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AIM: To investigate the effect of long non-coding RNA-HIF1A-AS1(lncRNA HIF1A-AS1)on the chemotherapy sensitivity of vincristine(VCR)-resistant in retinoblastoma(RB)cells by regulating the expression of hypoxia-inducible factor-1α(HIF-1α).METHODS: The human RB VCR-resistant cell line SO-RB50/VCR was established, expression of lncRNA HIF1A-AS1 in SO-RB50 and SO-RB50/VCR cells were detected by reverse transcription-quantitative real-time PCR(RT-qPCR); inhibition of lncRNA HIF1A-AS1 expression or simultaneous overexpression of HIF-1α in SO-RB50/VCR cells, and then median inhibitory concentration(IC50)of VCR and cell proliferation and apoptosis were detected in SO-RB50/VCR cells; the protein expressions of HIF-1α, multidrug resistance associate protein(MRP)and P-glycoprotein(P-gp)were measured by Western blot.RESULTS: Compared with SO-RB50 cells, the expression levels of lncRNA HIF1A-AS1 and HIF-1α protein in SO-RB50/VCR cells were increased(P<0.05); after inhibiting the expression of lncRNA HIF1A-AS1 in SO-RB50/VCR cells, the apoptosis rate was significantly increased(P<0.05), optical density(OD450), the IC50 value of VCR on cells and the expression levels of HIF-1α, MRP and P-gp proteins were significantly reduced(P<0.05); overexpression of HIF-1α attenuates the inhibitory effect of down-regulated lncRNA HIF1A-AS1 expression on drug resistance in SO-RB50/VCR cells.CONCLUSION: The lncRNA HIF1A-AS1 was highly expressed in SO-RB50/VCR cells, and inhibition of lncRNA HIF1A-AS1 expression reduced VCR resistance in SO-RB50/VCR cells by down-regulating HIF-1α expression.
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AIM: To investigate the expression and diagnostic value of long non-coding RNA(LncRNA)hypoxia-inducible factor 1 alpha antisense RNA 1(HIF1A-AS1)in serum of patients with proliferative diabetic retinopathy(PDR).METHODS: A total of 160 patients with diabetic retinopathy(DR)admitted to our hospital from July 2019 to July 2021 were selected as the research objects. According to the degree of disease, they were divided into PDR group(80 cases)and nonproliferative diabetic retinopathy(NPDR)group(80 cases). At the same time, 100 healthy cases in our hospital were selected as the control group. Detect and compare serum triglyceride(TG), total cholesterol(TC), high-density lipoprotein cholesterol(HDL-C), low-density lipoprotein cholesterol(LDL-C), fasting blood glucose(FBG)and the level of glycosylated hemoglobin A1c(HbA1c); The expression level of LncRNA HIF1A-AS1 in serum was detected by real-time fluorescence quantitative PCR(qRT-PCR)method; Logistic regression was used to analyze the risk factors that affected the occurrence of PDR; Receiver operating characteristic curve(ROC)was used to analyze the clinical value of LncRNA HIF1A-AS1 level in the diagnosis of PDR. RESULTS: The expression level of LncRNA HIF1A-AS1 in the serum of the patients in the PDR group was significantly higher than that in the NPDR group and the control group, and the NPDR group was higher than the control group(P<0.05); The course of disease, HbA1c, TC, TG, LDL-C, FBG levels in the PDR group and the NPDR group were significantly higher than those of the control group, the HDL-C level in the PDR group was significantly lower than that in the control group(P<0.05); The level of LncRNA HIF1A-AS1 was positively correlated with the course of disease, HbA1c, TC, TG, LDL-C and FBG(P<0.05), and negatively correlated with HDL-C(P<0.05); Logistic regression analysis showed that the LncRNA HIF1A-AS1, course of disease, FBG, HbA1c, TC, TG, LDL-C were all risk factors for PDR(P<0.05); ROC results showed that the area under the curve(AUC)of the LncRNA HIF1A-AS1 level predicting PDR was 0.766(95%CI: 0.692～0.829), the corresponding sensitivity was 66.25% and the specificity was 78.75%.CONCLUSION: The level of LncRNA HIF1A-AS1 in the serum of PDR patients is up-regulated, it is a risk factor for the occurrence of PDR and it can be used as a potential serological indicator for predicting the occurrence of PDR.
