ABSTRACT
Ischemia reperfusion injury (IRI) of organs is a major challenge for clinicians, but the mechanism is still not elucidated, and the clinical treatment effect is still unsatisfactory. PARP-1-dependent cell death (parthanatos) is a non-apototic programmed cell death pathway involved in the development of the occurrence of IRI of organs. At the same time, parthanatos is also a multi-step pathway. There are many molecules in the parthanatos cascade that can be exploited to create therapeutic interventions for IRI, including PARP1, apoptosis inducing factor (AIF), and macrophage migration inhibitory factor (MIF). These critical molecules are involved in DNA damage, energy depletion and homeostasis imbalance. Therefore, these molecular signals in the parthanatos cascade represent promising therapeutic targets for the treatment of IRI. In the following, we will elaborate on the mechanisms and molecular characteristics of the parthanatos pathway and the relation between parthanatos pathway and IRI of vital organs. It aims to explore the posibility of IRI mechanism research and clinical treatment and to provide new ideas for clinicians and researchers.
ABSTRACT
Objective:To investigate the changes and clinical significance of high mobility group protein B1 (HMGB1) in condyloma acuminatum (CA).Methods:Sixty four patients with initial CA(initial group) and 48 patients with recurrent CA(recurrent goup) treated in the Second Affiliated Hospital of PLA Air Force Military Medical University Hospital from January 2019 to November 2020 were included. In the same period, 31 patients who underwent circumcision and 19 female underwent sexual organ cosmetic plastic surgery were taken as the control group, and the normal foreskin and healthy vulva were collected. The expression of HMGB1 in wart was detected by real-time quantitative polymerase chain reaction(RT-PCR), and the expression of soluble apoptosis related factor ligand (sFasL), cell lymphoma-2 gene (Bcl-2), soluble apoptosis related factor (SFAS) and Survivin, caspase-3 were detected. At the same time, serum interleukin (IL) - 6, IL-17, IL-23 and tumor necrosis factor - α (TNF- α) were detected by enzyme-linked immunosorbent assay(ELISA).Results:The expression of HMGB1 mRNA in the warts of patients in the initial group, recurrence group and control group were 1.96 ± 0.20, 1.53 ± 0.14, 1.05 ± 0.11, there was statistical difference ( F = 15.20, P<0.05) ; the expression of HMGB1 mRNA in the warts of patients in the initial group was significantly higher than that in the recurrence group and the control group ( P<0.05), and the recurrence group was also significantly higher than that in the control group ( P<0.05). The mRNA expressions of sFas, Bcl-2, sFasL and caspase-3 in the warts of patients in the initial group were significantly lower than those in the recurrence group and the control group: 0.52 ± 0.08 vs. 0.82 ± 0.16, 1.10 ± 0.19; 0.50 ± 0.05 vs. 0.79 ± 0.13, 1.08 ± 0.21; 0.47 ± 0.06 vs. 0.81 ± 0.15, 1.01 ± 0.19; 0.35 ± 0.04 vs. 0.68 ± 0.09, 0.91 ± 0.16, P<0.05; and the recurrence group were also significantly lower than those in the control group ( P<0.05). The expression of Survivin mRNA in the warts of patients in the initial group was significantly higher than those in the recurrence group and the control group: 2.14 ± 0.40 vs. 1.60 ± 0.27, 0.99 ± 0.18, P<0.05, and the recurrence group was also significantly higher than that in the control group ( P<0.05). The serum levels of TNF-α and IL-6 in the initial group were significantly lower than that in the recurrence group and the control group: (20.08 ± 1.95) μg/L vs. (26.93 ± 2.74), (37.65 ± 3.83) μg/L; (31.05 ± 3.24) μg/L vs. (38.13 ± 3.76), (45.98 ± 4.69) μg/L, P<0.05; and the recurrence group were also significantly lower than those in the control group ( P<0.05). The serum levels of IL-17 and IL-23 in the primary group were significantly higher than those in the recurrence group and the control group: (423.71 ± 28.68) ng/L vs. (384.26 ± 21.70) and (335.43 ± 19.65) ng/L; (289.50 ± 18.53) ng/L vs. (251.07 ± 15.96) and (214.67 ± 13.20) ng/L, P<0.05; and the recurrence group were also significantly higher than those in the control group ( P<0.05). The results of correlation analysis showed that the mRNA expression of HMGB1 in the warts of CA patients were negatively correlated with the mRNA expression of caspase-3, sFas, Bcl-2 and sFasL in the warts ( r = - 0.602, - 0.734, - 0.692, - 0.717, P<0.05), and was positive correlation with Survivin mRNA expression ( r = 0.645, P<0.05); and were positive correlation with the contents of IL-17 and IL-23 in serum ( r = 0.673, 0.685, P<0.05), and negatively correlated with the contents of TNF-α and IL-6 ( r = - 0.698, - 0.764, P<0.05). Conclusions:HMGB1 is obviously abnormal in the warts of patients with condyloma acuminatum, and is closely related to apoptosis, immune and inflammation-related factors, and may be jointly involved in the occurrence and recurrence of CA.
