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1.
Chinese Journal of Biotechnology ; (12): 190-210, 2024.
Artigo em Chinês | WPRIM | ID: wpr-1008089

RESUMO

The Spt-Ada-Gcn5-acetyltransferase (SAGA) is an ancillary transcription initiation complex which is highly conserved. The ADA1 (alteration/deficiency in activation 1, also called histone H2A functional interactor 1, HFI1) is a subunit in the core module of the SAGA protein complex. ADA1 plays an important role in plant growth and development as well as stress resistance. In this paper, we performed genome-wide identification of banana ADA1 gene family members based on banana genomic data, and analyzed the basic physicochemical properties, evolutionary relationships, selection pressure, promoter cis-acting elements, and its expression profiles under biotic and abiotic stresses. The results showed that there were 10, 6, and 7 family members in Musa acuminata, Musa balbisiana and Musa itinerans. The members were all unstable and hydrophilic proteins, and only contained the conservative SAGA-Tad1 domain. Both MaADA1 and MbADA1 have interactive relationship with Sgf11 (SAGA-associated factor 11) of core module in SAGA. Phylogenetic analysis revealed that banana ADA1 gene family members could be divided into 3 classes. The evolution of ADA1 gene family members was mostly influenced by purifying selection. There were large differences among the gene structure of banana ADA1 gene family members. ADA1 gene family members contained plenty of hormonal elements. MaADA1-1 may play a prominent role in the resistance of banana to cold stress, while MaADA1 may respond to the Panama disease of banana. In conclusion, this study suggested ADA1 gene family members are highly conserved in banana, and may respond to biotic and abiotic stress.


Assuntos
Musa/genética , Filogenia , Proteínas Fúngicas , Núcleo Celular , Histonas , Estresse Fisiológico/genética
2.
Journal of Clinical Hepatology ; (12): 700-705, 2024.
Artigo em Chinês | WPRIM | ID: wpr-1016512

RESUMO

ObjectiveTo investigate the association of the polymorphisms of the acetyl-CoA acetyltransferase 1 (ACAT1) gene and the melatonin receptor 1B (MTNR1B) gene with the susceptibility to nonalcoholic fatty liver disease (NAFLD). MethodsA total of 164 healthy controls and 228 NAFLD patients were enrolled in this study. PCR and sequencing methods were used to determine the genotypes of the polymorphisms of the ACAT1 gene at the rs1044925 and rs1157651 loci and the MTNR1B gene at the rs10830963 locus, and fasting venous blood samples were collected for biochemical analysis. The t-test was used for comparison of normally distributed continuous data between groups, and the non-parametric Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups; the chi-square test was used for comparison of categorical data between groups. ResultsThere were no significant differences between the NAFLD group and the healthy control group in the genotype distribution of the ACAT1 gene at the rs1044925 and rs1157651 loci and the MTNR1B gene at the rs10830963 locus (all P>0.05). The carriers of AA genotype at the rs1044925 locus of the ACAT1 gene had a significantly higher level of low-density lipoprotein than the carriers of C allele (Z=-2.08, P=0.04), and the carriers of G allele at the rs10830963 locus of the MTNR1B gene had a significantly higher level of fasting blood glucose than the carriers of CC genotype (Z=-3.01, P<0.01). ConclusionThe polymorphisms of the ACAT1 gene at the rs1044925 and rs1157651 loci and the MTNR1B gene at the rs10830963 locus were not associated with the susceptibility to NAFLD. The rs1044925 locus of the ACAT1 gene and the rs10830963 locus of the MTNR1B gene are associated with the levels of low-density lipoprotein and fasting blood glucose, respectively.

3.
Chinese Pharmacological Bulletin ; (12): 71-76, 2023.
Artigo em Chinês | WPRIM | ID: wpr-1013880

RESUMO

Aim To study the effect of human urinary kallidinogenase(HUK)on the cognitive function of SAMP8 mouse model and its mechanism. Methods SAMP8 mice were divided intofive groups:SAMP8 group,treatment group(giving 8.75×10-3,1.75×10-2,3.5×10-2,7.0×10-2 HUK),and the SAMR1 vehicle group was used as blank control. Each group was performed Morris water maze to detect spatial cognition. Afterwards the group with the most obvious cognitive improvement(HUK group)was selected for the follow-up experiments. Immunohistochemical detection of ChAT expression in CA3 area was further verified by RtPCR. Western blot was used to detect the expression of PSD95,SYN,BDNF,and pCREB protein. The activity of MPO and the content of IL-1β and IL-18 were determined. Results The passing times in the SAMP8 group was less than that of the SAMR1 group(P<0.05). The passing times of treatment group increased compared with the SAMP8 group(P<0.05 or P<0.01),and the spatial probe time of the target quadrant was shorter(P<0.05 or P<0.01). We conducted follow-up experiments with group d(HUK group). The expression of ChAT positive cells in CA3 area of SAMP8 group was significantly lower than that of SAMR1 group; the expression of positive cells in HUK group significantly increased; RtPCR showed that ChAT expression in SAMP8 group was lower than that in SAMR1 group,and ChAT expression was significantly higher than that in SAMP8 group after HUK treatment. Compared with the SAMR1 group,the levels of IL-1β,IL-18 and MPO activity in the CA3 area of SAMP8 group significantly increased,and the protein expressions of PSD95,SYN,BNDF and pCREB decreased. After HUK treatment,the content of IL-1β,IL-18 and MPO activity decreased,and the expression of PSD95,SYN,BNDF and pCREB increased. Conclusions HUK can improve the spatial cognition of SAMP8 mice. The mechanism may be achieved by promoting the expression of ChAT in CA3 area,reducing the oxidative stress and increasing synapse-related proteins.

