Astragaloside IV plays a neuroprotective role by promoting PPARγ in cerebral ischemia-reperfusion rats.

BACKGROUND: Cerebral ischemia-reperfusion injury (CIRI) usually occurs during the treatment phase of ischemic disease, which is closely related to high morbidity and mortality. Promoting neurogenesis and synaptic plasticity are effective neural recovery strategies for CIRI. Astragaloside IV (AS-IV) has been shown to play a neuroprotective role in some neurological diseases. In the current study, we evaluated the effect and possible mechanism of AS-IV in CIRI rats. METHODS: The middle cerebral artery occlusion reperfusion (MCAO/R) model was established in rats to simulate the occurrence of human CIRI. First, we determined the cerebral injury on the 1st, 3rd, 5th and 7th day after cerebral ischemia-reperfusion (I/R) surgery by neurological deficit detection, TTC staining, TUNEL staining and Western blot analysis. Furthermore, rats were pre administered with AS-IV and then subjected to cerebral I/R surgery. Brains were collected on the 3rd day to evaluate the neuroprotective effect of AS-IV. RESULTS: Our results showed that on the 3rd day after I/R, the neurological impairment score and infarct volume were highest, the levels of apoptosis and expression of Caspase3 and Bax reached the peak. AS-IV treatment apparently attenuated neurological dysfunction, reduced infarct volume and pathological damage, promoted the neurogenesis, and alleviated the pathological damage caused by cerebral I/R involved in thickening and blurring of synaptic membranes, reduction of microtubules and synaptic vesicles, and loss of synaptic cleft. Our study also showed that AS-IV promoted the transcription and expression of the peroxisome proliferators-activated receptors γ (PPARγ) and brain-derived neurotrophic factor (BDNF), increased the expression of phosphorylation of tyrosine kinase receptor B (TrkB) and downstream PI3K/Akt/mTOR pathway proteins. Notably, when GW9662, an inhibitor of PPARγ was administered with AS-IV, the neuroprotective effect of AS-IV was reduced. CONCLUSIONS: These findings suggested that AS-IV has neuroprotective function in CIRI rats, and its molecular mechanism may depend on the phosphatidylinositide 3-kinase (PI3K)/protein kinase B (PKB)/Akt signalling pathway activated by PPARγ. AS-IV could be an effective therapeutic drug candidate for CIRI treatment.

Astragaloside IV mediates radiation-induced neuronal damage through activation of BDNF-TrkB signaling.

BACKGROUND: Electromagnetic radiation is relevant to human life, and radiation can trigger neurodegenerative diseases by altering the function of the central nervous system through oxidative stress, mitochondrial dysfunction, and protein degradation. Astragaloside IV (AS-IV) is anti-oxidative, anti-apoptotic, activates the BDNF-TrkB pathway and enhances synaptic plasticity in radiated mice, which can exert its neuroprotection. However, the exact molecular mechanisms are still unclear. PURPOSE: This study investigated whether AS-IV could play a neuroprotective role by regulating BDNF-TrkB pathway in radiation damage and its underlying molecular mechanisms. METHODS: Transgenic mice (Thy1-YFP line H) were injected with AS-IV (40 mg/kg/day body weight) by intraperitoneal injection daily for 4 weeks, followed by X-rays. PC12 cells and primary cortical neurons were also exposed to UVA after 24 h of AS-IV treatment (25 µg/ml and 50 µg/ml) in vitro. The impact of radiation on learning and cognitive functions was visualized in the Morris water maze assay. Subsequently, Immunofluorescence and Golgi-Cox staining analyses were utilized to investigate the structural damage of neuronal dendrites and the density of dendritic spines. Transmission electron microscopy was performed to examine how the radiation affected the ultrastructure of neurons. Finally, western blotting analysis and Quantitative RT-PCR were used to evaluate the expression levels and locations of proteins in vitro and in vivo. RESULTS: Radiation induced BDNF-TrkB signaling dysregulation and decreased the levels of neuron-related functional genes (Ngf, Bdnf, Gap-43, Ras, Psd-95, Arc, Creb, c-Fos), PSD-95 and F-actin, which subsequently led to damage of neuronal ultrastructure and dendrites, loss of dendritic spines, and decreased dendritic complexity index, contributing to spatial learning and memory deficits. These abnormalities were prevented by AS-IV treatment. In addition, TrkB receptor antagonists antagonized these neuroprotective actions of AS-IV. 7,8-dihydroxyflavone and AS-IV had neuroprotective effects after radiation. CONCLUSION: AS-IV inhibits morphological damage of neurons and cognitive dysfunction in mice after radiation exposure, resulting in a neuroprotective effect, which were mediated by activating the BDNF-TrkB pathway.

