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Hui Huang

Publications and source records attributed to Hui Huang.

At least 19 recordsLinked to original sources
PubMed2026

Schisantherin B mitigates cisplatin-induced ototoxicity by modulating the CNPY2-PERK/CHOP signaling axis.

Irreversible cisplatin-induced hearing loss (CIHL) is a refractory chemotherapy-related adverse effect with limited clinical treatments. Schisantherin B (STB), a lignan isolated from Schisandra chinensis, is widely recognized for its neuroprotective properties, while its role in auditory injury remains unclear. Herein, we found that STB alleviated cisplatin-induced ototoxicity in House Ear Institute Organ of Corti 1 (HEI-OC1) cells and guinea pig models, protecting cochlear hair cells, synaptic ribbons and spiral ganglion neurons, and partially restoring auditory brainstem response (ABR) thresholds. Furthermore, combined drug affinity responsive target stability (DARTS) assay, the cellular thermal shift assay (CETSA), and the surface plasmon resonance (SPR) assay, we confirmed STB directly binds to the canopy FGF signaling regulator 2 (CNPY2), a key initiator of endoplasmic reticulum (ER) stress. Notably, consistent dual in vitro and in vivo validation confirmed that STB exerts no regulatory effect on CNPY2 protein abundance, yet suppressed the downstream Protein kinase R-like endoplasmic reticulum kinase / C/EBP homologous protein (PERK/CHOP) signaling cascade and ER stress-mediated apoptosis. Moreover, molecular docking and co-immunoprecipitation (co-IP) validated the physical binding of STB to CNPY2 and the endogenous interaction between CNPY2 and PERK. Additionally, CNPY2 overexpression and shRNA knockdown further verified this functional relationship. Integrated proteomic and transcriptomic analyses showed STB partially reversed cisplatin-triggered inflammation and excessive ER stress. Collectively, our results suggest STB may serve as a potential otoprotective agent. The CNPY2-PERK/CHOP axis is closely linked to cisplatin-induced cochlear damage and offers a feasible target for intervention against CIHL. Abbreviations: CIHL, cisplatin-induced hearing loss; STB, Schisantherin B; HEI-OC1, house ear institute organ of corti 1; ABR, auditory brainstem response; DARTS, drug affinity responsive target stability; CETSA, cellular thermal shift assay; SPR, surface plasmon resonance; CNPY2, canopy FGF signaling regulator 2; ER, endoplasmic reticulum; PERK, protein kinase R-like endoplasmic reticulum kinase; CHOP, C/EBP homologous protein; co-IP, co-immunoprecipitation; STA, Schisantherin A; STC, Schisantherin C; dB SPL, decibels sound pressure level; EDTA, ethylenediaminetetraacetic acid; dB SPL, decibels sound pressure level; SGN, spiral ganglion neuron; IHCs, inner hair cells; OHCs, outer hair cells; CCK-8, Cell Counting Kit-8; OD, optical density; ODb, blank sample, ODc, control sample; NC, negative control; PVDF, polyvinylidene difluoride; RT, room temperature; LC-MS/MS, liquid chromatography tandem mass spectrometry; MS, mass spectrometry; DMSO, dimethyl sulfoxide; KDs, equilibrium dissociation constants; SP, standard precision; SEM, standard error of the mean; HSD, honestly significant difference; Ctrl, control group; CV, cell viability; Kd, dissociation rate constant; Ka, association rate constant; STS, sodium thiosulfate; UPR, unfolded protein response; BLB, blood-labyrinth barrier.

Apoptosis↗
PubMed2026

A novel PKHD1 missense variant disrupting splicing in a fetus with Caroli disease.