4.
Acta Pharmaceutica Sinica B ; (6): 4733-4747, 2023.
Artigo em Inglês | WPRIM | ID: wpr-1011203

RESUMO

Glioblastoma (GBM) is a highly aggressive and lethal brain tumor with an immunosuppressive tumor microenvironment (TME). In this environment, myeloid cells, such as myeloid-derived suppressor cells (MDSCs), play a pivotal role in suppressing antitumor immunity. Lipometabolism is closely related to the function of myeloid cells. Here, our study reports that acetyl-CoA acetyltransferase 1 (ACAT1), the key enzyme of fatty acid oxidation (FAO) and ketogenesis, is significantly downregulated in the MDSCs infiltrated in GBM patients. To investigate the effects of ACAT1 on myeloid cells, we generated mice with myeloid-specific (LyzM-cre) depletion of ACAT1. The results show that these mice exhibited a remarkable accumulation of MDSCs and increased tumor progression both ectopically and orthotopically. The mechanism behind this effect is elevated secretion of C-X-C motif ligand 1 (CXCL1) of macrophages (Mφ). Overall, our findings demonstrate that ACAT1 could serve as a promising drug target for GBM by regulating the function of MDSCs in the TME.

5.
Chinese Journal of Oncology ; (12): 540-549, 2022.
Artigo em Chinês | WPRIM | ID: wpr-940920

RESUMO

Objective: To observe the platinum drugs resistance effect of N-acetyltransferase 10 (NAT10) overexpression in breast cancer cell line and elucidate the underlining mechanisms. Methods: The experiment was divided into wild-type (MCF-7 wild-type cells without any treatment) group, NAT10 overexpression group (H-NAT10 plasmid transfected into MCF-7 cells) and NAT10 knockdown group (SH-NAT10 plasmid transfected into MCF-7 cells). The invasion was detected by Transwell array, the interaction between NAT10 and PARP1 was detected by co-immunoprecipitation. The impact of NAT10 overexpression or knockdown on the acetylation level of PARP1 and its half-life was also determined. Immunostaining and IP array were used to detect the recruitment of DNA damage repair protein by acetylated PARP1. Flow cytometry was used to detect the cell apoptosis. Results: Transwell invasion assay showed that the number of cell invasion was 483.00±46.90 in the NAT10 overexpression group, 469.00±40.50 in the NAT10 knockdown group, and 445.00±35.50 in the MCF-7 wild-type cells, and the differences were not statistically significant (P>0.05). In the presence of 10 μmol/L oxaliplatin, the number of cell invasion was 502.00±45.60 in the NAT10 overexpression group and 105.00±20.50 in the NAT10 knockdown group, both statistically significant (P<0.05) compared with 219.00±31.50 in wild-type cells. In the presence of 10 μmol/L oxaliplatin, NAT10 overexpression enhanced the binding of PARP1 to NAT10 compared with wild-type cells, whereas the use of the NAT10 inhibitor Remodelin inhibited the mutual binding of the two. Overexpression of NAT10 induced PARP1 acetylation followed by increased PARP1 binding to XRCC1, and knockdown of NAT10 expression reduced PARP1 binding to XRCC1. Overexpression of NAT10 enhanced PARP1 binding to LIG3, while knockdown of NAT10 expression decreased PARP1 binding to LIG3. In 10 μmol/L oxaliplatin-treated cells, the γH2AX expression level was 0.38±0.02 in NAT10 overexpressing cells and 1.36±0.15 in NAT10 knockdown cells, both statistically significant (P<0.05) compared with 1.00±0.00 in wild-type cells. In 10 μmol/L oxaliplatin treated cells, the apoptosis rate was (6.54±0.68)% in the NAT10 overexpression group and (12.98±2.54)% in the NAT10 knockdown group, both of which were statistically significant (P<0.05) compared with (9.67±0.37)% in wild-type cells. Conclusion: NAT10 overexpression enhances the binding of NAT10 to PARP1 and promotes the acetylation of PARP1, which in turn prolongs the half-life of PARP1, thus enhancing PARP1 recruitment of DNA damage repair related proteins to the damage sites, promoting DNA damage repair and ultimately the survival of breast cancer cells.


