RESUMO
Skeletal muscle atrophy is a common and serious complication of chronic kidney disease (CKD). Oxidative stress and autophagy are the primary molecular mechanisms involved in muscle atrophy. Calycosin, a major component of Radix astragali, exerts anti-inflammatory, anti-oxidative stress and anti-autophagy effects. We investigated the effects and mechanisms of calycosin on skeletal muscle atrophy in vivo and in vitro. 5/6 nephrectomy (5/6 Nx) rats were used as a model of CKD. We evaluated bodyweight and levels of serum creatinine (SCr), blood urea nitrogen (BUN) and serum albumin (Alb). H&E staining, cell apoptosis, oxidative stress biomarkers, autophagosome and LC3A/B levels were performed and evaluated in skeletal muscle of CKD rat. Calycosin treatment improved bodyweight and renal function, alleviated muscle atrophy (decreased the levels of MuRF1 and MAFbx), increased superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activity and reduced malondialdehyde (MDA) levels in skeletal muscle of CKD rats. Importantly, calycosin reduced autophagosome formation, down-regulated the expression of LC3A/B and ATG7 through inhibition of AMPK and FOXO3a, and increased SKP2, which resulted in decreased expression of CARM1, H3R17me2a. Similar results were observed in C2C12 cells treated with TNF-α and calycosin. Our findings showed that calycosin inhibited oxidative stress and autophagy in CKD induced skeletal muscle atrophy and in TNF-α-induced C2C12 myotube atrophy, partially by regulating the AMPK/SKP2/CARM1 signalling pathway.
Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Autofagia/efeitos dos fármacos , Isoflavonas/farmacologia , Músculo Esquelético/patologia , Atrofia Muscular/patologia , Estresse Oxidativo/efeitos dos fármacos , Proteína-Arginina N-Metiltransferases/metabolismo , Insuficiência Renal Crônica/patologia , Animais , Apoptose/efeitos dos fármacos , Arginina/metabolismo , Peso Corporal/efeitos dos fármacos , Linhagem Celular , Regulação para Baixo/efeitos dos fármacos , Fibrose , Histonas/metabolismo , Rim/efeitos dos fármacos , Rim/patologia , Rim/fisiopatologia , Masculino , Metilação , Camundongos , Fibras Musculares Esqueléticas/efeitos dos fármacos , Fibras Musculares Esqueléticas/metabolismo , Fibras Musculares Esqueléticas/patologia , Nefrectomia , Ratos Sprague-Dawley , Insuficiência Renal Crônica/fisiopatologia , Proteínas Quinases Associadas a Fase S/metabolismo , Transdução de Sinais , Fator de Necrose Tumoral alfaRESUMO
BACKGROUND: Cervical cancer (CC) is one of the most common cancers among females worldwide. Spindle and kinetochore-associated complex subunit 3 (SKA3), located on chromosome 13q, was identified as a novel gene involved in promoting malignant transformation in cancers. However, the function and underlying mechanisms of SKA3 in CC remain unknown. Using the Oncomine database, we found that expression of SKA3 mRNA is higher in CC tissues than in normal tissues and is linked with poor prognosis. METHODS: In our study, immunohistochemistry showed increased expression of SKA3 in CC tissues. The effect of SKA3 on cell proliferation and migration was evaluated by CCK8, clone formation, Transwell and wound-healing assays in HeLa and SiHa cells with stable SKA3 overexpression and knockdown. In addition, we established a xenograft tumor model in vivo. RESULTS: SKA3 overexpression promoted cell proliferation and migration and accelerated tumor growth. We further identified that SKA3 is involved in regulating cell cycle progression and the PI3K/Akt signaling pathway via RNA-sequencing (RNA-Seq) and gene set enrichment analyses. Western blotting results revealed that SKA3 overexpression increased levels of p-Akt, cyclin E2, CDK2, cyclin D1, CDK4, E2F1 and p-Rb in HeLa cells. Additionally, the use of an Akt inhibitor (GSK690693) significantly reversed the cell proliferation capacity induced by SKA3 overexpression in HeLa cells. CONCLUSIONS: We suggest that SKA3 overexpression contributes to CC cell growth and migration by promoting cell cycle progression and activating the PI3K-Akt signaling pathway, which may provide potential novel therapeutic targets for CC treatment.