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SUMMARY: Renal ischemia-reperfusion injury (IRI)is an unavoidable consequence in renal transplantation and multiple clinical settings. A debate has been raised about the particular role of hypoxia-inducible factor (HF-1α) in the renal injury pathogenesis and the renal cortex ultrastructural alterations. Also, we investigated the antioxidant/anti-inflammatory effect of thymoquinone and its modulatory role on HIF-1α in protection against renal IRI.Adult male Wister albino rats were assigned into 3 groups (n=16); 1) Sham-operated, 2) IRI model and 3) renal IRI pre-treated with thymoquinone 10 mg.kg-1.day-1 (TQ-IRI) for 10 days and at the reperfusion onset. Following the operation, 8 rats from each group were euthanized after 3 hours and the remaining 8 rats at 24 hours. Renal injury was assessed by the increased blood urea nitrogen, creatinine level, and the EGTI histological injury scoreat both 3 and 24h. HIF-1α was upregulated (p<0.01) and was correlated with renal tissue reactive oxygen species (ROS) production and total oxidant capacity (TAC) consumption. Elevated inflammatory markers (NFkB, MCP-1 and VCAM-1) were associated with renal IRI.Thymoquinone treatment inhibited the accumulation of HIF-1α (p<0.01), reduced renal oxidation/inflammation process and markedly diminished renal injury.
RESUMEN: La lesión por isquemia-reperfusión renal (IRR) es una consecuencia inevitable en el trasplante renal como también en múltiples contextos clínicos. Se ha suscitado una discusión referente a la relación particular del factor inducible por hipoxia (HF- 1α) en la patogénesis de la lesión renal y las alteraciones ultraestructurales de la corteza renal. Además, investigamos el efecto antioxidante / antiinflamatorio de la timoquinona y su papel modulador sobre HIF-1α en la protección contra IRR. Se utilizaron ratas albinas Wister macho adultas divididas en 3 grupos (n = 16); 1) Intervención simulada, 2) modelo IRR y 3) IRR pretratado con timoquinona 10 mg/kg-1. día-1 (TQ-IRR) durante 10 días y al inicio de la reperfusión. Posterior a la operación, 8 ratas de cada grupo fueron sacrificadas después de 3 horas y las 8 ratas restantes a las 24 horas. La lesión renal se evaluó por el aumento de nitrógeno ureico en sangre, el nivel de creatinina y la puntuación de lesión histológica EGTI tanto a las 3 como a las 24 horas. HIF-1α se incrementó (p <0,01) y se correlacionó con la producción de especies de oxígeno reactivo (ROS) del tejido renal y el consumo de capacidad oxidante total. Los marcadores inflamatorios elevados (NFkB, MCP-1 y VCAM-1) se asociaron con IRR. El tratamiento con timoquinona inhibió la acumulación de HIF-1α (p <0,01), redujo el proceso de oxidación / inflamación renal y disminuyó notablemente la lesión renal.