BACKGROUND: Caroli disease (CD) is a rare inherited disorder characterized by dilatation of intrahepatic bile ducts, and prenatal diagnosis of this disease is extremely rare. PKHD1 is the only known causative gene, yet the pathogenicity of most missense variants remains unclear. METHODS: Exome sequencing (ES) was performed on a fetus with clinical features of CD. Candidate variants were validated by Sanger sequencing in the family. The impact of the novel missense variant on pre-mRNA splicing was assessed using minigene assays, and structural modeling of the PKHD1 protein was conducted with AlphaFold 3. RESULTS: At 23 weeks of gestation, the fetus showed hepatic cysts on ultrasound and a "central dot" sign on MRI, suggesting a diagnosis of CD. The fetus also exhibited features of autosomal recessive polycystic kidney disease and oligohydramnios. ES identified and Sanger sequencing confirmed three PKHD1 variants: a paternal nonsense variant c.5323C>T; p.(Arg1775*), and two maternal missense variants c.6682G>C; p.(Glu2228Gln) and c.8012G>T; p.(Arg2671Leu). The variant c.6682G>C is novel and minigene assays demonstrated that it caused exon 40 skipping, leading to an in‑frame deletion (c.6491_6682del; p.(Gly2164_Arg2227del)). Structural modeling predicts that this deletion lies within a large β‑barrel domain and may compromise its structural stability. Conclusion We characterize a novel missense variant that causes aberrant splicing of PKHD1 in CD. This finding underscores the necessity of functional analysis for evaluating the pathogenicity of missense variants, especially those at the last nucleotide of an exon. Our study expands the mutation spectrum of PKHD1 and provides insights into genotype‑phenotype correlations.

Humans↗
PubMed2005

Renal 20-HETE inhibition attenuates changes in renal hemodynamics induced by L-NAME treatment in pregnant rats.

We previously reported that inhibition of nitric oxide (NO) synthesis by N-nitro-L-arginine methyl ester (L-NAME) during late pregnancy leads to increased production of renal vascular 20-hydroxyeicosatetraenoic acid (20-HETE), a cytochrome P-450 (CYP) 4A-derived vasoconstrictor, in pregnant rats. However, the effect of upregulation of vascular 20-HETE production on renal function after NO inhibition is not known. To test the hypothesis that increased gestational vascular 20-HETE synthesis after NO inhibition is involved in mediating blood pressure and renal functional changes, we first determined the IC(50) value of the effect of nitroprusside (SNP), a NO donor, on renal 20-HETE production in cortical microsomes. We then divided pregnant rats and age-matched virgin rats into a vehicle control group, an L-NAME treatment group (0.25 mg/ml in drinking water), and a group treated with L-NAME plus N-methylsulfonyl-12,12-dibromododec-11-enamide (DDMS; CYP4A-selective inhibitor, 10 mg.kg(-1).day(-1) iv). After 4 days of treatment, we measured blood pressure, renal blood flow (RBF), renal vascular resistance (RVR), and glomerular filtration rate (GFR) in each group. The addition of SNP (IC(50) = 22 microM) decreased renal cortical 20-HETE production. In pregnant rats, L-NAME treatment led to significantly higher mean arterial pressure (MAP) and RVR, and lower RBF and GFR. Combined treatment with DDMS and L-NAME significantly attenuated the increases in MAP and RVR and the decrease in GFR, but not the reduction in RBF induced by L-NAME treatment. L-NAME and L-NAME plus DDMS had no significant impact on renal hemodynamics in virgin rats. In addition, chronic treatment with DDMS selectively inhibited cortical 20-HETE production without a significant effect on CYP4A expression in L-NAME-treated pregnant rats. In conclusion, NO effectively inhibits renal cortical microsomal 20-HETE production in female rats. In pregnant rats, the augmentation of renal 20-HETE production after NO inhibition is associated with increased MAP and RVR, whereas decreased GFR is negated by treatment of a selective and competitive CYP4A inhibitor. These results demonstrate that the interaction between renal 20-HETE and NO is important in the regulation of renal function and blood pressure in pregnant rats.

Age Factors↗
PubMed2005

Three-dimensional architecture of phycobilisomes from Nostoc flagelliforme revealed by single particle electron microscopy.

Phycobilisomes are protein complexes that harvest light and transfer energy to the photo system. Here, the three dimensional structure of intact phycobilisomes from Nostoc flagelliforme is studied by a combination of negative stain electron microscopy and cryo-electron microscopy. Results show that the intact phycobilisomes are composed of a tricylindrical core and six rods. Each allophycocyanin cylinder presents a double-layered structure when viewed from the side and a triangular shape when viewed from the top. These characteristics indicate that allophycocyanin trimers in the intact phycobilisomes are arranged into hexameric oligomers in a parallel manner.

Cryoelectron Microscopy↗
PubMed2005

Notch1 modulates timing of G1-S progression by inducing SKP2 transcription and p27 Kip1 degradation.

