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Schistosomiasis is a neglected zoonotic parasitic disease. Currently, praziquantel is the drug of choice for the treatment of schistosomiasis, and is the only effective chemical for treatment of schistosomiasis japonica. Since its introduction in the 1970s, praziquantel has been used for large-scale chemotherapy of schistosomiasis for over 40 years. However, there have been reports pertaining to the resistance to praziquantel in schistosomes. Therefore, development of novel antischistosomal agents as alternatives of praziquantel, is of great need. Histone deacetylases and histone acetyltransferases have been recently reported to play critical roles in the growth, development and reproduction of schistosomes, and are considered as potential drug targets for the treatment of schistosomiasis. This review summarizes the latest advances of histone deacetylase and histone acetyltransferase inhibitors in the research on antischistosomal drugs, so as to provide insights into research and development of novelantischistosomal agents.
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Acute myeloid leukemia (AML) is a malignant clonal disease of hematopoietic stem cells, characterized by the proliferation of abnormal primordial cells of myeloid origin in bone marrow, blood and other tissues. At present, the standard induction therapy for AML mainly includes “3+7” standard treatment(anthracycline combined with cytarabine), allogeneic hematopoietic stem cell transplantation (Allo-HSCT) and targeted drug therapy. However, AML cells usually express high levels of P-glycoprotein, which mediates the efflux of chemotherapeutic drugs, which makes AML cells resistant to chemotherapy, resulting in many patients who are not sensitive to chemotherapy or relapse after complete remission. And some patients can not tolerate intensive therapy or lack of donors and can not use Allo-HSCT therapy. Therefore, it is of great clinical significance to find new drugs to improve the efficacy of AML patients. Epigenetic disorders play a key role in the pathogenesis of many diseases, especially cancer. Studies have shown that most AML patients have epigenetic regulatory gene mutations, such as DNMT3A, IDH and TET2, and these mutations are potentially reversible, which has become one of the therapeutic targets of AML. Histone deacetylase inhibitors (HDACi) can regulate the balance between histone acetylation and deacetylation, change the expression of proto-oncogenes or tumor suppressor genes that control cancer progression from epigenetics, and play an important role in many kinds of tumor therapy. At present, HDACi has shown the ability to induce differentiation, cell cycle arrest and apoptosis of AML cells. The mechanism may be mainly related to HDACi inducing chromatin conformation opening of tumor suppressor gene by inhibiting HDAC activity, promoting oncogene damage and preventing oncogene fusion protein from recruiting HDAC. Although the preclinical outcome of HDACi is promising, it is not as effective as the conventional therapy of AML. However, the combination strategy with various anticancer drugs is in clinical trials, showing significant anti-AML activity, improving efficacy through key targeting pathways in a typical synergistic or additive way, increasing AML sensitivity to chemotherapy, reducing tumor growth and metastasis potential, inhibiting cell mitotic activity, inducing cell apoptosis, regulating bone marrow microenvironment, which provides a good choice for the treatment of AML. Especially for those AML patients who are not suitable for intensive therapy and drug resistance to chemotherapy. This review introduces the relationship between HDAC and cancer; the classification of HDAC and its function in AML; the correlation between HDAC and AML; the clinical application of five types of HDACi; preclinical research results and clinical application progress of six kinds of HDACi in AML, such as Vrinota, Belinostat, Panobinostat, Valproic acid, Entinostat, and Chidamide, the mechanism of HDACi combined with other anticancer drugs in AML indicates that the current HDACi is mainly aimed at various subtypes of pan-HDAC inhibitors, with obvious side effects, such as fatigue, thrombocytopenia, nausea, vomiting, diarrhea. In recent years, the next generation of HDACi is mainly focused on the selectivity of analogues or isomers. Finding the best combination of HDACi and other drugs and the best timing of administration to balance the efficacy and adverse reactions is a major challenge in the treatment of AML, and the continued development of selective HDACi with less side effects and more accurate location is the key point for the development of this drug in the future. It is expected to provide reference for clinical treatment of AML.