Subject(s)
Animals , Male , Rats , Reperfusion Injury/drug therapy , Benzoquinones/therapeutic use , Acute Kidney Injury/drug therapy , Rats, Wistar , Oxidative Stress , Hypoxia-Inducible Factor 1/antagonists & inhibitors , Anti-Inflammatory Agents/therapeutic use , Antioxidants/therapeutic useABSTRACT
Animals exposed to hypobaric hypoxia triggers a physiological hypoxia response via Hypoxia Inducible Factor (HIF) proteins that functions as transcriptional complexes. As the South American camelids inhabit at high Andean altitudes we have asked if they have developed genetic adaptations to live at high altitudes. In the present study we investigate genetic structures of the HIF1A proteins carried by members of the superorder Cetartiodactyla. During our investigation we discovered the existence of a genetic event that caused the loss of most of the bHLH domain in the proteins borne by the Alpaca and other members of the Cetartiodactyla superorder; we designate them as bHLH short sequences. Further analysis at the nucleotide level revealed in the 12 short sequences included in the study the presence at the 5´end of the bHLH domains stop codons. Seven out of the 12 short HIF1A proteins, have an identical or almost identical nucleotide sequence at their 5´end with a same TAA stop codon and at the same position. As the mutations affects to both the Artiodactyls and Cetaceans, we postulate that the mutation(s) occurred before their divergence about 55 million years ago. The relevance of these findings for genetic adaptation of Alpacas to hypobaric hypoxia of high altitude conditions is discussed.
Los animales expuestos a hypoxia hipobárica generan una respuesta hipóxica fisiológica debido a unas proteinas de Factor-Hipoxia Inducible (HIF) que funcionan como complejos transcripcionales. Debido a que los camelidos Americanos habitan en las grandes alturas andinas, nos hemos preguntado si han desarrollado una adaptación genética para vivir a grandes alturas. Eneste estudio hemos investigado la estructura genética de las proteinas HIF1A que llevan consigo los miembros de la superorden de los cetartiodáctilos. Durante nuestra investigación, descubrimos la existencia de un evento genético que causó la perdida de la mayoría del dominio bHLH en las proteinas transmitidas por la alpaca y otros miembros de la superorden de los cetartiodáctilos; las hemos designado como secuencias cortas de bHLH. Análisis posteriores a nivel nucleótido revelaron que en la doceava secuencia corta incluida en el studio, hubo presencia de codones de terminación en el extreme 5' del dominio de bHLH. Siete de las doce proteinas cortas HIF1A, tiene una secuencia idéntica o casi idéntica de nucleotidos en su extremo 5', con el mismo codón de terminación TAA y en la misma posición. Debido a que la mutación afecta tanto a Artiodáctilos como Cetáceos, proponemos que la mutación(es) ocurrió antes de su divergencia hace unos 55 millones de años. Analizamos la relevancia de estos descubrimientos sobre la adaptación genética de las alpacas a la hipoxia hipobárica en condiciones de grandes alturas.
Subject(s)
Animals , Camelids, New World , Adaptation, Physiological/genetics , Basic Helix-Loop-Helix Transcription Factors/genetics , Hypoxia-Inducible Factor 1, alpha Subunit/genetics , HypoxiaABSTRACT
This paper explores the potential mechanism of microRNA-143–5p regulation effects on pulmonary arterysmooth muscle cells (PASMCs) functions in hypoxic pulmonary hypertension (HPH) via targeting HIF-1a,which may offer a new idea for HPH therapy. PASMCs were transfected with mimics control/miR-143–5pmimics or inhibitor control/miR-143–5p inhibitor. We used Western blotting and RT-qPCR to detect the proteinand mRNA expressions, CCK-8 assay to detect cellular viability, Annexin V-FITC/PI staining and caspase3/cleaved caspase-3 protein to evaluate cellular apoptosis, transwell migration experiment for cellularmigration measurement and Dual luciferase reporter gene assay to prove the target of miR-143–5p. Cells underhypoxic condition presented the decreased protein and mRNA expressions of a-smooth muscle actin (SM-aactin), Myocardin, smooth muscle myosin heavy chain (SMMHC), and smooth muscle-22a (SM22a),Calponin1 and Hypoxia-inducible factor-1a(HIF-1a), the increased cell viability and miR-143–5p level; Overexpression of miR-143–5p obviously reduced vascular smooth muscle-specific contraction marker proteinlevels and cellular apoptosis, increased cellular migration of PASMCs with hypoxia stimulation; Low-expression of miR-143–5p caused the opposite changes, while co-transfected with Si HIF-1a blocked thebeneficial effects of miR-143–5p inhibition on PASMCs under hypoxia. MicroRNA-143–5p can promote thephenotype conversion, proliferation and migration of pulmonary artery smooth muscle cells under hypoxiccondition through direct targeting of HIF-1a.
