Hypoxia inducible factor

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Pola mRNA Hypoxia Inducible Factor-1a (HIF-1a) dan Exkspresi Protein HIF-1a Ginjal Tikus pada Hipoksia Sistemik Kronik

Pola mRNA Hypoxia Inducible Factor-1a (HIF-1a) dan Exkspresi Protein HIF-1a Ginjal Tikus pada Hipoksia Sistemik Kronik

The aim of this study was to examine the pattern of Hypoxia Inducible Factor- 1α (HIF-1α) mRNA and HIF-1α protein expression in rat kidney in systemic chronic hypoxia using hypoxic chamber with 8% Oxygen and 92% Nitrogen and control group with normoxia condition. The pattern was observed based on RT-PCR result comparing the ratio of control group and the hypoxic group. Six groups of 8 male mice Spraque and Dawley each group were used in this experiment. The result showed that HIF-1α mRNA expression elevated gradually and reached the peak at 3 days in hypoxia group and started to decrease in 7 day hypoxia group. The HIF-1α protein expression was measured employing Western Blott method that showed an increase expression until 14 days of hypoxia which eventually started to decrease. Oksigen memegang peranan penting
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13 Lee mas

Ekspresi Relatif MRNA Hypoxia Inducible Factor-1? Pada Sel Glioma Penderita

Ekspresi Relatif MRNA Hypoxia Inducible Factor-1? Pada Sel Glioma Penderita

Keyword: Glioma, mRNA HIF-1 α, hypoxia ABSTRAK Glioma merupakan tumor otak primer yang cukup sering ditemukan di Indonesia di antara keganasan otak lainnya. Penderita glioma mempunyai angka harapan hidup yang rendah dikarenakan seringnya mengalami resistansi terapi. Hipoksia merupakan salah satu faktor yang mempengaruhi respons terapi radiasi. Penelitian ini menganalisis ekpresi mRNA HIF-1α, sebagai petanda jaringan hipoksia, pada glioma derajat rendah dibandingkan dengan glioma derajat tinggi. Sampel terdiri dari 15 jaringan glioma derajat rendah, 6 jaringan glioma derajat tinggi, dan 21 sel lekosit dari penderita glioma sebagai kontrol sel normal. Ekspresi relatif mRNA HIF-1α ditentukan secara kuantitatif dengan menggunakan Real Time RT-PCR. Hasil penelitian menunjukkan bahwa ekspresi mRNA HIF-1 α pada glioma derajat tinggi lebih tinggi dibandingkan dengan glioma derajat rendah, namun tidak bermakna secara statistik. Hal yang menarik yaitu ditemukan peningkatan ekspresi mRNA HIF-1α dengan nilai yang bervariasi pada 73 % sampel glioma derajat rendah dibandingkan dengan sel lekosit sebagai kontrol sel normal. Begitu juga pada glioma derajat tinggi ditemukan peningkatan mRNA HIF-1α dengan nilai yang bervariasi pada 100% sampel. Dengan demikian, dapat disimpulkan bahwa terdapat variasi derajat hipoksia pada glioma derajat rendah dan derajat tinggi yang mungkin mempengaruhi keberhasilan terapi radiasi.
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5 Lee mas

Korelasi antara Hypoxia Inducible Factor-1α dengan Aktivitas Enzim Creatine Kinase Jaringan Otot Jantung Tikus pada Aktivitas Fisik Aerobik dan Anaerobik

Korelasi antara Hypoxia Inducible Factor-1α dengan Aktivitas Enzim Creatine Kinase Jaringan Otot Jantung Tikus pada Aktivitas Fisik Aerobik dan Anaerobik