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Gliomas are commonly central nervous system tumors. The conventional treatment is surgical resection combined with chemoradiotherapy， but glioma patients often have a poor prognosis. Therefore， there is an urgent need to identify new potential targets in gliomas and develop more effective treatments. Valproic acid has the properties of histone deacetylase inhibitor， which has been proven to have inhibitory effects on various tumors. It is confirmed that valproic acid could promote apoptosis and cell arrest of glioma cells， inhibit cell invasion and glioma stem cells， increase the sensitivity of glioma cells to radiotherapy and chemotherapy by regulating ERK/Akt signaling pathway， Akt/mTOR signaling pathway， and regulating expression levels of RECK， MGMT， Nrf2， PON2， Smad4， GSK3β and other proteins. In addition， valproic acid can also enhance the effectiveness of anticancer drugs by inhibiting the growth of glioma stem cells and inducing their differentiation. In conclusion， valproic acid can inhibit glioma through multiple targeted actions， and may become a new targeted drug for the treatment of glioma.
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Epilepsy is a complex neurological disease and its pathogenesis has not yet been clear. More and more evidence shows that epilepsy is closely related to epigenetic mechanisms such as histone deacetylation. Histone deacetylase inhibitors, a kind of epilepsy targeting drug potentially, are involved in the protection of neurogenesis and the regulation of a variety of neural signal cascade reactions. This article reviews the research progress of histone deacetylation in the pathogenesis of epilepsy and application prospect of histone deacetylase inhibitors in treating epilepsy.
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Great progress has been made in the treatment of lymphoma in recent decades, but the prognosis for patients with relapsed or refractory lymphoma is often disappointing. Studies have found that the pathogenesis of non-Hodgkin lymphoma is associated with changes in histone acetylation. Histone deacetylase inhibitors can increase the level of histone acetylation in lymphoma cells, and exert anti-lymphoma effects through mechanisms such as cell cycle inhibition, induction of apoptosis, and immunomodulation. However, histone deacetylase inhibitors alone have limited therapeutic effects, and the combination with other antineoplastic drugs for the treatment of relapsing and refractory non-Hodgkin lymphoma has shown good efficacy. Summarizing basic research and clinical trials of histone deacetylase inhibitor containing regimens provides ideas for the treatment of lymphoma.
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Depression is a serious mental illness with a high incidence. At present, we do not fully understand the specific pathological mechanisms of depression, and the efficacy of drug treatments is very limited. Recent studies have shown that epigenetic changes that occur in specific brain regions may be a key mechanism by which environmental factors to interact with individuals to influence the risk of depression. Therefore, drugs that target epigenetic regulation may become a new direction for the development of antidepressants. Histone deacetylase inhibitors (HDACi) are a class of compounds that inhibit histone deacetylase activity, which has been reported to be associated with depression; this article addresses the use of HDACi in preclinical studies, and their potential therapeutic role and limitations of use in depression.
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Immunotherapy is one of the main strategies of anti-tumor therapy at present, in which immune checkpoint inhibitors (ICIs) are the most widely used drug. ICIs resistance is mediated by a variety of cytokines and immune cells, and the mechanism is complex, which is the main reason for the failure of immunotherapy in cancer patients. Histone deacetylase inhibitor (HDACi), as a class of epigenetic regulatory drugs, plays an important role in regulating cell cycle, proliferation, differentiation, and activity. In recent years, Studies have found that HDACi can not only regulate cell biological characteristics, but also closely related to the improvement of tumor ICIs drug resistance. Therefore, the study on how HDACi enhances the efficacy of ICIs is of great significance to tumor immunotherapy. This article will review the research progress of HDACi combined with ICIs in treating malignant tumors and their related mechanism. .