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OBJECTIVE@#To explore the inhibitory effects of silencing long non-coding RNA (LncRNA) HIF1A-AS2 on epithelialmesenchymal transition (EMT) and tumor stem cell-like phenotype in cervical cancer cells.@*METHODS@#We designed 3 shRNA constructs for silencing HIF1A-AS2 in CaSki cells, and the shRNA with the strongest interference effect was selected for subsequent experiment. CaSki cells were transfected with shRNA-NC or Sh-HIF1A-AS2, and the changes in cell viability, invasion ability, EMT, expressions of EMT-related proteins, formation of cell spheres and expressions of stem cell markers were detected.@*RESULTS@#Transfection with shRNA-NC and Sh-HIF1A-AS2 did not significantly affected the viability of CaSki cells (@*CONCLUSIONS@#Silencing HIF1A-AS2 can inhibit proliferation, invasion and migration of cervical cancer cells
Subject(s)
Female , Humans , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation/genetics , Epithelial-Mesenchymal Transition/genetics , Gene Expression Regulation, Neoplastic , Hypoxia-Inducible Factor 1, alpha Subunit , RNA, Long Noncoding/genetics , RNA, Small Interfering/genetics , Uterine Cervical Neoplasms/geneticsABSTRACT
Background: Hypoxia is a common feature of cancers. Hypoxia-inducible factor 1A (HIF1A) is a causative agent that changes the transcriptional response of tumors under hypoxia. Some alterations lead to an increase in HIF1A activity and this supports other critical pathways leading to angiogenesis, metabolic adaptation and tumor progression. This retrospective study was designed to evaluate the differences of tissue expressions of HIF1A in a spectrum of cervical neoplasms.Methods: Tissue expression of HIF1A was studied in a total of 107 formalin-fixed, paraffin-embedded uterine cervical tumors specimens and its association with different clinicopathologic parameters was evaluated.Results: In this series, there were 30 low and 29 high grade cervical intraepithelial neoplasms (CINs), 27 squamous cell carcinomas, 15 adenosquamous carcinomas and 6 adenocarcinomas. Strong and diffuse nuclear staining was evaluated as positive HIF1A expression. Positive HIF-1 alpha expression was detected in 7 (25.9%) of squamous cell carcinomas, 1 of adenocarcinomas (16.7%) and only 1 of HGSILs (3.4%). Statistically it was determined that the positivity rate of strong nuclear HIF1A expression was significantly higher in invasive carcinomas when compared with in non-invasive squamous cell carcinomas (p=0.07). Contrary, there was no statistically significant difference according to the subtypes of carcinomas due to scarce number of cases with adenocarcinoma (p=0.188).Conclusions: Our findings were demonstrated to link of nuclear HIF1A expression and the invasive characters of uterine neoplasms. As a result, HIF-1 alpha expression may be important in foreseeing of the invasion and tumor progression.
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Abstract Persistent apical periodontitis (AP) is a situation involving an inflammatory and immune response caused mainly by anaerobic polymicrobial infection of the root canal system and the outcome and follow-up of the root canal treatment has been reported as intimately related to host response. The apical periodontitis repair might be associated with genetic polymorphisms. This study aimed to evaluate the association between HIF1A genetic polymorphisms (rs2301113 and rs2057482) with PAP in Brazilian patients. Subjects with at least 1 year of follow-up after root canal therapy (RCT) were recalled. Sixty-four subjects with signs/symptoms of PAP and 84 subjects with root canal-treated teeth exhibiting healthy perirradicular tissues (healed) were included. Genomic DNA was extracted from saliva and used for HIF1A genotyping by real-time PCR. Genotype and allele frequencies were compared by c2 or Fisher's exact tests and odds ratio was implemented, using Epi Info 3.5.2. All tests were performed with an established alpha of 0.05. There was no association between allele and genotype distribution for HIF1As polymorphisms and PAP (p>0.05). The genetic polymorphisms in HIF1A were not associated with persistent apical periodontitis.