Aerobic and anaerobic physical activities performed for 10 days without any rest-day can lead to hypoxia and histopathological changes in the heart muscle. Hypoxic conditions correlated to histopathological changes in the heart muscle. To determine whether these hypoxic conditions also correlated with indicators of tissue damage to the heart muscle, further research needs to be done. This study aims to determine the correlation between hypoxia that occurs in the heart muscle due to the imposition of aerobic and anaerobic physical activity without a day of rest with CK enzyme activity of cardiac muscle tissue. Heart muscle tissue derived from rats fed aerobic and anaerobic physical activity using a treadmill for 1,3,7 and 10 days without a day of rest. Then measuring the levels of HIF-1α as a parameter hypoxia and CK enzyme activity as a parameter cardiac muscle cell damage. The results showed that there was a very strong correlation between the concentration of HIF-1α with tissue CK activity in anaerobic group (r = 0.91, p <0.05), whereas the aerobic group correlation being (r = 0, 48, p <0.05). The condition of hypoxia on the heart muscle caused by aerobic and anaerobic physical activity carried out for 10 days without a day of rest strongly correlate with indicators of tissue damage in groups of anaerobic and aerobic groups were correlated.
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Papel hipottico del factor 1 inducible por hipoxia en las adaptaciones a la microgravedad

Papel hipottico del factor 1 inducible por hipoxia en las adaptaciones a la microgravedad

Maintaining health in microgravity and overcoming environmental hazards such as cosmic radiation are essential for long-term space flight. Recent studies have focused on the involvement of hypoxia-inducible factor (HIF)-1 in altered gravity using cell-based or in vivo mouse model systems. HIF-1alpha and its target downstream gene expression are

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Influencia de la hipoxia sobre el metabolismo óseo. Rol central del factor inducible por hipoxia

Influencia de la hipoxia sobre el metabolismo óseo. Rol central del factor inducible por hipoxia

Knowledge of hypoxia influencing human physiology determined discovery of hypoxia-inducible factor (HIF). Regulation of molecules by this transcription factor is varied and complex and understanding continues on. Several processes like ossification, remodeling and repair occur in bone tissue physiology and pathophysiology. Decrease in available oxygen determines HIF stabilization in many cell groups, standing out osteogenic lineage cells. This factor regulates expression of multiple genes, the most important perhaps the vascular endothelial growth factor (VEGF) due to its implication in the osteogenic process, directly activating osteoblastic cells or indirectly through potent angiogenesis. In this article, we review HIF central role on bone physiology, detailing regulation of key molecular pathways dependent of this transcription factor.
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TítuloEffects of severe hypoxia on bone marrow mesenchymal stem cells differentiation potential

TítuloEffects of severe hypoxia on bone marrow mesenchymal stem cells differentiation potential

As known from the literature, several molecular pathways involved in cellular metabolism are altered under hypoxia [50]. MSCs are characterized by metabolic flexibility and are able to survive hypoxic conditions [34, 51]. Hypoxia stabilized hypoxia-inducible factor-1 𝛼 (HIF-1𝛼). HIF-1𝛼 is a transcriptional factor that activates more than 40 genes and plays essential roles in a variety of cellular and systemic homeostatic responses to hypoxia [50]. This transcriptional factor downregulates mitochondrial oxygen consumption [52] and upregulates key markers of stem cells like Oct-4, Rex-1, and SSEA4 [41]. We hypothesized that the relationship between HIF-1𝛼, cells proliferation, and stem cell markers could explicate our results and the absence of differentiation under severe hypoxic conditions. In addition, several papers support this hypothesis [16, 53, 54]. In this regard, Carrancio et al. [16] reported that hypoxic culture conditions compared to the standard protocols showed to improve MSCs yield and reduce cells expansion time. Also, Ohnishi et al. [54] analyzed the gene expression of rat BM-MSCs in hypoxia and observed an upregulation of genes involved in development, cell proliferation, and cell survival. Finally, Dos Santos et al. [53] reported that hypoxia promotes cell proliferation and expansion. During proliferation, the majority of the cells undergo cell division and cannot begin the cellular mech- anisms involved in differentiation processes. Therefore the use of low pO 2 levels to increase the in vitro survival or self-renewal of human mesenchymal stem cells represents an
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12 Lee mas