ABSTRACT
Objective:To explore the changes of serum B7 homolog 2 (B7-H2), tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), interleukin(IL)-37 and IL-17A levels in patients with primary immune thrombocytopenia (ITP), and to analyze the clinical guiding significance of each index level in the diagnosis and treatment of ITP.Methods:A total of 90 patients with ITP treated in Jining Hospital of Traditional Chinese Medicine from September 2018 to September 2020 were retrospectively selected as the research group, and 90 healthy patients during the same period were selected as the normal control group. The levels of serum B7-H2, TRAIL, IL-37, IL-17A and platelet count (PLT) in the two groups were compared, and the clinical guidance significance of each index level in the diagnosis and treatment of ITP were analyzed by receiver operating characteristic(ROC) curve.Results:The serum levels of B7-H2, TRAIL, IL-37 and IL-17A in the research group were higher than those in the normal control group, and PLT was lower than that in the normal control group: (25.12 ± 4.29) μg/L vs. (12.26 ± 3.10) μg/L, (0.92 ± 0.20) μg/L vs.(0.46 ± 0.13) μg/L, (145.02 ± 43.18) ng/L vs. (50.23 ± 14.07) ng/L, (21.63 ± 5.06) ng/L vs. (7.71 ± 2.04) ng/L, (16.12 ± 4.61) × 10 9/L vs. (200.86 ± 61.4) × 10 9/L, there were statistical differences ( P<0.05). After treatment for 3 months, 73 patients obtained remission, and 13 patients were non-remission. The levels of B7-H2, TRAIL, IL-37, IL-17A in the non-remission group before and after treatment were higher than those in the remission group, and PLT was lower than that in the remission group, there were statistical differences ( P<0.05). Pearson correlation analysis showed that the levels of serum B7-H2, TRAIL, IL-37 and IL-17A were negatively correlated with PLT ( P<0.05). The ROC curve analysis showed that the area under the curve of serum B7-H2, TRAIL, IL-37 and IL-17A in prognostic prediction was 0.916, the sensitivity and the specificity were 94.12% and 86.30%. Conclusions:The serum levels of B7-H2, TRAIL, IL-37 and IL-17A in patients with ITP are significantly elevated, and are closely related to the level of PLT. Combined detection can help clinically predict the direction of disease outcome.
ABSTRACT
@#Parthanatos is a form of programmed cell death,which is also known as poly(ADP-ribose)polymerase 1(PARP1)-mediated apoptosis-inducing factor(AIF)and macrophage migration inhibitory factor(MIF)-dependent cell death according to its molecular mechanism. Parthanatos is the main cause of a variety of neurodegenerative diseases,such as Parkinson's disease(PD),Alzheimer's disease(AD),motor neuron disease,and is also involved in the pathogenesis of some tumors,such as lung cancer and breast cancer. Therefore,a thorough understanding of the molecular mechanism of Parthanatos is crucial for the therapeutic strategies of related diseases. In recent years,studies have found that effective regulation of the occurrence of Parthanatos by regulating the key proteins PARP1,AIF and MIF is expected to become a therapeutic target for many diseases. Based on the specific molecular mechanism of Parthanatos,this paper reviewed the research progress of therapeutic strategies for related diseases from the aspects of inhibiting and promoting Parthanatos.
ABSTRACT
Auditory neuropathy spectrum disorder (ANSD) represents a variety of sensorineural deafness conditions characterized by abnormal inner hair cells and/or auditory nerve function, but with the preservation of outer hair cell function. ANSD represents up to 15% of individuals with hearing impairments. Through mutation screening, bioinformatic analysis and expression studies, we have previously identified several apoptosis-inducing factor (AIF) mitochondria-associated 1 (AIFM1) variants in ANSD families and in some other sporadic cases. Here, to elucidate the pathogenic mechanisms underlying each AIFM1 variant, we generated AIF-null cells using the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system and constructed AIF-wild type (WT) and AIF-mutant (mut) (p.T260A, p.R422W, and p.R451Q) stable transfection cell lines. We then analyzed AIF structure, coenzyme-binding affinity, apoptosis, and other aspects. Results revealed that these variants resulted in impaired dimerization, compromising AIF function. The reduction reaction of AIF variants had proceeded slower than that of AIF-WT. The average levels of AIF dimerization in AIF variant cells were only 34.5%‒49.7% of that of AIF-WT cells, resulting in caspase-independent apoptosis. The average percentage of apoptotic cells in the variants was 12.3%‒17.9%, which was significantly higher than that (6.9%‒7.4%) in controls. However, nicotinamide adenine dinucleotide (NADH) treatment promoted the reduction of apoptosis by rescuing AIF dimerization in AIF variant cells. Our findings show that the impairment of AIF dimerization by AIFM1 variants causes apoptosis contributing to ANSD, and introduce NADH as a potential drug for ANSD treatment. Our results help elucidate the mechanisms of ANSD and may lead to the provision of novel therapies.