Assuntos
Feminino , Humanos , Neoplasias da Mama/enzimologia , Linhagem Celular Tumoral , Resistencia a Medicamentos Antineoplásicos , Células MCF-7 , Acetiltransferases N-Terminal/metabolismo , Compostos Organoplatínicos/farmacologia , Oxaliplatina/farmacologia , Proteína 1 Complementadora Cruzada de Reparo de Raio-X
6.
Chinese Journal of Biochemistry and Molecular Biology ; (12): 1023-1032, 2022.
Artigo em Chinês | WPRIM | ID: wpr-1015774

RESUMO

Lysine acetyltransferase 5 (KAT5), a member of the MYST family, can participate in cellular processes such as transcription, DNA repair, differentiation and signal transduction by acetylating different substrates. The role of KAT5 cannot be replaced by other MYST family members, and the knockout of KAT5 can directly lead to apoptosis, indicating that KAT5 may be located in the upstream of physiological signaling pathways in cells and play an extremely important and unique role. Therefore, the changes in KAT5 expression are very likely to lead to the occurrence and development of tumors. Previous studies have found that KAT5 is downregulated in breast cancer, melanoma, and lung cancer, and is considered a tumor suppressor in these tumors. However, in recent years, studies have found that KAT5 can be either highly or lowly expressed in breast cancer, liver cancer, melanoma, prostate cancer, lung cancer and other tumors. On the premise of high KAT5 expression, KAT5 can play a tumor-promoting role. While on the premise of low KAT5 expression, KAT5 can also play as a tumor suppressor. With further decrease of KAT5 expression, its tumor suppressive effect is weakened, which may lead to the occurrence and development of tumors. In addition, KAT5 has also been found to be differentially expressed in osteosarcoma, thyroid cancer, glioblastoma, colorectal cancer and other tumors, and the differential expression of KAT5 is closely related to the proliferation, metastasis, apoptosis, drug and radiotherapy resistance of tumor cells. Therefore, KAT5 is one of the potential tumor therapeutic targets. Here, we summarize the expression of KAT5 in tumors and the tumor-suppressing or tumor-promoting signaling pathways involved in the corresponding expression in recent years, hoping to provide new inspiration and reference for tumor treatment and prognosis monitoring.

7.
Chinese Journal of Biochemistry and Molecular Biology ; (12): 603-613, 2022.
Artigo em Chinês | WPRIM | ID: wpr-1015706

RESUMO

N-acetyltransferase 10 (NAT10) is a key enzyme involved in the acetylation of mRNA, which regulates the expression of target genes and biological functions of various cancers via ac4C (N4-acetylcytidine) acetylation. However, whether NAT10 is involved in regulating the malignant behavior of cervical cancer is rarely reported. This study investigated the effects and specific mechanism of NAT10 on the growth activity, proliferation capacity, migration and invasion capacity of cervical cancer cells. First, we found a highly expression of NAT10 in cervical cancer and was associated with poor patient prognosis by TCGA database analysis. MTT assays and colony formation assays showed that overexpression of NAT10 promoted the growth activity (P<0. 05) and proliferation ability (P<0. 001) of cervical cancer cells; Transwell migration and invasion assays showed that overexpression of NAT10 promoted the migration (P<0. 01) and invasion (P<0. 05) ability of cervical cancer cells; Western blotting showed that overexpression of NAT10 increased the mesothelial cell marker vimentin and resulted in a decrease in the epithelial cell marker E-cadherin. Bioinformatics analysis exhibited that discoidin domain receptor 1 (DDR1) might be a downstream target gene of NAT10. RNA binding protein immunoprecipitation (RIP) assays showed that NAT10 could directly bind to DDR1 mRNA (P<0. 05), Western blotting and RT-qPCR experiments showed that overexpression of NAT10 significantly increased the expression and mRNA levels of DDR1 (P<0. 05). Furthermore, RNA acetylation co-immunoprecipitation experiments showed that overexpression of NAT10 promoted the acetylation level of DDR1 mRNA (P<0. 001), and EGFP reporter assays confirmed that NAT10 recognized the acetylation site of DDR1 mRNA. The results of mRNA half-life experiments showed that NAT10 increased the stability of DDR1 mRNA (P<0. 05). In conclusion, our research confirms that NAT10 promotes the growth activity, and migration and invasion ability of cervical cancer cells and its specific mechanism is to extend the stability of DDR1 by acetylation modification, thereby increasing its expression levels, which might provide new insights into the molecular mechanism of acetylation modification of mRNA on the pathogenesis of cervical cancers.