Carotid body and cardiorespiratory alterations in intermittent hypoxia: the oxidative link

Carotid body and cardiorespiratory alterations in intermittent hypoxia: the oxidative link

CB [12, 15], but nitric oxide (NO) and pro-inflammatory molecules may also be involved [8]. Since reactive oxygen (ROS) and nitrogen species are potential mediators of cardiovascular alterations in OSA patients [3–5] and CIH-exposed animals [15–17], we hypothesised that an antioxidant treatment may prevent both CB and cardiorespiratory altered functions. Thus, we studied the effects of ascorbic acid on the potentiation of CB chemosensory and ventilatory hypoxic responses, HRV alterations and the hyperten- sion induced by CIH in rats. Malondialdehyde (MDA) assay was used as a systemic oxidative stress marker, along with 3- nitrotyrosine (3-NT) and inducible nitric oxide synthase (iNOS) immunoreactivity as oxidative stress markers in the CB. This study was performed in anaesthetised rats, a necessary condition to record CB chemosensory neural activity.
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Inflammation and oxidative stress during intermittent hypoxia: the impact on chemoreception

Inflammation and oxidative stress during intermittent hypoxia: the impact on chemoreception

Chronic intermittent hypoxia increases the TNF- α - and IL-1β-immunoreactive levels in the CB, suggesting a potential role for these pro-inflammatory cytokines in modulating the enhanced CB chemosensory activity following CIH (Del Rio et al. 2011). Thus, we investigated whether these cytokines contribute to the CIH-induced CB chemosensory potentiation. We found that ibuprofen, which prevents the CIH-induced increased TNF-α and IL-1β in the CB, failed to impede the potentiation of the CB chemosensory responses to hypoxia, although it effectively prevented the enhanced ventilatory responses to hypoxia and the hypertension (Del Rio et al. 2012b). Figure 1 shows the effects of antioxidant (i.e. ascorbic acid) and anti-inflammatory (i.e. ibuprofen) treatments on the CB chemosensory discharge measured in normoxia and hypoxia from rats exposed to CIH for 21 days. Figure 1A and B illustrates representative recordings of carotid sinus nerve discharges induced by short hypoxic challenges in a sham rat (a rat exposed to air:air cycles instead of a rat exposed to intermittent hypoxia) and in a rat exposed to CIH for 21 days. As is shown in Fig. 1B, CIH increased baseline chemosensory discharges in normoxia and enhanced the chemosensory responses to hypoxia, although no differences were observed between the responses to hyperoxic stimuli. Figure 1C and D summarizes the effects of ascorbic acid and ibuprofen on CB chemosensory discharges measured in normoxia and in response to 10% O 2 .
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Expresión inducible de hPPARγ en el hígado de ratones C57BL/6 con diabetes mellitus tipo 2.

Expresión inducible de hPPARγ en el hígado de ratones C57BL/6 con diabetes mellitus tipo 2.

identificar los genes activados por PPARγ. Esto lo realizaron en células HepG2 y por medio de cotransfección de dos plásmidos no virales: pTet-Off vector y pBI-EGFP-hPPARγ (plásmido bicistrónico, para la expresión de simultánea del gen de interés, hPPAR γ, y del gen reportero, EGFP). En nuestro caso, también se empleó un sistema de dos plásmidos, con un vector respuesta que también cuenta con un gen reportero (ZsGreen1), aunque únicamente cuenta con un promotor y la expresión se da por medio de un único mRNA, con un elemento IRES entre los dos genes. Por otro lado, Liu et al. 2013 llevaron a cabo la expresión de hPPARγ y C/EBP por medio de un sistema inducible Tet-On de tipo viral (lentivirus) en líneas celulares de adipocitos y con diferentes elementos respuesta y proteínas activadoras. Además, no se tiene conocimiento de reportes previos en los que se haya llevado a cabo la expresión de PPARγ humano en modelos animales con sistemas de expresión inducible.
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Assessing gene action for hypoxia tolerance in
  cotton (Gossypium hirsutum L.)