Subject(s)
Humans , Apoptosis Inducing Factor/metabolism , NAD/metabolism , Dimerization , ApoptosisABSTRACT
The apoptosis-inducing factor, mitochondrion-associated 1(AIFM1)gene encodes an apoptosis-inducing factor(AIF)protein with apoptosis and redox function.AIF is widely expressed within cells in human tissues, and playing an important role in the mitochondria.Mutations in the AIFM1 gene are associated with severe X-linked mitochondrial encephalomyopathy, Cowchock syndrome, X-linked spondyloepimeta-physeal dysplasia with hypomyelinating leukodystrophy, auditory neuropathy and other diseases.AIFM1 gene mutations exhibit a wide range of clinical phenotypes, but the pathogenesis between mutations and phenotypes and phenotypic severity remains unclear.This paper summarizes the reported AIFM1 mutation-related loci, phenotypes, and possible pathogenesis mechanisms, and provide a brief review of AIFM1 mutation-related diseases and their progression.
ABSTRACT
Introduction: More than half of all worldwide deaths and disabilities were caused by stroke. Large artery atherosclerosis is identified as a high etiological risk factor because it accounts for 20% of ischemic stroke. Objectives: To identify the significance of TRAIL and adropin release and the relative changes related to S100B levels, as well as the relationship between these biomarkers and the final infarct core, the clinical outcome, and the presence of large artery atherosclerosis in acute stroke patients. Materials and methods: Over a one-year period, demographic, clinical, and neuroimaging findings of 90 consecutive patients with acute ischemic stroke were evaluated. Results: The mean age of participants was 69.28 ± 10 and 39 patients were female. The increased level of S100B and the decreased levels of sTRAIL with adropin were significantly associated with moderate to severe neurologic presentation (p=0.0001, p=0.002, p=0.002, respectively). On the control CT, a large infarct core was significantly associated with decreased serum levels of sTRAIL and adropin (p=0.001 and p=0.000, respectively); however, the levels of S100B were not significantly associated with good ASPECTS score (p=0.684). Disability and an unfavorable outcome were significantly related to the decreased level of sTRAIL and adropin (p=0.001 and p=0.000 for THRIVE score>5, respectively). Decreased sTRAIL and adropin levels and an increased S100B level were correlated with the presence of large artery atherosclerotic etiologic factors (p=0.000, p=0.000, p=0.036, respectively). Conclusion: TRAIL and adropin serum levels were associated with poor clinical outcomes and greater infarcted area in acute ischemic stroke patients.
Introducción. Más de la mitad de todas las muertes y discapacidades en todo el mundo fueron causadas por accidentes cerebrovasculares. La aterosclerosis de las grandes arterias se identifica como un factor de alto riesgo etiológico debido a que representa el 20 % de los accidentes cerebrovasculares isquémicos. Objetivo. Determinar la importancia de la liberación de TRAIL y adropina y los cambios relativos relacionados con los niveles de S100B, así como la relación entre estos biomarcadores y el núcleo final del infarto, el resultado clínico y la presencia de aterosclerosis de arterias grandes en pacientes con accidente cerebrovascular agudo. Materiales y métodos. Durante un año, se evaluaron los hallazgos demográficos, clínicos y de neuroimágenes de 90 pacientes con accidente cerebrovascular isquémico agudo. Resultados. La edad media de los pacientes fue de 69,28 ± 10 y 39 eran mujeres. El aumento del nivel de S100B y la disminución de los niveles de sTRAIL y adropina se asociaron significativamente con una presentación neurológica moderada a grave en los pacientes (p=0,0001, p=0,002 y p=0,002, respectivamente). En la TC de control, un gran núcleo de infarto se asoció significativamente con una disminución del nivel sérico de sTRAIL y adropina (p=0,001 y p=0,000, respectivamente); sin embargo, los niveles de S100B no se asociaron significativamente con una buena puntuación en el ASPECT (p=0,684). La discapacidad y el resultado desfavorable se relacionaron significativamente con la disminución de los niveles de sTRAIL y adropina (p=0,001 y p=0,000 para una puntuación >5 en el THRIVE, respectivamente). La disminución de los niveles de sTRAIL y adropina y el aumento del nivel de S100B, se correlacionaron con la presencia de un factor etiológico aterosclerótico de arterias grandes entre la población de estudio (p=0,000, p=0,000 y p=0,036, respectivamente). Conclusiones. Los niveles séricos de TRAIL y adropina se asociaron con un resultado clínico deficiente y una mayor área infartada en pacientes con ataque cerebrovascular isquémico agudo.