RESUMO
While many previous studies have reported an association between the single-nucleotide polymorphisms (SNPs) of the podocin and proteinuria occurred, a conclusive relationship has not been defined in every oligoallelic state of amino acid (AA) mutations in podocin. In this study, we performed a meta-analysis of the published data to investigate the impact of the oligoallelic AA mutations of the podocin on proteinuria; a total 16 AA mutations were investigated for oligoallelic pathogenicity. Despite significant heterogeneity within some of the comparisons, the results revealed significantly higher risks of proteinuria in early-onset (onset age <16) individuals for five mutations (P118L, R138Q, R168H, V180M, and V260E), and in all onset ages individuals for five mutations (R138Q, G140X, R229Q, V260E, and V290M) compared to non-variant individuals. We also tested the steroid response in individuals with R229Q and E237Q. No statistically significant differences in the two mutations carrier rate were observed between steroid resistance patients and controls. No AA mutation was selected for meta-analysis on the recurrence of proteinuria after renal transplantation as lack of control data. In conclusion, our meta-analysis tested the pathogenicity of the oligoallelic AA mutations in podocin and suggested the potential causative mutations, and the alleles showing an association with protein susceptibility. The sensitivity and specificity of each causative mutation are pending further testing.
Assuntos
Aminoácidos/genética , Peptídeos e Proteínas de Sinalização Intracelular/genética , Proteínas de Membrana/genética , Síndrome Nefrótica/genética , Proteinúria/genética , Alelos , Heterozigoto , Humanos , Mutação , Polimorfismo de Nucleotídeo Único , Viés de PublicaçãoRESUMO
Ketoacids (KA) are known to improve muscle mass among patients with chronic kidney disease (CKD) on a low-protein diet (CKD-LPD), but the mechanism of its preventive effects on muscle atrophy still remains unclear. Since muscle atrophy in CKD may be attributable to the down-regulation of the Wnt7a/Akt/p70S6K pathway and the activation of the ubiquitin-proteasome system (UPS) and the apoptotic signalling pathway, a hypothesis can readily be drawn that KA supplementation improves muscle mass by up-regulating the Wnt7a/Akt/p70S6K pathway and counteracting the activation of the UPS and caspase-3-dependent apoptosis in the muscle of CKD-LPD rats. Rats with 5/6 nephrectomy were randomly divided into three groups, and fed with either 22 % protein (normal-protein diet; NPD), 6 % protein (LPD) or 5 % protein plus 1 % KA for 24 weeks. Sham-operated rats with NPD intake were used as the control. The results demonstrated that KA supplementation improved protein synthesis and increased related mediators such as Wnt7a, phosphorylated Akt and p70S6K in the muscle of CKD-LPD rats. It also inhibited protein degradation, withheld the increase in ubiquitin and its ligases MAFbx (muscle atrophy F-box) and MuRF1 (muscle ring finger-1) as well as attenuated proteasome activity in the muscle of CKD-LPD rats. Moreover, KA supplementation gave rise to a reduction in DNA fragment, cleaved caspase-3 and 14 kDa actin fragment via the down-regulation of the Bax:Bcl-2 ratio in the muscle of CKD-LPD rats. The beneficial effects unveiled herein further consolidate that KA may be a better therapeutic strategy for muscle atrophy in CKD-LPD.
Assuntos
Suplementos Nutricionais , Modelos Animais de Doenças , Cetoácidos/uso terapêutico , Músculo Esquelético/metabolismo , Atrofia Muscular/prevenção & controle , Proteínas Proto-Oncogênicas/agonistas , Insuficiência Renal Crônica/dietoterapia , Proteínas Wnt/agonistas , Animais , Apoptose , Dieta com Restrição de Proteínas/efeitos adversos , Regulação para Baixo , Masculino , Proteínas Musculares/biossíntese , Músculo Esquelético/patologia , Atrofia Muscular/etiologia , Nefrectomia/efeitos adversos , Complexo de Endopeptidases do Proteassoma/metabolismo , Proteólise , Proteínas Proto-Oncogênicas/metabolismo , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Insuficiência Renal Crônica/metabolismo , Insuficiência Renal Crônica/patologia , Insuficiência Renal Crônica/fisiopatologia , Transdução de Sinais , Ubiquitinação , Regulação para Cima , Proteínas Wnt/metabolismoRESUMO
Diabetic nephropathy (DN) is one of the most common and serious chronic complications of diabetes mellitus, however, no efficient clinical drugs exist for the treatment of DN. We selected and synthesized several sesquiterpene lactones (SLs), and then used the MTT assay to detect rat mesangial cells (MCs) proliferation, ELISA to measure the expression level of monocyte chemoattractant protein-1 (MCP-1), transforming growth factor beta (TGF-ß1) and fibronectin(FN), real-time fluorescent quantitative PCR analysis to measure the MCP-1 and TGF-ß1 gene expression, western blot to detect the level of IκBα protein and EMSA to measure the activation of nuclear factor kappa B (NF-κB). We discovered that SLs, including parthenolide (PTL), micheliolide (MCL), arglabin, and isoalantolactone (IAL), as well as several synthetic analogs of these molecules, could effectively attenuate the high glucose-stimulated activation of NF-κB, the degradation of IκBα, and the expression of MCP-1, TGF-ß1 and FN in rat mesangial cells (MCs). These findings suggest that SLs and their derivatives have potential as candidate drugs for the treatment of DN.
