1.Bioinformatic analysis of differentially expressed proteins in the dorsal raphe nucleus of rats after continuous treatment with olanzapine.
Ping HUANG ; Jian Cheng ZHU ; Hua LI ; Yan Zhao WANG ; Yi Min TANG ; Qiang LIU
Journal of Southern Medical University 2022;42(8):1221-1229
OBJECTIVE:
To analyze the differentially expressed proteins in the dorsal raphe nucleus of rats treated with olanzapine and explore the possible mechanism of metabolic disorders in the early stage of olanzapine treatment.
METHODS:
Twenty male and 20 female SD rats were both randomized equally into olanzapine group and control group for daily treatment with olanzapine and saline for 4 weeks, respectively. One hour after the last treatment, the dorsal raphe nucleus of the rats was dissected for proteomic analysis using iTRAQ combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS). GO, KEGG pathway, COG, pathways and protein interaction network analyses of the differentially expressed proteins were performed. Several target genes were selected from the proteomic list, and their expression levels in the dorsal raphe nucleus of another 24 mice with identical grouping and treatment using real time real-time quantitative PCR and Western blotting.
RESULTS:
A total of 214 differentially expressed proteins were identified in the dorsal raphe nucleus of olanzapine-treated mice, including 72 unregulated and 142 downregulated proteins. GO analyses showed that the differentially expressed proteins were enriched in cellular process, biological regulation, metabolic process, response to stimulus, multicellular organismal process, bindings, catalytic activity, molecular function regulator and transcription regulator activity. KEGG analysis suggested that these proteins were enriched in fluid shear stress and atherosclerosis, serotonergic synapse, butanoate metabolism, thyroid hormone synthesis and IL-17 signaling pathway. The differentially expressed proteins Cav1, Hsp90b1, Canx, Gnai1, MAPK9, and LOC685513 were located at the nodes of the protein-protein interaction network in close relation with metabolic disorders. In olanzapine-treated mice, the expression of Hmgcs2, a negative regulator of apoptosis, was significantly down-regulated in the dorsal raphe nucleus, where the expressions of Pla2g4e, Slc6a4 and Gnai1 involved in serotonergic synapse were significantly upregulated.
CONCLUSION
In the early stage of treatment, olanzapine may contribute to the occurrence of metabolic disorders in rats by regulating the expressions of Cav1, Hsp90b1, Canx, Gnai1, MAPK9, LOC685513 (Gng14) and 5-HTR2 synapse-related proteins in the dorsal raphe nucleus.
Animals
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Chromatography, Liquid
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Computational Biology
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Dorsal Raphe Nucleus
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Female
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GTP-Binding Protein alpha Subunits, Gi-Go
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Male
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Mice
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Olanzapine/adverse effects*
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Proteomics
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Rats
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Rats, Sprague-Dawley
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Tandem Mass Spectrometry
2.Icaritin promotes chondrogenic differentiation of BMSCs by Wnt/β-catenin signaling pathway.
Jian-Yang WANG ; Chang-Chang YIN ; Cui-Cui WU ; Shu-Guo GENG ; Ming YIN
China Journal of Chinese Materia Medica 2016;41(4):694-699
To investigate the effect of icaritin (ICT) combined with GDF-5 on chondrogenic differentiation of bone marrow stromal cells (BMSCs), and discuss the action of Wnt signaling pathway, full bone marrow adherent method was used to isolate and culture SD rats BMSCs, and the cells at P3 generation were taken and divided into 6 groups: BMSCs group, ICT group, GDF-5 group, GDF-5+ICT group, GDF-5+ICT+SB216763 group, and GDF-5+ICT+ XAV-939 group. The cells were induced and cultured for 14 days. The morphology change was observed by inverted microscope. Alcian blue staining method was used to detect the changes of proteoglycans. RT-PCR was used to detect the mRNA expressions of aggrecan, Col2, Sox9, Dvl1, Gsk3β, and β-catenin. The protein expressions of collagen 2 (COL2) and β-catenin were detected by Western blot. The results indicated that, compared with the BMSCs group, gradual increase was present in proteoglycan Alcian blue staining; mRNA expressions of cartilage differentiation marker genes aggrecan, COL2, Sox9 and the protein expression of COL2, as well as mRNA and protein expressions of Wnt signaling pathway-related gene β-catenin, but with gradual decrease in Gsk3β mRNA expressions in GDF-5 group, GDF-5+ICT group and GDF-5+ICT+SB216763 group. On the contrary, compared with GDF-5+ICT group, there was a decrease in expressions of Dvl1, and β-catenin related to chondrogenic differentiation and Wnt signaling pathway, a increase in Gsk3β mRNA expression, and also a decrease in protein expressions of COL2 and β-catenin in GDF-5+ICT+XAV-939 group, with statistically significant difference between two groups. GDF-5 in combination with icaritin can induce chondrogenic differentiation of BMSCs in rats, and icaritin (ICT) can promote the chondrogenic differentiation. ICT can promote the chondrogenic differentiation of BMSCs in vitro probably by activating the Wnt/β-catenin signaling pathway.