Suppression of NLRP3 inflammasome activation by astragaloside IV via promotion of mitophagy to ameliorate radiation-induced renal injury in mice.

Background: Irradiation (IR) promotes inflammation and apoptosis by inducing oxidative stress and/or mitochondrial dysfunction (MD). The kidneys are rich in mitochondria, and mitophagy maintains normal renal function by eliminating damaged mitochondria and minimizing oxidative stress. However, whether astragaloside IV (AS-IV) can play a protective role through the mitophagy pathway is not known. Methods: We constructed a radiation injury model using hematoxylin and eosin (HE) staining, blood biochemical analysis, immunohistochemistry, TdT-mediated dUTP nick end labeling (TUNEL) staining, ultrastructural observation, and Western blot analysis to elucidate the AS-IV resistance mechanism for IR-induced renal injury. Results: IR induced mitochondrial damage; the increase of creatinine (SCr), blood urea nitrogen (BUN) and uric acid (UA); and the activation of NOD-like receptor thermal protein domain-associated protein 3 (NLRP3) inflammasome and apoptosis in renal tissue. AS-IV administration attenuated the IR-induced MD and reactive oxygen species (ROS) levels in the kidney; enhanced the levels of mitophagy-associated protein [PTEN-induced putative kinase 1 (PINK1)], parkin proteins, and microtubule-associated protein 1 light 3 (LC3) II/I ratio in renal tissues; diminished NLRP3 inflammasome activation-mediated proteins [cleaved cysteinyl aspartate-specific proteinase-1 (caspase-1), interleukin-1ß (IL-1ß)] and apoptosis-related proteins [cleaved caspase-9, cleaved caspase-3, BCL2-associated X (Bax)]; reduced SCr, BUN, and UA levels; and attenuated the histopathological alterations in renal tissue. Conversely, mitophagy inhibitor cyclosporin A (CsA) suppressed the AS-IV-mediated protection of renal tissue. Conclusions: AS-IV can strongly diminish the activation and apoptosis of NLRP3 inflammasome, thus attenuating the renal injury induced by radiation by promoting the PINK1/parkin-mediated mitophagy. These findings suggest that AS-IV is a promising drug for treating IR-induced kidney injury.

Astragaloside IV ameliorates radiation-induced nerve cell damage by activating the BDNF/TrkB signaling pathway.

Radiation can induce nerve cell damage. Synapse connectivity and functionality are thought to be the essential foundation of all cognitive functions. Therefore, treating and preventing damage to synaptic structure and function is an urgent challenge. Astragaloside IV (AS-IV) is a glycoside extracted from Astragalus membranaceus (Fisch.). Bunge is a widely used traditional Chinese medicine in China with various pharmacological properties, including protective effects on the central nervous system (CNS). In this study, the effect of AS-IV on synapse damage and BDNF/TrkB signaling pathway in radiated C57BL/6 mice with X-rays was investigated. PC12 cells and primary cortical neurons were exposed to UVA in vitro. Open field test and rotarod test were used to observe the effects of AS-IV on the motor and explore the abilities of radiated mice. The pathological changes in the brain were observed by hematoxylin and eosin and Nissl staining. Immunofluorescence analysis was used to detect the synapse damage. The expressions of the BDNF/TrkB pathway and neuroprotection-related molecules were detected by Western blotting and Quantitative-RTPCR, respectively. The results showed that AS-IV could improve the motor and explore abilities of radiated mice, reduce pathological damage to the cortex, enhance neuroprotection functions, and activate BDNF/TrkB pathway. In conclusion, AS-IV could relieve radiation-induced synapse damage, at least partly through the BDNF/TrkB pathway.

