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1.
Gene ; 766: 145128, 2021 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-32911026

RESUMEN

BACKGROUND: The pathogenesis of osteonecrosis of the femoral head (ONFH) is unclear. Our previous study demonstrated that upregulated miR-335 in bone microvascular endothelial cells (BMECs) might be associated with the disease of steroid-induced ONFH. Here, we study the preventive effect of ICA on steroid-induced ONFH in rats. METHOD: 90 rats were separated into three groups: control group, methylprednisolone (MPS) group, and MPS + Icariin (ICA) group. Four weeks later, histological analyses were performed. Thrombomodulin (TM) and vascular endothelial growth factor (VEGF) were tested. MiRNA-335 expression was screened in the three groups using Agilent Gene Spring GX software. Target genes of miRNA-335 were detected by bioinformatics analysis. The functions of BMECs were analyzed by scratch, angiogenesis and cell survival rate. RESULTS: ICA can prevent the occurrence of steroid-associated ONFH in rats and reduce the amount of TM and VEGF in serum induced by glucocorticoids. ICA could regulate the overexpression of miRNA-335 induced by glucocorticoids. We predicted the Gene ontology (GO) and signaling pathways of target genes. At 24 hours, we found that ICA significantly promoted BMECs migration abilities. We also found that ICA could promote the angioplasty ability of BMECs. ICA could improve the survival rate of BMECs after steroid-induced injury. CONCLUSIONS: ICA is effective to prevent the occurrence of steroidinduced ONFH. ICA has a protective effect against steroid-induced BMECs injury. ICA regulated the imbalance of miRNA-335 expression induced by the glucocorticoid in BMECs, which provides a new viewpoint to explore the mechanism of ICA in preventing steroid-induced ONFH.


Asunto(s)
Células Endoteliales/efectos de los fármacos , Necrosis de la Cabeza Femoral/tratamiento farmacológico , Cabeza Femoral/efectos de los fármacos , Flavonoides/farmacología , MicroARNs/metabolismo , Neovascularización Patológica/tratamiento farmacológico , Sustancias Protectoras/farmacología , Adipogénesis/efectos de los fármacos , Animales , Movimiento Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Endoteliales/metabolismo , Femenino , Cabeza Femoral/metabolismo , Necrosis de la Cabeza Femoral/inducido químicamente , Necrosis de la Cabeza Femoral/metabolismo , Glucocorticoides/metabolismo , Metilprednisolona/farmacología , Neovascularización Patológica/metabolismo , Osteocitos/efectos de los fármacos , Osteogénesis/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacos , Esteroides/farmacología , Factor A de Crecimiento Endotelial Vascular/metabolismo
2.
Chemosphere ; 262: 127792, 2021 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-32805656

RESUMEN

Tebuconazole is a triazole fungicide, used in agriculture to treat phytopathogenic fungi, and as a biocide, has been reported to be related to reproductive and developmental toxicity. The purpose of this study was to investigate the effect of tebuconazole exposure on rat fetal Leydig cells and fetal testis during pregnancy. Pregnant Sprague-Dawley rats were randomly divided into 4 groups, daily gavaged with corn oil (as a control), 25, 50, and 100 mg/kg body weight tebuconazole for 10 days (from the 12th day of pregnancy). Tebuconazole increased fetal serum testosterone and progesterone levels at a dose of 100 mg/kg. Exposure to 100 mg/kg tebuconazole significantly caused an increase in the number of fetal Leydig cells per testis without inducing cell aggregation. Tebuconazole up-regulated the expression of Star, Cyp11a1, Hsd17b3, and Fshr and their proteins. Further investigation found that tebuconazole caused increased phosphorylation of AKT1, ERK1/2, and mTOR, the level of BCL2, as well as the decrease of Beclin1, LC3B, and BAX, which may contribute to the fetal Leydig cell autophagy and proliferation. In conclusion, in utero exposure of tebuconazole causes the proliferation of fetal Leydig cells.


Asunto(s)
Fungicidas Industriales/toxicidad , Células Intersticiales del Testículo/efectos de los fármacos , Efectos Tardíos de la Exposición Prenatal/inducido químicamente , Reproducción/efectos de los fármacos , Triazoles/toxicidad , Animales , Femenino , Células Intersticiales del Testículo/metabolismo , Masculino , Fosfoproteínas/genética , Fosforilación , Embarazo , Efectos Tardíos de la Exposición Prenatal/patología , Distribución Aleatoria , Ratas , Ratas Sprague-Dawley , Testículo/efectos de los fármacos , Testículo/embriología , Testículo/patología , Testosterona/sangre , Regulación hacia Arriba
3.
Food Chem ; 338: 127826, 2021 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-32810815

RESUMEN

This study aimed to evaluate the mutagenicity and oral acute toxicity of winter mushroom powder (PW) treated by atmospheric non-thermal plasma (ANP). Winter mushroom powder without plasma treatment (CW) containing an equivalent amount of sodium nitrite as PW was used as a control. The Ames test revealed that the number of revertant colonies did not significantly increase compared to that in the control. Acute toxicity was assessed in rats that were fed a single dose of winter mushroom powder (5000 mg/kg body weight). Results of the acute toxicity test revealed no remarkable clinical symptoms in any of the rats. No significant difference was observed in of the serum biochemical parameters between the treatments. Regardless of the ANP treatment, mild histological changes were observed in few rats in all groups. Therefore, it is concluded that ANP treatment did not cause any mutagenicity or acute toxicity in the winter mushroom.