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Objective To investigate the role of histone deacetylase 3 ( HDAC3 ) inhibitor (HDAC3I) in hypoxia-reoxygenation( H/R) injury of PC12 cells. Methods H/R cell injury model was established by using PC 12 cells for 4 hours hypoxia and then reoxygenation for 24 hours. HDAC3I treatment group was pretreated with RGFP966 for 1 hour and then subjected to hypoxia-reoxygenation injury. The experiment was divided into three groups: normal control group, model group and HDAC3I treatment group, and 3 repetitions for each group. Cell viability was determined using MTT. Cellulose dehydrogenase (LDH) was detected by colorimetry. Flow cytometry was used to detect apoptosis and intracellular reactive oxygen species ( ROS ), respectively. The activity of superoxide dismutase ( SOD ) was determined by xanthine oxidase method. The malondialdehyde ( MDA ) content was determined by the thiobarbituric acid method. Western blotting was used to detect the expression of Bax, Bcl-2, cleaved-Caspase-3 and HDAC3 proteins. Results Compared with the control group, the cell viability of the model group and HDAC3I treatment group decreased significantly (P<0. 05), and the cell LDH (P<0. 05) and apoptosis (P<0. 05) increased significantly. The cell viability of HDAC3I treatment group was significantly higher than that of the model group (P<0. 05), while the LDH (P<0. 05) and apoptosis of HDAC3I treatment group were lower than the model group (P<0. 05). In addition, compared with the control group, the ROS and MDA (P< 0. 05 ) of the model group and the HDAC3I treatment group increased significantly, and the SOD decreased significantly (P<0. 05). ROS and MDA in the HDAC3I treatment group (P<0. 05) were significantly lower than the model group, while the SOD level was higher than the model group (P< 0. 05). Western blotting analysis showed that compared with the control group, Bax and cleaved-Caspase-3 in the model group and HDAC3I treatment group increased significantly, and Bcl-2 decreased significantly (P<0. 05). The Bax and cleaved-Caspase-3 in the HDAC3I treatment group were significantly lower than the model group, and Bcl-2 was significantly higher than the model group (P<0. 05). Compared with the control group, the expression of HDAC3 protein in the model group increased significantly (P<0. 05), while the HDAC3 protein in the HDAC3I treatment group decreased significantly (P<0. 05). Conclusion HDAC3I reduces PC 12 cell apoptosis induced by hypoxia/reoxygenation by reducing oxidative stress.
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Objective: To investigate the effect of combination treatments of cisplatin and KK4 and ICG15042 peanut testa extracts against cholangiocarcinoma cells in vitro.Methods: The growth inhibition, cell cycle arrest and apoptosis of cholangiocarcinoma cells were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and flow cytometry analysis, respectively. The levels of proteins involved in apoptosis were assessed using Western blotting assays. The caspase activity was assessed using a colorimetric caspase activity assay. Results: Cisplatin and peanut (KK4 and ICG15042) testa extracts inhibited the growth of cholangiocarcinoma cell lines (KKU-M214 and KKU-100 cells) in a dose- and time-dependent manner. The combination treatments reduced cell viability and induced apoptosis of cholangiocarcinoma cells more efficiently than single-drug treatments. Cancer cell death synergistically mediated by cisplatin and peanut testa extracts was observed in KKU-M214 cells (combination index < 1.0) but not in KKU-100 cells (combination index > 1.0). The combination treatments also increased the sub-G1 population and caused KKU-M214 cell cycle arrest at S and G2/M phases, which were the combined effects of cisplatin (S phase arrest) and peanut testa extracts (G2/M phase arrest). In addition, pERK1/2, Ac-H3, Bcl-2 and proteins related to apoptosis, including Bax and caspases 3, 8, 9, exhibited enhanced expression in KKU-M214 cells. The combination treatments caused down-regulation of p53, whereas the expression of p21 was fairly constant when compared with cisplatin single drug treatment. Conclusions: Peanut testa extracts in combination with cisplatin synergistically reduce cell viability and induce apoptosis through stimulation of caspases 3, 8 and 9 in KKU-M214 cells.
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Objective: To investigate the effect of combination treatments of cisplatin and KK4 and ICG15042 peanut testa extracts against cholangiocarcinoma cells in vitro. Methods: The growth inhibition, cell cycle arrest and apoptosis of cholangiocarcinoma cells were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and flow cytometry analysis, respectively. The levels of proteins involved in apoptosis were assessed using Western blotting assays. The caspase activity was assessed using a colorimetric caspase activity assay. Results: Cisplatin and peanut (KK4 and ICG15042) testa extracts inhibited the growth of cholangiocarcinoma cell lines (KKU-M214 and KKU-100 cells) in a dose- A nd time-dependent manner. The combination treatments reduced cell viability and induced apoptosis of cholangiocarcinoma cells more efficiently than singledrug treatments. Cancer cell death synergistically mediated by cisplatin and peanut testa extracts was observed in KKU-M214 cells (combination index 1.0). The combination treatments also increased the sub-G1 population and caused KKU-M214 cell cycle arrest at S and G2/M phases, which were the combined effects of cisplatin (S phase arrest) and peanut testa extracts (G2/M phase arrest). In addition, pERK1/2, Ac-H3, Bcl-2 and proteins related to apoptosis, including Bax and caspases 3, 8, 9, exhibited enhanced expression in KKU-M214 cells. The combination treatments caused down-regulation of p53, whereas the expression of p21 was fairly constant when compared with cisplatin single drug treatment. Conclusions: Peanut testa extracts in combination with cisplatin synergistically reduce cell viability and induce apoptosis through stimulation of caspases 3, 8 and 9 in KKU-M214 cells.