Resumo A periodontite apical persistente (PAP) é uma condição que envolve uma resposta inflamatória e imunológica causada principalmente por infecções polimicrobianas de origem anaeróbia no sistema de canais radiculares, tornando o resultado e o acompanhamento do tratamento do canal radicular intimamente relacionados à resposta do hospedeiro. O reparo da periodontite apical pode estar associado a polimorfismos genéticos. Este estudo teve como objetivo avaliar a associação entre os polimorfismos genéticos do HIF1A (rs2301113 e rs2057482) com a PAP em pacientes brasileiros. Indivíduos com pelo menos 1 ano de acompanhamento após o tratamento do canal radicular (TCR) foram agendados para consulta de acompanhamento. Sessenta e quatro indivíduos com sinais/sintomas de PAP e 84 indivíduos com dentes tratados endodonticamente e tecidos perirradiculares saudáveis (cicatrizados) foram incluídos no presente estudo. O DNA genômico foi extraído da saliva e utilizado para a genotipagem do HIF1A por PCR em tempo real. O genótipo e as frequências alélicas foram comparados por teste c2 ou exato de Fisher e odds-ratio foi implementado por meio do software Epi Info 3.5.2. Todos os testes realizados foram estabelecidos com a=0,05. Não houve associação entre alelo e distribuição genotípica para polimorfismos do HIF1A e PAP (p> 0,05). Os polimorfismos genéticos em HIF1A não foram associados à periodontite apical persistente.
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Periapical Periodontitis/genetics , Polymorphism, Genetic , Bone Remodeling/genetics , Hypoxia-Inducible Factor 1, alpha Subunit/genetics , Neovascularization, Pathologic/genetics , Periapical Periodontitis/pathology , Root Canal Therapy , Brazil , DNA/genetics , Real-Time Polymerase Chain Reaction , Gene Frequency , GenotypeABSTRACT
Objective To observe the effect of HIF-1a/iNOS signaling pathway on myocardial ischemia-reperfusion injury in rat heart transplantation and the protective mechanism of N-acetylcysteine (NAC) on donor heart after cardiac transplantation in rats.Methods Eighty healthy male Lewis rats were randomly divided into 3 groups,the control group (0.3 ml saline was infused via inferior vena cava 30 min before donor harvest or implantation),NAC donor pretreatment group [NAC (30 mg/kg.w) was injected into the vena cava of donor rat 30 min defore donor harvest],and the NAC receptor pretreatment group (NAC 300 mg/kg.w was injected into the vena cava of the recipient rats 30 min before transplantation.The 30 min was injected into the vena cava of the recipient rats).A transplant model was established and the graft was obtained after 24 h transplantation.The expression of iNOS,HIF-1a and mRNA in cardiac muscle tissue was detected by immunohistochemistry and Real time-PCR.Results HIF-1a protein expression in graft myocardial tissue was significantly lower in NAC donor pretreatment and recipient pretreatment group compared with control group (P ＜0.05),the differences were statistically significant (2.72±0.17 vs.2.24±0.23 vs.3.14.±0.16,F=56.26,P =0.000).The iNOS protein expression in NAC donor pretreatment group,and NAC recipient pretreatment group were lower than that in the control group (1.52±0.18 vs.1.61±0.19 vs.3.30±0.18,F=232.345,P =0.000),the differences were statistically significant (P ＜ 0.05).24 h after transplantation,the differences in graft myocardial tissue HIF-1a and iNOS mRNA among the three groups were statistically significant (F=7.467,16.490,P=0.003,0.000).The expression of iNOS mRNA in the NAC receptor pretreatment group was significantly lower than that in the control group (P＜0.05).Conclusion HIF-1a/iNOS signaling pathway can regulate ischemia reperfusion injury in rat heart transplantation,and the protective effect of NAC on donor heart maybe mediated via this pathway.
