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1.
J Ethnopharmacol ; 331: 118237, 2024 Sep 15.
Article in English | MEDLINE | ID: mdl-38688355

ABSTRACT

ETHNOPHARMACOLOGICAL RELEVANCE: Bu-Zhong-Yi-Qi Decoction(BZYQD) is a traditional formula commonly used in China, known for its effects in tonifying Qi and raising Yang. It can relieve symptoms of cognitive impairment such as forgetfulness and lack of concentration caused by qi deficiency, which is common in aging and debilitating. However, much of the current research on BZYQD has been focused on its impact on the digestive system, leaving its molecular mechanisms in improving cognitive function largely unexplored. AIM OF THE STUDY: Cognitive decline in the aging central nervous system is intrinsically linked to oxidative damage. This study aims to investigate the therapeutic mechanism of BZYQD in treating mild cognitive impairment caused by qi deficiency, particularly through repair of mitochondrial oxidative damage. MATERIALS AND METHODS: A rat model of mild cognitive impairment (MCI) was established by administering reserpine subcutaneously for two weeks, followed by a two-week treatment with BZYQD/GBE. In vitro experiments were conducted to assess the effects of BZYQD on neuronal cells using a H2O2-induced oxidative damage model in PC12 cells. The open field test and the Morris water maze test evaluated the cognitive and learning memory abilities of the rats. HE staining and TEM were employed to observe morphological changes in the hippocampus and its mitochondria. Mitochondrial activity, ATP levels, and cellular viability were measured using assay kits. Protein expression in the SIRT3/MnSOD/OGG1 pathway was analyzed in tissues and cells through western blotting. Levels of 8-OH-dG in mitochondria extracted from tissues and cells were quantified using ELISA. Mitochondrial morphology in PC12 cells was visualized using Mito Red, and mitochondrial membrane potential was assessed using the JC-1 kit. RESULTS: BZYQD treatment significantly improved cognitive decline caused by reserpine in rats, as well as enhanced mitochondrial morphology and function in the hippocampus. Our findings indicate that BZYQD mitigates mtDNA oxidative damage in rats by modulating the SIRT3/MnSOD/OGG1 pathway. In PC12 cells, BZYQD reduced oxidative damage to mitochondria and mtDNA in H2O2-induced conditions and was associated with changes in the SIRT3/MnSOD/OGG1 pathway. CONCLUSION: BZYQD effectively counteracts reserpine-induced mild cognitive impairment and ameliorates mitochondrial oxidative stress damage through the SIRT3/MnSOD/OGG1 pathway.


Subject(s)
Cognitive Dysfunction , Drugs, Chinese Herbal , Mitochondria , Oxidative Stress , Rats, Sprague-Dawley , Sirtuin 3 , Superoxide Dismutase , Animals , Cognitive Dysfunction/drug therapy , Cognitive Dysfunction/metabolism , Oxidative Stress/drug effects , Drugs, Chinese Herbal/pharmacology , Mitochondria/drug effects , Mitochondria/metabolism , Rats , PC12 Cells , Male , Sirtuin 3/metabolism , Superoxide Dismutase/metabolism , Signal Transduction/drug effects , Disease Models, Animal , Hippocampus/drug effects , Hippocampus/metabolism , Neuroprotective Agents/pharmacology , Sirtuins
2.
Aging Dis ; 2024 Jan 05.
Article in English | MEDLINE | ID: mdl-38270115

ABSTRACT

Alzheimer's disease (AD) is the most prevalent form of dementia and is characterized by progressive degeneration of brain function. AD gradually affects the parts of the brain that control thoughts, language, behavior and mental function, severely impacting a person's ability to carry out daily activities and ultimately leading to death. The accumulation of extracellular amyloid-ß peptide (Aß) and the aggregation of intracellular hyperphosphorylated tau are the two key pathological hallmarks of AD. AD is a complex condition that involves both non-genetic risk factors (35%) and genetic risk factors (58-79%). The glymphatic system plays an essential role in clearing metabolic waste, transporting tissue fluid, and participating in the immune response. Both non-genetic and genetic risk factors affect the glymphatic system to varying degrees. The main purpose of this review is to summarize the underlying mechanisms involved in the deregulation of the glymphatic system during the progression of AD, especially concerning the diverse contributions of non-genetic and genetic risk factors. In the future, new targets and interventions that modulate these interrelated mechanisms will be beneficial for the prevention and treatment of AD.

