum with regard to decreasing ROS generation, and increasing the expression of the antiapoptotic gene Bcl-2 and decreasing the expression of the proapoptotic genes Bax and P53. In conclusion, the addition of 40 µg/mL(-1) Vc can maintain a certain physiological level of ROS, trigger the expression of the antiapoptosis gene Bcl-2, suppress the proapoptotic gene P53 and Bax pathway, and further promote the proliferation of caprine SSCs in vitro.


Subject(s)
Ascorbic Acid/pharmacology , Cell Culture Techniques/veterinary , Cell Differentiation/physiology , Goats/metabolism , Spermatogonia/metabolism , Stem Cells/metabolism , Alkaline Phosphatase/analysis , Animals , Apoptosis/physiology , Cell Culture Techniques/methods , Cell Differentiation/drug effects , Flow Cytometry/veterinary , GATA4 Transcription Factor/analysis , Immunohistochemistry/veterinary , Integrins/analysis , Male , Octamer Transcription Factor-3/analysis , Spermatogonia/cytology , Spermatogonia/ultrastructure , Stem Cells/cytology , Stem Cells/ultrastructure , Thy-1 Antigens/analysis , Vimentin/analysis
14.
Zhong Yao Cai ; 36(11): 1748-51, 2013 Nov.
Article in Chinese | MEDLINE | ID: mdl-24956812

ABSTRACT

OBJECTIVE: Based on the important medicinal applications of artemisinic acid and the superiority of Thin Layer Chromagraphy (TLC), the spot area method of TLC was presented to determine the content changes of artemisinic acid of Artemisia annua at different growing stages. METHODS: The separation conditions including chromatographic solutions and chromogenic agent were optimized. The detection limit and the linear concentration range were analyzed. And the content changes of artemisinic acid of Artemisia annua at different growing stages were detected. RESULTS: The results showed that artemisinic acid extracted from Artemisia annua could be separated completely by the chromatographic solutions composed by petroleum ether,acetone and ethyl acetate (80: 19: 1). The artemisinic acid was clearly colored using the chromogenic agent consisting by ethanol, bromophenol blue and sulfuric acid. The detection limit of TLC was 0.05 mg/mL. The spot area of TLC had a good linear relationship within the range of 0.05-0.6 mg/mL, accorded with regression equation of y = 11.162 x + 0.0823. The results showed that the content of artemisinic acid at 0.041 mg/g in April which below the detection limit of TLC had no color spot. Contrarily, the spots of artemisinic acid were obvious in materials growing from May to September, and content was about 0.7, 1.2, 2.1, 2.4 and 2.7 mg/g, respectively corresponding to results by HPLC. CONCLUSION: The method can be applied to the quantitative analysis of artemisinic acid in Artemisia annua.


Subject(s)
Artemisia annua/chemistry , Artemisia annua/growth & development , Artemisinins/analysis , Artemisinins/isolation & purification , Chromatography, High Pressure Liquid , Chromatography, Thin Layer , Plant Extracts/analysis , Plant Extracts/isolation & purification , Plants, Medicinal/chemistry , Plants, Medicinal/growth & development , Reproducibility of Results , Seasons , Sensitivity and Specificity , Solvents/chemistry
15.
Zhong Yao Cai ; 35(12): 1914-7, 2012 Dec.
Article in Chinese | MEDLINE | ID: mdl-23705352

ABSTRACT

OBJECTIVE: To study the variation of content of Artemisic acid of Artemisia annua from eight areas of four provinces around Wuling Mountain. METHODS: Artemisic acid of plants were extracted by organic solvent method. Qualitative and quantitative analysis of Artemisic acid were measured by thin layer chromatography (TLC) and high performance thin layer chromatography (HPLC), respectively. RESULTS: The results showed the average levels of Artemisic acid in May and August changed from 0.964 to 2.288 mg/g and from 1.837 to 3.737 mg/g, respectively. The average level in August was 1.5 times as that in May. The Artemisic acid in cultured plants was higher than the levels in wild plants, and Artemisic acid in plant collected below 300 m altitude was higher than that of the plant collected above 300 m altitude. CONCLUSION: The biosynthesis of Artemisic acid depends on the plant growth stage,which is mainly accumulated in plant at the mature stage.