8.
Rev. ANACEM (Impresa) ; 16(2): 101-107, 2022. ilus
Artigo em Espanhol | LILACS | ID: biblio-1525495

RESUMO

Las N-terminal acetiltransferasas (NaT) son fundamentales en el desarrollo, funcionamiento y vida media celular, acetilando gran parte del proteoma humano. Entre las ocho NaT identificadas, N-terminal acetiltransferasa A (NaTA) acetila a un mayor número de sustratos, teniendo además un rol fundamental en el neurodesarrollo. Previamente, estudios han demostrado que mutaciones en la subunidad catalítica de NaTA, NAA10, se asocian con trastornos del neurodesarrollo. Sin embargo, nuevas líneas investigativas sugieren que mutaciones de la subunidad auxiliar, NAA15, también tendrían un rol importante en el desarrollo de estos trastornos. Esta revisión se realiza con el objetivo de recopilar evidencia sobre variantes de NAA15 relacionadas con Discapacidad Intelectual (DI) y Trastorno de Espectro Autista (TEA). Se consultaron fuentes actualizadas sobre acetilación N-terminal, NaT, DI y TEA y mutaciones reportadas de NAA15 y sus expresiones fenotípicas, publicadas entre 2011 y 2022. Se concluye que, aun cuando existe relación entre mutaciones de NAA15, DI y TEA, todavía es necesario esclarecer los mecanismos fisiopatológicos de estos trastornos, el rol de NaTA y el impacto de variantes de sus subunidades en las vías moleculares y el fenotipo, lo que se dificulta por razones que van desde la complejidad de estas vías hasta el elevado costo de análisis genéticos. Se sugiere continuar la investigación en esta área, para comprender las bases moleculares subyacentes a estos trastornos y el rol de las mutaciones en subunidades de NaTA, con el fin último de estudiar potenciales tratamientos que mejoren la calidad de vida de las personas con estos trastornos y sus familias.


Nt-acetyltransferases (NaT) are essential in cell development, function and half-life, catalyzing most of the human proteome. Among the eight NaTs identified, N-terminal acetyltransferase A (NaTA) acetylates a greater number of substrates, also having a fundamental role in neurodevelopment. Previously, studies have shown that mutations in the catalytic subunit of NaTA, NAA10, are associated with neurodevelopmental disorders. However, new research lines suggest that mutations of the NAA15 helper subunit also plays an important role in the development of these disorders. This review is carried out with the objective of gathering evidence on NAA15 variants related to Intellectual Disability (ID) and Autism Spectrum Disorder (ASD). Updated sources on N-terminal acetylation, N-acetyltransferases, DI and TEA and reported mutations of NAA15 and their phenotypic expressions, published between 2011 and 2022 were consulted. It is concluded that even though there is a relationship between mutations of NAA15, ID and ASD exists, it is still necessary to clarify the pathophysiological mechanisms of these disorders, the role of NaTA and the impact of variants of its subunits in the molecular pathways and in the phenotype, for reasons ranging from the complexity of these pathways to the high cost of genetic testing. It is suggested to continue research in this area, to understand the molecular bases underlying these disorders and the role of mutations in NatA subunits, with the ultimate aim of studying potential treatments that improve the quality of life of people with these disorders and their families.


Assuntos
Humanos , Acetiltransferase N-Terminal A/genética , Transtorno do Espectro Autista/genética , Deficiência Intelectual/genética , Variação Genética , Acetiltransferase N-Terminal A/metabolismo , Mutação/genética
9.
Mem. Inst. Oswaldo Cruz ; 117: e210328, 2022. tab, graf
Artigo em Inglês | LILACS-Express | LILACS | ID: biblio-1375902

RESUMO

BACKGROUND Distinct N-acetyltransferase 2 (NAT2) slow acetylators genotypes have been associated with a higher risk to develop anti-tuberculosis drug-induced hepatotoxicity (DIH). However, studies have not pointed the relevance of different acetylation phenotypes presented by homozygotes and compound heterozygotes slow acetylators on a clinical basis. OBJECTIVES This study aimed to investigate the association between NAT2 genotypes and the risk of developing DIH in Brazilian patients undergoing tuberculosis treatment, focusing on the discrimination of homozygotes and compound heterozygotes slow acetylators. METHODS/FINDINGS The frequency of NAT2 genotypes was analysed by DNA sequencing in 162 patients undergoing tuberculosis therapy. The mutation analyses revealed 15 variants, plus two new NAT2 mutations, that computational simulations predicted to cause structural perturbations in the protein. The multivariate statistical analysis revealed that carriers of NAT2*5/*5 slow acetylator genotype presented a higher risk of developing anti-tuberculosis DIH, on a clinical basis, when compared to the compound heterozygotes presenting NAT2*5 and any other slow acetylator haplotype [aOR 4.97, 95% confidence interval (CI) 1.47-16.82, p = 0.01]. CONCLUSION These findings suggest that patients with TB diagnosis who present the NAT2*5B/*5B genotype should be properly identified and more carefully monitored until treatment outcome in order to prevent the occurrence of anti-tuberculosis DIH.