3.
Int J Biol Macromol ; 201: 93-103, 2022 Mar 15.
Article in English | MEDLINE | ID: mdl-34973980

ABSTRACT

Hyperbranched polysaccharides (HBPSs) are the main components in cell wall and exopolysaccharide (EPS) of Pleurotus tuber-regium. To enhance the yield of these macromolecules, corn oil at 4% addition exhibited the best effect for production of mycelial biomass at 20.49 g/L and EPS at 0.59 g/L, which was 2.56 folds and 1.90 folds of the control, respectively. The treated hyphae were much thicker with smooth surface, while its cell wall content (43.81 ± 0.02%) was 1.96 times of the control (22.34 ± 0.01%). Moreover, a large number of lipid droplets could be visualized under the view of confocal laser scanning microscopy (CLSM). RNA-seq analysis revealed that corn oil could enter the cells and result in the up-regulation of genes on cell morphology and membrane permeability, as well as the down-regulation on expression level of polysaccharide hydrolase and genes involved in the MAPK pathway, all of which probably contribute to the increase of polysaccharides production.


Subject(s)
Corn Oil , Pleurotus , Biomass , Mycelium/metabolism , Pleurotus/metabolism , Polysaccharides/metabolism
4.
Zhongguo Zhong Yao Za Zhi ; 46(23): 6216-6223, 2021 Dec.
Article in Chinese | MEDLINE | ID: mdl-34951248

ABSTRACT

This study aims to explore the effect of extract of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma, and Chuanxiong Rhizoma(hereinafter referred to as GNS) on the SIRT1-autophagy pathway of endothelial cell senescence induced by hydrogen peroxide(H_2O_2). To be specific, vascular endothelial cells were classified into the blank control group(control), model group(model), model + DMSO group(DMSO), resveratrol group(RESV), and GNS low-dose(GNS-L), medium-dose(GNS-M), and high-dose(GNS-H) groups. They were treated with H_2O_2 for senescence induction except the control. After intervention of cells in each group with corresponding drugs for 24 h, cell growth status was observed under an inverted microscope, and the formation of autophagosome under the transmission electron microscope. In addition, the changes of microtubule-associated protein 1 light chain 3ß(LC3 B) were detected by immunofluorescence staining. The autophagy flux was tracked with the autophagy double-labeled adenovirus(mRFP-GFP-LC3) fusion protein. Dansylcadaverine(MDC) staining was employed to determine the autophagic vesicles, and Western blot the expression of sirtuin 1(SIRT1), ubiquitin-binding protein p62, and LC3Ⅱ. After H_2O_2 induction, cells demonstrated slow growth, decreased adhesion ability, raised number of SA-ß-gal-stained blue ones, a certain number of autophagosomes with bilayer membrane and secondary lysosomes in the cytoplasm, and slight rise of autophagy flux level. Compared with the model group, GNS groups showed improved morphology, moderate adhesion ability, complete and smooth membrane, decreased SA-ß-gal-stained blue cells, many autophagosomes, autophagic vesicles, and secondary lysosomes in the cytoplasm, increased autophagolysosomes, autophagy flux level, and fluorescence intensity of LC3 B and MDC, up-regulated expression of SIRT1 and LC3Ⅱ, and down-regulated expression of p62, suggesting the improvement of autophagy level. GNS can delay the senescence of vascular endothelial cells. After the intervention, the autophagy flux and related proteins SIRT1, LC3Ⅱand p62 changed significantly, and the autophagy level increased significantly. However, EX527 weakened the effect of Chinese medicine in delaying vascular senescence. GNS may delay the senescence of vascular endothelial cells through the SIRT1 autophagy pathway.


Subject(s)
Autophagy , Drugs, Chinese Herbal/pharmacology , Endothelial Cells/drug effects , Panax , Cells, Cultured , Cellular Senescence , Hydrogen Peroxide , Panax/chemistry , Sirtuin 1/genetics
5.
Zhongguo Zhong Yao Za Zhi ; 45(12): 2938-2946, 2020 Jun.
Article in Chinese | MEDLINE | ID: mdl-32627470