Cyclin-dependent kinase inhibitors (CKIs) and Notch receptor activation have been shown to influence adult stem cells and progenitors by altering stem cell self-renewal and proliferation. Yet, no interaction between these molecular pathways has been defined. Here we show that ligand-independent and ligand-dependent activation of Notch1 induces transcription of the S phase kinase-associated protein 2 (SKP2), the F-box subunit of the ubiquitin-ligase complex SCF(SKP2) that targets proteins for degradation. Up-regulation of SKP2 by Notch signaling enhances proteasome-mediated degradation of the CKIs, p27 Kip1 and p21 Cip1, and causes premature entry into S phase. Silencing of SKP2 by RNA interference in G1 stabilizes p27 Kip1 and p21 Cip1 and abolishes Notch effect on G1-S progression. Thus, SKP2 serves to link Notch1 activation with the cell cycle machinery. This novel pathway involving Notch/SKP2/CKIs connects a cell surface receptor with proximate mediators of cell cycle activity, and suggests a mechanism by which a known physiologic mediator of cell fate determination interfaces with cell cycle control.

3T3 Cells↗
PubMed2005

A new dynamic model of CD8+ T effector cell responses via CD4+ T helper-antigen-presenting cells.

A long-standing paradox in cellular immunology has been the conditional requirement for CD4(+) Th cells in priming of CD8(+) CTL responses. We propose a new dynamic model of CD4(+) Th cells in priming of Th-dependent CD8(+) CTL responses. We demonstrate that OT II CD4(+) T cells activated by OVA-pulsed dendritic cells (DC(OVA)) are Th1 phenotype. They acquire the immune synapse-composed MHC II/OVAII peptide complexes and costimulatory molecules (CD54 and CD80) as well as the bystander MHC class I/OVAI peptide complexes from the DC(OVA) by DC(OVA) stimulation and thus also the potential to act themselves as APCs. These CD4(+) Th-APCs stimulate naive OT I CD8(+) T cell proliferation through signal 1 (MHC I/OVAI/TCR) and signal 2 (e.g., CD54/LFA-1 and CD80/CD28) interactions and IL-2 help. In vivo, they stimulate CD8(+) T cell proliferation and differentiation into CTLs and induce effective OVA-specific antitumor immunity. Taken together, this study demonstrates that CD4(+) Th cells carrying acquired DC Ag-presenting machinery can, by themselves, efficiently stimulate CTL responses. These results have substantial implications for research in antitumor and other aspects of immunity.

Amino Acid Sequence↗
PubMed2005

Carbon nanotubes as a secondary support of a catalyst layer in a gas diffusion electrode for metal air batteries.

In this paper, we report the use of binary carbon supports (carbon nanotubes (CNTs) and active carbon) as a catalyst layer for fabricating gas diffusion electrodes. The electrocatalytic properties for the oxygen reduction reaction (ORR) were evaluated by polarization curves and electrochemical impedance spectroscopy (EIS) in an alkaline electrolyte. The binary-support electrode exhibits better performance than the single-support electrode, and the best performance is obtained when the mass ratio of carbon nanotubes and active carbon is 50:50. The results from the electrode kinetic parameters indicate that the introduction of carbon nanotubes as a secondary support provides high accessible surface area, good electronic conductivity, and fast ORR kinetics. Furthermore, the effect of CNT support on the electrocatalytic properties of Pt nanoparticles for binary-support electrodes was also investigated by different loading-reduction methods. The electrocatalytic activity of the binary-support electrodes is improved dramatically by Pt loading on CNT carbon support, even at very low Pt loading. Additionally, the EIS analysis results indicate that the process of ORR may be controlled by diffusion of oxygen in the electrode thin film for binary-support electrodes with or without Pt catalyst.

Journal Article↗
PubMed2005

[Changes in neuropeptide substance P and calcitonin gene-related peptide in scarless wound healing in fetal rabbit skin].

OBJECTIVE: To explore the role of the neuropeptide in fetal scarless wound healing. METHODS: The animal models were replicated in which a full-thickness wound was inflicted on the back of rabbit fetuses at 22 days gestation (term=31 days). The progress of wound healing was observed on micro-and macro-levels at the 1 st, 2 nd, 3 rd, 5 th, 7 th, 14 th post-injury days respectively. The expression level of substance P(SP) and calcitonin gene-related peptide (CGRP) during wound healing was assessed by immunohistochemistry staining. RESULTS: The cutaneous wounds in fetal rabbits healed more rapidly than that in adult and without the formation of scar. The expression of SP and CGRP of wound decreased in fetal rabbit at the early stage post-injury, down to the lowest levels on the 2 nd post-injury day (75%, 80%, vs. fetal control, both P<0.01), then increased gradually, up to the normal levels. The expression of SP and CGRP of wound decreased in adult rabbit at the early stage post-injury, down to the lowest levels on the post-injury 1 st day (60%, 76%, vs. adult control, both P<0.01), then increased quickly, up to a higher level than normal group, reaching the peak on the post-injury 7th day(168%, 126%, vs. adult control, both P<0.01), then returned to the normal levels. CONCLUSION: The low level of neuropeptide may contribute to scarless wound healing in fetal rabbit.

