Participation of lncRNAs in the development of diabetic complications: Systematic review and meta-analysis. I. Rat.

AIMS: We evaluated the involvement of lncRNAs in the development of pathologies associated with chronic hyperglycaemia in rat models in a model of type 1, type 2 and gestational diabetes. METHODS: Reports were searched in Dialnet, Scielo, HINARI, Springer, ClinicalKey, OTseeker, PubMed and different grey literature databases with any restrictions. Bibliography databases will be searched from their inception to December 2022. RESULTS: Thirty-seven studies met our criteria, and they had the following characteristics: original experimental studies on diabetes, the lncRNAs were extracted or measured from tissues of specific areas and the results were expressed in terms of standard measures by RT-PCR. In most studies, both primary and secondary outcomes were mentioned. On the other hand, we found a total of nine diabetic complications, being retinopathy, nephropathy and neuropathy the most representatives. Additionally, it was found that MALAT1, H19, NEAT1 and TUG1 are the most studied lncRNAs about these complications in rats. On the other hand, the lncRNAs with the highest rate of change were MSTRG.1662 (17.85; 13.78, 21.93), ENSRNOT00000093120_Aox3 (7.13; 5.95, 8.31) and NONRATG013497.2 (-5.55; -7.18, -3.93). CONCLUSIONS: This review found a significant involvement of lncRNAs in the progression of pathologies associated with chronic hyperglycaemia in rat models, and further studies are needed to establish their potential as biomarkers and therapeutic targets for diabetes.

Verbascoside-enriched fraction from Buddleja cordata Kunth ameliorates the effects of diabetic embryopathy in an animal model.

BACKGROUND: The deleterious effects of diabetes mellitus (DM) over development are apparently due to an increase in oxidative stress. Some antioxidants could prevent developmental alterations produced by diabetic state. Extracts of plants of the genus Buddleja are used traditionally for Mexican indigens to ameliorate some diseases. The purpose of this work was to evaluate the effect of the extract of Buddleja cordata over diabetic embryopathy. METHODS: Two experimental approaches were used: an in vivo study and an in vitro model. In the first, rats were treated with streptozotocin, streptozotocin plus methanolic extract of B. cordata, or none. Females were sacrificed at gestational day (GD) 19, and biochemical clinical parameters were measured; also, the fetuses were obtained and morphologically analyzed. In the in vitro model, a verbascoside-enriched fraction (VEF) of the extract was used in whole embryo culture in order to search for the mechanisms for embryoprotection effect over hyperglycemia-induced malformations. RESULTS: In the in vivo experiments, B. cordata extract reduces the frequency and severity of fetal malformations produced by chemically induced diabetes, and additionally partially ameliorates the diabetic condition; in the in vitro model, both severity and frequency of embryo dysmorphogenesis were reduced by the VEF; also, this fraction reduces lipoperoxidation without affecting the activity of the antioxidant enzymes. CONCLUSION: The results suggest that verbascoside of methanolic extract and enriched fraction can directly affect the redox state, and thus, prevents the embryotoxicity mediated by oxidative stress, in embryos of diabetic pregnancy.

Bcl-2 expression in a diabetic embryopathy model in presence of polyamines.

The frequency of congenital malformations is 3-5 times higher in mothers with pregestational diabetes mellitus than in general population. Apparently, this problem is due to change in the expression of apoptotic and antiapoptotic genes induced by the oxidative stress derived from the diabetes/hyperglycemia. One of these genes is Bcl-2, which is associated with the control and inhibition of apoptosis. The purpose of the present work was to study the effect of polyamine addition over expression of Bcl-2 gene in a model of diabetic embryopathy. For this, gestational day 10.5 (GD10.5) rat embryos were incubated at 37°C for 24 h in control medium, medium with high glucose, or medium with high glucose and supplemented with spermidine or spermine. Post-cultured embryos were harvested and observed to obtain morphological scores; some of them were subjected to molecular biology studies: DNA isolation plus conventional PCR or RNA isolation plus RT-PCR; other embryos were fixed with paraformaldehyde and used for immunohistochemical detection of Bcl-2 protein. Although Bcl-2 mRNA was similarly expressed in all rat embryo treatments, Bcl-2 protein was found only in control-incubated embryos. In conclusion, it seems that the inhibition of Bcl-2 gene expression induced by glucose was not reversed by polyamines.