10.
Braz. J. Pharm. Sci. (Online) ; 58: e19221, 2022. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1374557

RESUMO

Abstract The purpose of the current work was to assess a possible role of cytochrome P450 1A2 (CYP1A2) and N-acetyltransferase 2 (NAT2) in the metabolic activation of 2,6-dimethylaniline (2,6-DMA) and also clarify the function of DNA repair in affecting the ultimate mutagenic potency. Two cell lines, nucleotide excision repair (NER)-deficient 5P3NAT2 and proficient 5P3NAT2R9 both expressing CYP1A2 and NAT2, were treated with 2,6-DMA for 48 h or its metabolites for 1 h. Cell survival determined by trypan blue exclusion and MTT assays, and 8-azaadenine-resistant mutants at the adenine phosphoribosyltransferase (aprt) gene locus were evaluated. 5P3NAT2 and 5P3NAT2R9 cells treated with 2,6-DMA and its metabolites showed a dose-dependent increase in cytotoxicity and mutant fraction; N-OH-2,6-DMA and 2,6-DMAP in serum-free α-minimal essential medium (MEM) are more potent than 2,6-DMA in complete MEM. 5P3NAT2 cells was more sensitive to the cytotoxic and mutagenic action than 5P3NAT2R9 cells. H2DCFH-DA assay showed dose-dependent ROS production under 2,6- DMAP treatment. These findings indicate that the genotoxic effects of 2,6-DMA are mediated by CYP1A2 activation via N-hydroxylation and the subsequent esterification by the phase II conjugation enzyme NAT2, and through the generation of ROS by hydroxylamine and/or aminophenol metabolites. NER status is also an important contributor


Assuntos
Células/classificação , Citocromo P-450 CYP1A2/análise , Genotoxicidade , Linhagem Celular/classificação , Hidroxilamina/agonistas , Reparo do DNA
11.
Acta Pharmaceutica Sinica B ; (6): 838-852, 2022.
Artigo em Inglês | WPRIM | ID: wpr-929329

RESUMO

Parkin, an E3 ubiquitin ligase, plays a role in maintaining mitochondrial homeostasis through targeting damaged mitochondria for mitophagy. Accumulating evidence suggests that the acetylation modification of the key mitophagy machinery influences mitophagy level, but the underlying mechanism is poorly understood. Here, our study demonstrated that inhibition of histone deacetylase (HDAC) by treatment of HDACis activates mitophagy through mediating Parkin acetylation, leading to inhibition of cervical cancer cell proliferation. Bioinformatics analysis shows that Parkin expression is inversely correlated with HDAC2 expression in human cervical cancer, indicating the low acetylation level of Parkin. Using mass spectrometry, Parkin is identified to interact with two upstream molecules, acetylase acetyl-CoA acetyltransferase 1 (ACAT1) and deacetylase HDAC2. Under treatment of suberoylanilide hydroxamic acid (SAHA), Parkin is acetylated at lysine residues 129, 220 and 349, located in different domains of Parkin protein. In in vitro experiments, combined mutation of Parkin largely attenuate the interaction of Parkin with PTEN induced putative kinase 1 (PINK1) and the function of Parkin in mitophagy induction and tumor suppression. In tumor xenografts, the expression of mutant Parkin impairs the tumor suppressive effect of Parkin and decreases the anticancer activity of SAHA. Our results reveal an acetylation-dependent regulatory mechanism governing Parkin in mitophagy and cervical carcinogenesis, which offers a new mitophagy modulation strategy for cancer therapy.

12.
Acta Pharmaceutica Sinica B ; (6): 3322-3334, 2021.
Artigo em Inglês | WPRIM | ID: wpr-922797

RESUMO

10-Deacetylbaccatin III (10-DAB) C10 acetylation is an indispensable procedure for Taxol semi-synthesis, which often requires harsh conditions. 10-Deacetylbaccatin III-10-

13.
Chinese Journal of Biochemistry and Molecular Biology ; (12): 339-346, 2021.
Artigo em Chinês | WPRIM | ID: wpr-1015971

RESUMO

Mitochondrial pyruvate dehydrogenase complex (PDC) is crucial for glucose homeostasis in mammalian cells‚ decarboxylation of glycolytic intermediate pyruvate to acetyl coenzyme A (acetyl-CoA) in mitochondria. Dihydrolipoyl acetyltransferase (DLAT) is a subunit of the pyruvate dehydrogenase complex. Here‚ we reported that DLAT was commonly increased in lung cancer and its expression was associated with worse clinical outcomes. We found that suppression of DLAT in lung cancer cells resulted in reduced nucleic acid biosynthesis and attenuated cancer cell proliferation through controlling acetylation level of 6-phosphogluconate dehydrogenase (6PGD) ‚ the third enzyme in the oxidative pentose phosphate pathway (PPP) ‚ in which ribulose-5-phosphate (Ru-5-P) is produced for nucleic acid biosynthesis. Together‚ our study contributes to recent interest and discussion cross talk in cancer metabolism‚ which contributes to tumor growth. Future mechanistic studies should lead to the elucidation of the mode of action of DLAT in human lung cancer and establish DLAT as a viable drug target.