Assessing gene action for hypoxia tolerance in cotton (Gossypium hirsutum L.)

All higher plants require water, but excess water in roots can cause injury and can be lethal due to low oxygen diffusion through water coupled with its limited solubility leads to low oxygen availability (Sairam et al., 2008). Cotton (Gossypium hirsutum L.) is a widely studied crop and is the main source of natural fiber worldwide (Riaz et al., 2013); occupies a pivotal position as fiber crop of masses of the world in general and of Pakistan in particular (Imran et al., 2012). Periodic flooding or waterlogged conditions reduces crop yields of cotton in developing countries like Pakistan, India, and China (Pang et al., 2004). The yield penalty resulting from waterlogging may vary between 15% to 80%, depending on the species, soil type and duration of the stress (Zhou, 2010). Cotton is sensitive to waterlogging which results in young square shedding at reproductive stage which is found to be the result of hormonal imbalance (de-Brito et al., 2013). Waterlogging sensitivity in cotton is linked with photosynthetic inhibition (Najeeb et al., 2015). Plant tolerance to low O 2 varies dramatically between species, suggesting the existence of distinct, species-specific molecular mechanisms of plant adaptation to hypoxia (Zhou, 2010). It is, therefore, developing waterlogging tolerant varieties has long been a top priority for plant breeders. The agronomic definition of waterlogging tolerance is the maintenance of relatively high grain yields under waterlogging relative to non-waterlogged conditions (Setter and Waters, 2003). Accordingly, numerous phenotype-based indices have been used both in genetic studies (Parelle et al., 2010) and in breeding programs (Zhou, 2010).
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Clonación y expresión de LcrE de Chlamydia trachomatis en Escherichia coli usando un plásmido inducible con arabinosa

Clonación y expresión de LcrE de Chlamydia trachomatis en Escherichia coli usando un plásmido inducible con arabinosa

El gen lcrE fue clonado mediante la siguiente estrategia, figura 3, el producto de PCR fue ligado con la T4 DNA Ligasa al vector TA pCR 2.1 (Invitrogen), el plásmido recombinante se denominó pCR-lcrE. La mezcla de ligación fue usada para transformar Escherichia coli, cepa DH5α; y las colonias transformantes, fueron seleccionadas de placas de agar Luria Bertani (LB) suplementadas con 50 mg/ml de ampicilina. El fragmento extraído con EcoRI fue purificado en gel de agarosa de bajo punto de fusión al 1% por fenol/cloroformo, y ligado al plásmido pBAD gIII, el plásmido recombinante fue denominado pBAD-lcrE, e introducido en Escherichia coli TOP 10 inducible con arabinosa. La orientación correcta y contenido de nucleótidos del fragmento introducido fue probado por secuenciación.
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Carotid body potentiation during chronic intermittent hypoxia: implication for hypertension

Carotid body potentiation during chronic intermittent hypoxia: implication for hypertension

Chemoreflexes and baroreflexes play a main role in the control of the cardiorespiratory function in health and disease. Several stud- ies reported interactions between peripheral chemoreceptors and arterial baroreceptors (Heistad et al., 1975; Mancia et al., 1976; Somers et al., 1991). It has been shown that hypoxic chemoreflex activation elicits baroreflex inhibition mainly attributed to central nervous system modulation at the level of the nucleus of the soli- tary tract (Miura and Reis, 1972). The available evidence suggests that an enhanced CB chemoreflex drive contributes to the car- diorespiratory alteration induced by CIH (Mitchell et al., 2001; McGuire et al., 2003; Peng et al., 2003; Peng and Prabhakar, 2004; Rey et al., 2004; Del Rio et al., 2010). Indeed, CIH enhanced CB chemoreceptor response to hypoxia (Peng and Prabhakar, 2004; Rey et al., 2004; Del Rio et al., 2012), which in turn produces sympathetic activation and hypertension. Accordingly, a blunted baroreflex response to the subsequent hypertension during CIH, may contribute to the further deterioration of the reflex con- trol of BP. Indeed, we found that rats exposed to CIH displayed CB chemosensory potentiation and BRS reduction with the same time-course. Then, our results suggest that repetitive activation of the peripheral hypoxic chemoreflex during CIH result in alter- ations of the baroreflex control. Nevertheless, we cannot rule out that exposure to CIH per se may reduce BRS, or reset the barore- flex set-point to operate at a higher level of BP. Further studies are needed to prove that CB potentiation leads to baroreflex impairment following CIH.
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Rabbit Ventilatory Responses to Peripheral Chemoexcitators: Effects of Chronic Hypoxia