Liver mitochondrial membrane fluidity at early development of diabetes and its correlation with the respiration.

The biological membranes are important in cell function but, during development of diseases such as diabetes, they are impaired. Consequently, membrane-associated biological processes are impaired as well. The mitochondria are important organelles where oxidative phosphorylation takes place, a process closely related with the membranes. In general, it is accepted that the development process of diabetes decreases membrane fluidity. However, in some cases, it has been found to increase membrane fluidity of mitochondria but to decrease the Respiratory Control (RC) index. In this study we found an increase of membrane fluidity and an increase of the RC at an early phase of the development of a type 2 diabetes model. We measured the lipoperoxidation, analyzed the fatty acids composition by gas chromatography, and assessed membrane fluidity using three fluorescent monitors located at different depths inside the bilayer, dipyrenilpropane (DPyP), diphenylhexatriene (DPH), and trimethylammonium diphenylhexatriene (TMA-DPH). Our findings indicate that in the initial stage of diabetes development, when lipoperoxidation still is not significant, the membrane fluidity of liver mitochondria increases because of the increment in the unsaturated to saturated fatty acids ratio (U/S), thus producing an increase of the RC. The membrane fluidity is not the same at all depths in the bilayer. Contrary to the results obtained in mitochondria, the diabetes induced a decrease in the U/S fatty acids ratio of liver total lipids, indicating that the mitochondria might have an independent mechanism for regulating its fatty acids composition.

Biphasic effect of alpha-linolenic acid on glucose-induced dysmorphogenesis and lipoperoxidation in whole rat embryo in culture.

Type 1 diabetes mellitus complicated with pregnancy, know as diabetic embryopathy, is the cause of neonatal malformations and low for gestational age neonates. With the use of the whole-embryo culture system, it has been demonstrated that high glucose causes embryo dysmorphogenesis, and that oxidative stress appears to be the main mechanism. In recent years, beneficial effect of omega-3 fatty acids has been demonstrated in various diabetic models, and in diabetic complications. Since diabetic embryopathy is mediated probably through membrane lipoperoxidation, This study was designed to find if omega-3 fatty acids could ameliorate the effect of high glucose over the dysmorphogenesis of whole rat embryo in culture. Postimplantational rat embryos were cultured in hyperglycemic media, with addition of alpha-linolenic acid, and morphologic and morphometric parameters were registered. Also, lipoperoxidation and fatty acids composition were measured in cultured embryos. Growth of embryos cultured in presence of glucose was very affected, whereas lipoperoxidation was increased, and it was found that Triton X-100 causes similar results than glucose. Addition of low micromolar doses of alpha-linolenic acid overcome the effect of high glucose or Triton X-100, but higher doses does not ameliorates the effects of the carbohydrate or the detergent. Paradoxically, there are not significant changes in fatty acids composition, although the U/S fatty acids ratio shows an increasing tendency by high glucose and a normalizing tendency by omega-3 fatty acids. In conclusion, glucose and Triton X-100 induces in vitro dysmorphogenesis in post-implantational rat embryos associated with increased lipoperoxidation; and this nocive effect could be ameliorated by low micromolar doses of ALA.

Even a Chronic Mild Hyperglycemia Affects Membrane Fluidity and Lipoperoxidation in Placental Mitochondria in Wistar Rats.