14.
Chinese Journal of Biochemistry and Molecular Biology ; (12): 475-486, 2021.
Artigo em Chinês | WPRIM | ID: wpr-1015955

RESUMO

Lysine acetylation has emerged as one of the most important post-translational modifications that participates in various biological and pathological processes. Histone acetyltransferase 1 (HAT1) as the first identified protein ε-amino lysine acetyltransferase is able to regulate the acetylation of histones and non-histone proteins. However‚ the acetylation substrates and sites mediated by HAT1 in liver cancer are poorly understood. In this study‚ we demonstrated that HAT1 was highly expressed in the liver cancer tissues‚ which was negatively associated with the prognosis of patients. Based on the establishment of the HAT1-knockout HepG2 cell line‚ we employed a quantitative proteomics approach to study the profiling of acetylation mediated by HAT1 in HepG2 cells. Interestingly‚ we identified a total of 858 Kac sites on 547 proteins in the HepG2 cell line‚ in which HAT1 mediated the levels of Kac of 74 sites on 68 proteins. The pathways and metabolic processes that were affected by HAT1-dependent acetylation modification were analyzed by bioinformatics. The results show that Kac regulates disease development‚ RNA biology‚ spliceosome and nucleosome assembly‚ oxidative stress‚ various signaling pathways and metabolic pathways‚ etc.. Moreover‚ we verified that the HAT1-mediated acetylation modification could promote abnormal lipid metabolism. CCK8 assays‚ clone formation and Edu assays revealed that HAT1 could remarkably enhance the cell proliferation of liver cancer in vitro. Thus‚ our finding explored the profiling of HAT1-mediated protein acetylation in HepG2 cells‚ which provides new insights into the underlying mechanism by which HAT1 mediates the development of liver cancer. Clinically‚ the HAT1-mediated acetylation sites could be used for the precise targets of drug development.

15.
Chinese Journal of Clinical Pharmacology and Therapeutics ; (12): 1186-1192, 2021.
Artigo em Chinês | WPRIM | ID: wpr-1014963

RESUMO

Reversible post-translational modification of proteins is an important process in the physiological regulation of all tissues, including the heart. Lysine acetylation occurs in all organisms, including prokaryotes, and is regulated by a balance between lysine acetyltransferase (adding acetyl to the ε-amino group of lysine) and deacetylase (acetyl group that removes lysine ε-amino group). The heart is an organ rich in acetylated lysine, but the role of acetylated lysine in the heart remains to be elucidated. Therefore, in this paper, we systematically reviewed the gene list of acetyltransferase and deacetylase in mammalian genome and indicated their mRNA expression. The purpose of this study is to discover the research progress of dynamic regulation of lysine acetylation in heart disease and to provide a theoretical basis for the discovery of molecular targets.

16.
Chinese Journal of Anesthesiology ; (12): 965-969, 2021.
Artigo em Chinês | WPRIM | ID: wpr-911309

RESUMO

Objective:To evaluate the relationship between choline acetyltransferase (ChAT) positive neurons in parabrachial nucleus and development of fear memory in mice.Methods:Eighteen healthy male ChAT-ires-cre mice, aged 8-9 weeks, weighing 22-25 g, were divided into 3 groups ( n=6 each) using a random number table method: Cre-dependent AAV-DIO-hM 3Dq-mcherry (Gq) virus/clozapine-N-oxide (CNO) group (group Gq/CNO), Gq/normal saline (NS) group (group Gq/NS) and Cre-dependent AAV-DIO-mcherry (mc) virus/CNO group (group mc/CNO). Gq virus was injected into parabrachial nucleus, and CNO 2 mg/kg was injected intraperitoneally 3 weeks later in group Gq/CNO.Gq virus was injected into parabrachial nucleus, and the equal volume of normal saline was injected intraperitoneally 3 weeks later in group Gq/NS.In group mc/CNO, mc virus was injected into parabrachial nucleus, and CNO 2 mg/kg was injected intraperitoneally 3 weeks later.The fear conditioning test was performed at 30 min after intraperitoneal injection in all the 3 groups.The brains were then removed and sliced.The virus expression and areas of the brain projected by ChAT positive neurons were observed. Results:Compared with group Gq/CNO, the percentage of freezing time was significantly increased during testing phase in Gq/NS and mc/CNO groups ( P<0.05). Gq/mc virus carrying fluorescent protein mcherry was expressed in parabrachial nucleus and was co-expressed with mcherry-ChAT.The fibers of ChAT positive neurons projected to the red nucleus, substantia nigra, central amygdala, anterodorsal thalamic nucleus and bed nucleus of stria terminalis. Conclusion:The ChAT positive neurons in parabrachial nucleus are involved in the regulation of the development of fear memory in mice, which can impair fear memory, and the regulation is carried out probably through central amygdala.