Asunto(s)
Flammulina/química , Industria de Procesamiento de Alimentos/métodos , Polvos/toxicidad , Administración Oral , Animales , Masculino , Pruebas de Mutagenicidad , Gases em Plasma , Polvos/administración & dosificación , Polvos/química , Ratas Sprague-Dawley , Pruebas de Toxicidad Aguda
4.
Gene ; 766: 145154, 2021 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-32949699

RESUMEN

CircRNA serves a crucial role in the development of heart failure (HF). Nevertheless, the regulatory mechanisms of circ_0062389 in HF are unknown. This study aims to examine the effect and mechanism of circ_0062389 on cardiomyocyte apoptosis in HF rats and H9C2 cells. Rats were divided into 5 groups (n = 8/group): the Control group, Sham group, HF group, HF + si-NC group, and HF + si-circRNA group. The echocardiography was used to examine the cardiac function, including LVIDd, LVIDs, IVSd, and IVSs. The apoptosis of myocardial tissue was detected through TUNEL method. H9C2 cells were randomly assigned into Control group (untransfected H9C2 cells), H/R group (untransfected H/R H9C2 cells), H/R + si-NC group (transfected si-NC) and H/R + si-circRNA group (transfect si-circ_0062389). Cell apoptosis was assessed through flow cytometry. The expression of circ_0062389 in myocardial tissues of HF rats was significantly higher than that of Control group and Sham group. Silencing circ_0062389 significantly reduced the levels of LVIDd, LVIDs, IVSd, and IVSs. Additionally, silencing circ_0062389 could significantly reduce the apoptosis rate of rat cardiomyocytes. Besides, silencing circ_0062389 significantly reduced the expression of TGF-ß1 and Smad3 protein. Silencing circ_0062389 could alleviate cardiomyocyte apoptosis in HF rats via modulating TGF-ß1/Smad3 signaling pathway, which might be a promising target for the treatment of HF.


Asunto(s)
Apoptosis/genética , Insuficiencia Cardíaca/genética , Miocitos Cardíacos/patología , Interferencia de ARN/fisiología , ARN Circular/genética , Transducción de Señal/genética , Proteína smad3/genética , Factor de Crecimiento Transformador beta1/genética , Animales , Línea Celular , Corazón/fisiología , Insuficiencia Cardíaca/patología , Masculino , Miocardio/patología , Ratas , Ratas Sprague-Dawley
5.
Gene ; 766: 145153, 2021 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-32950633

RESUMEN

AIM: Acute lung injury (ALI) is the mild form of acute respiratory distress syndrome (ARDS) which is a common lung disease with a high incidence and mortality rate. Recent studies manifested that some circular RNAs were associated with ALI. In this study, we aimed to uncover the effect of circular RNA circ_0054633 on ALI initiation and progression and proposed a new mechanism related to ALI. METHODS: The lipopolysaccharides (LPS)-induced acute lung injury model were build both in vivo of rat and in vitro of primary murine pulmonary microvascular endothelial cells (MPVECs). Hematoxylin and eosin (H&E) was employed to observe the tissue morphology and estimate the degree of lung damage. We used real-time quantitative polymerase chain reaction (RT-qPCR) to measure the expression level of circ_0054633. The expression levels of inflammatory cytokines IL-17A and tumor necrosis factor-α (TNF-α) were detected by ELISA. The effects of circ_0054633 on MPVECs proliferation and apoptosis were detected with the help of CCK-8 and apoptosis assay, separately. The expression level of NF-κB p65 protein was measured by Western blot. RESULTS: circ_0054633, IL-17A, TNF-α and NF-κB p65 were all overexpressed in LPS-treated rat and MPVECs, and LPS enhanced the proliferation and apoptosis of MPVECs. While circ_0054633 silencing reversed the above promotion effects of LPS on IL-17A, TNF-α expression and MPVECs proliferation and apoptosis. CONCLUSIONS: Quietness of circ_0054633 alleviated LPS-induced ALI via NF-κB signaling pathway, implicating circ_0054633 may be a potential biomarker for diagnose and therapy of ALI.


Asunto(s)
Lesión Pulmonar Aguda/metabolismo , Proliferación Celular/fisiología , Células Endoteliales/metabolismo , Inflamación/metabolismo , FN-kappa B/metabolismo , ARN Circular/metabolismo , Lesión Pulmonar Aguda/inducido químicamente , Animales , Apoptosis/efectos de los fármacos , Apoptosis/fisiología , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Inflamación/inducido químicamente , Interleucina-17/metabolismo , Lipopolisacáridos/farmacología , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Masculino , Ratones , Ratas , Ratas Sprague-Dawley , Factor de Transcripción ReIA/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo
6.
Sci Total Environ ; 750: 141404, 2021 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-33182165

RESUMEN

The toxic effect of high-dose of short-chain chlorinated paraffins (SCCPs) has been extensively studied, however the possible health risks induced by SCCPs at low-dose remain largely unknown. In this study, a comprehensive toxicology analysis of SCCPs was conducted with the exposure levels from the environmental dose to the Lowest Observed Adverse Effect Level (LOAEL) of 100 mg/kg/day. General toxicology analysis revealed inconspicuous toxicity of the environmental dose of SCCPs, high dose SCCP exposure inhibited the growth rate and increased the liver weight of rat. Metabolomics analysis indicated that SCCP-induced toxicity was triggered at environmentally relevant doses. First, inhibition of energy metabolism was observed with the decrease in blood glucose and the dysfunction of TCA cycle, which may have contributed to lower body weight gain in rats exposed to a high dose of SCCPs. Second, the increase of free fatty acids indicated the acceleration of lipid metabolism to compensate for the energy deficiency caused by hypoglycemia. Lipid oxidative metabolism inevitably leads to oxidative stress and stimulates the up-regulation of antioxidant metabolites such as GSH and GSSH. The up-regulation of polyunsaturated fatty acids (PUFAs) and phospholipids composed of arachidonic acid indicates the occurrence of inflammation. Dysfunction of lipid metabolism can be an indicator of SCCP-induced liver injury.