It is known the deleterious effects of diabetes on embryos, but the effects of diabetes on placenta and its mitochondria are still not well known. In this work we generated a mild hyperglycemia model in female wistar rats by intraperitoneal injection of streptozotocin in 48 hours-old rats. The sexual maturity onset of the female rats was delayed around 6-7 weeks and at 16 weeks-old they were mated, and sacrificed at day 19th of pregnancy. In placental total tissue and isolated mitochondria, the fatty acids composition was analyzed by gas chromatography, and lipoperoxidation was measured by thiobarbituric acid reactive substances. Membrane fluidity in mitochondria was measured with the excimer forming probe dipyrenylpropane and mitochondrial function was measured with a Clark-type electrode. The results show that even a chronic mild hyperglycemia increases lipoperoxidation and decreases mitochondrial function in placenta. Simultaneously, placental fatty acids metabolism in total tissue is modified but in a different way than in placental mitochondria. Whereas the chronic mild hyperglycemia induced a decrease in unsaturated to saturated fatty acids ratio (U/S) in placental total tissue, the ratio increased in placental mitochondria. The measurements of membrane fluidity showed that fluidity of placenta mitochondrial membranes increased with hyperglycemia, showing consistency with the fatty acids composition through the U/S index. The thermotropic characteristics of mitochondrial membranes were changed, showing lower transition temperature and activation energies. All of these data together demonstrate that even a chronic mild hyperglycemia during pregnancy of early reproductive Wistar rats, generates an increment of lipoperoxidation, an increase of placental mitochondrial membrane fluidity apparently derived from changes in fatty acids composition and consequently, mitochondrial malfunction.

[Antiteratogenic capacity of resveratrol in streptozotocin-induced diabetes in rats]. / Capacidad antiteratogénica del resveratrol en diabetes inducida por estreptozotocina en ratas.

OBJECTIVES: To evaluate the anti-hyperglycemic and anti-teratogenic capacity of resveratrol in streptozotocin-induced diabetic rats. MATERIAL AND METHODS: Experimental study. There were three groups, of five rats each; two groups were treated with a single dose of 50 mg/Kg of streptozotocin (STZ), in citrate buffer, at the 4th day of gestation, and the third group was considered the control, and were administered with citrate buffer only. One of the two STZ induced groups was administered with 100 mg/Kg resveratrol the days 8th to 12th, when neurulation occurs. Fetuses were obtained the 19th gestational day, and were subjected to morphologic analysis; and in fetal liver the activities of scavenging enzymes catalase, superoxide dismutase and gluthathione peroxidase were measured. RESULTS: Resveratrol can decrease the teratogenic effect of STZ-induced diabetes, and scavenging activities were beneficed by the administration of this antioxidant. CONCLUSION: Resveratrol shown embryoprotective properties mediated by amelioration of oxidative stress produced by maternal hyperglycemia.

Capacidad antiteratogénica del resveratrol en diabetes inducida por estreptozotocina en ratas / Antiteratogenic capacity of resveratrol in streptozotocin-induced diabetes in rats

Evaluar la capacidad antihiperglucémica y antiteratogénica del resveratrol en ratas inducidas a diabetes por estreptozotocina. Materiales y métodos. Estudio de tipo experimental. Se tuvieron tres grupos, de cinco ratas Wistar preñadas cada uno, dos de los cuales fueron tratados el cuarto día de gestación con una dosis de estreptozotocina de 50 mg/kg, disuelta en tampón de citratos, y el otro fue considerado como control, y solo se le administró el tampón de citratos. A uno de los grupos inducidos con estreptozotocina se le administró resveratrol a dosis de 100 mg/kg durante los días 8 al 12 de gestación, cuando sucede la neurulación. Los fetos se obtuvieron el día 19 de gestación y se les realizó un análisis morfológico, y en el hígado fetal se determinó la actividad de las enzimas depuradoras de especies reactivas catalasa, superóxido dismutasa y glutatión peroxidasa. Resultados. La administración de resveratrol (DM+R) revierte los parámetros a valores similares a los del grupo control. Las actividades de catalasa y de glutatión peroxidasa, se vieron incrementadas en el grupo tratado con resveratrol con respecto al grupo diabético, en cuanto a la frecuencia de malformaciones en el grupo control y en el grupo tratado con resveratrol no presentaron malformaciones, mientras que en las ratas con diabetes inducida, se encontró una elevada frecuencia de malformaciones. Conclusiones. El resveratrol muestra propiedades antiteratogénicas a través de la disminución del estrés oxidativo que se presenta a causa de la hiperglucemia materna...