Animals↗
PubMed2005

[In vitro induction of mature dendritic cells from malignant pleural effusions].

BACKGROUND & OBJECTIVE: Dendritic cells (DCs) can activate immunologic naive T cells to initiate antigen-specific immune responses. This study was to in vitro induce mature DCs from malignant pleural effusions of patients with lung cancer. METHODS: Malignant pleural effusions (500-1 000 ml) were collected from 16 patients with primary lung cancer. Precursory DCs were obtained through density gradient centrifugation and magnetic cell sorting from malignant pleural effusions, and cultured with interleukin-4 (IL-4), granulocyte-macrophage colony-stimulating factor (GM-CSF), and tumor necrosis factor-alpha (TNF-alpha). Morphology of DCs was observed under invert optical microscope and electronic microscope; phenotype of DCs was analyzed by flow cytometry. Effect of DCs on proliferation of tumor infiltrative lymphocytes (TILs) was observed through mixed lymphocyte reaction (MLR). RESULTS: Mature DCs with typical morphology (elongated dendritic processions observed under invert optical microscope and electronic microscope) were induced from malignant pleural effusions. Expressions of surface phenotypes were higher in DCs induced for 48 h than in DCs induced for 0, 24, 96, and 192 h. When activated with DCs, proliferation of TILs was enhanced. CONCLUSION: Mature DCs could be induced from malignant pleural effusions of patients with lung cancer.

Adenocarcinoma↗
PubMed2004

SED, a normalization free method for DNA microarray data analysis.

BACKGROUND: Analysis of DNA microarray data usually begins with a normalization step where intensities of different arrays are adjusted to the same scale so that the intensity levels from different arrays can be compared with one other. Both simple total array intensity-based as well as more complex "local intensity level" dependent normalization methods have been developed, some of which are widely used. Much less developed methods for microarray data analysis include those that bypass the normalization step and therefore yield results that are not confounded by potential normalization errors. RESULTS: Instead of focusing on the raw intensity levels, we developed a new method for microarray data analysis that maps each gene's expression intensity level to a high dimensional space of SEDs (Signs of Expression Difference), the signs of the expression intensity difference between a given gene and every other gene on the array. Since SED are unchanged under any monotonic transformation of intensity levels, the SED based method is normalization free. When tested on a multi-class tumor classification problem, simple Naive Bayes and Nearest Neighbor methods using the SED approach gave results comparable with normalized intensity-based algorithms. Furthermore, a high percentage of classifiers based on a single gene's SED gave good classification results, suggesting that SED does capture essential information from the intensity levels. CONCLUSION: The results of testing this new method on multi-class tumor classification problems suggests that the SED-based, normalization-free method of microarray data analysis is feasible and promising.

Central Nervous System Neoplasms↗
PubMed2004

Evolutionary conservation and selection of human disease gene orthologs in the rat and mouse genomes.

BACKGROUND: Model organisms have contributed substantially to our understanding of the etiology of human disease as well as having assisted with the development of new treatment modalities. The availability of the human, mouse and, most recently, the rat genome sequences now permit the comprehensive investigation of the rodent orthologs of genes associated with human disease. Here, we investigate whether human disease genes differ significantly from their rodent orthologs with respect to their overall levels of conservation and their rates of evolutionary change. RESULTS: Human disease genes are unevenly distributed among human chromosomes and are highly represented (99.5%) among human-rodent ortholog sets. Differences are revealed in evolutionary conservation and selection between different categories of human disease genes. Although selection appears not to have greatly discriminated between disease and non-disease genes, synonymous substitution rates are significantly higher for disease genes. In neurological and malformation syndrome disease systems, associated genes have evolved slowly whereas genes of the immune, hematological and pulmonary disease systems have changed more rapidly. Amino-acid substitutions associated with human inherited disease occur at sites that are more highly conserved than the average; nevertheless, 15 substituting amino acids associated with human disease were identified as wild-type amino acids in the rat. Rodent orthologs of human trinucleotide repeat-expansion disease genes were found to contain substantially fewer of such repeats. Six human genes that share the same characteristics as triplet repeat-expansion disease-associated genes were identified; although four of these genes are expressed in the brain, none is currently known to be associated with disease. CONCLUSIONS: Most human disease genes have been retained in rodent genomes. Synonymous nucleotide substitutions occur at a higher rate in disease genes, a finding that may reflect increased mutation rates in the chromosomal regions in which disease genes are found. Rodent orthologs associated with neurological function exhibit the greatest evolutionary conservation; this suggests that rodent models of human neurological disease are likely to most faithfully represent human disease processes. However, with regard to neurological triplet repeat expansion-associated human disease genes, the contraction, relative to human, of rodent trinucleotide repeats suggests that rodent loci may not achieve a 'critical repeat threshold' necessary to undergo spontaneous pathological repeat expansions. The identification of six genes in this study that have multiple characteristics associated with repeat expansion-disease genes raises the possibility that not all human loci capable of facilitating neurological disease by repeat expansion have as yet been identified.