3. Thromboelastography parameters predict early neurological deterioration in patients with acute ischemic stroke
Xiaoping SHEN ; Shilie WANG ; Jianping LIU ; Nianchun LI
International Journal of Cerebrovascular Diseases 2019;27(12):891-895
Objective:
To investigate the predictive value of thromboelastography (TEG) for early neurological deterioration (END) in patients with acute ischemic stroke.
Methods:
Patients with acute ischemic stroke admitted to the Department of Neurology, Jiujiang Hospital Affiliated to Nanchang University from January 2018 to May 2019 were included as case group, and the healthy physical examinees in the same period were selected as control group. END was defined as an increase of ≥2 of the National Institutes of Health Stroke Scale score from baseline within 7 d after the onset of acute ischemic stroke. All subjects were routinely tested for traditional coagulation function, including prothrombin time, activated partial thromboplastin time, thrombin time, and plasma fibrinogen level. The reaction time (R value), coagulation time (K value), coagulation angle (α) and maximum amplitude (MA value) were monitored by TEG. Univariate analysis was used to compare the differences in clinical and laboratory results between the END group and the non-END group, and then multivariate
4.FOXO4 maintains senescence in human umbilical cord mesenchymal stem cells by repressing apoptosis.
Ping-Ping WU ; Wen-Long HU ; Chang-Chang YIN ; Jiu-Wang FEI
Acta Physiologica Sinica 2020;72(4):426-432
The purpose of the present study was to investigate the effects of forkhead box O4 (FOXO4) on the senescence of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs). The hUC-MSCs were induced to senescence by natural passage, and FOXO4 expression was inhibited by lentiviral shRNA transfection. The hallmark of cell senescence was analyzed by β-galactosidase staining, and the cell viability was assayed by CCK-8 method. Flow cytometry was used to investigate the apoptosis of hUC-MSCs. The expression levels of Bcl-2, Bax, FOXO4, interleukin 6 (IL-6) and cleaved Caspase-3 were detected by qPCR and Western blot. Immunofluorescence staining was used to detect FOXO4 expression. The amount of IL-6 secreted by hUC-MSCs was detected by ELISA. The results showed that, compared with the passage 1, senescent hUC-MSCs showed up-regulated expression levels of Bax and FOXO4, down-regulated expression levels of Bcl-2 and cleaved Caspase-3, and increased IL-6 mRNA expression and secretion. FOXO4 inhibition in senescent hUC-MSCs promoted cell apoptosis, reduced cell viability, and inhibited the mRNA expression and secretion of IL-6. These results suggest that FOXO4 maintains viability and function of senescent hUC-MSCs by repressing their apoptosis response, thus accelerating senescence of the whole cell colony.