To evaluate the anti-hyperglycemic and anti-teratogenic capacity of resveratrol in streptozotocin-induced diabetic rats. Material and methods. Experimental study. There were three groups, of five rats each; two groups were treated with a single dose of 50 mg/Kg of streptozotocin (STZ), in citrate buffer, at the 4th day of gestation, and the third group was considered the control, and were administered with citrate buffer only. One of the two STZ induced groups was administered with 100 mg/Kg resveratrol the days 8th to 12th, when neurulation occurs. Fetuses were obtained the 19th gestational day, and were subjected to morphologic analysis; and in fetal liver the activities of scavenging enzymes catalase, superoxide dismutase and gluthathione peroxidase were measured. Results. Resveratrol can decrease the teratogenic effect of STZ-induced diabetes, and scavenging activities were beneficed by the administration of this antioxidant. Conclusion. Resveratrol shown embryoprotective properties mediated by amelioration of oxidative stress produced by maternal hyperglycemia...

Change in lipoperoxidation but not in scavenging enzymes activity during polyamine embryoprotection in rat embryo cultured in hyperglycemic media.

DM1 complicated with pregnancy is the cause of neonatal malformations and low-for-gestational-age neonates. With the use of the whole-embryo culture system, it has been demonstrated that high glucose causes embryo dysmorphogenesis. Previously, our group has found that spermidine or spermine addition reverts almost fully the severity and frequency of dysmorphogenesis, whereas the effect of arginine and putrescine it is only partial. A hypothesis for polyamine mechanism is the amelioration of oxidative stress caused by high glucose. The purpose of this work was to evaluate the effect of polyamines over the activity of scavenging enzymes and lipoperoxidation in whole-embryo rat in culture. Post-implantation (gestational day 10.5) rat embryos were cultured for 24 h in normal medium or hyperglycemic medium, alone or supplemented with L-arginine or polyamine. Embryos were recovered and visualized, and morphologic parameters were registered. Cultured embryos were homogenized, and superoxide dismutase and glutathione-reductase activities, as well as lipoperoxidation, were measured. The activity of superoxide dismutase and glutathione peroxidase were not affected by the treatment, but lipoperoxidation was increased in embryos cultured in hyperglycemic medium; spermidine or spermine supplementation restore lipoperoxidation to near-normal values, and putrescine and L-arginine reverts only partially the glucose effect. Taken together, these results pointed out that spermidine and spermine embryoprotection could be mediated by direct antioxidant activity. However, further studies are needed to support this hypothesis.

The protective effect of testosterone on streptozotocin-induced apoptosis in beta cells is sex specific.

OBJECTIVES: To investigate the protective role of steroid hormones on streptozotocin (STZ)-induced apoptosis in rat pancreatic beta cells. METHODS: Two sets of experiments were performed. In the first, male rats were orchidectomized and substituted 72 hours later with testosterone, estradiol, or progesterone, and 24 hours later, administered with STZ. Subjects were killed 6 hours later, and apoptosis was determined in sections of the pancreas. In the second experiment, male or female rats were gonadectomized, were further substituted with testosterone, and then administered STZ. Six hours later, the animals were killed, and apoptosis, as well as immunoreactive expression of insulin, catalase, or Cu/Zn superoxide dismutase, was determined in sections of the pancreas. In addition, gonadectomized male or female subjects were substituted with testosterone and administered STZ, and 24 hours later, serum glucose and insulin were measured. RESULTS: It was found that the cytoprotective effect was only shown in testosterone-treated male rats but not progesterone- or estradiol-treated male rats. In addition, the effect was seen in male rats but not in female rats, and there was an inverse correlation between apoptotic index and antioxidant enzyme immunoreactivity. CONCLUSIONS: The cytoprotective effect of testosterone is sex specific and is related to the induction of antioxidant enzyme activities in pancreatic beta cells.