17.
Journal of International Oncology ; (12): 415-419, 2021.
Artigo em Chinês | WPRIM | ID: wpr-907554

RESUMO

Objective:To investigate the expression and clinical significance of histone acetyltransferase P300 in hepatocellular carcinoma.Methods:From January 2013 to December 2017, surgical specimens of 100 patients with hepatocellular carcinoma were collected from the Department of General Surgery of Sichuan Mianyang 404 Hospital. The expressions of P300, CD90, alpha fetoprotein (AFP), Ki-67 and CD34 in hepatocellular carcinoma tissue were detected. At the same time, 42 hepatic hemangioma specimens and 56 liver tissue specimens with moderate to severe liver cirrhosis were collected, and the positive expression rate of P300 in tissues was detected. The correlations between the expression of P300 and clinicopathological features and prognosis of patients with hepatocellular carcinoma were analyzed.Results:The positive expression rates of P300 in normal liver tissue, liver cirrhosis tissue and hepatocellular carcinoma tissue increased gradually, which were 11.9% (5/42), 32.1% (18/56) and 57.0% (57/100) respectively, with a statistically significant difference ( χ2=27.192, P<0.001). Tumor grade ( χ2=9.337, P=0.009), T stage ( χ2=8.794, P=0.032), clinical TNM stage ( χ2=6.121, P=0.013), AFP ( χ2=11.040, P=0.001), CD90 ( χ2=9.903, P=0.002), CD34 ( χ2=4.066, P=0.044) significantly affected the expression of P300. Spearman rank correlation analysis showed that the abnormal expression of P300 was positively correlated with the expression of AFP ( r=0.335, P=0.001), CD90 ( r=0.328, P=0.002) and CD34 ( r=0.264, P=0.047) , but had no significant correlation with the expression of Ki-67 ( P>0.05). Survival analysis showed that the 5-year survival rate of patients with P300 positive expression was 17.6%, and that of patients with P300 negative expression was 62.5%, and there was a statistically significant difference ( χ2=10.596, P<0.001). Cox multivariate analysis showed that P300 positive expression ( RR=2.554, 95% CI: 1.261-4.502, P=0.009), CD90 positive expression ( RR=3.574, 95% CI: 1.021-11.980, P=0.030) and TNM Ⅱ-Ⅳ stage ( RR=0.332, 95% CI: 0.105-0.596, P=0.002) were independent risk factors for poor prognosis of patients with hepatocellular carcinoma. Conclusion:The positive expression of P300 is closely related to the occurrence of hepatocellular carcinoma and can be used as an independent factor to judge the poor prognosis of patients with hepatocellular carcinoma.

18.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 47-52, 2021.
Artigo em Chinês | WPRIM | ID: wpr-906329

RESUMO

Objective:To observe the effects of Cnidii Fructus hypnotic active components (CHC) on the behaviors of rats with p-chlorophenylalanine (PCPA)-induced insomnia and melatonin (MT) synthesis rate-limiting enzyme arylalkylamine <italic>N</italic>-acetyltransferase (AANAT), and explore the protective mechanism of CHC on the pineal gland. Method:Male SD rats of SPF grade were randomly divided into a blank control group, a model group, a MT group, and high-, medium-, and low-dose CHC groups with 10 rats in each group. Except for the blank control group, other groups received 4.5% PCPA suspension at 10 mL·kg<sup>-1</sup>, intragastric administration, for two consecutive days. After PCPA model of insomnia was established, normal and model groups were gavaged at the same volume of 2% Tween-80, MT control group (10 mg·kg<sup>-1</sup>), CHC was high, medium and low (60, 30, 15 mg·kg<sup>-1</sup>), 10 mL·kg<sup>-1</sup>, once a day, for consecutive 7 days. Four days after administration, open field, elevated cross maze, and pentobarbital sodium-induced sleep tests were conducted, respectively. Serum MT was detected by enzyme-linked immunosorbent assay. The mRNA expression level of AANAT was determined by real-time fluorescence-based quantitative polymerase chain reaction (Real-time PCR). The expression of AANAT protein in the pineal gland was detected by Western blot. Result:Compared with the results in the blank control group, the total distance of open field activity and standing times and duration in the central area were increased (<italic>P</italic><0.05, <italic>P</italic><0.01), the proportions of open arm entry (OE%) and open arm time (OT%) were decreased (<italic>P</italic><0.05), and the sleep latency was prolonged (<italic>P</italic><0.01) in the model group. Compared with the model group, no significant difference was observed in the low-dose CHC group, while other groups exhibited reduced total distance of activity (<italic>P</italic><0.05, <italic>P</italic><0.01), elevated OE% (<italic>P</italic><0.05), shortened sleep latency, and prolonged sleep time (<italic>P</italic><0.05, <italic>P</italic><0.01). Compared with the serum MT in the blank control group, that in the model group was decreased (<italic>P</italic><0.01). Compared with the model group, no significant difference was observed in the low-dose CHC group, while other groups displayed increased serum MT (<italic>P</italic><0.05). The mRNA and protein expression of AANAT was decreased in the model group as compared with that in the blank control group (<italic>P</italic><0.01). Compared with the model group, the MT group and the high-dose CHC group showed up-regulated expression (<italic>P</italic><0.05). Conclusion:CHC improved the behavioral indexes of PCPA-induced insomnia, increased the synthesis and secretion of MT in pineal cells, and elevated the serum MT level, which was related to the up-regulation of the mRNA and protein expression of AANAT in the pineal gland.