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Objective To investigate the effect of histone deacetylase inhibitor trichostatin A (TSA) on the migration of human esophageal squamous carcinoma cells(ESCC) and the possible mechanism. Methods KYSE-150 cells and EC9706 cells were cultured and Transwell assay was performed to detect the role of TSA alone and combined with protein kinase C (PKC) inhibitor AEB071 on cell migration; the images of morphology after cells treatment with TSA or combination of AEB071 with TSA; Western blotting was conducted to examine the protein level of epithelial-mesenchymal transition (EMT) related signaling molecules. Results TSA promoted the migration of ESCC cells significantly, and PKC inhibitor AEB071 partly inhibited the effect of TSA-promoted ESCC cells migration. Treatment with TSA resulted in the cell morphology transitioned from epithelia oval-like to mesenchymal spindle-like, indicating the EMT. AEB071 partially rescued ESCC cells morphological changes which TSA induced. Western blotting showed that TSA reduced the expression of E-cadherin and augmented the expression of vimentin, β-catenin, Slug and acH3, whereas AEB071 obviously blocked the EMT-related protein level changes which induced by TSA. Conclusion TSA promotes ESCC cells migration via inducing EMT process and the mechanism may be mediated by PKC signaling pathway.
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Objective@#Our study aimed to investigate the effect of histone deacetylase inhibitor trichostatin A（TSA）on the cisplatin（CDDP） resistance of ovarian cancer cell lines and its molecular mechanism.@*Methods@#Cisplatin-resistant ovarian cancer cell line C13* and its parental line OV2008 were incubated with TSA（200 nmol/L） or/and CDDP（20 μmol/L），the inhibitory rate of tumor cells was determined by MTT assay. Flow cytometry and Western blotting were used to detect apoptosis of tumor cells. Western blotting was also used to detect STAT3 expression in C13* and OV2008 cells. After down-regulation of STAT3 by transfection of STAT3 siRNA in C13* cells，cisplatin-induced apoptosis was evaluated by flow cytometry.@*Results@#MTT assay showed that the proliferation inhibitory rates of the combination group after 48 or 72 h treatment were significantly higher than those of TSA and CDDP groups. The level of STAT3 protein was much higher in C13* cells than in OV2008 cells. Flow cytometry and Western blotting showed that TSA combined with CDDP significantly enhanced the apoptotic rate of C13* cells. STAT3 expression level was significantly higher in C13* cells than in OV2008. Downregulation of STAT3 can significantly improve CDDP-induced apoptosis of C13* cells. @*Conclusion@#Down-regulation of STAT3 by TSA endows cisplatin-resistant cells C13* with increased sensitivity to cisplatin.
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Objective@#To explore effects of histone deacetylase inhibitor Belinostat on the immunologic function of dendritic cells (DC) and its possible mechanism.@*Methods@#Cultured mouse bone marrow-derived DC from C57BL/6 mouse in vitro. The experiments were divided into 0, 50, 100 nmol/L Belinostat + immature DC (imDC) group, and 0, 50, 100 nmol/L Belinostat mature DC (mDC). The changes of the ultrastructure of DC were observed by transmission electron microscope (TEM). Immunophenotype and CCR7 expression rate were detected by FCM, and the migration rate was observed by chemotaxis assay. The proliferation of lymphocytes stimulated by different DC was detected by mixed lymphocyte culture reaction. The cytokines in the culture supernatant, including TNF-α, IL-12 and IL-10, were examined by ELISA. RQ-PCR was used to examine the relative expression of mRNA in RelB.@*Results@#Successful cultured and identified the qualified imDC and mDC. Belinostat decreased the expression of CCR7 on imDC [(25.82±7.25)% vs (50.44±5.61)% and (18.71±2.00)% vs (50.44±5.61)%], meanwhile increased the rate on mDC [(71.14±1.96)% vs (64.90±1.47)%]. Chemotaxis assay showed that the migration rate of Belinostat+imDC and Belinostat+mDC group were both decreased, but the difference in imDC was not significant. T lymphocyte proliferation rate stimulated by 100 nmol/L Belinostat+imDC group was lower than imDC group in condition irritation cell∶reaction cell=1∶2 [(227.09±13.49)% vs (309.49±53.69)%]. Belinostat significantly suppressed the secretion of cytokines TNF-α, IL-12 and IL-10 (all P<0.01). The relative expression of mRNA in RelB was slightly decreased in Belinostat+imDC and Belinostat+mDC group (all P<0.05).@*Conclusion@#Belinostat could effectly suppress DC maturation and regulate immune tolerance of DC, which may be due to the down-regulation of mRNA level of RelB in DC.