Apoptosis
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Cell Cycle Proteins
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Cell Survival
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Cellular Senescence
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Forkhead Transcription Factors
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Humans
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Mesenchymal Stem Cell Transplantation
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Mesenchymal Stem Cells
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Transcription Factors
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Umbilical Cord
5. Study on active components and targets of Ginseng in Prevention and treatment of PTSD based on Network Pharmacology
Zhong-Li WANG ; Li-Jun GAN ; Xiu-Xiu LIU ; Xiu-Li LI ; Ben-Yi YI
Chinese Pharmacological Bulletin 2021;37(8):1159-1163
Aim To obtain the active components and targets of ginseng in the prevention and treatment of traumatic stress disorder(PTSD) through the method of network pharmacology. Methods The active components and target information of ginseng with medicinal value were obtained by TCMSP research platform, and the gene information closely related to the pathogenesis of PTSD was obtained by searching GeneCard and OMIM database. The two were matched to obtain the medicinal components and target genes of ginseng in the prevention and treatment of PTSD. The drug-dis- ease-target network diagram was drawn by R and Perl computer languages, and the target genes were analyzed by PPI network analysis, gene ontology ( GO ) and signal transduction pathway ( KEGG) enrichment analysis. Results According to the general pharmacological research methods of traditional Chinese medi cine, the screening parameters of active components were set, and nine kinds of high value medicinal ingredients of Panax ginseng were obtained. There was a drug-target relationship between the nine medicinal components and sixteen target genes related to PTSD disease. Through PPI, GO and KEGG analysis, it was found that the target genes were mainly enriched in physiological functions such as neurotransmitters, syn-aptic plasticity, ion channels and so on. Conclusions Ginseng has the pharmacological effect of preventing and treating PTSD, which may play a role in regulating the metabolism and receptor activity of monoamine neurotransmitters.
6.Regulation of SIRT1 by SREBP-1 in Adipogenic Differentiation of Mesenchymal Stem Cells.
Wen-Long HU ; Jun CAO ; Chang-Chang YIN ; Ping-Ping WU ; Chen YAN ; Zi-Yao WANG
Journal of Experimental Hematology 2016;24(2):590-595
OBJECTIVETo study the regulation of SIRT1 by transcription factor SREBP-1 in adipogeneic differentiation of bone marrow mesenchymal stem cells (BMMSC).
METHODSOil red O staining was used to identify the adipogenic differentiation of BMMSC; the mRNA transcription levels of AP2, LPL, SREBF-1, SIRT1 gene were detected by RT-PCR; the expession level of SREBP-1 was determined by Western-blot. The chromatin immunoprecipitation (ChIP) assay was used to investigate the binding of SREBP-1 to SIRT1 promoter.
RESULTSBMMSC exposed to adipogenesis inducing medium become mature adipocytes at day 14; the mRNA transcription levels of AP2, LPL, SREBF-1, SIRT1 genes were up-regulated in adipocyte differentiation of BMMSC; the protein level of SREBP-1 was higher obviously; SIRT1 gene sequences was succesfully amplified from the genomic DNA immunoprecipitated by SREBP-1 antibody.
CONCLUSIONSREBP-1 can bind to the promoter region of the SIRT1 gene in adipogenesis of BMMSC, and may be involved in the transcriptional regulation of the SIRT1 gene.
Adipocytes ; cytology ; Adipogenesis ; Cell Differentiation ; Cells, Cultured ; Chromatin Immunoprecipitation ; Gene Expression Regulation ; Humans ; Mesenchymal Stromal Cells ; cytology ; Promoter Regions, Genetic ; Sirtuin 1 ; metabolism ; Sterol Regulatory Element Binding Protein 1 ; metabolism ; Up-Regulation
7.Effect of ABCE1 gene silencing on proliferation and migration of human bladder cancer cell line T24
Qiang LI ; Jiujiang WANG ; Shuifa LUO ; Junjie ZHU
Chinese Journal of Pathophysiology 2015;(5):929-933
AIM:To investigate the effect of small interfering RNA ( siRNA)-mediated ABCE1 knockdown on the survival, cell cycle and invasion of human bladder cancer cell line T24.METHODS:The siRNA against ABCE1 was constructed and transfected into the T24 cells with LipofectamineTM 2000.The expression of ABCE1 was detected by RT-PCR and Western blot.Flow cytometry was used to detect the cell cycle.The effects of ABCE1 gene silencing on prolifera-tion, migration and invasion of T24 cells were evaluated by CCK-8 assay, wound-healing assay and Transwell invasion as-say, respectively.RESULTS: Compared with control group and blank group, the mRNA and protein levels of ABCE1 were significantly decreased in experimental group after transfected with ABCE1 siRNA.The cell cycle was arrested at G0/G1 phase and the cell number in S phase was decreased in the T24 cells.Compared with control group and blank group, the proliferation of T24 cells in experimental group was inhibited significantly, and the migration and invasion abilities of T24 cells in experimental group decreased significantly.CONCLUSION:Knockdown of ABCE1 gene may decrease migration, invasion and proliferation abilities in T24 cells.