ABSTRACT

The aim of this paper was to observe the changes of intestinal flora in vascular aging mice, in order to explore the relationship between vascular aging and intestinal flora and the effects of extracts of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma on intestinal flora of vascular aging mice. A model of vascular aging in mice was induced through intrape-ritoneal injection with streptozotocin(STZ) combined with high-fat diet. Biochemical detection was performed on serum cholesterol(CHO), triglyceride(TG), high-density liptein cholesterol(HDL-C), low-density liptein cholesterol(LDL-C) and blood glucose(GLU). HE staining was used to detect mice thoracic aorta morphology, and the expressions of cyclin-dependent kinase inhibitor 2 A(p16) and cyclin-dependent kinase inhibitor 1 A(p21) protein in mice thoracic aorta were detected by Western blot. The 16 S rDNA gene of mice intestinal flora was detected by Illumina MiSeq high-throughput sequencing technology to explore the changes of intestinal flora in each group. The results demonstrated that the GLU level in low-dose and high-dose TCM groups decreased, but with unobvious changes in blood lipid indexes. Metformin could significantly decrease the levels of GLU(P<0.01), CHO and LDL-C in mice(P<0.05). Intravascular injury was not obvious in each drug group, and the expressions of p16 and p21 protein were significantly decreased(P<0.05). The intestinal flora of each group was mainly composed of Firmicutes(F) and Bacteroidetes(B) at the level of the phylum, but the B/F ratio was different from that of the youth group and the blank control group. The B/F ratio of the model group was significantly lower(P<0.01), and compared with the model group, the B/F ratio of the high-dose group and the metformin group was signi-ficantly higher(P<0.05). There were dominant and differential floras in the intestine of each group of mice. The results showed that extracts of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma could improve the intestinal flora structure and create a good intestinal environment by increasing the B/F ratio, which provides a new possible pathway for lowering blood glucose and blood lipids and delaying vascular aging.


Subject(s)
Drugs, Chinese Herbal , Gastrointestinal Microbiome , Panax , Aging , Animals , Glucose , Lipids , Mice
6.
Biomed Environ Sci ; 32(3): 231-233, 2019 Mar.
Article in English | MEDLINE | ID: mdl-30987699

ABSTRACT

On December 14, 2017, a faculty member of a university in Hunan Province reported that an anthrax vaccine strain might have recovered virulence during an undergraduate experiment and potential exposure could not be ruled out for the students involved. Upon receiving the case report, the CDC, health bureaus, and local governments at the county, prefectural, and provincial levels promptly organized experts in different fields (including epidemiologists, biosafety experts, and laboratory testing experts) for case investigation, evaluation, and response. As the investigation results showed, no virulence recovery was identified in the involved anthrax vaccine strain; and no contamination of Bacillus anthracis was detected at the involved areas. Thus, the university returned to normal functioning.


Subject(s)
Anthrax Vaccines/analysis , Bacillus anthracis/pathogenicity , Containment of Biohazards , China , Humans , Laboratories/statistics & numerical data , Virulence
7.
Article in English | MEDLINE | ID: mdl-29277917

ABSTRACT

Ericerus pela Chavannes (Hemiptera: Coccoidae) is an economically important scale insect because the second instar males secrete a harvestable wax-like substance. In this study, we report the molecular cloning of a fatty acyl-CoA reductase gene (EpFAR) of E. pela. We predicted a 520-aa protein with the FAR family features from the deduced amino acid sequence. The EpFAR mRNA was expressed in five tested tissues, testis, alimentary canal, fat body, Malpighian tubules, and mostly in cuticle. The EpFAR protein was localized by immunofluorescence only in the wax glands and testis. EpFAR expression in High Five insect cells documented the recombinant EpFAR reduced 26-0:(S) CoA and to its corresponding alcohol. The data illuminate the molecular mechanism for fatty alcohol biosynthesis in a beneficial insect, E. pela.


Subject(s)
Aldehyde Oxidoreductases/genetics , Hemiptera/genetics , Aldehyde Oxidoreductases/metabolism , Amino Acid Sequence , Animals , Hemiptera/enzymology , Male , Phylogeny , Sequence Analysis, DNA
8.
International Eye Science ; (12): 2252-2255, 2017.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-669407

ABSTRACT

·Cataract refers to the degenerative changes in lens quality caused by various causes of reduced transparency or color change. Surgical treatment is the main treatment modality at present. Among them, phacoemulsification has become the main surgical treatment for cataract because of its advantages such as short operation time, small incision and quicker healing. Today, cataract surgery has gradually shift to refractive surgery, and is no longer simply as cataract surgery. But after cataract phacoemulsification, the symptoms and visual quality are different. The main causes include refractive error, postoperative dry eyes and postoperative corneal astigmatism. This article reviews the factors that influence the visual quality of cataract phacoemulsification and its future trends.