Subject(s)
Stroke , Infarction, Posterior Cerebral Artery , TNF-Related Apoptosis-Inducing LigandABSTRACT
The rapidly developing resistance of cancers to chemotherapy agents and the severe cytotoxicity of such agents to normal cells are major stumbling blocks in current cancer treatments. Most current chemotherapy agents have significant cytotoxicity, which leads to devastating adverse effects and results in a substandard quality of life, including increased daily morbidity and premature mortality. The death receptor of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) can sidestep p53-dependent pathways to induce tumor cell apoptosis without damaging most normal cells. However, various cancer cells can develop resistance to TRAIL-induced apoptosis via different pathways. Therefore, it is critical to find an efficient TRAIL sensitizer to reverse the resistance of tumor cells to TRAIL, and to reinforce TRAIL's ability to induce tumor cell apoptosis. In recent years, traditional Chinese medicines and their active ingredients have shown great potential to trigger apoptotic cell death in TRAIL-resistant cancer cell lines. This review aims to collate information about Chinese medicines that can effectively reverse the resistance of tumor cells to TRAIL and enhance TRAIL's ability to induce apoptosis. We explore the therapeutic potential of TRAIL and provide new ideas for the development of TRAIL therapy and the generation of new anti-cancer drugs for human cancer treatment. This study involved an extensive review of studies obtained from literature searches of electronic databases such as Google Scholar and PubMed. "TRAIL sensitize" and "Chinese medicine" were the search keywords. We then isolated newly published studies on the mechanisms of TRAIL-induced apoptosis. The name of each plant was validated using certified databases such as The Plant List. This study indicates that TRAIL can be combined with different Chinese medicine components through intrinsic or extrinsic pathways to promote cancer cell apoptosis. It also demonstrates that the active ingredients of traditional Chinese medicines enhance the sensitivity of cancer cells to TRAIL-mediated apoptosis. This provides useful information regarding traditional Chinese medicine treatment, the development of TRAIL-based therapies, and the treatment of cancer.
ABSTRACT
【Objective】 To explore the establishment methods of transgenic human umbilical cord mesenchymal stem cells (hUC-MSCs) overexpressing tumor necrosis factor(TNF)-related apoptosis-inducing ligand (TRAIL) based on the transposons, and attempt to apply it on the nude mice mode with glioma. 【Methods】 PiggyBac transposon system specially designed by us was used to prepare non-targeting and Her2-targeting hUC-MSCs that can stably express TRAIL through puromycin screening. The glioma cells expressing firefly luciferase (U87MG-LUC) were injected into the skull of the immunodeficient mice (BALB/c-nu/nu) with 1×106 cells per mouse. After 7 days of injection, the mice transplanted with U87MG were detected with a small animal living imager to determine the size and location of the tumors in skull. Then we injected the glioma-transplantation nude mouse with two kinds of transgenic hUC-MSCs expressing TRAIL (named as untarget-TRAIL and target-TRAIL, respectively), or the non-transgenic hUC-MSCs (all 1×106 cells per mouse) or PBS (named as WT-MSCs and PBS for negative control) respectively, and then monitored the changes of tumor signals by a small animal living imager every week for 3~4 weeks. 【Results】 After six passages to expand the cells, the both transgenic cell lines can stably express TRAIL gene. Their ratio of green fluorescent protein (GFP) positive cells can reach 93%-97%, and the positive ratio of their MSC-specific surface markers still maintained normal (CD34+, CD45+, and HLA-DR+ all <0.1%, CD90>99%, CD73>88%, and CD105 >60%). The median survival time (d) of U87MG-transplanted nude mice in the groups of untarget-TRAIL, target-TRAIL, WT-MSCs, and PBS was 41 vs 39 vs 24 vs 23(P<0.05). 【Conclusion】 The transgenic hUC-MSCs overexpressing TRAIL gene can significantly prolong the survival time of nude mice with brain glioma.