Asunto(s)
Hidrocarburos Clorados , Parafina , Animales , China , Monitoreo del Ambiente , Hidrocarburos Clorados/análisis , Hidrocarburos Clorados/toxicidad , Metabolismo de los Lípidos , Masculino , Metabolómica , Parafina/análisis , Parafina/toxicidad , Ratas , Ratas Sprague-Dawley
7.
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue ; 32(10): 1189-1193, 2020 Oct.
Artículo en Chino | MEDLINE | ID: mdl-33198861

RESUMEN

OBJECTIVE: To explore the mechanism of resveratrol on ameliorating the cognitive dysfunction induced by sepsis associated encephalopathy (SAE) in rats. METHODS: The 12 weeks old male Sprague-dawley (SD) male rats were randomly divided into sham group, sepsis group and resveratrol group, with 30 rats in each group. The rat model of sepsis was made by injecting LPS (10 mg/kg) into tail vein. The rats in sham group was given the same amount of normal saline (NS). After LPS injection, resveratrol (8 mg×kg-1×d-1) was intraperitoneally injected once daily for 2 days in the resveratrol group; the same amount of NS was given to the sepsis group and sham group. At 24 hours after model establishment, the cognitive function of the experimental rats was assessed by the Morris water maze test. The blood-brain barrier (BBB) permeability was evaluated by the brain water content (BWC) and Evans blue (EB) test. The protein expressions of matrix metalloproteinase 9 (MMP-9), Occludin and Claudin-5 in cortical tissue were detected by Western Blot. Double immunofluorescence was used to verify the co-localization of MMP-9 protein and the marker protein of astrocyte GFAP in the cortical tissue of rats. RESULTS: Compared with the sham group, the escape latency in the sepsis group was significantly longer [48-hour escape latency (s): 56.56±6.43 vs. 36.62±3.32, 72-hour escape latency (s): 57.72±7.23 vs. 26.46±4.24, both P < 0.01], the BWC and extravasation of EB were increased [BWC: (84.56±2.03)% vs. (76.82±2.22)%, EB (µg/g): 17.56±2.28 vs. 6.25±1.36, both P < 0.01], the expression of MMP-9 protein was increased (MMP-9/ß-actin: 0.73±0.01 vs. 0.24±0.01, P < 0.01), the protein expressions of Occludin and Claudin-5 were decreased (Occludin/ß-actin: 0.45±0.02 vs. 0.86±0.04, Claudin-5/ß-actin: 0.62±0.03 vs. 0.96±0.05, both P < 0.01). At the same time, the co-localization expression of MMP-9 protein and the astrocytes of the cortical were increased [MMP-9 fluorescence intensity (AU): 38.66±4.26 vs. 17.23±3.04, MMP-9 positive cells: (26.92±1.77)% vs. (12.82±1.46)%, both P < 0.01]. Compared with the sepsis group, the escape latency in resveratrol group was significantly shorter [48-hour escape latency (s): 41.42±6.27 vs. 56.56±6.43, 72-hour escape latency (s): 33.46±7.17 vs. 57.72±7.23, both P < 0.01], the BWC and extravasation of EB were decreased [BWC: (77.15±2.27)% vs. (84.56±2.03)%, EB (µg/g): 7.74±1.88 vs. 17.56±2.28, both P < 0.01], the expression of MMP-9 protein was decreased (MMP-9/ß-actin: 0.25±0.01 vs. 0.73±0.01, P < 0.01), the protein expressions of Occludin and Claudin-5 were increased (Occludin/ß-actin: 0.82±0.03 vs. 0.45±0.02, Claudin-5/ß-actin: 0.92±0.04 vs. 0.62±0.03, both P < 0.01). At the same time, the co-localization expression of MMP-9 protein and the astrocytes of the cortical were decreased [MMP-9 fluorescence intensity (AU): 19.44±4.37 vs. 38.66±4.26, MMP-9 positive cells: (13.11±1.29)% vs. (26.92±1.77)%, both P < 0.01]. CONCLUSIONS: Resveratrol can inhibit the expression of MMP-9 protein in the astrocytes of the cortical cortex of rats, and then reduce the degradation of tight junction proteins of Occludin and Claudin-5, thereby reducing BBB permeability and eventually ameliorate the cognitive dysfunction induced by SAE.


Asunto(s)
Disfunción Cognitiva , Encefalopatía Asociada a la Sepsis , Animales , Barrera Hematoencefálica , Claudina-5/metabolismo , Disfunción Cognitiva/tratamiento farmacológico , Disfunción Cognitiva/etiología , Masculino , Ratas , Ratas Sprague-Dawley , Resveratrol/farmacología , Encefalopatía Asociada a la Sepsis/tratamiento farmacológico
8.
J Biomed Nanotechnol ; 16(6): 899-909, 2020 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-33187585

RESUMEN

A well-studied subject of epigenetics, the histone methylation located at lysine and arginine is overseen via methyltransferases and demethylases. Lysine-specific demethylase 4A (KDM4A) comprises a lysine demethylase and possesses specificity for H3K9me3 and H3K36me3, which is capable of being used in order to activate histone transcription. Our team examined the expression of KDM4A within Sprague Dawley (SD) rats and further investigated the mechanism via which this phenomena regulates osteogenic variation within the present study. The overexpression of KDM4A facilitated the process of osteoblast differentiation in bone mesenchymal stem cells (BMSC), while the knocking down differentiation via osteoblast was restrained via the suppression of the expression of Runx2, Osterix, alkaline phosphatase (ALP), and osteocalcin (OCN). Knocking down KDM4A lowered levels of the promoter expression of Runx2, osterix, and OCN, and raised levels of H3K27me3 expression. The results demonstrated that KDM4A possesses a crucial role within the differentiation of osteoblasts and furthermore regulates the expression of Runx2, Osterix, and OCN via H3K9me3. The present research may provide new insights into the treatment of bone healing.


Asunto(s)
Lisina , Osteogénesis , Animales , Diferenciación Celular , Subunidad alfa 1 del Factor de Unión al Sitio Principal/genética , Osteoblastos/metabolismo , Osteocalcina/genética , Osteogénesis/genética , Ratas , Ratas Sprague-Dawley , Factores de Transcripción/genética , Factores de Transcripción/metabolismo
9.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 36(10): 911-917, 2020 Oct.
Artículo en Chino | MEDLINE | ID: mdl-33148386

RESUMEN

Objective To observe the effect of serum of SD rats lavaged by modified Zuojin decoction on the apoptosis and proliferation of human gastric cancer cells and its mechanism. Methods SD rats were gavaged with modified Zuojin decoction to prepare their sera. Human SGC-7901 and MKN-45 cells were cultured and treated with the sera (0, 25, 50, 100, 200, 400) mL/L. MTT assay was used to observe the effect of drug-containing serum on the proliferation of human gastric cancer cells. Immunofluorescence method was used to detect the expression of ki67 after treatment with the drug-containing serum. The effect of drug-containing serum on the apoptosis of gastric cancer cells was detected by flow cytometry. Western blot analysis was used to detect the protein levels of apoptosis-associated cleaved caspase-3, cleaved caspase-8, cleaved caspase-9, BAX and Bcl2 in SGC-7901 and MKN-45 cells. Results The drug-containing serum significantly inhibited the proliferation and induced the apoptosis of SGC-7901 and MKN-45 cells, and the positive rate of ki67 expression was significantly reduced. The levels of cleaved caspase-3, cleaved caspase-9 and BAX proteins in SGC-7901 and MKN-45 cells increased and the levels of Bcl2 protein decreased. Conclusion The drug-containing serum can significantly inhibit the proliferation and induce the apoptosis of human gastric cancer SGC-7901 and MKN-45 cells, and the mechanism may be related to the activation of mitochondrial pathways.