9.
Plant Cell Rep ; 34(9): 1629-46, 2015 Sep.
Article in English | MEDLINE | ID: mdl-26021845

ABSTRACT

KEY MESSAGE: Silicon enhances root water uptake in salt-stressed cucumber plants through up-regulating aquaporin gene expression. Osmotic adjustment is a genotype-dependent mechanism for silicon-enhanced water uptake in plants. Silicon can alleviate salt stress in plants. However, the mechanism is still not fully understood, and the possible role of silicon in alleviating salt-induced osmotic stress and the underlying mechanism still remain to be investigated. In this study, the effects of silicon (0.3 mM) on Na accumulation, water uptake, and transport were investigated in two cucumber (Cucumis sativus L.) cultivars ('JinYou 1' and 'JinChun 5') under salt stress (75 mM NaCl). Salt stress inhibited the plant growth and photosynthesis and decreased leaf transpiration and water content, while added silicon ameliorated these negative effects. Silicon addition only slightly decreased the shoot Na levels per dry weight in 'JinYou 1' but not in 'JinChun 5' after 10 days of stress. Silicon addition reduced stress-induced decreases in root hydraulic conductivity and/or leaf-specific conductivity. Expressions of main plasma membrane aquaporin genes in roots were increased by added silicon, and the involvement of aquaporins in water uptake was supported by application of aquaporin inhibitor and restorative. Besides, silicon application decreased the root xylem osmotic potential and increased root soluble sugar levels in 'JinYou 1.' Our results suggest that silicon can improve salt tolerance of cucumber plants through enhancing root water uptake, and silicon-mediated up-regulation of aquaporin gene expression may in part contribute to the increase in water uptake. In addition, osmotic adjustment may be a genotype-dependent mechanism for silicon-enhanced water uptake in plants.


Subject(s)
Cucumis sativus/physiology , Plant Roots/metabolism , Salt Tolerance/drug effects , Silicon/pharmacology , Water/metabolism , Aquaporins/genetics , Aquaporins/metabolism , Biological Transport , Biomass , Carbohydrates/analysis , Cucumis sativus/drug effects , Cucumis sativus/genetics , Cucumis sativus/growth & development , Dithiothreitol/pharmacology , Gene Expression Regulation, Plant/drug effects , Genes, Plant , Mercuric Chloride/pharmacology , Osmosis/drug effects , Photosynthesis/drug effects , Plant Leaves/drug effects , Plant Leaves/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Plant Roots/drug effects , Plant Roots/genetics , Plant Stomata/drug effects , Plant Stomata/physiology , Plant Transpiration/drug effects , Quantitative Trait, Heritable , Salt Tolerance/genetics , Seedlings/drug effects , Seedlings/physiology , Sodium/metabolism , Solubility , Xylem/drug effects , Xylem/physiology
10.
Int J Ophthalmol ; 7(1): 38-43, 2014.
Article in English | MEDLINE | ID: mdl-24634861

ABSTRACT

AIM: To investigate the protective effects of the natural medicinal monomer ecdysterone (ECR) with estrogenic activity against oxidative damage in human lens epithelial cells B3 (HLE-B3) caused by hydrogen peroxide 21(H2O2) and to pursue the possible mitochondrial proteomic regularity of the protective effects. METHODS: HLE-B3 cells were treated with H2O2 (300µmol/L), ß-estuarial (E2; 10(-8)mol/L) and H2O2, ECR (10(-6)mol/L) and H2O2, or left untreated. Altered expression of all mitochondrial proteins was analyzed by protein array and surface-enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS). The mass/charge (M/Z) ratios of each peak were tested by the Kruskal-Wallis rank sum test, and the protein peak value of the M/Z ratio for each treatment by pair comparison was analyzed with the Nemenyi test. RESULTS: H2O2 up-regulated expression of two protein spots (with M/Z of 6 532 and 6 809). When E2 mitigated the oxidative damage, the expression of one protein spot (M/Z 6 532) was down-regulated. In contrast, ECR down-regulated both of protein spots (M/Z 6 532 and 6 809). CONCLUSION: ECR could effectively inhibite H2O2 induced oxidative damage in HLE-B3 cells. The protein spot at M/Z of 6 532 might be the target spot of ECR against oxidative damage induced by H2O2.

11.
J Insect Sci ; 142014.
Article in English | MEDLINE | ID: mdl-25826465

ABSTRACT

To explore the function of small heat shock protein genes (shsps) and hsp70 in Ericerus pela, we cloned the full-length cDNA sequences of hsp21.5, hsp21.7, hsp70, and hsc70 and the genomic sequence of hsc70. Open reading frames of the four hsps were 570, 564, 1,908, and 1,962 base pairs (bp), respectively, which encode proteins with calculated molecular mass of 21.5, 21.7, 69.8, and 71.6 kDa. Amino acid sequence analysis revealed the presence of the conserved Hsp motifs in all four proteins. The genomic DNA of hsc70 had four introns. ep-hsp21.5 was orthologous and ep-hsp21.7 was species specific. Expression of all four transcripts during heat or cold stress and development was examined by quantitative real-time polymerase chain reaction. All four hsps were upregulated during heat or cold stress in female adults, indicating a correlation between the four hsps and heat or cold-stress tolerance in female adults. ep-hsp21.7 and ep-hsp70 were upregulated during heat stress in male larvae, implying a correlation between the two hsps and heat-stress tolerance in male larvae. The four ep-hsps were also upregulated during the developmental process in males, and ep-hsp21.5, ep-hsp70, and ep-hsc70 were upregulated in females, which indicates their possible role in the developmental regulation of E. pela.