The Protective Effects of Peroxisome Proliferator-Activated Receptor Gamma in Cerebral Ischemia-Reperfusion Injury.

Cerebral ischemia-reperfusion injury (CI/RI) is a complex pathological process that often occurs secondary to trauma, surgery, and shock. Peroxisome proliferator activated receptor gamma (PPARγ) is a subunit of the PPAR and is a ligand-activated nuclear transcription factor. After being activated by its ligand, PPARγ can combine with specific DNA response elements to regulate the transcription and expression of genes. It has a wide range of biological functions, such as regulating lipid metabolism, improving insulin sensitivity, modulating anti-tumor mechanisms, and inhibiting inflammation. In recent years, some studies have shown that PPARγ exerts a protective effect during CI/RI. This article aims to summarize the research progress of studies that have investigated the protective effects of PPARγ in CI/RI and the cellular and molecular mechanisms through which these effects are modulated, including inhibition of excitatory amino acid toxicity, reduced Ca2+ overload, anti-oxidative stress, anti-inflammation, inhibition of microglial activation, maintain the BBB, promotion of angiogenesis, and neurogenesis and anti-apoptotic processes.

Preliminary study on the anti-apoptotic mechanism of Astragaloside IV on radiation-induced brain cells.

With multiple targets and low cytotoxicity, natural medicines can be used as potential neuroprotective agents. The increase in oxidative stress levels and inflammatory responses in the brain caused by radiation affects cognitive function and neuronal structure, and ultimately leads to abnormal changes in neurogenesis, differentiation, and apoptosis. Astragaloside Ⅳ (AS-Ⅳ), one of the main active constituents of astragalus, is known for its antioxidant, antihypertensive, antidiabetic, anti-infarction, anti-inflammatory, anti-apoptotic and wound healing, angiogenesis, and other protective effects. In this study, the mechanism of AS-IV against radiation-induced apoptosis of brain cells in vitro and in vivo was explored by radiation modeling, which provided a theoretical basis for the development of anti-radiation Chinese herbal active molecules and brain health products. In order to study the protective mechanism of AS-IV on radiation-induced brain cell apoptosis in mice, the paper constructed a radiation-induced brain cell apoptosis model, using TUNEL staining, flow cytometry, Western blotting to analyze AS-IV resistance mechanism to radiation-induced brain cell apoptosis. The results of TUNEL staining and flow cytometry showed that the apoptosis rate of radiation group was significantly increased. The results of Western blotting indicated that the expression levels of p-JNK, p-p38, p53, Caspase-9 and Caspase-3 protein, and the ratio of Bax to Bcl-2 in radiation group were significantly increased. There was no significant difference in the expression levels of JNK and p38. After AS-IV treatment, the apoptosis was reduced and the expression of apoptosis related proteins was changed. These data suggested that AS-IV can effectively reduce radiation-induced apoptosis of brain cells, and its mechanism may be related to the phosphorylation regulation of JNK-p38.

Astragaloside IV ameliorates radiation-induced senescence via antioxidative mechanism.