Asunto(s)
Apoptosis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Medicamentos Herbarios Chinos/farmacología , Mitocondrias/efectos de los fármacos , Neoplasias Gástricas/patología , Animales , Línea Celular Tumoral , Humanos , Ratas , Ratas Sprague-Dawley , Neoplasias Gástricas/tratamiento farmacológico
10.
Zhongguo Zhong Yao Za Zhi ; 45(19): 4705-4711, 2020 Oct.
Artículo en Chino | MEDLINE | ID: mdl-33164436

RESUMEN

To explore the effect of Fuke Qianjin Capsules on anti-endometrial fibrosis in intrauterine adhesion(IUA) rats through TGF-ß1-PI3 K/Akt signaling pathway. With female SD rats as the object, IUA rat models were established through mechanical injury and infection, and they were randomly divided into normal group, sham operation group, Bujiale group(0.63 mg·kg~(-1)·d~(-1)), and high-dose Fuke Qianjin Capsules group(1.008 g·kg~(-1)·d~(-1)), medium-dose Fuke Qianjin Capsules group(0.504 g·kg~(-1)·d~(-1)), low-dose Fuke Qianjin Capsules group(0.252 g·kg~(-1)·d~(-1)). The rats were sacrificed 21 days after drug administration, and the uterus and liver were removed after blood collection from the abdominal aorta. The morphology of the uterus was observed with the naked eyes; the pathological and morphological changes of the uterine tissue and liver were observed by HE staining; the degree of fibrosis of the uterine tissue was observed by Masson staining; the expressions of TGF-ß1, TNF-α and IL-6 in serum were detected; the expressions of TGF-ß1, PI3 K, Akt, p-Akt protein in uterine tissue were detected by Western blot. The results showed that Fuke Qianjin Capsules could improve the pathological changes of uterine tissues in IUA rats, without damage to liver tissues, and reduce the expressions of TGF-ß1, TNF-α and IL-6 in serum(P<0.01); significantly reduce TGF-ß1, PI3 K, p-Akt protein expression in uterine tissues(P<0.05, P<0.01). It is indicated that Fuke Qianjin Capsules could exert the anti-endometrial fibrosis effect by regulating the TGF-ß1-PI3 K/Akt signal pathway, so as to achieve the effect in treating IUA rats, especially with the best effect in medium-dose Fuke Qianjin Capsules group.


Asunto(s)
Medicamentos Herbarios Chinos , Factor de Crecimiento Transformador beta1 , Animales , Cápsulas , Femenino , Fosfatidilinositol 3-Quinasas/genética , Proteínas Proto-Oncogénicas c-akt/genética , Ratas , Ratas Sprague-Dawley , Transducción de Señal , Factor de Crecimiento Transformador beta1/genética
11.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi ; 34(11): 1438-1445, 2020 Nov 15.
Artículo en Chino | MEDLINE | ID: mdl-33191703

RESUMEN

Objective: To investigate the effects of silencing P75 neurotrophin receptor (P75NTR) and nerve growth factor (NGF) overexpression on the proliferative activity and ectopic osteogenesis ability of bone marrow mesenchymal stem cells (BMSCs) combined with demineralized bone matrix for heterotopic osteogenesis. Methods: BMSCs of Sprague Dawley (SD) rats were cultured and passaged by adherent isolation method. The third generation BMSCs were transfected with lentivirus mediated P75NTR gene silencing (group B), NGF overexpression gene (group C), P75NTR silencing and NGF overexpression double genes (group D), respectively, and untransfected cells as control (group A). After 7 days of transfection, the expression of fluorescent protein of the target gene was observed by fluorescence microscope; cell counting kit 8 method was used to detect the cells activity for 8 days after transfection; the expressions of P75NTR and NGF proteins in each group were detected by Western blot. The adhesion of BMSCs to demineralized bone matrix (DBM) was observed by inverted phase contrast microscope and scanning electron microscope after transfection of p75NTR silencing and NGF overexpression double genes. After transfection, BMSCs and DBM were co-cultured to prepare 4 groups of tissue engineered bone, which were respectively placed in the dorsal subcutaneous tissue of 8-week-old SD rats to construct subcutaneous ectopic osteogenesis model ( n=6). HE staining was performed at 4 and 8 weeks after operation. ALP staining was used to observe the formation of calcium nodules at 8 weeks after operation. The expressions of Runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP), and osteocalcin (OCN) were detected by real-time fluorescent quantitative PCR. Results: At 7 days after transfection, there was no fluorescence expression in group A, red fluorescence expression was seen in group B, green fluorescence expression in group C, and red-green compound fluorescence expression in group D. The fluorescence expression rate of target gene was about 70%. Western blot detection showed that the relative expression of P75NTR protein in groups A and C was significantly higher than that in groups B and D, and the relative expression of NGF protein in groups C and D was significantly higher than that in groups A and B ( P<0.05). With the passage of time, the cell proliferation activity increased in all groups, especially in group D, which was significantly higher than that in group A at 3-8 days ( P<0.05). The results of inverted phase contrast microscope and scanning electron microscope showed that BMSCs could adhere well to DBM. In the subcutaneous ectopic osteogenesis experiment, HE staining showed that at 4 and 8 weeks after operation, the more bone tissue was formed in group D than in the other 3 groups. ALP staining showed that group D had the highest ALP activity and better osteogenic expression. Compared with group A, the relative expressions of Runx2, ALP, and OCN mRNAs in group D were significantly higher than those in group A ( P<0.05). Conclusion: Silencing P75NTR and NGF overexpression double genes co-transfected BMSCs with DBM to construct tissue engineered bone has good ectopic osteogenic ability. By increasing NGF level and closing P75NTR apoptosis channel, it can not only improve cell activity, but also promote bone tissue regeneration.