Subject(s)
Heat-Shock Proteins/genetics , Hemiptera/genetics , Insect Proteins/genetics , Amino Acid Sequence , Animals , Base Sequence , Cloning, Molecular , Female , Gene Expression Profiling , HSC70 Heat-Shock Proteins/genetics , HSC70 Heat-Shock Proteins/metabolism , HSP70 Heat-Shock Proteins/genetics , HSP70 Heat-Shock Proteins/metabolism , Heat-Shock Proteins/metabolism , Hemiptera/growth & development , Hemiptera/metabolism , Insect Proteins/metabolism , Introns , Larva , Male , Molecular Sequence Data , Real-Time Polymerase Chain Reaction , Sequence Alignment , Stress, Physiological , Temperature
12.
Chin J Integr Med ; 18(7): 529-33, 2012 Jul.
Article in English | MEDLINE | ID: mdl-22772916

ABSTRACT

OBJECTIVE: To investigate the protective effects of the natural medicinal monomer isopsoralen (ISR) with estrogenic activity against oxidative damage in human lens epithelial cells B3 (HLE-B3) caused by hydrogen peroxide (H(2)O(2)) and to pursue the possible mitochondrial proteomic regularity of the protective effects. METHODS: HLE-B3 cells were treated with H(2)O(2) (300 µ mol/L), ß-estradiol (E(2): 10(-8) mol/L) and H(2)O(2), ISR (10(-5) mol/L) and H(2)O(2), or left untreated. Altered expressions of all mitochondrial proteins were analyzed by protein array and surfaceenhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS). The mass/charge (m/z) ratios of each peak were tested by the Kruskal-Wallis rank sum test, and the protein peak value of the m/z ratio for each treatment by pair comparison was analyzed with the Nemenyi test. RESULTS: H(2)O(2) up-regulated the expressions of two protein spots (with m/z of 6532 and 6809). E(2) mitigated the oxidative damage, and the expression of one protein spot (m/z 6532) was down-regulated. In contrast, ISR down-regulated both of protein spots (m/z 6532 and 6809). CONCLUSIONS: ISR could effectively inhibit H(2)O(2)-induced oxidative damage in HLE-B3 cells. The protein spot at m/z of 6532 might be the target spot of ISR against oxidative damage induced by H(2)O(2).


Subject(s)
Epithelial Cells/metabolism , Furocoumarins/pharmacology , Lens, Crystalline/pathology , Mitochondria/metabolism , Oxidative Stress/drug effects , Protective Agents/pharmacology , Proteomics/methods , Cell Line , Epithelial Cells/drug effects , Epithelial Cells/pathology , Estradiol/pharmacology , Humans , Hydrogen Peroxide/toxicity , Oxidation-Reduction/drug effects , Proteome/metabolism
13.
J Zhejiang Univ Sci B ; 13(5): 402-7, 2012 May.
Article in English | MEDLINE | ID: mdl-22556179

ABSTRACT

OBJECTIVE: The incidence of after-cataracts [also known as posterior capsular opacification (PCO)] is between 30% and 50% three years following cataract surgery. Suppressing the proliferation of lens epithelial cells (LECs) is a primary goal in preventing PCO. Here, we investigated the proteomic regulation of the inhibitory effects of curcumin (Cur) on the proliferation of human lens epithelial B3 (HLE-B3) cells. METHODS: Recombinant human basic fibroblast growth factor (rhbFGF) was used to induce proliferation of HLE-B3 cells, which were incubated with 20 mg/L Cur in a CO(2) incubator for 24 h. RESULTS: We found that the absorbance (A) value of rhbFGF group was significantly higher than the A value of the control group. Furthermore, the A value of the Cur group was significantly lower compared to the rhbFGF group, with an inhibition of 53.7%. Five different protein spots were obtained from proliferative HLE-B3 cells induced by rhbFGF. Eight different protein spots were obtained in HLE-B3 cells incubated with Cur. There were the common variational protein spots at mass/charge (m/z) ratios of 8093 and 13767 between rhbFGF group and control group as well as between the Cur group and rhbFGF group. CONCLUSIONS: These results show that Cur effectively inhibited HLE-B3 cell proliferation induced by rhbFGF. The protein spots at m/z of 8093 and 13767 may be the targets of Cur-induced inhibition of HLE-B3 cell proliferation. Cur may be a reliable and effective drug for prevention and treatment of polymerase chain reaction (PCR).