Polyamines protect rat embryo in vitro from high glucose-induced developmental delay and dysmorphogenesis.

BACKGROUND: Pregnancy in mammals with diabetes mellitus results in low birth weight, malformations, and intrauterine death. Parenteral application of natural polyamines or their precursor, L-arginine, to diabetic pregnant rats partially prevents the alterations of development caused by diabetes mellitus. This experiment has been designed to understand if this preventive action also occurs in rat whole embryo in culture. MATERIALS AND METHODS: Rat embryos of gestational day 10 were cultured for 24 h in normal medium, high glucose medium, or high glucose medium supplemented with polyamines or L-arginine, and furthermore embryo growth and development were evaluated. RESULTS: L-arginine and putrescine partially prevents the dysmorphogenic effects of high glucose, whereas spermidine and spermine prevent these effects almost completely. CONCLUSIONS: Polyamines directly protect the embryo from the toxic effect of high glucose concentration on growth and development, although the mechanism remains to be elucidated.

Effect of L-arginine on arginase activity in male accessory sex glands of alloxan-treated rats.

Arginase activity has been identified in the prostate, and may be important in the synthesis of polyamines in accessory sex glands in the male. Polyamines in turn may mediate the action of androgens. Diabetic patients have disordered androgen synthesis. The purpose of this work was to evaluate the effect of L-arginine on arginase activity in accessory sex glands of male rats under normal and diabetic conditions (alloxan 120 mg/kg, i.p.). Normal and diabetic male rats were untreated or were treated with insulin or L-arginine for 96 h, and sacrificed. Arginase activity was measured in serum and in accessory sex glands. Arginase activity in accessory glands did not change significantly with induction of diabetes. Arginase activity was increased in diabetic insulin-treated rats, but there was no arginase response to L-arginine administration in diabetic animals. These findings stand in contrast to beneficial effects of L-arginine previously observed when this amino acid was administered for a long time (at least 10 days). We suspect that altered arginase activity in accessory sex glands may play a role in the reproductive dysfunction caused by diabetes, inasmuch as arginase activity can be increased in experimentally diabetic rats by the administration of insulin.

Slight influence of the estrous cycle stage on the mucosal and systemic specific antibody response induced after vaginal and intraperitoneal immunization with protoxin Cry1Ac from Bacillus thuringiensis in mice.

Since the immune response appears to be variable according to the hormonal stage of the mammalian female, the aim of this study was to determine whether estrous cycle stage modifies the mucosal and systemic immune responses induced by intraperitoneal and vaginal immunization of mice with protoxin Cry1Ac. We tested the influence of three immunizations on the specific antibody response elicited at estrus and diestrus, that were the same estrous cycle stages at which the antigen was applied. Both intraperitoneal and vaginal immunization of mice with Cry1Ac either at estrus or diestrus induces specific antibody responses at serum, vagina and large intestine. The stage of the estrous cycle have little or non influence in the magnitude of the response induced, since only at serum the IgM was slightly higher at estrus than at diestrus by both routes. At the large intestine only the IgA response elicited via the intraperitoneal route changed, being higher at diestrus, whereas at the vagina IgA response induced did not change significantly due to the cycle stage. Present results suggest that Cry1Ac may be used as an antigen carrier as it can elicit antibody responses at systemic level and at several mucosal sites including the vagina that are not modified significantly throughout the reproductive cycle.