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Objective To observe the regulatory role of long non-coding RNA HIF1A-AS1 on the autophagy of pancreatic cancer PANC1 cells induced by hypoxia.Methods The pancreatic cancer PANC1 cells were cultured in a three-gas incubator filled with hypoxic gas mixture (94% N2,5% CO2,1% O2) for 3, 6, 12, 24, 36 and 48 h.HIF1A-AS1 overexpression and low expression PANC1 cells were obtained by the infection of recombinant adenovirus carrying HIF1A-AS1 and the transfection of HIF1A-AS1 targeting siRNA by liposome, and routinely cultured PANC1 cells served as control.The expression of HIF1A-AS1 of PANC1 cells was detected by real-time quantitative PCR after being cultured in hypoxia-induced condition for 24 h.The apoptosis rate was detected by flow cytometry.The autophagy related proteins Beclin 1 were detected by western blot.Results The expression of HIF1A-AS1 in hypoxic cells was increased as the hypoxic time increased since 6 h and peaked at 36 h, which was significantly higher than that in control group (P<0.01).HIF1A-AS1 relative expression in HIF1A-AS1 overexpression and low expression PANC1 cells was 4.49±0.53 and 0.49±0.07, which were normalized to that of control group with the relative expression of 1.Control group had lower HIF1A-AS1 expression than HIF1A-AS1 overexpression PANC1 cells but higher HIF1A-AS1 in HIF1A-AS1 low expression PANC1 cells, and the differences were statistically significant (P<0.01).The cell apoptosis rate of control, HIF1A-AS1 overexpression and low expression PANC1 cells was (8.27±1.28)%, (6.56±1.49)% and (19.9±2.34)% after 24 h hypoxic culture.Control group had higher HIF1A-AS1 expression than HIF1A-AS1 overexpression PANC1 cells but lower HIF1A-AS1 in HIF1A-AS1 low expression PANC1 cells, and the differences were statistically significant (P<0.01).The expression of Beclin 1 protein was protein 1.05±0.11, 1.29±0.19 and 0.38±0.18, respectively.Control group had lower Beclin 1 expression than HIF1A-AS1 overexpression PANC1 cells but higher Beclin 1 in HIF1A-AS1 low expression PANC1 cells, and the differences were statistically significant (P<0.01).Conclusions HIF1A-AS1 can promote autophagy of pancreatic cancer PANC1 cells induced by hypoxia and participate in the pathogenesis and metastasis of pancreatic cancer.
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It has been demonstrated that hypoxia retards the growth of fish, reduces the survival of their larvae, deforms their vertebral column, but despite this teleost fish have the ability to completely regenerate many of their tissues, particularly the retina. As we do not have enough information about the effects of hypoxia on the eyeball, orbit and retina of Atlantic salmon (Salmo salar), we propose the following objectives: 1) Compare the morphological changes of the eyeball of fish subject to hypoxia and normoxia. 2) Determine changes in the orbit structure. 3) Describe the retina of salmon alevins. 4). Recognize hypoxic cells using the anti-Hif1a antibody in the retina of alevins as a sensor. 5) Determine the Shh morphogenic expression in alevins exposed to different times of hypoxia. Around 1,000 Salmo salar alevins were placed in a continuous water flow of 9 °C at 100 % SatO2 and alevins maintained at a hypoxia of 60 % SatO2. The latter were transferred to normoxia (at days two, four, and eight after hatching). A control group maintained at continuous normoxia and another at continuous hypoxia was also considered. All the alevins were euthanized at 950 UTAs (±2 months after hatching). Diaphonization (double-stain) according to the Hanken & Wassersug technique was undertaken to describe the morphology of the periocular cartilage and to measure the ocular diameter. The HIF-1a factor antibody 1:50, and the anti-Shh antibody dilution of 1:100 were used. The alevins after hatching had large eyeballs with the optic cup having an embryonic shape, even a choroidal fissure. The greatest thickness was observed in the nasal ventral zone which corresponds to a zone of pluripotent cells. The optic cup aspect with embryonic characteristics has only been reported in salmonids. The central retina of the alevins those were cultivated with a 60 % saturation of O2 for two, four or eight days had positive immunostaining when analyzed with the anti-HIF1a antibody hypoxia sensor. The inner ganglion and nuclear layers had immunopositive cells, with the highest in the alevins that were two days in hypoxia and the lowest when the hypoxia was chronic. Nevertheless, in the latter case the alevins had anatomical deformation of the eyeball and periocular cartilage. The anti-Shh antibody clearly shows a gradient that is expressed in the germinative zone and in the cells of the inner ganglion and nuclear layers. The eyeball and particularly the retina in salmon alevins are an example of neuronal plasticity and neurogenesis.