OBJECTIVES: Ageing is a universal and gradual process of organ deterioration. Radiation induces oxidative stress in cells, which leads to genetic damage and affects cell growth, differentiation and senescence. Astragaloside (AS)-IV has antioxidative, anti-apoptotic and anti-inflammatory properties. METHODS: To study the protective mechanism of AS-IV on radiation-induced brain cell senescence, we constructed a radiation-induced brain cell ageing model, using biochemical indicators, senescence-associated galactosidase (SA-ß-gal) senescence staining, flow cytometry and Western blotting to analyse the AS-IV resistance mechanism to radiation-induced brain cell senescence. KEY FINDINGS: Radiation reduced superoxide dismutase (SOD) activity and expressions of cyclin-dependent kinase (CDK2), CDK4, cyclin E and transcription factor E2F1 proteins, and increased expressions of p21, p16, cyclin D and retinoblastoma (RB) proteins, malondialdehyde (MDA) activity, SA-ß-gal-positive cells and cells stagnating in G1 phase. After treatment with AS-IV, the level of oxidative stress in cells significantly decreased and expression of proteins related to the cell cycle and ageing significantly changed. In addition, SA-ß-gal-positive cells and cells arrested in G1 phase were significantly reduced. CONCLUSIONS: These data suggest that AS-IV can antagonize radiation-induced brain cells senescence; and its mechanism may be related to p53-p21 and p16-RB signalling pathways of ageing regulation.

Histomorphometric adaptation of yak (Bos grunniens) abomasum to the Qinghai-Tibetan plateau environment / Adaptación histomorfométrica del abomaso de yak (Yos grunniens) al medio ambiente de la meseta tibetana Qinghai

Six abomasums of yaks (Bosgrunniens) were studied with gross dissection and histological methods. It was found that the mucosa of the yak abomasum was covered with simple columnar epithelium. There were lots of spiral folds (10) in the fundic glandular area. The developed membrane of lamina propria was occupied by high density glands. According to the morphological characteristics of the glands, the abomasum was divided into the cardiac, gastric and pyloric glands. Cardiac glands were curved tubular glands with the intumescent bottom and small glandular cavity. Fundic glands were simple tubular glands or branched tubular glands, where the chief, parietal and mucous neck cells can be observed clearly. Pyloric glands were curled tubular glands, the closer to the deep of the lamina propria, the more obvious the glands curl. Staining of glycoconjugate revealed that the mucosal epithelium of the cardiac gastric and pyloric glands and gastric pits epithelium mainly secreted neutral glycoconjugate, but other portions of cardiac and gastric glands secreted mixed and acid glycoconjugate respectively. By Gordon-Sweet's reticular fiber staining, it was found that the mucous neck cells possessed the characteristic of argyrophilic phenomenon. There was a large number of argyrophilic granules in the supranuclear cytoplasm in contrast with the chief cells. Furthermore, there were isolated lymphoid nodules and diffuse lymphoid tissue in the abomasum glands, especially in corpus abomasi. Grimelius silver staining showed that the argyrophil cells were located in the glandular epithelium and lamina propria of glands, which can also be observed in connective tissue. These endocrine cells dispersed individually in epithelial cells, occasionally in 3­5 cell groups. Therefore, the yaks were grazed throughout the year on diverse natural grassland and had evolved morphological characteristics of the abomasum enabling them to consume a wide variety of plant species, thereby better adapting them to harsh plateau environment.