8.Correlation Analysis of Myelosuppression after Chemotherapy for Triple-Negative Breast Cancer and Its Prognosis
Kunjian XIA ; Lin WANG ; Zhenhua SUN
Journal of Sun Yat-sen University(Medical Sciences) 2024;45(3):475-483
ObjectiveTo investigate whether severe myelosuppression after chemotherapy is associated with prognosis in patients with breast cancer. MethodsTriple negative breast cancer (TNBC) patients who received chemotherapy at the Second Affiliated Hospital of Nanchang University from May 2, 2013 to May 2, 2018 were divided into a control group (no/mild myelosuppression) and a case group (severe myelosuppression). In this study, 251 patients with TNBC met the inclusion and exclusion criteria, including 125 patients in the control group (20 patients with grade 0 myelosuppression, 43 patients with grade I myelosuppression, 62 patients with grade Ⅱ myelosuppression), 126 patients in the case group (114 patients with grade Ⅲ myelosuppression, 12 patients with grade Ⅳ myelosuppression). The general clinicopathological data of the patients in the two groups, including age, pathological type of tumor, tumor T stage, tumor N stage, tumor Nottingham grade, intravascular cancer thrombus, were analyzed using the χ2 test. The disease-free survival (DFS) and overall survival (OS) of the two groups were analyzed using the Kaplan-Meier method. A Cox proportional hazards regression model with multiple factors was used to analyze the impact of post-chemotherapy severe myelosuppression on disease-free survival (DFS) and overall survival (OS) in patients with TNBC. ResultsThe differences in general clinicopathologic data between the two groups of patients were not statistically significant (all P>0.05). The 5-year disease-free survival (DFS) rate was significantly lower in the control group compared with the case group (75.2% vs. 85.7%, P=0.027). However, there was no statistically significant difference in the 5-year overall survival (OS) rate between the two groups (88.8% vs. 95.2%, P=0.057). The analysis of the multifactorial Cox proportional hazards regression model revealed that post-chemotherapy severe myelosuppression was an independent protective factor for disease-free survival (DFS) (HR=0.332, 95% CI: 0.173-0.638, P=0.001) and overall survival (OS) (HR=0.193, 95% CI: 0.062-0.602, P=0.005) in TNBC patients. ConclusionOur results show that TNBC patients with severe myelosuppression after chemotherapy have longer disease-free survival (DFS) than those with no/mild myelosuppression, and overall survival (OS) also tend to be prolonged compared with those with no/mild myelosuppression, and severe myelosuppression after chemotherapy can be used as an independent predictor of a good prognosis in breast cancer.
9.Effects of Hydrogen Sulfide on Inflammatory Factors and Mitochondrial Energy Metabolic Disorders After Reperfusion Injury in Rats.