Asunto(s)
Células Madre Mesenquimatosas , Proteínas del Tejido Nervioso , Receptores de Factores de Crecimiento , Animales , Células de la Médula Ósea , Matriz Ósea , Diferenciación Celular , Células Cultivadas , Silenciador del Gen , Lentivirus , Factor de Crecimiento Nervioso/genética , Factor de Crecimiento Nervioso/metabolismo , Proteínas del Tejido Nervioso/genética , Proteínas del Tejido Nervioso/metabolismo , Osteogénesis , Ratas , Ratas Sprague-Dawley , Receptor de Factor de Crecimiento Nervioso , Receptores de Factores de Crecimiento/genética , Receptores de Factores de Crecimiento/metabolismo , Transfección
12.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi ; 34(11): 1429-1437, 2020 Nov 15.
Artículo en Chino | MEDLINE | ID: mdl-33191702

RESUMEN

Objective: To study the local vascular remodeling, inflammatory response, and their correlations following acute spinal cord injury (SCI) with different grades, and to assess the histological changes in SCI rats. Methods: One hundred and sixteen adult female Sprague Dawley rats were randomly divided into 4 groups ( n=29). The rats in sham group were received laminectomy only. A standard MASCIS spinal cord compactor was applied with drop height of 12.5, 25.0, or 50.0 mm to establish the mild, moderate, or severe SCI model, respectively. Quantitative rat endothelial cell antigen 1 (RECA1) and CD68 positive areas and the correlations were studied by double immunofluorescent (DIF) staining at 12 hours, 24 hours, 3 days, 7 days, and 28 days following SCI. Moreover, qualitative neurofilament-H (NF-H) and glial fibrillary acidic protein (GFAP) positive glial cells were studied by DIF staining at 28 days. ELISA was used to detect the levels of tumor necrosis factor α (TNF-α), interleukin 1ß (IL-1ß), and IL-6 in spinal cord homogenates at 12 hours, 24 hours, and 3 days, and the correlations between TNF-α, IL-1ß, or IL-6 levels and microvascular density (RECA1) were accordingly studied. Moreover, the neural tissue integrity and neuron damage were assessed by HE staining at 12 hours, 24 hours, 3 days, 7 days, and 28 days, and Nissl's staining at 28 days following SCI, respectively. Results: DIF staining revealed that the ratio of RECA1 positive area was the highest in moderate group, higher in mild and severe groups, and the lowest in sham group with significant differences between groups ( P<0.05). The ratio of CD68 positive area was the highest in severe group, higher in moderate and mild groups, and the lowest in sham group with significant differences between groups ( P<0.05), except the comparisons between mild and moderate groups at 24 hours and 28 days after SCI ( P>0.05). There was no significant correlation between the RECA1 and CD68 expressions in sham group at different time points ( P>0.05). At 12 and 24 hours after SCI, the RECA1 and CD68 expressions in mild and moderate groups showed significant positive correlations ( P<0.05), while no significant correlation was found in severe group ( P>0.05). No significant correlations between the RECA1 and CD68 expressions was shown in all SCI groups at 3 days and in severe group at 7 days ( P>0.05), while the negative correlations were shown in mild and moderate groups at 7 days, and in all SCI groups at 28 days ( P<0.05). In mild, moderate, and severe groups, the axons became disrupted, shorter and thicker rods-like, or even merged blocks with increased injury, while the astrocytes decreased in number, unorganized and condensed in appearance. ELISA studies showed that TNF-α, IL-1ß, and IL-6 levels in sham group were significantly lower than those in other 3 groups at different time points ( P>0.05). The differences in TNF-α, IL-1ß, and IL-6 levels between SCI groups at different time points were sinificant ( P<0.05), except IL-1ß levels between the mild and moderate groups at 12 hours ( P>0.05). Three inflammatory factors were all significantly correlated with the microvascular density grades ( P<0.05). Histological analysis indicated that the damage to spinal cord tissue structure correlated with the extent of SCI. In severe group, local hemorrhage, edema, and infiltration of inflammatory cells were found the most drastic, the grey/white matter boundary was disappeared concurrently with the formation of cavity and shortage of normal neurons. Conclusion: In the acute stage following mild or moderate SCI, progressively aggravated injury result in higher microvessel density and increased inflammation. However, at the SCI region, the relation between microvessel density and inflammation inverse with time in the different grades of SCI. Accordingly, the destruction of neural structures positively relate to the grades of SCI and severity of inflammation.


Asunto(s)
Traumatismos de la Médula Espinal , Remodelación Vascular , Animales , Femenino , Ratas , Ratas Sprague-Dawley , Médula Espinal , Factor de Necrosis Tumoral alfa
13.
J Pharmacol Sci ; 144(4): 237-244, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-33070843

RESUMEN

Hypoxic pulmonary hypertension (HPH) is a progressive and irreversible disease that reduces survival. Echinacoside is a phenylethanoid glycoside from Tibetan herbs known for its vasorelaxant effect and for inhibiting the proliferation of rat pulmonary arterial smooth muscle cells. This study aimed to investigate the effect of echinacoside on HPH. Sprague Dawley rats were housed in a hypobaric hypoxia chamber (4500 m) for 28 days to obtain the HPH model. Echinacoside (3.75, 7.5, 15, 30 and 40 mg/kg) was administered by intraperitoneal injection from the 1st to the 28th day. The mean pulmonary artery pressure (mPAP), right ventricular hypertrophy index, hemoglobin, hematocrit, red blood cell concentration and morphological change of pulmonary arteries were evaluated. Vascular perfusion assay was used to assess the pulmonary artery function. Echinacoside reduced mPAP, hemoglobin, hematocrit, right ventricular hypertrophy index and mean wall thickness% of pulmonary arteries in HPH rats. It significantly increased maximum vasoconstriction percentage of pulmonary arteries induced by noradrenaline in a dose-dependent manner. In addition, it improved the responsiveness of pulmonary arteries to acetylcholine and sodium nitroprusside. Therefore, Echinacoside might be an effective treatment against HPH, since it regulated pulmonary artery endothelium and smooth muscle layer function and improved the remodeling of pulmonary artery.