Animals↗
PubMed2004

Study on reactions of 2-(dinitromethylene)-4,5-imidazolidinedione.

Some new reactions of 2-(dinitromethylene)-4,5-imidazolidinedione (1) with water, alcohols, carboxylic acids, and alkalis were discovered. By reaction of 1 with carboxylic acids, large particle size 1,1-diamino-2,2-dinitroethylene (2) was prepared. By reaction of 1 with methanol, the methanol adduct (4) was synthesized and characterized. By reaction of 1 with water, the synthetic pathway of 2-methylimidazole to 2 could be achieved in a continuous process. By reaction of 1 with KOH, 2 and potassium dinitromethane (6) could be formed at different temperature, respectively. Compounds 1 and 4 decomposed into parabanic acid (5), losing nitrogen oxides and carbon oxides. Some explosive properties of 1 were studied. The mechanisms of synthesis of 1, 2, and 5 are discussed.

Journal Article↗
PubMed2004

Structures of the neuronal and endothelial nitric oxide synthase heme domain with D-nitroarginine-containing dipeptide inhibitors bound.

In a continuing effort to unravel the structural basis for isoform-selective inhibition of nitric oxide synthase (NOS) by various inhibitors, we have determined the crystal structures of the nNOS and eNOS heme domain bound with two D-nitroarginine-containing dipeptide inhibitors, D-Lys-D-Arg(NO)2-NH(2) and D-Phe-D-Arg(NO)2-NH(2). These two dipeptide inhibitors exhibit similar binding modes in the two constitutive NOS isozymes, which is consistent with the similar binding affinities for the two isoforms as determined by K(i) measurements. The D-nitroarginine-containing dipeptide inhibitors are not distinguished by the amino acid difference between nNOS and eNOS (Asp 597 and Asn 368, respectively) which is key in controlling isoform selection for nNOS over eNOS observed for the L-nitroarginine-containing dipeptide inhibitors reported previously [Flinspach, M., et al. (2004) Nat. Struct. Mol. Biol. 11, 54-59]. The lack of a free alpha-amino group on the D-nitroarginine moiety makes the dipeptide inhibitor steer away from the amino acid binding pocket near the active site. This allows the inhibitor to extend into the solvent-accessible channel farther away from the active site, which enables the inhibitors to explore new isoform-specific enzyme-inhibitor interactions. This might be the structural basis for why these D-nitroarginine-containing inhibitors are selective for nNOS (or eNOS) over iNOS.

Amino Acid Substitution↗
PubMed2004

Synergistic effect of lymphotactin and interferon gamma-inducible protein-10 transgene expression in T-cell localization and adoptive T-cell therapy of tumors.