Subject(s)
Curcumin/pharmacology , Epithelial Cells/drug effects , Epithelial Cells/physiology , Lens, Crystalline/drug effects , Lens, Crystalline/physiology , Proteome/metabolism , Cell Line , Cell Proliferation/drug effects , Epithelial Cells/cytology , Humans , Lens, Crystalline/cytology
14.
Int J Ophthalmol ; 4(3): 255-8, 2011.
Article in English | MEDLINE | ID: mdl-22553656

ABSTRACT

AIM: To study the effects of elemene (Ele) on proliferation and cell cycle of human lens epithelial cells B3 (HLE-B3) and the mechanisms of its signal transduction. METHODS: Recombinant human basic fibroblast growth factor (rhbFGF) was used to induce proliferation of HLE-B3 cells, which were incubated with 80mg/L Ele for 24 hours. The inhibitory effects of Ele on the proliferation of HLE-B3 cells were evaluated by MTT method. The effect of Ele on HLE-B3 cell cycle was analyzed by flow cytometry(FCM). The expressions of protein kinase A (PKA) and protein kinase G (PKG) of HLE-B3 were also analyzed by FCM. RESULTS: Ele altered the cell cycle of HLE-B3 and effectively inhibited HLE-B3 cell proliferation induced by rhbFGF. Ele up-regulated PKA and down-regulated the expression of PKG in HLE-B3 cell. CONCLUSION: Ele inhibits HLE-B3 proliferation, making it an attractive potential agent in regimens to treat after-cataracts.

15.
Zhonghua Yan Ke Za Zhi ; 46(5): 427-31, 2010 May.
Article in Chinese | MEDLINE | ID: mdl-20654217

ABSTRACT

OBJECTIVE: To investigate the inhibitory effects of natural medicinal monomer elemene (Ele) on proliferation of human lens epithelial cells B3 (HLE-B3) inducing by recombinant human basic fibroblast growth factor(rhbFGF) and to pursue the proteomics regularity of the inhibitory effects of Ele on proliferation of HLE-B3. METHODS: Experimental study. This study is divided into three group: control group, rhbFGF group and Ele group. Using 10 microg/L rhbFGF to induce proliferation of HLE-B3. Proliferative HLE-B3 were incubated with 80 mg/L Ele in CO2 incubator for 24 hours. Then the inhibitory effects of Ele on proliferation of HLE-B3 was detected by methyl thiazolyl tetrazolium (MTT). The change of expressions of all protein in HLE-B3 was assayed and analyzed by protein array and surface-enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS) proteomics technology. RESULTS: MTT test showed that the A values of rhbFGF (0.599+/-0.053) group were higher than that of control group (0.409+/-0.042) remarkably. The A values of Ele group (0.450+/-0.061) decreased obviously compared to rhbFGF group, the inhibition rates were 24.90% (F=28.886, P=0.000). Five different protein spots were obtained in proliferative HLE-B3 induced by rhbFGF. The expressions were up-regulated in two of the five protein spots at the ratios of mass/charge (m/z) of 8093 and 9516, while the expressions were down-regulated in three of the five protein spots at m/z of 5361, 9666 and 13 767. Ten different protein spots were obtained in HLE-B3 incubated with Ele. The expressions were up-regulated in four of the ten protein spots at m/z of 2487, 4392, 8566 and 11 600, while the expressions were down-regulated in six of the ten protein spots at m/z of 3679, 4826, 6861, 9516, 9557 and 9672. CONCLUSIONS: Ele could effectively inhibit HLE-B3 proliferation induced by rhbFGF. The protein spot at m/z of 9516 might be the target of proliferative inhibition in HLE-B3 by Ele.


Subject(s)
Cell Proliferation/drug effects , Epithelial Cells/drug effects , Lens, Crystalline/cytology , Proteome/analysis , Sesquiterpenes/pharmacology , Cells, Cultured , Epithelial Cells/metabolism , Fibroblast Growth Factor 2/pharmacology , Humans , Lens, Crystalline/drug effects , Lens, Crystalline/metabolism , Proteomics , Recombinant Proteins/pharmacology
16.
Guang Pu Xue Yu Guang Pu Fen Xi ; 29(8): 2199-203, 2009 Aug.
Article in Chinese | MEDLINE | ID: mdl-19839338