Asunto(s)
Glicósidos/administración & dosificación , Glicósidos/farmacología , Hipertensión Pulmonar/tratamiento farmacológico , Hipertensión Pulmonar/etiología , Hipoxia/complicaciones , Fitoterapia , Arteria Pulmonar/efectos de los fármacos , Arteria Pulmonar/fisiopatología , Remodelación Vascular/efectos de los fármacos , Vasoconstricción/efectos de los fármacos , Animales , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Glicósidos/uso terapéutico , Hipertensión Pulmonar/fisiopatología , Hipertensión Pulmonar/prevención & control , Técnicas In Vitro , Inyecciones Intraperitoneales , Masculino , Ratas Sprague-Dawley , Vasodilatadores
14.
Int J Nanomedicine ; 15: 6945-6960, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-33061361

RESUMEN

Background: Natural clay nanomaterials are an emerging class of biomaterial with great potential for tissue engineering and regenerative medicine applications, most notably for osteogenesis. Materials and Methods: Herein, for the first time, novel tissue engineering scaffolds were prepared by 3D bioprinter using nontoxic and bioactive natural attapulgite (ATP) nanorods as starting materials, with polyvinyl alcohol as binder, and then sintered to obtain final scaffolds. The microscopic morphology and structure of ATP particles and scaffolds were observed by transmission electron microscope and scanning electron microscope. In vitro biocompatibility and osteogenesis with osteogenic precursor cell (hBMSCs) were assayed using MTT method, Live/Dead cell staining, alizarin red staining and RT-PCR. In vivo bone regeneration was evaluated with micro-CT and histology analysis in rat cranium defect model. Results: We successfully printed a novel porous nano-ATP scaffold designed with inner channels with a dimension of 500 µm and wall structures with a thickness of 330 µm. The porosity of current 3D-printed scaffolds ranges from 75% to 82% and the longitudinal compressive strength was up to 4.32±0.52 MPa. We found firstly that nano-ATP scaffolds with excellent biocompatibility for hBMSCscould upregulate the expression of osteogenesis-related genes bmp2 and runx2 and calcium deposits in vitro. Interestingly, micro-CT and histology analysis revealed abundant newly formed bone was observed along the defect margin, even above and within the 3D bioprinted porous ATP scaffolds in a rat cranial defect model. Furthermore, histology analysis demonstrated that bone was formed directly following a process similar to membranous ossification without any intermediate cartilage formation and that many newly formed blood vessels are within the pores of 3D-printed scaffolds at four and eight weeks. Conclusion: These results suggest that the 3D-printed porous nano-ATP scaffolds are promising candidates for bone tissue engineering by osteogenesis and angiogenesis.


Asunto(s)
Regeneración Ósea/fisiología , Células Madre Mesenquimatosas/citología , Ingeniería de Tejidos/métodos , Andamios del Tejido/química , Animales , Huesos/fisiología , Calcio/metabolismo , Chlorocebus aethiops , Condrogénesis , Fuerza Compresiva , Regulación de la Expresión Génica , Humanos , Compuestos de Magnesio/química , Masculino , Ensayo de Materiales , Nanotubos/química , Osteogénesis/fisiología , Alcohol Polivinílico/química , Porosidad , Impresión Tridimensional , Ratas Sprague-Dawley , Compuestos de Silicona/química , Células Vero , Microtomografía por Rayos X
15.
Int J Nanomedicine ; 15: 6975-6991, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-33061363

RESUMEN

Purpose: Small extracellular vesicles (sEV) are a heterogeneous group of vesicles that consist of proteins, lipids and miRNA molecules derived from the cell of origin. Although xenogeneic sEV have been applied for soft tissue regeneration successfully, the regeneration effect of allogeneic and xenogeneic sEV has not been compared systematically. Methods: Our previous study has shown that sEV derived from rat adipose tissue successfully induced neoadipose regeneration. In this study, sEV were isolated from rat adipose tissue (r-sEV-AT) and porcine adipose tissue (p-sEV-AT), the morphology, size distribution and marker proteins expression of r-sEV-AT and p-sEV-AT were characterized. Besides, the sEV/AT ratio was evaluated and compared between r-sEV-AT and p-sEV-AT. Rat adipose-derived stromal/stem cells (rASCs) and rat aorta endothelial cells (rECs) were adopted to test the cellular response to allogeneic and xenogeneic sEV-AT. The effects of allogeneic and xenogeneic sEV-AT on host cells migration and neoadipose formation were evaluated in a subcutaneous custom-designed model. A full-thickness skin wound healing model was used to further compare the ability of allogeneic and xenogeneic sEV-AT in inducing complex soft tissue regeneration. Results: p-sEV-AT showed similar morphology and size distribution to r-sEV-AT. Marker proteins of sEV were detected in both r-sEV-AT and p-sEV-AT. The sEV/AT ratio of porcine was slightly higher than that of rat. The effects of r-sEV-AT and p-sEV-AT on the differentiation of rASCs and rECs showed no significant difference. When allogeneic and xenogeneic sEV-AT were subcutaneously implanted into the back of SD rats, the host cells chemotactic infiltration was observed in 1 week and neoadipose tissue formation was induced in 8 weeks; no significant difference was observed between allogeneic and xenogeneic sEV-AT. For complex soft tissue regeneration, both allogeneic and xenogeneic sEV-AT significantly promoted wound re-epithelialization, granulation tissue formation and hair follicle regeneration and then accelerated skin wound healing. Conclusion: Our results demonstrated that sEV derived from the same tissues of different species might be loaded with similar therapeutic substance benefitting tissue repair and regeneration, and paved the way for future research aimed at xenogeneic sEV application.