ABSTRACT
Objective:To explore the mechanism of circular permuted tumor necrosis factor-related apoptosis-inducing ligand (CPT) reversing the resistance to imatinib in chronic myeloid leukemia (CML) cells.Methods:Five patients with CML in the Affiliated Hospital of Inner Mongolia Medical University from 2016 to 2020 were selected, and heparinized bone marrow blood samples were collected at the first diagnosis and imatinib resistance phase, and mononuclear cells were isolated. The mononuclear cells collected at the first diagnosis were named A1-E1, and the mononuclear cells collected after imatinib resistance were named A2-E2. Human CML wild-type K562 cell line (K562-W) was given gradually increasing small doses of low-concentration imatinib to obtain imatinib-resistant K562 cells (K562-R). K562-R cells were cultured with 20 μg/L CPT and these cells were set as CPT-K562-R group. The CCK-8 method was used to detect the half inhibitory concentration ( IC50) of cells for imatinib. K562-W and K562-R cells were used to establish CML xenografts nude mice models, then the nude mice were divided into K562-W, K562-R and CPT-K562-R xenograft groups. Imatinib was perfused orally in all three groups, and CPT was injected subcutaneously in the CPT-K562-R group at the same time. The tumor volume of the three groups of nude mice before and 4 weeks after treatment with imatinib, and the survival time of the three groups of nude mice were compared. Western blot was used to detect the changes of tyrosine protein kinase receptor B4 (EphB4) and myeloid cell leukemia protein 1 (Mcl-1) protein levels in bone marrow mononuclear cells, K562 cell line and transplanted tumor tissues of CML patients. Results:The expressions of EphB4 protein in A2-E2 cells of 5 patients with CML were higher than those in A1-E1 cells (all P < 0.01). The IC50 of K562-W, K562-R and CPT-K562-R cells for imatinib were (0.160±0.015) mg/L, (5.450±0.460) mg/L, (0.300±0.035) mg/L, and the difference was statistically significant ( F = 390.65, P < 0.01). In cells of K562-W group, EphB4 and Mcl-1 proteins were expressed at low levels (0.54±0.02 and 0.70±0.08); in cells of K562-R group, the expressions of EphB4 and Mcl-1 proteins were enhanced (3.04±0.11 and 2.88±0.04); in cells of CPT-K562-R group, the expressions of EphB4 and Mcl-1 proteins decreased (0.57±0.03 and 0.38±0.04). Before imatinib treatment, there was no statistically significant difference in the tumor volumes of nude mice among the K562-W, K562-R and CPT-K562-R xenograft groups ( F = 0.39, P = 0.68), suggesting the transplanted tumors formed in nude mice were balanced; after imatinib treatment, the difference in the tumor volumes among the three groups were statistically significant ( F = 26.16, P < 0.01). The survival time of nude mice in the K562-W, K562-R and CPT-K562-R xenograft groups was (18.5±3.3) d, (10.0±2.4) d and (17.5±1.6) d, and the difference was statistically significant ( F = 20.45, P < 0.01). In K562-W xenograft group, both EphB4 and Mcl-1 proteins were expressed at low levels (0.55±0.06 and 0.67±0.06); in K562-R xenograft group, the expressions of EphB4 and Mcl-1 proteins were enhanced (1.95±0.08 and 6.21±0.53); the expressions of EphB4 and Mcl-1 in CPT-K562-R xenograft group decreased (0.59±0.04 and 0.37±0.04) and were close to their expressions in K562-W xenograft group. Conclusion:CPT may enhance the sensitivity of CML to imatinib by inhibiting the expressions of EphB4 and Mcl-1, and this may be a targeted pathway for imatinib therapy.
ABSTRACT
Acetaminophen (APAP) is a widely used analgesic and antipyretic drug, which is safe at therapeutic doses but can cause severe liver injury and even liver failure after overdoses. The mouse model of APAP hepatotoxicity recapitulates closely the human pathophysiology. As a result, this clinically relevant model is frequently used to study mechanisms of drug-induced liver injury and even more so to test potential therapeutic interventions. However, the complexity of the model requires a thorough understanding of the pathophysiology to obtain valid results and mechanistic information that is translatable to the clinic. However, many studies using this model are flawed, which jeopardizes the scientific and clinical relevance. The purpose of this review is to provide a framework of the model where mechanistically sound and clinically relevant data can be obtained. The discussion provides insight into the injury mechanisms and how to study it including the critical roles of drug metabolism, mitochondrial dysfunction, necrotic cell death, autophagy and the sterile inflammatory response. In addition, the most frequently made mistakes when using this model are discussed. Thus, considering these recommendations when studying APAP hepatotoxicity will facilitate the discovery of more clinically relevant interventions.