ABSTRACT

The binding reaction of rutin-Sm with serum albumin (SA) was investigated by the fluorescence method in physiological condition. The authors studied mainly the quenching mechanism of the fluorescence of SA by rutin-Sm, and calculation of the binding constants K(LB) of human serum albumin (HSA) and bovine serum albumin (BSA) with rutin-Sm by Lineweaver-Burk equation at different temperatures respectively, then obtained the thermodynamic parameters of HSA and BSA with rutin-Sm according to the calculated binding constants K(LB) at different temperature, meanwhile the type of binding forces of HSA and BSA with rutin-Sm was determined. The results showed that the emission spectra of BSA (HSA) in the presence and absence of rutin-Sm are different. The emission spectra of BSA (HSA) in the presence of rutin-Sm can be quenched. The quenching mechanism of rutin-Sm to SA was static quenching with non-radiation energy transfer for new complex of SA and rutin-Sm. The binding constants K(LB) (L x moL(-1)) were 6.540 x 10(5) and 3.265 x 10(5) for BSA, and 6.830 x 10(5) and 4.665 x 10(5) for HSA at 295 K and 310 K respectively. And the type of bonding forces was estimated by the calculation of thermodynamic parameters of the reaction of rutin-Sm with SA at different temperatures, and the result showed that the binding forces were mainly H-bond and Van der Waals between BSA and rutin-Sm due to the deltaH < 0 and deltaS < 0, and the main electrostatic interaction of rutin-Sm and HSA because of deltaH < 0 and deltaS > 0. The effect of rutin-Sm on the conformation of serum albumin was also studied by using synchronous fluorescence spectroscopy. Results indicated that rutin-Sm could be deposited and transported by serum protein in vivo.


Subject(s)
Rutin/chemistry , Serum Albumin/chemistry , Animals , Cattle , Humans , Protein Binding , Serum Albumin, Bovine , Spectrometry, Fluorescence , Thermodynamics
17.
Zhongguo Zhong Yao Za Zhi ; 32(8): 725-8, 2007 Apr.
Article in Chinese | MEDLINE | ID: mdl-17608231

ABSTRACT

OBJECTIVE: To investigate the effect of Rhizoma Curcumae (RC), arsenite trioxide (As2O3) on proliferation ana signal transduction molecule in lens epithelial cell (LEC), in order to provide experiment evidence for prevention and treatment of after cataract. METHOD: Proliferation of cultured bovine LEC were induced by induced by recombinant human basic fibroblast growth factor (rhbFGF); Inhibitory rates of LEC proliferation induced by RC, As2O3 were detected by methyl thiazolyl tetrazolium (MTT); Inhibitory effects of expression of proliferating cell nuclear antigen (PCNA) induced by RC, As2O3 in LEC were assayed via flow cytometer (FCM); Concentrations of LEC calcium ([Ca2+]i) were determined by spectrofluoremeter, intracellular concentrations of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) of LEC were measured by radioimmunoassay. RESULT: Inhibitory rates of RC, As2O3 on LEC proliferation induced by rhbFGF increased significantly, showing dose-dependent (P < 0.01). PCNA expression of LEC proliferation induced by rhbFGF were down regulated obviously by RC, As2O3, showing dose-dependent (P < 0.01). Concentrations of [Ca2+]and cAMP increased and cGMP decreased significantly in LEC of proliferation inhibited by RC, As2O3 (P < 0.01). CONCLUSION: RC, As2O3 can inhibit LEC proliferation obviously. Signal transductions of [Ca2+]i, cAMP, cGMP may be the important molecular mechanism. There are broad prospect for RC, As2O3 on prevention and treatment of after cataract.


Subject(s)
Arsenicals/pharmacology , Cell Proliferation/drug effects , Curcuma/chemistry , Drugs, Chinese Herbal/pharmacology , Epithelial Cells/drug effects , Oxides/pharmacology , Rhizome/chemistry , Animals , Arsenic Trioxide , Calcium/metabolism , Cattle , Cells, Cultured , Cyclic AMP/metabolism , Cyclic GMP/metabolism , Dose-Response Relationship, Drug , Drugs, Chinese Herbal/isolation & purification , Epithelial Cells/cytology , Epithelial Cells/metabolism , Fibroblast Growth Factor 2/genetics , Fibroblast Growth Factor 2/pharmacology , Flow Cytometry , Growth Inhibitors/pharmacology , Lens, Crystalline/cytology , Lens, Crystalline/drug effects , Lens, Crystalline/metabolism , Proliferating Cell Nuclear Antigen/metabolism , Radioimmunoassay , Recombinant Proteins/pharmacology , Signal Transduction/drug effects
18.
Gene Expr Patterns ; 6(7): 730-7, 2006 Oct.
Article in English | MEDLINE | ID: mdl-16495162

ABSTRACT

We have examined the expression of three paralogous Hox genes from E11.5 through E15.5 in the mouse spinal cord. These ages coincide with major phases of spinal cord neurogenesis, neuronal differentiation, cell migration, gliogenesis, and motor neuron cell death. The three genes, Hoxa10, Hoxc10, and Hoxd10, are all expressed in the lumbar spinal cord and have distinct expression patterns. Mutations in these three genes are known to affect motor neuron patterning. All three genes show lower levels of expression at the rostral limits of their domains, with selective regions of higher expression more caudally. Hoxa10 and Hoxd10 expression appears confined to postmitotic cell populations in the intermediate and ventral gray, while Hoxc10 is also expressed in proliferating cells in the dorsal ventricular zone. Hoxc10 and Hoxd10 expression is clearly excluded from the lateral motor columns at rostral lumbar levels but is present in this region more caudally. Double labeling demonstrates that Hoxc10 expression is correlated with ventrolateral LIM gene expression in the caudal part of the lumbar spinal cord.