Subject(s)
Artemisia annua/chemistry , Artemisinins/analysis , Plants, Medicinal/chemistry , Altitude , Artemisia annua/growth & development , Artemisinins/isolation & purification , Chromatography, High Pressure Liquid , Chromatography, Thin Layer , Plants, Medicinal/growth & development , Seasons
16.
Nanotechnology ; 22(35): 355603, 2011 Sep 02.
Article in English | MEDLINE | ID: mdl-21828895

ABSTRACT

In this paper, a pH-inductive protein-scaffold biosynthesis of shape-tunable crystalline gold nanoparticles at room temperature has been developed. By simple manipulation of the reaction solution's pH, anisotropic gold nanoparticles including spheres, triangles and cubes could be produced by incubating an aqueous solution of sodium tetrachloroaurate with Dolichomitriopsis diversiformis biomasses after immersion in ultrapure Millipore water overnight. A moss protein with molecular weight of about 71 kDa and pI of 4.9 was the primary biomolecule involved in the biosynthesis of gold nanoparticles. The secondary configuration of the proteins by CD spectrum implied that the moss protein could display different secondary configurations including random coil, α-helix and intermediate conformations between random coil and α-helix for the experimental pH solution. The growth process of gold nanoparticles further showed that the moss protein with different configurations provided the template scaffold for the shape-controlled biosynthesis of gold nanoparticles. The constrained shape of the gold nanoparticles, however, disappeared in boiled moss extract. The gold nanoparticles with designed morphology were successfully reconstructed using the moss protein purified from the gold nanoparticles. Structural characterizations by SEM, TEM and SAED showed that the triangular and cubic gold nanoparticles were single crystalline.


Subject(s)
Crystallization/methods , Gold/chemistry , Metal Nanoparticles/chemistry , Plant Extracts/chemistry , Plant Proteins/chemistry , Bryopsida/chemistry , Bryopsida/metabolism , Chlorides/chemistry , Chlorides/metabolism , Circular Dichroism , Color , Electrophoresis, Polyacrylamide Gel , Gold/metabolism , Gold Compounds/chemistry , Gold Compounds/metabolism , Hydrogen-Ion Concentration , Isoelectric Focusing , Metal Nanoparticles/ultrastructure , Microscopy, Electron , Particle Size , Plant Extracts/metabolism , Plant Proteins/metabolism , Protein Conformation , Spectrophotometry, Ultraviolet , Time Factors
17.
Colloids Surf B Biointerfaces ; 73(1): 75-9, 2009 Oct 01.
Article in English | MEDLINE | ID: mdl-19481910

ABSTRACT

The design, synthesis, characterization and application of biologically synthesized nanomaterials have become an important branch of nanotechnology. In this paper, we report the extracellular synthesis of gold nanoparticles using Barbated Skullcup (BS) herb (a dried whole plant of Scutellaria barbata D. Don) as the reducing agent. After exposing the gold ions to BS herb extract, rapid reduction of gold ions is observed leading to the formation of gold nanoparticles in solution. UV-vis spectrum of the aqueous medium containing gold nanoparticles showed a peak at around 540 nm. Transmission electron microscopy (TEM) micrograph analysis of the gold nanoparticles indicated that they were well-dispersed and ranged in size 5-30 nm. When the gold nanoparticles were modified on the glassy carbon electrode (GCE), it could enhance electronic transmission rate between the electrode and the p-nitrophenol.


Subject(s)
Electrochemistry/methods , Gold/chemistry , Metal Nanoparticles/chemistry , Plant Extracts/pharmacology , Electrodes , Metal Nanoparticles/ultrastructure , Microscopy, Electron, Transmission , Nitrophenols/chemistry , Oxidation-Reduction/drug effects , Particle Size , Scutellaria/chemistry , Spectrophotometry, Ultraviolet
18.
J Asian Nat Prod Res ; 7(2): 145-9, 2005 Apr.
Article in English | MEDLINE | ID: mdl-15621617

ABSTRACT

From the rhizoma of Cimicifuga foetida L. (Ranunculaceae) a new chromone, 6'-hydroxylangelicain (18), has been isolated together with 20 known compounds. The structure of 18 has been elucidated on the basis of spectroscopic and chemical evidence.


Subject(s)
Chromones/isolation & purification , Cimicifuga/chemistry , Chromatography, Gel , Chromones/chemistry , Drugs, Chinese Herbal/chemistry , Drugs, Chinese Herbal/isolation & purification , Molecular Structure , Nuclear Magnetic Resonance, Biomolecular , Optical Rotation , Periodic Acid/chemistry , Rhizome/chemistry , Spectrometry, Mass, Electrospray Ionization , Spectrophotometry, Infrared , Spectrophotometry, Ultraviolet
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