Se ha demostrado que la hipoxia retarda el crecimiento de los peces, reduce la supervivencia de sus larvas, deforma su columna vertebral, pero a pesar de esto, este pez teleósteo tiene la capacidad de regenerar completamente muchos de sus tejidos, en particular la retina. Como no existe suficiente información sobre los efectos de la hipoxia en el bulbo ocular, la órbita y retina del salmón del Atlántico (Salmo salar), los objetivos de este trabajo fueron: 1) Comparar los cambios morfológicos del bulbo ocular del pescado sujetos a hipoxia y normoxia; 2) Determinar los cambios en la estructura de la órbita; 3) Describir la retina de los alevines de salmón; 4) Reconocer las células hipóxicas utilizando el anticuerpo anti-Hif1a en la retina de alevines como un sensor; 5) Determinar la expresión morfogenética de Shh en alevines expuestos a diferentes momentos de hipoxia. Alrededor de 1.000 alevines Salmo salar se colocaron en un flujo continuo de agua a 9 °C, con 100 % de SatO2 y otros alevines se mantuvieron con una hipoxia de 60 % SatO2. Estos últimos fueron trasladados a normoxia (en los días dos, cuatro y ocho después de la eclosión). Un grupo control se mantuvo a normoxia continua y otro grupo a hipoxia continua. Todos los alevines se sacrificaron a 950 UTA (+ dos meses después de la eclosión). Se realizcón una diafonización (doble tinción), de acuerdo con la técnica de Hanken & Wassersug, para describir la morfología del cartílago periocular y para medir el diámetro ocular. Se utilizaron el anticuerpo anti-Hif1a a una dilución 1:50, y el anticuerpo anti-Shh a una dilución de 1:100. Los alevines después de la eclosión presentaron grandes bulbos oculares, con la copa óptica con forma embrionaria, incluso una fisura coroidea. El mayor espesor se observó en la zona ventral nasal que corresponde a una zona de células pluripotentes. El aspecto de la copa óptica con características embrionarias sólo se ha informado en los salmónidos. La retina central de los alevines fueron cultivadas con una saturación de 60 % de O2 para dos, cuatro y ocho días, y presentó inmunotinción positiva cuando se analizó con el sensor de hipoxia, el anticuerpo anti-HIF1a. El ganglio interior y las capas nucleares presentaron células immunopositivas, con los niveles más altos en los alevines con dos días de hipoxia y niveles más bajos en hipoxia crónica. Sin embargo, en éste último caso los alevines presentaron una deformación anatómica del bulbo ocular y el cartílago periocular. El anticuerpo anti-Shh mostró claramente un gradiente expresado en la zona germinativa y en las células del ganglio interior y las capas nucleares. El bulbo ocular y en particular la retina en alevines de salmón son un ejemplo de plasticidad neuronal y neurogénesis.