Cambios en la actividad de arginasa pancreática en ratas púberes y adultas inducidos por la castración / Activity pancreatic arginase changes inducted by castration in pubescent and adult rats

La actividad de arginasa en riñón e intestino delgado en algunas especies de mamíferos es sensible a la castración, un hallazgo que puede sugerir su dependencia de testosterona. Sin embargo, hasta donde conocemos, la información sobre la regulación de la actividad de arginasa pancreática es escasa. En este trabajo se estudió el efecto de la orquidectomía sobre la actividad de arginasa pancreática en ratas púberes y adultas. Las ratas púberes y adultas, de 21 días y 4 meses de edad, respectivamente, fueron orquidectomizadas y sacrificadas a varios tiempos después de la cirugía. Grupos de ratas intactas sirvieron como controles. La actividad de arginasa y las proteínas fueron determinadas en tejido pancreático. En suero, la actividad de la enzima fue determinada en adición a la glucosa, los triglicéridos y las proteínas totales. En las ratas la actividad de arginasa pancreática y sérica mostró un pico el día 5 poscirugía, mientras que en las adultas, se observó un aumento en la actividad de arginina el día 20. Se observaron cambios en las proteínas séricas y pancreáticas en ratas púberes castradas, pero no en las adultas. Estos resultados sugieren que la arginasa pancreática es dependiente de andrógenos, y que hay diferencia de edades, probablemente debido a patrones distintos de secreción hormonal en ratas adultas y púberes.

La hipoglucemia como factor de riesgo sobre el desarrollo embrionario en diabetes mellitus. Revisión bibliográfica / Hypoglycemia as a risk factor on the embrionary development in diabetes mellitus

La hipoglucemia, o disminución de la concentración normal de glucosa en sangre, es una secuela importante en el control estricto en pacientes diabéticos tipo 1. En mujeres diabéticas embarazadas, la hipoglucemia al parecer no afecta en gran medida el desarrollo embrionario o fetal; mientras que en roedores, es una importante causa de teratogénesis y/o embriotoxicidad, cuando se presenta durante la organogénesis. A pesar de las diferencias metabólicas, endocrinas y temporales de la gestación en humano y roedor, debe considerarse un posible daño al producto por la hipoglucemia durante el embarazo diabético en humanos

Sistemas moleculares en las interacciones celulares I: fecundación en mamíferos / Molecular systems in the cellular interactions I: fecundation in mammals

Las células de los orgnismos multicelulares requieren de diversos mecanismos de reconocimiento entre si, a fin de funcionar de manera integral. En el presente trabajo se revisan los sistemas moleculares de reconocimiento y adhesión celulares en general, y en particular aquellos que participan en la fecundación del óvulo en los mamíferos. Varias proteínas de la superficie del espermatozoide con actividad catalítica (galactosiltransferasa, proteasas y glicosidasas) o de lectinas, reconocen y unen de manera específica a componentes glicoprotéicos de la zona pelúcida del óvulo, como un requisito previo a la fecundación.

Mecanismo de acción de hormonas esteroides I: estrógenos / Mechanism of action of steroid hormones I: estrogens

Las hormonas esteroides son moléculares muy versátiles: a pesar de estar relacionada entre sí por su estructura química, cumplen funciones muy diversas, e incluso antagónicas. Su mecanismo de acción no esta aclarado totalmente. Los estrógenos participan en la regulación de prácticamente todos los eventos reproductivos y sexuales de la hembra, sin embargo, las acciones intracelulares por las que se llevan a cabo no se conocen adecuadamente las interrogantes. Se acepta en la actualidad la existencia de un receptor citoplásmico y/o nuclear, sin aclarar satisfactoriamente cómo las hormonas llegan hasta el núcleo. Los eventos endócrinos que se expresan rápidamente (segundos) se deben a una posible interacción con la membrana celular. El propósito de esta revisión es analizar y conciliar los datos reportados sobre el mecanismo de acción de estrogenos.