Yong Hua YANG ; Wei WANG ; Bin HU ; Hai Long YANG ; Xi Chun WANG
Acta Academiae Medicinae Sinicae 2019;41(2):234-241
Objective To explore the effect of hydrogen sulfide on inflammatory factors and energy metabolism of mitochondria after limbs reperfusion injury in rats. Methods Sixty rats were divided into three groups:sham operation group,control group(ischemia-reperfusion injury + saline group),and experimental group(ischemia-reperfusion injury + HS group).Wistar rat models of limb ischemia-reperfusion injury were established.Skeletal muscle samples were collected to determine the levels of necrosis decomposition products [including myoglobin(MB),lipoprotein complex(LPC)and lipid peroxide(LPO)];blood samples were collected to determine the levels of interleukin(IL)-1,IL-6 and tumor necrosis factor-α(TNF-α);mitochondria were extracted for mitochondrial transmembrane potential measurement and ATP content detection.Statistical analysis was made on the test results. Results After ischemia reperfusion injury,the levels of MB,LPO,and LPC in skeletal muscle,liver,lung and renal tissues of the control group were significantly increased(MB:P =0.003,P =0.001,P =0.001,P =0.001;LPO:P =0.001,P =0.001,P =0.001,P =0.002;LPC:P =0.000,P =0.002,P =0.002,P =0.003),and hydrogen sulfide treatment during ischemia reperfusion significantly inhibited the production of MB,LPO,and LPC(MB:P =0.021,P =0.036,P =0.005;LPO:P =0.003,P =0.008,P =0.010,P =0.015;LPC:P =0.002,P =0.026,P =0.007,P =0.006).Ischemia/reperfusion of lower extremity in rats resulted in increased levels of IL-1,IL-6,and TNF-α in the serum of rats,and the levels of IL-1,IL-6,and TNF-increased over time,with statistically significant differences in IL-1,IL-6,and TNF-α among groups at 3 h(IL-1:P =0.019,P =0.011,P =0.009,$P_{12_{h}}$=0.008,and P =0.002;IL-6:P =0.026,P =0.009,P =0.002, $P_{12_{h}}$=0.002,P =0.003;TNF-α:P =0.002,P =0.002,P =0.005,$P_{12_{h}}$=0.002,P =0.003).The levels of IL-1,IL-6,and TNF-α in serum were significantly inhibited during ischemia reperfusion(IL-1:P =0.035,P =0.039,P =0.012,$P_{12_{h}}$=0.005,P =0.006;IL-6:P =0.042,P =0.025,P =0.023,$P_{12_{h}}$=0.006,P =0.005;TNF-α:P =0.005,P =0.003,P =0.022,$P_{12_{h}}$=0.005,P =0.005),and such inhibitory effects became even more obvious over time.After limb ischemia and reperfusion in the control group,the mitochondrial transmembrane potential of skeletal muscle cells significantly decreased compared with that of the sham group(t=6.698;P=0.001).After hydrogen sulfide treatment,the mitochondrial membrane potential energy of the experimental group was significantly higher than that of the control group(t=7.507,P = 0.000).The ATP level in the mitochondria of ischemia reperfusion rats in the control group was significantly lower than that in the sham group(t=7.526,P = 0.000).The content of mitochondrial ATP in the experimental group was significantly higher than that in the control group after hydrogen sulfide treatment(t=8.604,P = 0.000). Conclusions Hydrogen sulfide can alleviate the injury of skeletal muscle and distal organs after limb ischemia-reperfusion and reduce local inflammatory reaction.In addition,it is valuable in alleviating mitochondrial transmembrane potential and energy metabolism disorders during reperfusion injury.
Animals
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Energy Metabolism
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Hydrogen Sulfide
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pharmacology
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Inflammation
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metabolism
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Interleukin-6
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metabolism
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Mitochondrial Diseases
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pathology
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Rats
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Rats, Wistar
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Reperfusion Injury
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Tumor Necrosis Factor-alpha
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metabolism
10.Correlation between FOXP3, CD11c Protein Expression and Prognosis of Patients with Diffuse Large B-cell Lymphoma.
Yan-Bing LI ; Zhi WANG ; Hong-Bo ZUO ; Li-Bo LI
Journal of Experimental Hematology 2021;29(1):104-108
OBJECTIVE:
To investigate the correlation between FOXP3, CD11c protein expression and the prognosis of patients with diffuse large B-cell lymphoma (DLBCL).
METHODS:
This study included 48 patients with DLBCL who were admitted to Jiujiang No.1 People's Hospital and TCM-Integrated Hospital of Southern Medical University from January 2015 to January 2019. The DLBCL tissues removed during the operation were collected as test specimens. The expression of FOXP3 and CD11c protein were detected by immunohistochemistry. The deadline for postoperative follow-up was December 31, 2019, and the patient's short-term efficacy (complete remission, partial remission) and progression-free survival were recorded.
RESULTS:
FOXP3 protein was positively expressed in the nucleus, mostly focally or diffusely distributed, the FOXP3
CONCLUSION
In some patients with DLBCL, FOXP3 and CD11c expresse positively, and the positive expression rate is related to the clinical stage and international prognostic index score. The positive expression of FOXP3 and CD11c indicate a good prognosis.
Forkhead Transcription Factors
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Humans
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Immunohistochemistry
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Lymphoma, Large B-Cell, Diffuse
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Prognosis
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Proteomics