Subject(s)
Animals , Eye/anatomy & histology , Hypoxia , Orbit/anatomy & histology , Retina/anatomy & histology , Salmo salar/anatomy & histologyABSTRACT
Objective To study the regulatory effects of long non-coding RNA HIF1A-AS1 on the myocardial ischemia reperfusion (I/R) injury and the related mechanism. Methods Myocardial I/R injury model was established with SD rats, and hypoxia reoxygenation (H/R) model was established with rat cardiac myocytes. si-HIF1A-AS1 was used to inhibit HIF1A-AS1 expression in the cardiac myoctyes. Then the mRNA expression of HIF1A-AS1 was detected by real-time PCR, the growth vitality of cardiac myocytes was investigated by MTT assay, the concentration of lactate dehydrogenase (LDH) in the culture media was detected by ELISA, and the autophagy-associated protein Beclin-1 expression was observed by Western blotting analysis. Results HIF1A-AS1 expression was increased in cardiac muscle of rat I/R model and rat cardiac myocytes of H/R model. Inhibition of HIF1A-AS1 by siRNA protected the cardiomyocytes against H/R injuries, reversing the decreased growth vitality of cardiac myoctyes, increased LDH level in the culture media, and increased expression of autophagy-related protein Beclin-1 induced by H/R stimulation. Conclusion Inhibition of long non coding RNA HIF1A AS1 might play a protective role in I/R injury of cardiac myoctyes by inhibiting the excessive autophagy of cardiomyocytes.
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BACKGROUND: Growing tumors adapt to a hypoxic environment and increase anaerobic glycolysis. This metabolic switch is related to aggressive behavior. We investigated the relationship between glycolytic metabolism biomarkers associated with hypoxia-inducible factor (HIF)-1alpha and prognosis. METHODS: We performed immunohistochemical staining of HIF-1alpha, pyruvate dehydrogenase kinase (PDK) 1 and lactate dehydrogenase (LDH) 5 in 74 patients with oral squamous cell carcinoma (SCC) who had received curative radical resection. RESULTS: High reactivity of HIF-1alpha, PDK 1 and LDH 5 was observed in 29 (39.2%), 32 (43.2%) and 54 (73.0%) patients, respectively. Expression levels of the three biomarkers were significantly correlated. All three markers were highly expressed in 16 (21.6%) patients. Elevated expression of the three markers was associated with increased invasiveness (p = 0.043) and recurrence (p = 0.017) of tumors. In survival analysis, upregulation of the three markers was additionally associated with shorter disease free survival (DFS, p = 0.001) and overall survival (OS, p = 0.002). High expression of all three markers was a strong independent prognostic factor for DFS (p = 0.030) and OS (p = 0.026). CONCLUSIONS: Oral SCC with altered glycolytic metabolism exhibits a more invasive and aggressive phenotype. Our results indicate that glycolytic metabolism biomarkers related to HIF-1alpha may be independent prognostic factors in patients with oral SCC.
Subject(s)
Humans , Biomarkers , Carcinoma, Squamous Cell , Disease-Free Survival , Glycolysis , Hypoxia-Inducible Factor 1, alpha Subunit , Isoenzymes , L-Lactate Dehydrogenase , Oxidoreductases , Phenotype , Phosphotransferases , Prognosis , Protein Serine-Threonine Kinases , Pyruvic Acid , Recurrence , Up-RegulationABSTRACT
Objective To investigate the correlation between the expression of HIF-1a (hypoxia inducible factor 1,HIF-1a) in perihematomal brain issue and formation of brain edema in patients with hypertensive cerebral hemorrhage.Method Perihematomal brain issue was collected in the course of hematoma elimination in 32 patients with hypertemive intraeerebral hemorrhage.Expressions of HIF-1a and vascular endothelial growth factor (VEGF) were observed by immunohistochemistry.The volume of perihematomal brain edema on computed tomographie scan was determined by computed tomographic scan before surgery.The results of staining and the volume of perihematomal brain edema were analyzed in double blind fashion.Results HIF-1a protein immunohistochemical staining positive cells (2.8?0.8/HP) were identified dispersedly from 4 hours after acute hemorrhagic stroke in perihematomal brain issue,and reached the peak at 24～48 hours (12.5?3.9/HP).High expression of HIF-1a progressed at 48～72 hours (12.2?1.8/HP) after acute hemorrhagic stroke.There was a positive correlation between the expression of HIF-1a and VEGF (r=0.76,t=6.37,P