ABSTRACT
OBJECTIVE: To observe the effect of acupuncture (Acupunct) on cerebral infarction volume and expression of poly ADP ribose polymerase 1 (PARP1), apoptosis-inducing factor (AIF) and endonuclease G (Endo-G) in the cerebral cortex tissue at different time-points after cerebral ischemia (CI) in acute cerebral infarction rats, so as to explore its underlying mechanisms in prolonging time window of thrombolysis. METHODS: Forty-eight SD rats were randomly divided into sham operation, model, intravenous thrombolysis (IVT)-4.5 h, IVT-6 h, IVT-9 h, Acupunct＋IVT-4.5 h, Acupunct ＋IVT-6 h, Acupunct＋IVT-9 h groups (n＝6 in each group). The CI model was established by using modified autologous thromboembolism via the right common carotid artery. Two hours after modeling, rats of the Acupunct groups received Acupunct stimulation of "Shuigou" (GV26) and bilateral "Neiguan" (PC6) for 30 min. Thrombolysis was conducted by injection of recombinant human tissue-type plasminogen activator (rt-PA, 10 mg/kg) via caudal vein. The neurological deficit was assessed with reference to Bederson's methods. 2，3，5-triphenyltetrazolium chloride (TTC) staining was used to assess the cerebral infarction volume, and the expression of cerebral PARP1, AIF and Endo-G proteins detected by Western blot. RESULTS: Compared with the sham operation group, the neurological score and percentage of cerebral infarction volume, expression levels of PARP1, AIF and Endo-G proteins were significantly increased in the model group (P0.05). The levels of neurological score, percentage of cerebral infarction volume, and AIF expression were significantly lower in both the Acupunct＋IVT 4.5 h and Acupunct＋IVT-6 h groups than in the simple IVT-4.5 h and simple IVT-6 h groups, respectively (P<0.05), and the expression levels of PARP1 and Endo-G proteins were obviously lower in the Acupunct＋IVT-4.5 h group than in the IVT-4.5 h group (P<0.05). Endo-G proteins were obviously lower in the Acupunct＋IVT-9 h group than in the IVT-9 h group (P<0.05). CONCLUSION: Acupuncture may improve neurological function, reduce cerebral infarction volume and prolong the time window of thrombolysis in CI rats, which may be associated with its effect in suppressing AIF/PARP1/ Endo-G signaling.
ABSTRACT
Objective: To explore the possibility of promoting tumor necrosis factor-related apoptosis inducing ligand (TRAIL)-induced apoptosis in prostate cancer PC-3 cell by inhibiting Krüppel-like factor 5 (KLF5). Methods: MTT assay, flow cytometry, Western blot assay and qRT-PCR assay were deployed to detect the cell viability, apoptosis and apoptotic markers in KLF5-inhibited and TRAIL-induced PC-3 cells. Results: After KLF5 was inhibited in TRAIL-induced PC-3 cells, cell viability reduced, apoptosis enhanced, the expressions of DR4 and DR5 increased while the expression of cellular fas-associated death domain-like interleukin-1β converting enzyme inhibitory protein (c-FLIP) decreased. Conclusion: Inhibiting KLF5 suppresses cell viability by promoting TRAIL-induced apoptosis in prostate cancer cell PC-3. It may be a potential means to treat hormone-insensitive prostate cancer.
ABSTRACT
Objective: To investigate whether piperlongumine can sensitize prostate cancer cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and trigger apoptosis in prostate cells.Methods: Human prostate cancer cell lines PC3, LNCaP, and VCaP were cultured with piperlongumine and TRAIL. Then, cell proliferation, migration, caspase activation, apoptotic protein expressions, and death receptor expressions were measured. Results: Piperlongumine inhibited cell proliferation at low doses (<10 μM) alone and in combination with TRAIL (25 ng/mL), induced apoptosis, and suppressed cyclooxygenase activation. Additionally, piperlongumine induced expression of death receptors which potentiated TRAIL-induced apoptosis in cancer cells but did not affect decoy receptors. Piperlongumine also downregulated tumor cell-survival pathways, inhibited colony formation and migration of cancer cells alone or in combination with TRAIL. The combination of piperlongumine with TRAIL was found to be synergistic. Conclusions: Our findings indicate that piperlongumine can sensitize cancer cells to TRAIL through the upregulation of death receptors and can trigger apoptosis with the downregulation of anti-apoptotic proteins.
ABSTRACT
OBJECTIVE@#To explore the effect of pretreatment of neuroblastoma cells with hot water extract of Korean ginseng on MNNG-induced parthanatos and its mechanism.@*METHODS@#Neuroblastoma SH-SY5Y cells were pretreated with 1 mg/L hot water extract of Korean ginseng before induction with 250 μmol/L MNNG for 1 h or 4 h. CCK-8 and cell flow cytometry were used to detect cell survival rate. Western blotting was used to detect the changes in poly(ADP-ribose) (PAR) expression in the treated cells. Immunofluorescence assay was used to detect nuclear distribution of apoptosis-inducing factor (AIF), and flow cytometry was used to detect the level of reactive oxygen species (ROS) in the cells.@*RESULTS@#Compared with the blank control cells, MNNG-treated SH-SY5Y cells showed significantly decreased survival rate as the concentration of MNNG and the stimulation time increased ( < 0.05). Stimulation with MNNG also resulted in significantly increased expression of PAR protein in the cells ( < 0.05). Pretreatment of the cells with hot water extract of Korean ginseng obviously inhibited MNNG-induced cell death and significantly reduced AIF expression and nucleation in the cells ( < 0.05). MNNG stimulation significantly increased ROS level in the cells, which was decreased significantly by pretreatment of the cells with the extract ( < 0.05).@*CONCLUSIONS@#Pretreatment with hot water extract of Korean ginseng reduces MNNG-induced parthanatos and ROS production in SH-SY5Y cells.