Asunto(s)
Tejido Adiposo/fisiología , Vesículas Extracelulares/trasplante , Trasplante Heterólogo , Trasplante Homólogo , Adipocitos , Tejido Adiposo/citología , Animales , Diferenciación Celular , Movimiento Celular , Células Cultivadas , Células Endoteliales/citología , Espacio Extracelular , MicroARNs , Ratas Sprague-Dawley , Regeneración , Porcinos , Trasplante Heterólogo/métodos , Trasplante Homólogo/métodos , Cicatrización de Heridas
16.
Artículo en Chino | MEDLINE | ID: mdl-33036524

RESUMEN

Objective: To study the expression of high mobility group protein 1 (HMGB1) in the brain of rats after hyperbaric oxygen (HBO) treatment of acute carbon monoxide poisoning (DEACMP) , and to explore the mechanism of HBO in the prevention and treatment of DEACMP pathological process by regulating HMGB1. Methods: 108 SD rats were randomly divided into control group (NC group) and co group (CO group) . HBO treatment group (HBO group) , 48 rats in each group. Co group and HBO group were used to establish CO poisoning model, HBO group were treated with hyperbaric oxygen once a day. Water maze test was used to detect and analyze the memory retention ability of three groups of rats in 3 d, 7 d, 14 d. ELISA was used to detect the plasma concentration of HMGB1、IL-6、TNF-α in three groups of rats on the 1 d, 3 d, 7 d, 14 d, 21 d Concentration. Western blotting was used to detect the expression of HMGB1 and Caspase-3 in the brain of the three groups on the 1 d, 3 d, 7 d, 14 d, 21 d. TUNEL staining was used to detect the apoptosis of hippocampal neurons in the three groups. Results: Compared with NC group, the average escape latency of rats in CO group and HBO group was significantly prolonged, and the activity time of platform quadrant in CO group was significantly shortened on 14 d and 21 d (P<0.05) ; compared with CO group, the average escape latency of HBO group on 7 d, 14 d and 21 d was significantly shortened (P<0.05) . Compared with NC group, plasma HMGB1 in CO group and HBO group were significantly increased (P<0.05) ; after 3 days, HBO group was significantly lower than co group, the difference was statistically significant (P<0.05) . The levels of IL-6 and TNF-α in HBO group and co group increased rapidly and then decreased gradually. The increased levels of IL-6 and TNF-α in HBO group were significantly lower than those in CO group (P<0.05) . Compared with NC group, the expression of HMGB1 and Caspase-3 in CO group was significantly increased on 3 d, 7 d and 14 d (P<0.05) ; the expression of HMGB1 and Caspase-3 in HBO group was significantly increased on 3 d, 7 d, 14 d and 21 d (P<0.05) ; compared with CO group, the expression of HMGB1 and Caspase-3 in HBO group decreased significantly on 3 d, 7 d, 14 d and 21 d (P<0.05) . The apoptotic index of nerve cells in CO group began to increase at 3 days, which was significantly different from that of NC group (P<0.05) , and the difference was still statistically significant on 21 d (P<0.05) ; the apoptotic index of nerve cells in HBO group was slightly increased, but there was no significant difference compared with NC group (P>0.05) , and the apoptotic index of 3 d, 7 d, 14 d and 21 d in HBO group was significantly lower than that in CO group (P<0.05) . Conclusion: acute CO poisoning can induce the release of HMGB1 and a variety of inflammatory factors. HMGB1 can promote the apoptosis of nerve cells after acute CO poisoning by up regulating the expression of caspase-3 protein, and participate in the pathological process of DEACMP. HBO can down regulate the expression of HMGB1, IL-6, TNF-α and caspase-3 protein, inhibit the apoptosis of nerve cells, and play a protective role on nerve cells.


Asunto(s)
Encefalopatías , Intoxicación por Monóxido de Carbono , Proteína HMGB1 , Oxigenación Hiperbárica , Animales , Encefalopatías/terapia , Intoxicación por Monóxido de Carbono/terapia , Ratas , Ratas Sprague-Dawley
17.
Fa Yi Xue Za Zhi ; 36(4): 502-506, 2020 Aug.
Artículo en Inglés, Chino | MEDLINE | ID: mdl-33047534

RESUMEN

Abstract: Objective To study the characteristics of positive expression of integrin ß1 in the rat brain tissue of two kinds of traumatic brain injury models and to explore the feasibility of inferring the mode of traumatic brain injury using the positive expression of integrin ß1. Methods The occipital region of rats was hit by hydraulic impact method and pendulum striking method to produce two closed brain injury models of linear and rotation acceleration respectively, then 120 SD rats were randomly divided into linear acceleration injury group, rotation acceleration injury group, sham operation group and normal control group. Immunohistochemistry staining and Western blotting method were used to detect the positive expression of integrin ß1 in different parts of the brain tissue at 30 min, 3 h, 6 h, 12 h, 3 d and 7 d after rat injury. The data was processed statistically by SPSS 18.0 software. Results The positive expression of integrin ß1 was detected 30 min after brain injury and reached the peak 6 h after brain injury. With the extension of injury time, the expression tended to enhance. At the same time points after injury, the differences in the positive expression of integrin ß1 between the linear acceleration injury group and the rotation acceleration injury group in the occipital strike point and thalamus had no statistical significance ( P>0.05), but the differences in the expression of integrin ß1 in the frontal lobe and brain stem had statistical significance (P<0.05). Conclusion The characteristics of positive expression of integrin ß1 in brain tissue can be used to infer the strike point and the manner of injury and has application value for the reconstruction of craniocerebral injury process.


Asunto(s)
Lesiones Traumáticas del Encéfalo , Lesiones Encefálicas , Animales , Encéfalo/metabolismo , Integrina beta1/genética , Integrina beta1/metabolismo , Ratas , Ratas Sprague-Dawley
18.
Nat Commun ; 11(1): 5318, 2020 10 21.
Artículo en Inglés | MEDLINE | ID: mdl-33087709

RESUMEN

Synaptic vesicles (SVs) can be pooled across multiple synapses, prompting questions about their dynamic allocation for neurotransmission and plasticity. We find that the axonal traffic of recycling vesicles is not supported by ubiquitous microtubule-based motility but relies on actin instead. Vesicles freed from synaptic clusters undergo ~1 µm bouts of active transport, initiated by nearby elongation of actin filaments. Long distance translocation arises when successive bouts of active transport were linked by periods of free diffusion. The availability of SVs for active transport can be promptly increased by protein kinase A, a key player in neuromodulation. Vesicle motion is in turn impeded by shutting off axonal actin polymerization, mediated by nitric oxide-cyclic GMP signaling leading to inhibition of RhoA. These findings provide a potential framework for coordinating post-and pre-synaptic strength, using retrograde regulation of axonal actin dynamics to mobilize and recruit presynaptic SV resources.