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Objective: To explore effects of histone deacetylase inhibitor Belinostat on the immunologic function of dendritic cells (DC) and its possible mechanism. Methods: Cultured mouse bone marrow-derived DC from C57BL/6 mouse in vitro. The experiments were divided into 0, 50, 100 nmol/L Belinostat + immature DC (imDC) group, and 0, 50, 100 nmol/L Belinostat mature DC (mDC). The changes of the ultrastructure of DC were observed by transmission electron microscope (TEM). Immunophenotype and CCR7 expression rate were detected by FCM, and the migration rate was observed by chemotaxis assay. The proliferation of lymphocytes stimulated by different DC was detected by mixed lymphocyte culture reaction. The cytokines in the culture supernatant, including TNF-α, IL-12 and IL-10, were examined by ELISA. RQ-PCR was used to examine the relative expression of mRNA in RelB. Results: Successful cultured and identified the qualified imDC and mDC. Belinostat decreased the expression of CCR7 on imDC [(25.82±7.25)% vs (50.44±5.61)% and (18.71±2.00)% vs (50.44±5.61)%], meanwhile increased the rate on mDC [(71.14±1.96)% vs (64.90±1.47)%]. Chemotaxis assay showed that the migration rate of Belinostat+imDC and Belinostat+mDC group were both decreased, but the difference in imDC was not significant. T lymphocyte proliferation rate stimulated by 100 nmol/L Belinostat+imDC group was lower than imDC group in condition irritation cell∶reaction cell=1∶2 [(227.09±13.49)% vs (309.49±53.69)%]. Belinostat significantly suppressed the secretion of cytokines TNF-α, IL-12 and IL-10 (all P<0.01). The relative expression of mRNA in RelB was slightly decreased in Belinostat+imDC and Belinostat+mDC group (all P<0.05). Conclusion: Belinostat could effectly suppress DC maturation and regulate immune tolerance of DC, which may be due to the down-regulation of mRNA level of RelB in DC.
Sujet(s)
Animaux , Souris , Cellules cultivées , Cellules dendritiques , Inhibiteurs de désacétylase d'histone , Acides hydroxamiques , Souris de lignée C57BL , SulfonamidesRÉSUMÉ
Objective We aimed to investigate the expression of IRF4 and apoptosis of the histone deacetylase inhibitor LBH589 against MM cells in vitro,and study the mechanism of apoptosis when LBH589 alone or/and combined with lenalidomide in RPMI8226 cell so as to provide a new strategy for the treatment of multiple myeloma.Methods The cell viability on the growth of RPMI8226 cell were assessed by CCK8 assay.Apoptosis were measured by flow cytometry,The Grandpad software analyzes the statistical significance and evaluates the synergistic effect of the drug.The expression level of the related transcription factor and apoptotic gene protein were determined by western blot.The cell viability on the growth of RPMI8226-R cell were assessed by CCK8.Results LBH589 combined with lenalidomide have significant effect on inhibit cell proliferation and induce apoptosis in a dose-dependent manner.Apoptosis induced by LBH589/lenalidomide alone or combination was shown to involved PARP activation,decreased Bcl-2 and Bcl-xl expression.significantly down regulated transcriptional related factors of IRF4 and c-MYC expression compared with either agent alone.Conclusion LBH589 and lenalidomide alone or combination decrease the expression of transcription factor IRF4 and c-MYC,and have a significant synergistic effect,and highly expression of IRF in RPMI8226-R reduce proliferation inhibition.