Assuntos
Quimiocina CCL2/metabolismo , Glucose/fisiologia , Lactonas/farmacologia , Células Mesangiais/metabolismo , NF-kappa B/metabolismo , Sesquiterpenos/farmacologia , Fator de Crescimento Transformador beta1/metabolismo , Animais , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Quimiocina CCL2/genética , Nefropatias Diabéticas/tratamento farmacológico , Avaliação Pré-Clínica de Medicamentos , Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Lactonas/síntese química , Células Mesangiais/efeitos dos fármacos , Ligação Proteica , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Espécies Reativas de Oxigênio/metabolismo , Sesquiterpenos/síntese química , Fator de Crescimento Transformador beta1/genéticaRESUMO
OBJECTIVE: To investigate the effect of Shenshuai Yangzhen Capsule (SYC) on hypothalamic leptin-neuropeptide Y (NPY) and proopiomelanocortin (POMC) axes in chronic renal failure (CRF) rats with malnutrition (MN). METHODS: Forty-two male SD rats of SPF grade were established into CRF-MN model by 5/6 nephrectomy and 4% casein diet, the happening time of MN in them was recorded. Rats successfully modeled were randomized into three groups, 11 rats in Group A treated with SYC, 11 in group B treated with composite alpha-keto acid and 12 in Group C was untreated. Besides, a normal control group was set up with 8 healthy rats. After being treated for 4 weeks, the renal function related indices, including serum creatinine (Scr), blood urea nitrogen (BUN), 24 hour urine protein (24 h Upro), albumin (ALB), haemoglobin (Hb) insulin like growth factor-1 (IGF-1), total cholesterol (TC) and triglyeride (TG) were measured, and body weight, food intake in rats were observed dynamically, blood leptin and NPY level in rats were determined by radioimmunoassay; mRNA expressions of OB-Rb, NPY and POMC in hypothalamus were detected with RT-PCR. RESULTS: CRF rats revealed MN at the end of 10th week after modeling. Compared with Group C, the condition of MN in Group A was significantly improved, showing increase of food intake and body weight (P < 0.05), marked improvement of renal function (P < 0.05), decrease of LP and NPY levels in plasma (P < 0.05), as well as up-regulated NPY mRNA expression and down-regulated mRNA expressions of OB-Rb and POMC in hypothalamus (P < 0.01). CONCLUSION: SYC can improve the malnutrition condition in rats with CRF, which is possibly by way of depressing OB-Rb and POMC mRNA expression and upgrading NPY mRNA expression in hypothalamus.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Hipotálamo/metabolismo , Falência Renal Crônica/fisiopatologia , Desnutrição/fisiopatologia , Neuropeptídeo Y/metabolismo , Pró-Opiomelanocortina/metabolismo , Animais , Falência Renal Crônica/complicações , Falência Renal Crônica/metabolismo , Leptina/genética , Leptina/metabolismo , Masculino , Desnutrição/etiologia , Desnutrição/metabolismo , Neuropeptídeo Y/genética , Pró-Opiomelanocortina/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
Endothelial dysfunction plays important roles in vascular dysfunction under diabetic conditions. The generation of advanced glycation end products (AGEs), which can induce inflammation and oxidative stress, is pivotal in endothelial dysfunction. Salidroside, a major active compound in Rhodiola rosea, exerts protective effects against vascular diseases. To study the effects and mechanism of salidroside in diabetes-induced vascular endothelial dysfunction, an in vitro model was established with AGEs-induced human umbilical vein endothelial cells (HUVECs). Then, cell viability, cell apoptosis, pro-inflammatory cytokines and oxidative biomarkers were tested to determine the effects of salidroside at 10, 50 and 100 µM doses on AGEs induced HUVECs. Additionally, RNA-Seq and bioinformatics analyses were used to search for the underlying mechanism of salidroside. The results showed that salidroside promoted cell viability and significantly alleviated cell apoptosis in AGEs-induced HUVECs. Furthermore, salidroside remarkably decreased the levels of the pro-inflammatory cytokines TNF-α, IL-1ß and IL-6 and impeded the expression of VCAM-1 and ICAM-1 induced by AGEs. Additionally, salidroside promoted superoxide dismutase (SOD) activity and increased catalase (CAT) and glutathione peroxidase (GSH-Px) levels while inhibiting the intracellular generation of reactive oxygen species (ROS) and malondialdehyde (MDA) in AGEs-induced HUVECs. Importantly, salidroside alleviated endothelial inflammation and oxidative stress by activating AMPK phosphorylation and inhibiting NF-ĸB p65 and NLRP3 inflammasome activation. Therefore, we used compound C, an accepted AMPK inhibitor, to further demonstrate the mechanism. Interestingly, the phenomenon produced by salidroside was abolished. Our findings suggest that salidroside ameliorates AGEs-induced endothelial inflammation and oxidative stress, partially via the AMPK/NF-κB/NLRP3 signaling pathway.