ABSTRACT
Solute carrier (SLC) transporters meditate many essential physiological functions, including nutrient uptake, ion influx/efflux, and waste disposal. In its protective role against tumors and infections, the mammalian immune system coordinates complex signals to support the proliferation, differentiation, and effector function of individual cell subsets. Recent research in this area has yielded surprising findings on the roles of solute carrier transporters, which were discovered to regulate lymphocyte signaling and control their differentiation, function, and fate by modulating diverse metabolic pathways and balanced levels of different metabolites. In this review, we present current information mainly on glucose transporters, amino-acid transporters, and metal ion transporters, which are critically important for mediating immune cell homeostasis in many different pathological conditions.
ABSTRACT
Background: Imbalance of Th17/Treg cells might play a key role in the initiation of Crohn's disease (CD). Moreover, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) pathway was suggested to be implicated in the pathogenesis of CD and other autoimmune diseases. Aims: To investigate the influence of TRAIL gene deletion on the balance of Th17 and Treg cells in mice with experimental colitis. Methods: TRAIL-/- C57BL/6 mice were obtained by CRISPR/Cas9 method. Then 20 TRAIL-/- mice and 20 wild-type (WT) mice were randomly divided into WT group, TRAIL-/- group, WT trinitrobenzenesulfonic acid (TNBS)-induced colitis group and TRAIL-/- TNBS-induced colitis group, with 10 mice in each group. The severity of colonic inflammation was assessed by disease activity index (DAI) and histology activity index (HAI). Peripheral blood and colonic tissue were collected to determine the expression levels of Th17 and Treg cell-associated transcription factors (RORγt, Foxp3) and cytokines [interleukin-17 (IL-17), IL-10] by real-time PCR, Western blotting and ELISA method, respectively. Results: Compared with untreated mice, mice with experimental colitis showed decreased body weight, shortened colon length as well as increased DAI and HAI (P<0.05), and the manifestations of colitis in TRAIL-/- mice were more serious than that in WT mice (P<0.05). In WT mice with experimental colitis, the expression levels of RORγt and IL-17 in peripheral blood and colonic tissue were significantly increased than those in untreated WT mice (P<0.05), while the expression levels of Foxp3 and IL-10 were significantly decreased (P<0.05). Furthermore, the expression levels of all four Th17 and Treg cell-associated molecules were higher in TRAIL-/- colitis mice than in WT colitis mice (P<0.05). In addition, the ratio of RORγt to Foxp3 were higher in TRAIL-/- colitis mice than in WT colitis mice (P<0.05). Conclusions: Deletion of TRAIL gene may aggravate the severity of colonic inflammation via up-regulating Th17/Treg ratio in mice with experimental colitis.
ABSTRACT
Objective: To investigate whether piperlongumine can sensitize prostate cancer cells to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and trigger apoptosis in prostate cells. Methods: Human prostate cancer cell lines PC3, LNCaP, and VCaP were cultured with piperlongumine and TRAIL. Then, cell proliferation, migration, caspase activation, apoptotic protein expressions, and death receptor expressions were measured. Results: Piperlongumine inhibited cell proliferation at low doses (<10 μM) alone and in combination with TRAIL (25 ng/mL), induced apoptosis, and suppressed cyclooxygenase activation. Additionally, piperlongumine induced expression of death receptors which potentiated TRAIL-induced apoptosis in cancer cells but did not affect decoy receptors. Piperlongumine also downregulated tumor cell-survival pathways, inhibited colony formation and migration of cancer cells alone or in combination with TRAIL. The combination of piperlongumine with TRAIL was found to be synergistic. Conclusions: Our findings indicate that piperlongumine can sensitize cancer cells to TRAIL through the upregulation of death receptors and can trigger apoptosis with the downregulation of anti-apoptotic proteins.