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Objective To investigate the therapeutic effects of givinostat , a histone deacetylase inhibitor (HDACI), on the development of chronic asthma with airway inflammation , airway remodeling and airway hyperresponsiveness ( AHR) .Methods BALB/C mice were randomly divided into control group , asthma group, dexamethasone group and givinostat group (n=12 per group).AHR was assessed.Total cell numbers and differential counts , interleukin-4 ( IL-4 ) , interleukin-5 ( IL-5 ) and interferon-γ( IFNγ) levels in the bronchoalveolar lavage fluid ( BALF) were measured in the above 4 groups.The pathology of lung tissue was evaluated .Immunohistochemical ( IHC) staining and Western blot were used to detect αsmooth muscle actin(α-SMA) and transforming growth factor-β1(TGFβ1).Results Compared with the asthma only group, givinostat treatment relieved airway resistance (2.96 ±1.01 vs 6.50 ±0.79,P0.05] was enhanced in BALF.Inflammatory cell infiltration around the airway was reduced , with decreased inflammatory cell score [(1.60 ±0.69) points vs (3.40 ±0.68) points, P <0.01] and inflammatory cell number (111.65 ±31.41 vs 601.25 ±186.85, P<0.01).The goblet cell metaplasia [(26.36 ±2.33)%vs (57.21 ±11.56)%] and collagen deposition area [(52.77 ±7.58)μm2/μm vs (111.81 ±12.40)μm2/μm] were obviously reduced (P<0.01).The expressions of α-SMA and TGFβ1 in the lung tissue were both significantly decreased ( P<0.01 ) .Conclusion Givinostat treatment can reduce airway inflammation , airway remodeling and airway hyperresponsiveness in chronic asthma .Its effect is comparable to that of glucocorticoid hormone treatment .
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OBJECTIVE: To establish a LC-MS/MS method for determining F1 in rat plasma and study the pharmacokinetic properties of F1. METHODS: Ten healthy SD rats were enrolled in this study. They were randomly divided into two groups and received intragastric(10 mg·kg-1) and intravenous administration(5 mg·kg-1) of F1. After receiving F1, the concentrations of F1 in plasma were determined. Blood samples(0.1 mL)were immediately collected into heparinized tubes before injection and at 0, 0.08, 0.25, 0.5, 0.75, 1, 2, 4, 6, 8, 10, 12, 24 h after injection. The pharmacokinetic parameters were determined by DAS2.0 software, absolute bioavailability of F1 was calculated based on AUC and dose administered. RESULTS: The main pharmacokinetic parameters after intragastric and intravenous administration of F1 were as follows: ACU0-t(27.052±10.068), (153.878±88.777)ng·h·mL-1;AUC0-∞(31.425±9.261), (179.054±116.794)ng·h·mL-1;MRT0-t(10.722±4.335), (2.398±1.344)h; MRT0-∞ (15.651±5.917), (6.925±7.013)h;t1/2(4.294±1.534), (6.052±3.633)h;ρmax(18.394±17.856), (219.079±142.207)ng·mL-1, respectively. Absolute bioavailability value was 8.79%. CONCLUSION: This method can be used to determine the content of F1 in rat plasma. The experimental results can guide the structural optimization of F1, improve the pharmacokinetics of F1 in vivo and provide experimental basis for improving bioavailability of F1.
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Objective To investigate the impact of vorinostat(SAHA) on the erlotinib-resistance of human lung adenocarcinoma cell line PC-9/ER.Methods PC-9/ER cells were treated with erlotinib and SAHA,alone or in combination.The effects of proliferation inhibition were assayed by MTT method,the apoptosis ratios of cells were analyzed by flow cytometry.Results The results showed that SAHA inhibited the proliferation of PC-9/ER cells in a dose-dependent manner alone.The non-toxic doses of SAHA (1 μmol/L) significantly improved the sensitivity of PC-9/ER to Erlotinib,and induce apoptosis.Conclusion SAHA could partially improve PC-9/ER sensitivity to erlotinib.