Subject(s)
Homeodomain Proteins/genetics , Spinal Cord/embryology , Transcription Factors/genetics , Animals , Body Patterning/genetics , Cell Proliferation , Female , Gene Expression Regulation, Developmental , Homeobox A10 Proteins , In Situ Hybridization , Lumbosacral Region/innervation , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Motor Neurons/chemistry , Spinal Cord/metabolism
19.
Zhong Xi Yi Jie He Xue Bao ; 4(1): 39-42, 2006 Jan.
Article in Chinese | MEDLINE | ID: mdl-16409968

ABSTRACT

OBJECTIVE: To investigate the signal transduction mechanism of curcumin in inhibiting the proliferation of bovine lens epithelial cell (LEC) induced by recombinant human epidermal growth factor (rhEGF). METHODS: There were three groups in this experiment, which were normal control group, untreated group and curcumin-treated group. Proliferation of LEC was induced by rhEGF (50 microg/L). The concentration of intracellular Ca(2+) ([Ca(2+)](i)) in LEC was measured with Fura-2/AM by spectrofluorimetry. The contents of intracellular cAMP and cGMP were assayed by radioimmunoassay. RESULTS: The [Ca(2+)]i in LEC was obviously increased in the untrated group as compared with that in the normal control group (P<0.01), and the [Ca(2+)](i) in LEC in the curcumin-treated group was highest among three groups (P<0.01). The content of intracellular cAMP in LEC was decreased while the content of intracellular cGMP was obviously increased in the untreated group as compared with those in the normal control group (P<0.01). The content of intracellular cAMP in LEC was higher in the curcumin-treated group than that in the untreated group, while the content of intracellular cGMP was lower than that in the untreated group (P<0.01). CONCLUSION: The antiproliferation effects of curcumin on LEC may relate to the regulations of multiple processes of signal transduction.


Subject(s)
Curcumin/pharmacology , Epidermal Growth Factor/antagonists & inhibitors , Epithelial Cells/cytology , Lens, Crystalline/cytology , Signal Transduction , Animals , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Cattle , Cell Proliferation/drug effects , Cells, Cultured , Recombinant Proteins/antagonists & inhibitors
20.
J Neuroimmunol ; 152(1-2): 57-66, 2004 Jul.
Article in English | MEDLINE | ID: mdl-15223237

ABSTRACT

Protein kinase C (PKC) plays a critical role in signal transduction controlling T lymphocyte activation. Both positive and negative regulation of signal transduction is needed for proper control of T lymphocyte activation. We have found that a golli product of the myelin basic protein (MBP) gene can serve as a negative regulator of signaling pathways in the T lymphocyte, particularly the PKC pathway. Increased expression of golli BG21 in Jurkat T cells strongly inhibits anti-CD3-induced IL-2-luciferase activity, an indicator of T lymphocyte activation. Golli BG21 can be phosphorylated by PKC in vitro and its phosphorylation increases in PMA-activated Jurkat cells. BG21 inhibits the PMA-induced increase in AP-1 or NF-kappaB activation, consistent with golli acting in a PKC-mediated cellular event. Golli BG21 inhibition of the PKC pathway is not due to a direct action on PKC activation but in the cascade following PKC activation, since BG21 neither reduces PKC enzyme activity nor blocks the membrane association of PKCtheta brought on by T lymphocyte activation. The inhibitory function of BG21 is independent of its phosphorylation by PKC because a mutant BG21, in which the PKC sites have been mutated, is as effective as the wild type BG21 in inhibiting the PMA-induced AP-1 activation. Structure-function assays indicate that BG21 inhibitory activity resides in the golli domain rather than in MBP domain of the molecule. These results reveal a novel role for MBP gene products in T lymphocytes within the immune system.


Subject(s)
Lymphocyte Activation/immunology , Myelin Basic Protein/immunology , Nerve Tissue Proteins/immunology , Signal Transduction/immunology , T-Lymphocytes/immunology , Transcription Factors/immunology , Amino Acid Sequence , Blotting, Western , Flow Cytometry , Humans , Jurkat Cells , Microscopy, Fluorescence , Molecular Sequence Data , Myelin Basic Protein/genetics , Phosphorylation , Protein Kinase C/metabolism , Receptors, Antigen, T-Cell/immunology , Reverse Transcriptase Polymerase Chain Reaction , Transfection
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