ABSTRACT
Objective: To observe the effects of ginsenoside Rg1 pretreatment on the expression of survivin protein and apoptosis after spinal cord ischemia-reperfusion injury (SCII) in rats so as to explore the possible mechanism of ginsenoside Rg1 on motor function recovery after SCII in rats. Methods: We selected 120 adult healthy SD rats to construct the model of SCII and randomly divided them into four groups: sham operation group, ischemia group, ischemia-reperfusion group, and drug group. Basso Beattie and Bresnahan score (BBB score) was used to evaluate the motor function of the hind limbs of the rats. The expressions of survivin protein and apoptosis-inducing factor (AIF) was observed by immunohistochemistry. The expression and activity of survivin protein and Caspase-9 in each group were observed and analyzed by Western blot and RT-PCR. Results: The intervention of ginsenoside Rg1 could increase the score of the motor function of the rat hind limbs. It could decrease the number of AIF positive cells, but increase the number of survivin protein positive cells. Ginsenoside Rg1 could decrease the expressions of survivin and Caspase-9, and decrease the apoptosis of nerve cells in SCII. Conclusion: Ginsenoside Rg1 could inhibit the expression of Caspase-9 by promoting the expression of survivin protein and decrease the apoptosis of rat SCII induced by the level of cytoplasmic AIF.
ABSTRACT
Objective To investigate the influence of tumor necrosis factor-related apoptosis-inducing ligant (TRAIL) deficiency on mice colitis and the gut microbiota composition by inclding the expermental colitis model in tumor necrosis factor-related apoptosis-inducing ligand gene knockout ( TRAIL-/-) mice. Methods C57BL/6 TRAIL-/-mice and wild type (WT) mice were selected and assigned into TRAIL-/-control group (eight mice), TRAIL-/-colitis group (16 mice), WT control group (eight mice) and WT colitis group (16 mice).The mice of two colitis groups were oral administrated with 3.5% dextran sulphate sodium (DSS) in drinking water for seven consecutive days to induce experimental colitis model .The severity of colitis was evaluated by clinical appearance and histopathological examination .The colonic tissue samples of mice were collected and microbiota profile was analyzed by 16S rDNA sequencing method.USEARCH software and R language were used to analyze the difference of gut microbiota among TRAIL-/-control group, TRAIL-/-colitis group, WT control group and WT colitis group .T test and Mann-Whitney U test were used for statistical analysis . Results After modeling, the disease activity index (DAI) of WT colitis mice and TRAIL-/-colitis mice both gradually increased over time .Furthermore, compared with colitis mice, TRAIL-/-colitis mice developed body weight loss, diarrhea and hemafecia earlier .On the seventh day after modeling , the percentage of body weight loss of TRAIL-/-colitis mice and WT colitis mice was (28.98 ±2.84)%and (17.87 ±3.70)%, respectively; and the difference was statistically significant (t＝9.53, P?0.01).The length of colon of TRAIL-/-colitis mice was shorter than that of WT colitis mice ((4.63 ±0.28) cm vs.(6.02 ±0.41) cm), and the difference was statistically significant (t＝11.20, P?0.01).The DAI of TRAIL-/-colitis mice was higher than that of WT colitis mice (3.00 ±0.00 vs.2.32 ±0.05), and the difference was statistically significant (t ＝54.40, P? 0.01).The histological score of TRAIL-/-colitis mice was higher than that of WT colitis mice (6.19 ±0.25 vs. 3.87 ±0.22), and the difference was statistically significant (t ＝27.87, P?0.01).Under the microscope, colonic mucosal epithelial injury , crypt structure destruction and inflammatory cell infiltration were more obvious in TRAIL-/-colitis mice than in WT colitis mice.The alpha diversity of colonic flora was more significant in TRAIL-/-colitis group compared with that of WT colitis group .At the family level, the relative richness of Deferribacteraceae, Ruminococcaceae, Rikenellaceae, F16 and Paraprevotellaceae significantly increased in TRAIL-/-colitis group, but the relative richness of Enterococcaceae obviously reduced ((19.839 ±19.991)%vs. (7.224 ±11.241)%, (3.564 ±2.543)% vs.(2.861 ±3.821)%, (0.123 ±0.066)% vs.(0.068 ± 0.049)%, (0.032 ±0.033)% vs.(0.006 ±0.011)%, (0.153 ±0.098)% vs.(0.062 ±0.054)% and (0.013 ±0.027)%vs.(0.054 ±0.121)%, respectively; U＝51, 69, 53, 35, 49 and 69, respectively; P? 0.01 and 0.05, respectively).In addition, at the genus level the relative richness of Oscillospira, Mucispirillum and Cytophaga in TRAIL-/-colitis group remarkably elevated , and the relative richness of Enterococcus significantly decreased ((2.363 ±2.147)% vs.(1.813 ±2.847)%, (19.839 ±19.991)% vs.(7.223 ± 11.241)%, (0.104 ±0.153)%vs.(0.046 ±0.069)% and (0.076 ±0.049)% vs.(0.135 ±0.074)%, respectively; U＝70, 51, 66 and 65, respectively; P ?0.05 and 0.01, respectively).Conclusion TRAIL deficiency aggravate DSS-induced colitis, and increase the alpha diversity of colonic microbiota in colitis mice .