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Objective To investigate the potential protective effects of valproic acid (VPA) on gut barrier function after major burn injury in rats and its mechanism.Methods Forty male Sprague-Dawley (SD) rats were divided into sham + normal saline (NS),sham + VPA,scald + NS,and scald + VPA groups,with 10 rats in each group.Rat with 55％ total body surface area (TBSA) third-degree severe-bums model was reproduced by immersing into 80 ℃ water,and the rats in sham groups were given sham-bums by immersing into 37 ℃ water.The rats after severebums were immediately treated with 0.25 mL of 300 mg/kg VPA or NS by subcutaneous injection.Rats were sacrificed at 2 hours and 6 hours after injury,and abdominal aortic blood and ileal tissue were harvested.The levels of vascular endothelial growth factor (VEGF) were determined by enzyme-linked immunosorbent assay (ELISA).The intestinal permeability was evaluated by fluorescein isothiocyanate-dextran (FITC-dextran) determination.The histomorphological changes in gut barrier were evaluated by Chiu grading system.Levels of acetylated lysine at the ninth position of histone 3 protein (Ac-H3K9),hypoxia-inducible factor 1α (HIF-1α),zona occludens 1 (ZO-1) and myosin light chain kinase (MLCK) were determined by immunofluorescence staining and Western Blot.Results Compared with sham + NS group,rats in scald + NS group showed intestinal mucosal damage 2 hours after bum injury,as well as increased mucosal permeability,protein expression levels of HIF-1 α,VEGF,MLCK,and lowered levels of AC-H3K9 and ZO-1.These changes were much more prominent at 6 hours after injury.VPA treatment significantly attenuated the bum-induced intestinal damage.Compared with scald + NS group,the protective effects in scald + VPA group was not evident at 2 hours after injury;however,intestinal damage was much less severe at 6 hours after injury (Chiu score:2.03 ± 0.27 vs.3.12 ± 0.15),intestinal permeability was significantly decreased [FITC-dextran (μg/L):709 ± 76 vs.1138 ± 75],histone acetylation was enhanced [Ac-H3K9 (gray value):1.55 ± 0.12 vs.0.48±0.12],ZO-1 degradation was significantly inhibited (gray value:0.69 ± 0.12 vs.0.43 ± 0.16),the protein expression levels of VEGF and MLCK were significantly down-regulated [VEGF (ng/mg):51.7±3.7 vs.71.2±4.3,MLCK (gray value):1.98±0.20 vs.2.80±0.24],while the HIF-1 α protein expression levels were significantly reduced at both 2 hours and 6 hours after injury (gray value:2.50±0.39 vs.3.88±0.42 at 2 hours,1.83±0.42 vs.4.42±0.41 at 6 hours,all P ＜ 0.05).Conclusions Severe bum injury can induce histone deacetylation,ZO-1 degradation and intestinal barrier dysfunction.VPA can improve the levels of histone acetylation and ZO-1,and protect intestinal epithelial barrier function.These may probably be mediated through inhibiting HIF-1α and its downstream gene VEGF and MLCK.
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OBJECTIVE:To study the mechanism of histone deacetylase inhibitor RGFP109 in reversing resistance of glioma U251 cells. METHODS:TR/U251 cells resistance to temozolomide(TMZ)was extrablished. The test was divided into normal con-trol group,TMZ group(40 μmol/L)and TMZ(40 μmol/L)+RGFP109(0-120 μmol/L)different concentrations groups. After 24 h of adding into related medicines,CCK-8 was used to detect the cell survival rate and calculate the half inhibitory concentration (IC50). TUNEL and Annexin V/PI were used to detect the cell apoptosis in normal control group,TMZ group and TMZ+RGFP109 (42μmol/L)group. Immunoblotting was used to detect the O6-methyl guanine-DNA methyltransferase(MGMT),Survivin,B lym-phoma 2(Bcl-2),B lymphoma xL(Bcl-xL)protein expression;and gel migration test was used to detect the p65 acetylation level and its binding capacity with κB-DNA. RESULTS:Compared with normal control group,cell survival rate in TMZ+RGFP109 dif-ferent concentrations groups was obviously decreased (P<0.05),showing a concentration-dependent manner. When the RGFP109 concentration was 42 μmol/L,the sensitivity of TMZ to TR/U251 cells was the same with U251 cells. Compared with normal con-trol group,MGMT,Survivin,Bcl-2,Bcl-xL protein expressions in cells of TMZ groups were enhanced(P<0.01);p65 acetyla-tion level had no obvious changes,while the binding capacity of p65 and κB-DNA was strengthened (P<0.01). Compared with TMZ group,MGMT,Survivin,Bcl-2,Bcl-xL protein expressions in cells of TMZ groups were weakened(P<0.01);p65 acetyla-tion level was enhanced (P<0.01);and the binding capacity of p65 and κB-DNA was weakened (P<0.01). CONCLUSIONS:RGFP109 can reverse the resistance of U251 cells to TMZ by down-regulating the anti-apoptotic protein expressions adjusted by transcription factorκB(NF-κB)and weakening the binding of p65 andκB-DNA.