Asunto(s)
Citoesqueleto de Actina/fisiología , Proteínas Quinasas Dependientes de AMP Cíclico/fisiología , Óxido Nítrico/fisiología , Vesículas Sinápticas/fisiología , Animales , Transporte Axonal/fisiología , Transporte Biológico Activo , Células Cultivadas , GMP Cíclico/fisiología , Femenino , Hipocampo/citología , Hipocampo/fisiología , Proteínas Luminiscentes/metabolismo , Masculino , Neuronas/fisiología , Nocodazol/farmacología , Ratas , Ratas Sprague-Dawley , Transmisión Sináptica/fisiología , Vesículas Sinápticas/efectos de los fármacos
19.
Nan Fang Yi Ke Da Xue Xue Bao ; 40(10): 1390-1398, 2020 Oct 30.
Artículo en Chino | MEDLINE | ID: mdl-33118509

RESUMEN

OBJECTIVE: To investigate the effect of miR-133b on cardiomyocyte apoptosis induced by myocardial ischemia-reperfusion (I/R) and explore the mechanism. METHODS: Thirty-six adult SD rats were randomized into sham-operated group, I/R group, AdmiR-NC group and AdmiR-133b group, and rat models of myocardial I/R were established in the latter 3 groups with myocardial injections of saline or recombinant adenoviruses in the left ventricle. The expression of MiR-133b was detected using RT-qPCR, and cardiac function of the rats was determined using FDP 1 HRV and BRS analysis system. Serum CK-MB and cTnI levels were determined by ELISA, myocardial injury was evaluated with HE staining, cardiomocyte apoptosis was detected by flow cytometry, and ROS content was determined using a DCFH-DA probe. In the in vitro experiment, H9C2 myocardial cells with hypoxia/reoxygenation (H/R) treatment were transfected with Mir-NC or MiR-133b mimic, and the cellular expression of MiR-133b, cell apoptosis, and ROS content were determined. Dual luciferase reporter assay was performed to verify the targeting relationship between miR-133b and YES1. The effects of pc-YES1 or miR-133b mimic transfection on YES1 expression, apoptosis, and ROS content in H9C2 cells were evaluated. RESULTS: Compared with those in I/R group, miR-133b expression was obviously up-regulated, LVEDP, cTnI and CK-MB levels were significantly decreased, and LVSP, +dp/dt, -dp/dt, HR and CF levels were increased in admiR-133b group (P < 0.01). The rats in admiR-133b group showed obviously reduced pathological damage, cell apoptosis and ROS content compared with those in I/ R group (P < 0.01). In H9C2 cells exposed to H/R, transfection with miR-133b mimic significantly up-regulated miR-133b expression and decreased cell apoptosis and ROS content (P < 0.01). The results of dual luciferase reporter assay suggested a direct targeting relationship between miR-133b and YES1, and MiR-133b mimic transfection significantly down-regulated YES1 protein expression in cells with H/R exposure (P < 0.01). Co-transfection with pc-YES1 reversed the effect of miR-133b overexpression on myocardial cell apoptosis and ROS accumulation. CONCLUSIONS: miR-133b can inhibit I/R-induced myocardial cell apoptosis and ROS accumulation by targeting YES1 to reduce myocardial I/R injury in rats.


Asunto(s)
Daño por Reperfusión Miocárdica , Animales , Apoptosis , MicroARNs/genética , Miocitos Cardíacos , Ratas , Ratas Sprague-Dawley , Especies Reactivas de Oxígeno
20.
Nan Fang Yi Ke Da Xue Xue Bao ; 40(10): 1406-1414, 2020 Oct 30.
Artículo en Chino | MEDLINE | ID: mdl-33118513

RESUMEN

OBJECTIVE: To study the effects of high-fat (HF) diet and exercise on the expressions of asprosin and CTRP6 in adipose tissues in different regions of rats during mid-gestation. METHODS: Pregnant SD rats were fed on a standard chow diet or a high-fat (60% fat content) diet for 14 days starting on gestation day (GD) 1. Starting from GD3, the rats fed either on normal or high-fat diet in the exercise groups (CH-RW and HF-RW groups) were allowed access to the running wheels for voluntary running, and those in sedentary groups (CH-SD and HF-SD groups) remained sedentary. At the end of the 14 days, adipose tissues were sampled from different regions of the rats for detecting the mRNA and protein expressions of asprosin and CTRP6 using RT-qPCR and Western blotting. RESULTS: The mRNA expression of asprosin in retroperitoneal adipose tissues was significantly higher in HF-RW group than in the other 3 groups (P < 0.0001). Asprosin mRNA expression in subcutaneous adipose tissues was significantly higher in HF-SD group than in CH-SD group (P=0.0234) and comparable between HF-RW and CH-SD groups (P=0.0494). CTRP6 mRNA expression in retroperitoneal adipose tissues was also significantly higher in HF-RW group than in the other groups (P < 0.0001), and CTRP6 protein expression was signficiantly higher in HF-RW group than in CH-RW and HF-SD groups (P < 0.05). In subcutaneous adipose tissues, CTRP6 mRNA expression was significantly higher in CH-RW group than in HF-SD and HF-RW groups (P < 0.05). The protein expression level of CTRP6 in subcutaneous adipose tissues showed a significant negative correlation with blood glucose (r=-0.6038, P=0.0172), while its expression in retroperitoneal adipose tissues was positively correlated with blood glucose (r=0.5305, P= 0.0285); the mRNA expression levels of asprosin and CTRP6 were significantly lower in subcutaneous than in retroperitoneal adipose tissues (P < 0.0001). CONCLUSIONS: High-fat diet and exercise during mid-gedtation can affect the expression levels of asprosin and CTRP6 in adipose tissues of rats in a site-specific manner.


Asunto(s)
Dieta Alta en Grasa , Grasa Intraabdominal , Adipoquinas , Animales , Glucemia , Dieta Alta en Grasa/efectos adversos , Femenino , Embarazo , Ratas , Ratas Sprague-Dawley
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