Seis abomaso yak (Bosgrunniens) fueron estudiados con disección y métodos histológicos. Se encontró que la mucosa del abomaso yak estaba cubierta de epitelio columnar simple. Se observaron pliegues en espiral (10) en la zona glandular fúndica. La membrana desarrollada de la lámina propia contenía glándulas de alta densidad. De acuerdo con las características morfológicas de las glándulas, el abomaso se dividió en las glándulas cardíacas, gástricas y pilórica. Las glándulas cardíacas se curvan en glándulas tubulares con la parte inferior intumescente y una pequeña cavidad glandular. Las glándulas fúndicas eran glándulas tubulares simples o glándulas tubulares ramificadas, donde se pueden observar con claridad las células principales, parietales y mucosas del cuello. Las glándulas pilóricas fueron glándulas tubulares curvadas, cuanto más cercanas a la lámina propia, más evidente fue su forma ondulada. La tinción glucoconjugada reveló que el epitelio de la mucosa de las glándulas gástricas cardiacas, pilóricas y el epitelio de las fosas gástricas secretaron principalmente un glucoconjugado neutro, pero otras porciones cárdicas y de las glándulas gástricas secretaron un glucoconjugado mixto y ácido, respectivamente. A la tinción de fibras reticulares, se encontró que las células mucosas del cuello poseían características argirófilas. Se observó un gran número de gránulos en el citoplasma supranuclear en contraste con las células principales. Además, no fueron aislados los nódulos linfoides y presentaba tejido linfoide difuso en las glándulas de abomaso, especialmente en el cuerpo del abomaso. La tinción Gordon Sweet indicó que las células argirofílicas se localizaron en el epitelio y lámina propia glandular, lo que también se observó en el tejido conectivo. Estas células endocrinas se dispersan individualmente en las células epiteliales, de vez en cuando en grupos celulares de 3-5. De esta forma, los yak pastorean durante todo el año, en diversos pastizales naturales, y han evolucionado sus características morfológicas que les permiten consumir una amplia variedad de especies de plantas, con lo que se adaptan mejor a las condiciones inhóspitas.

Histomorphometric characterization of forestomach of yak (Bos grunniens) in the Qinghai-Tibetan Plateau / Caracterización histomorfométrica del preestómago del yak (Bos grunniens) en la Meseta Qinghai-Tíbet

Six forestomachs of yaks (Bos grunniens) were studied with gross dissection and histological methods. It was found that the forestomach of yak consisted of the following three parts, rumen, reticulum and omasum, which were composed of the mucosa, submucosa, muscularis and serosa. In addition, the mucosal epithelium was covered with stratified squamous epithelium, with part of keratinized the shallow cells. Rumen, the mucosa of which formed ligulate papillae varying in size and shape, was no muscularis mucosa. Reticulum, consisted of a surface epithelium that invaginated to various extent into the lamina propria, formed various folds in shape, namely, grid-like small rooms. Furthermore, there are many secondary folds densely covered with keratinized papillae. The most striking feature of the omasum was to be formed the laminae omasi varying in length, with short and rough papillae distributing on both sides. Taken together, there was no glands within the mucosa and lamina propria of forestomach of yak, where diffuse lymphoid tissues can be observed clearly. It is, therefore, believed that the yak forestomach may have evolved those specific structural characteristics in response to the unique living environment and dietary habits impose on the Qinghai-Tibetan Plateau.

Seis preestómagos de yaks (Bos grunniens) fueron estudiados mediante disección macroscópica y métodos histológicos. Se encontró que el preestómago del yak constaba de tres partes: rumen, retículo y omaso, compuestas de mucosa, submucosa, muscular y serosa. Además, el epitelio de la mucosa se conformó con epitelio escamoso estratificado, con parte de células cornificadas superficiales. En el rumen, la mucosa formó papilas linguladas que variaron en tamaño y forma. El retículo, consistió en una superficie epitelial que se invaginó en distinta medida en la lámina propia, conformando varias formas de pliegues, es decir, cuadrículas como pequeños cubículos. Además, existían muchos pliegues secundarios densamente cubiertos con papilas cornificadas. La característica más llamativa del omaso, fue formar láminas que variaron en longitud, con papilas cortas y ásperas distribuidas en ambos lados. Tomados en conjunto, no hubo glándulas dentro de la mucosa y la lámina propia del preestómago del yak, donde los tejidos linfoides difusos se pueden observar claramente. Por lo tanto, creemos que esas características estructurales específicas del preestómago del yak pudieron haber evolucionado en respuesta a las condiciones de vida únicas y hábitos dietéticos que se presentan en la meseta de Qinghai-Tíbet.