Rabbit Ventilatory Responses to Peripheral Chemoexcitators: Effects of Chronic Hypoxia

The ventilatory and cardiovascular responses to acute hypoxia (Chalmers et al. 1967 ; Korner and Edwards 1960 ) and NaCN (Docherty and McQueen 1979 ; Matsumoto 1986 ) were similar to those previously reported. However, CB denervation reduced the responses that were abolished only after total peripheral chemodenervation. In non-anesthetized rabbits, the hypoxic ventilatory responses are completely dependent on carotid afferents with negligible participation of the aortic chemoreceptors (Bouverot et al. 1973 ; Chalmers et al. 1967 ; Korner and Edwards 1960 ) . The observed differences could be partly explained by the use of anesthetic.
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Aplicación de un promotor inducible por campos electromagnéticos para manipular la expresión de luciferasa como gene reportero

Aplicación de un promotor inducible por campos electromagnéticos para manipular la expresión de luciferasa como gene reportero

Se ha informado sobre los efectos de los campos electromagnéticos (CEM) en los sistemas biológicos y se ha encontrado que éstos pueden afectar actividades metabólicas celulares tales c[r]

130 Lee mas

Transcitosis apical basolateral inducible del receptor de opioides tipo delta en células MDCK

Transcitosis apical basolateral inducible del receptor de opioides tipo delta en células MDCK

A este compartimento entran también proteínas que se endocitan ya sea desde el polo apical como del polo basolateral, que luego son destinadas a una ruta de degradación lisosomal o recic[r]

136 Lee mas

Inhibition of rat carotid body glomus cells TASK like channels by acute hypoxia is enhanced by chronic intermittent hypoxia

Inhibition of rat carotid body glomus cells TASK like channels by acute hypoxia is enhanced by chronic intermittent hypoxia

observed in cell from CIH-treated animals ( − 52.5 ± 1.5 mV; n = 28) with respect to control cells ( − 54.5 ± 1.9; n = 58). However, TASK- like channel conductance appears not to be modified by CIH, since the current amplitude distribution histogram in CIH and control glomus cells remain unchanged, suggesting that CIH has no direct effect on channel conductance. On the other hand, acute hypoxia in control glomus cells increased the inter-event interval to longer durations and reduced the dwell-time for the main open state of the channel, suggesting that acute hypoxia decreases channel activ- ity by destabilizing the open state. Thus, CIH exposure may affect some regulatory factors involved in the hypoxic control of TASK- like channel activity, rather than having a direct effect on channel number or expression.
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8 Lee mas

Hypoxia-induced autophagy as an additional mechanism in human osteosarcoma radioresistance

Hypoxia-induced autophagy as an additional mechanism in human osteosarcoma radioresistance

It is established that hypoxia is common in most solid tumors due to limited blood supply in the microenvironment. This low oxygen condition and cellular adaptive responses often cause tu- mor aggressiveness and resistance to treatments including irra- diation. Osteosarcoma (OS) is commonly known to be radio- resistant. To determine whether radioresistance of this solid tumor could possibly involve hypoxia, we recruited osteosarcoma tissues from 89 cases to stain the typical hypoxia marker, HIF-1 α , by immunohistochemistry, using 28 control samples from os- teochondroma (OC), the most common benign bone tumor.
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Contribution of inflammation on carotid body chemosensory potentiation induced by intermittent hypoxia