19.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 97-107, 2021.
Artigo em Chinês | WPRIM | ID: wpr-906306

RESUMO

Objective:To investigate the relationship between the single nucleotide polymorphism(SNP)of function genes and effective components of <italic>Salvia miltiorrhiza</italic> and the molecular mechanism of specific quality formation of <italic>S. miltiorrhiza</italic>. Method:The fingerprints of components in <italic>S. miltiorrhiza</italic> from eight different habitats and varieties were obtained by high-performance liquid chromatography (HPLC). The full-length cDNA of three functional genes<italic> </italic>acetyl-CoA C-acetyltransferase(<italic>SmAACT</italic>),4-diphosphocytidyl-2-C-methyl-<italic>D</italic>-erythritol kinase(<italic>SmCMK</italic>) and isopentenyl diphosphate isomerase(<italic>SmIPPI</italic>) in tanshinone metabolic pathway were amplified by polymerase chain reaction(PCR),cloned, and sequenced,followed by bioinformatics analysis. Result:The full-length cDNA sequences of three functional genes <italic>SmAACT</italic>,<italic>SmCMK</italic>, and <italic>SmIPPI</italic> in tanshinone metabolic pathway were obtained from 23 strains of <italic>S. miltiorrhiza</italic> from eight different habitats and varieties. As revealed by the analysis of SNP and amino acid polymorphisms of three functional genes,18,16, and 14 SNP sites were found respectively. HPLC results showed the samples from Beijing,Hubei,Shandong (No. SDB),Shanxi,Henan, and Shandong (No. SDZ) were clustered into one branch,and those from Hebei and Inner Mongolia were clustered into another branch, which suggested that the variation trend of <italic>S. miltiorrhiza</italic> components had little correlation with geographical distance,but the variety was a critical factor for the quality. Conclusion:There was an obvious genetic differentiation trend in <italic>S. miltiorrhiza</italic> from different habitats,and different origin-specific genotypes were formed. The molecular mechanism of the formation of the specific quality of <italic>S. miltiorrhiza</italic> from different habitats was discussed,which laid a foundation for the stability and effectiveness of clinical medication,and guided the breeding of excellent varieties of <italic>S. miltiorrhiza</italic>.

20.
Electron. j. biotechnol ; 48: 53-61, nov. 2020. ilus, graf
Artigo em Inglês | LILACS | ID: biblio-1254710

RESUMO

BACKGROUND: Alzheimer's disease (AD) is a chronic, progressive neurodegenerative disease. Recent studies have reported the close association between cognitive function in AD and purinergic receptors in the central nervous system. In the current study, we investigated the effect of CD73 inhibitor α, ß-methylene ADP (APCP) on cognitive impairment of AD in mice, and to explore the potential underlying mechanisms. RESULTS: We found that acute administration of Aß1­42 (i.c.v.) resulted in a significant increase in adenosine release by using microdialysis study. Chronic administration of APCP (10, 30 mg/kg) for 20 d obviously mitigated the spatial working memory impairment of Aß1­42-treated mice in both Morris water maze (MWM) test and Y-maze test. In addition, the extracellular adenosine production in the hippocampus was inhibited by APCP in Aß-treated mice. Further analyses indicated expression of acetyltransferase (ChAT) in hippocampus of mice of was significantly reduced, while acetylcholinesterase (AChE) expression increased, which compared to model group. We observed that APCP did not significantly alter the NLRP3 inflammasome activity in hippocampus, indicating that anti-central inflammation seems not to be involved in APCP effect. CONCLUSIONS: In conclusion, we report for the first time that inhibition of CD73 by APCP was able to protect against memory loss induced by Aß1­42 in mice, which may be due to the decrease of CD73-driven adenosine production in hippocampus. Enhancement of central cholinergic function of the central nervous system may also be involved in the effects of APCP.


Assuntos
Animais , Masculino , Camundongos , Difosfato de Adenosina/análogos & derivados , Doenças Neurodegenerativas/prevenção & controle , Hipocampo , Nucleotidases/antagonistas & inibidores , Acetilcolinesterase , Difosfato de Adenosina/administração & dosagem , Doença de Alzheimer/prevenção & controle , Teste do Labirinto Aquático de Morris , Camundongos Endogâmicos C57BL
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