The lack of efficient T-cell infiltration of tumors is a major obstacle to successful adoptive T-cell therapy. We have previously demonstrated that adenovirus (AdV)-mediated transgene lymphotactin (Lptn) or IP-10 expression in tumors can significantly enhance T-cell tumor infiltration. In this study, active OVA-specific CD8+ T cells were prepared by coculturing naive OVA-specific CD8+ T cells from transgenic OT I mice with OVA-I peptide-pulsed dendritic cells in vitro. These XCR-1- and CXCR3-expressing T cells predominantly secreted IFN-gamma and displayed significant killing activity (84% at effector:target cell ratio of 1.5) against OVA-expressing EG7 tumor cells through perforin-mediated pathway. Our data also showed that chemokine Lptn and IP-10 not only can chemoattract, but also stimulate proliferation of CD8+ T cells in vitro, and that a mixture of Lptn and IP-10 can more efficiently chemoattract CD8+ T cells than either one of them. Furthermore, we demonstrated that the transferred CD8+ T cells detected in group of tumors treated with both AdVLptn and AdVIP-10 (group a) are around 4 and 2 times more than that in groups of tumors treated with control AdVpLpA (group b) and either AdVIP-10 (group c) or AdVLptn (group d), respectively. Around 87.5% of mice in group a were tumor-free compared to the aggressive tumor growth in all 8 mice of group b and 25% or 37.5% cured mice seen in groups c and d (p<0.05). Thus, our results indicate that enhancement of adoptive T-cell therapy can be obtained by double tranmsgene Lptn and IP-10 expression, which facilitates CD8+ T-cell tumor localization through proliferation and chemoattraction of the transferred CD8+ T cells by in situ chemokine transgene expressions in the tumors. Collectively, our data provide solid evidence of a potent synergy between adoptive T-cell therapy and adenovirus-mediated Lptn and IP-10 gene transfer into tumor tissues, which culminated in the T-cell tumor localization and eradication of well-established tumor masses.

Adenoviridae↗
PubMed2004

New briaranes from the South China Sea gorgonian Junceella juncea.

Three new briarane diterpenes, juncins O-Q (1-3), along with five known briaranes, praelolide, junceellin A, gemmacolide A, gemmacolide B, and junceellolide D, were isolated from the EtOH/CH(2)Cl(2) extracts of the South China Sea gorgonian coral Junceella juncea. The structures of 1-3 were established by extensive spectroscopic analysis, including 1D and 2D NMR data.

Animals↗
PubMed2004

Early detection of disease and scheduling of screening examinations.

Special examinations exist for many chronic diseases, which can diagnose the disease while it is asymptomatic, with no signs or symptoms. The earlier detection of disease may lead to more cures or longer survival. This possibility has led to public health programs which recommend populations to have periodic screening examinations for detecting specific chronic diseases, for example, cancer, diabetes, cardiovascular disease and so on. Such examination schedules when embedded in a public health program are invariably costly and are ordinarily not chosen on the basis of possible trade-offs in costs and benefits for different screening schedules. The possible candidate number of examination schedules is so large that it is not feasible to carry out clinical trials to compare different schedules. Instead, this problem can be investigated by developing a theoretical model which can predict the eventual disease specific mortality for different examination schedules. We have developed such a model. It is a stochastic model which assumes that i) the natural history of the disease is progressive and ii) any benefit from earlier diagnosis is due to a change in the distribution of disease stages at diagnosis (stage shift). The model is general and can be applied to any chronic disease which satisfies our two basic assumptions. We discuss the basic ideas of schedule sensitivity and lifetime schedule sensitivity and its relation to the reduction in disease specific mortality. Our theory is illustrated by applications to breast cancer screening. The investigation of schedules compares not only examination schedules with equal intervals between examinations but also staggered schedules using the threshold method. (Examinations are carried out when an individual's risk status reaches a preassigned threshold value.).

Adult↗
PubMed2004

[Brain-derived neurotrophic factor induces rat bone marrow stromal cells to differentiate into neuron-like cells in vitro].

OBJECTIVE: To investigate brain-derived neurotrophic factor (BDNF)-induced differentiation of rat bone marrow stromal cells (MSCs) into neuron-like cells in vitro and observe its neuroprotective effect of BNDF on the differentiated cells, which might provide the better seed cells for treatment of nervous system diseases. METHODS: The fifth-passage MSCs were induced by BDNF and 2-mercapto ethanol beta-ME respectively, 1, 3 and 6 h after which the induced neuron-like cells were counted and compared. At 3 h, the neuron-like cells were identified by the immunocytochemical staining, reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting. RESULTS: The two induced cells both displayed neuronal morphologies with long and multipolar cell projections, but BDNF-induced cells survived for a longer time than beta-ME-induced ones. The results of immunocytochemical staining showed that the two neuron-like cells expressed nestin, neuron-specific enolase (NSE), neurofilament (NF), microtubule-associated protein (MAP-2), but not glial fibrillary acidic protein (GFAP). RT-PCR detected mRNA-positive NSE, NF and MAP-2 in the induced cells, with also mild positive GFAP mRNA. Western blotting identified also NSE expression in these neuron-like cells. CONCLUSION: BDNF alone may induce rat MSCs to differentiate into neuron-like cells in vitro, which have longer lifetime to better serve the purpose of transplantation and gene therapy for nervous system diseases.

Animals↗