Contribution of inflammation on carotid body chemosensory potentiation induced by intermittent hypoxia

et al. 2009 ) , but it is matter of debate if ROS per se may increase the CB chemosensory discharges (Gonzalez et al. 2007 ) . Thus, it is likely that molecules downstream of the ROS signals may mediate the CIH-induced effects of ROS on CB chemoreception. Among other molecules upregulated in the CB by CIH, such as ET-1 and iNOS (Iturriaga et al. 2009 ; Del Rio et al. 2010, 2011 ) , pro-in fl ammatory cytokines has been proposed as mediators of the CB chemosensory potentiation induced by CIH (Iturriaga et al. 2009 ; Del Rio et al. 2011 ) . Recently, we found that CIH induced a ROS-dependent increases of TNF- a and IL-1 b within the CB, suggesting that these pro-in fl ammatory cytokines may mediate the ROS-induced potentiation (Del Rio et al. 2011 ) . To test this hypothesis, we study the effects of the anti-in fl ammatory drug ibuprofen on the increased immunoreactive levels of TNF- a and IL-1 b in the rat CB, and the potentiation of the carotid chemosensory responses to hypoxia.
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Efecto de la infección con el virus del dengue en la modulación de la óxido nítrico sintasa inducible (iNOS) y su asociación con diabetes mellitus

Efecto de la infección con el virus del dengue en la modulación de la óxido nítrico sintasa inducible (iNOS) y su asociación con diabetes mellitus

secretan citocinas antiinflamatorias como IL-10 y TGF-β. Las citocinas estimuladoras de macrófagos M1 son IFN-γ y lipopolisacárido (LPS), mientras que de los macrófagos M2 son IL-4 IL-13. En la obesidad,se ha encontrado un incremento importante de los macrófagos M1 en el tejido adiposo; esto, aunado a una reducción en los niveles de moléculas antiinflamatorias como IL-10 y arginasa, con un consecuente incremento de citocinas proinflmatorias como TNF- α, que ha sido ampliamente descrito como factor proinflamatorio en obesidad, resistencia a la insulina y DM2. El mecanismo por el cual se incrementan los macrófagos M1 no ha sido descrito completamente, pero se ha propuesto que el aumento de peso induce inflamación local y producción de citocinas que promueven el reclutamiento de monocitos proinflamatorios circulantes los cuales en el tejido adiposo se diferencian a macrófagos M1 y su acumulación produce un desbalance entre las poblaciones de M1 y M2 . Los macrófagos M1 inducen un estado inflamatorio y de resitencia a la insulina a través de la inhibición de la señalización de la insulina, producido indirectamente por las citocinas TNF-α e IL-6 122 , mientras que los macrófagos M2 protegen de la resistencia a la insulina a través de un mecanismo dependiente de IL-10; a este respecto se ha descrito que en adipocitos 3T3-L1, la IL-10 bloquea la sobrerregulación de genes proinflamatorios inducidos por TNF-α, incluyendo IL-6 y RANTES (Regulated upon activation, normal T cell expressed and secreted) 123 . Por otro lado, la IL-10 también reduce la expresión de MCP-1 y protege contra la resistencia a la insulina en el hígado 124 .
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Expresión del gen de la hormona del crecimiento (hGH N) bajo el control de un promotor inducible con doxiciclina, en células hipofisiarias

Expresión del gen de la hormona del crecimiento (hGH N) bajo el control de un promotor inducible con doxiciclina, en células hipofisiarias

ubicándolo bajo la regulación del promotor bidireccional modulado por doxiciclina.. Obtención de la unidad transcripcional del gen hGH-N. A)- Esquema de la clona pBShGH- N(pro-) con la u[r]

94 Lee mas

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