Assuntos
Angiopatias Diabéticas/tratamento farmacológico , Endotélio Vascular/efeitos dos fármacos , Glucosídeos/farmacologia , Produtos Finais de Glicação Avançada/imunologia , Inflamação/tratamento farmacológico , Fenóis/farmacologia , Proteínas Quinases Ativadas por AMP/antagonistas & inibidores , Proteínas Quinases Ativadas por AMP/metabolismo , Linhagem Celular , Angiopatias Diabéticas/imunologia , Angiopatias Diabéticas/metabolismo , Endotélio Vascular/citologia , Endotélio Vascular/imunologia , Endotélio Vascular/metabolismo , Glucosídeos/uso terapêutico , Produtos Finais de Glicação Avançada/metabolismo , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/imunologia , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Inflamassomos/antagonistas & inibidores , Inflamassomos/imunologia , Inflamassomos/metabolismo , Inflamação/imunologia , Inflamação/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/imunologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/imunologia , Fenóis/uso terapêutico , RNA-Seq , Espécies Reativas de Oxigênio/imunologia , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologia , Fator de Transcrição RelA/imunologia , Fator de Transcrição RelA/metabolismoRESUMO
OBJECTIVE: To investigate the effects of capsule of Shenshuai Yangzhen, a preparation of traditional Chinese medicine, on malnutrition rats with chronic renal failure (CRF). METHODS: SD rats received 5/6 nephrectomy for preparation of CRF models, and fed 4% casein at the same time. Observed when malnutrition began. Those consistents with malnutrition of CRF condition were randomized into model control group, Ketosteril group, Shenshuai Yangzhen group, and normal control group. After 4-weeks treatment as indicated, The blood parameters, like blood serum albumin (ALB), type-1 insulin like growth factor (IGF-1), total cholesterol (TC), triglyeride (TG), urea nitrogen (BUN), serum creatinine (Scr), haemoglobin (Hb), 24 hour urineprotein (24hUpr) and weight were determined. Nephrotic tissue was observed by microscope (included HE and PAS). RESULTS: Malnutrition situation in CRF rats began at the end of 10-weeks. After 4-weeks treatment, weight in Shenshuai Yangzhen group were higher significantly (P < 0.05). Compared with model control group, blood serum BUN (P < 0.05), SCr (P < 0. 05) and 24h Upr (P < 0.001) in Shenshuai Yangzhen group were significantly lower with substantially elevated blood serum ALB, Hb, IGF-1 (P < 0.01; P < 0.001; P < 0.001, respectively). Pathology of Shenshuai Yangzhen group was a meliorated significantly after treated. CONCLUSION: Capsule of Shenshuai Yangzhen has a possible therapic effect on improving malnutrition in rats with renal insufficiency.
Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Falência Renal Crônica/complicações , Desnutrição/tratamento farmacológico , Fitoterapia , Plantas Medicinais/química , Animais , Nitrogênio da Ureia Sanguínea , Cápsulas , Colesterol/sangue , Creatinina/sangue , Combinação de Medicamentos , Medicamentos de Ervas Chinesas/farmacologia , Hemoglobinas/análise , Rim/patologia , Masculino , Desnutrição/sangue , Desnutrição/etiologia , Nefrectomia , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Albumina Sérica/análise , Triglicerídeos/sangueRESUMO
Chronic kidney disease (CKD) is a global public health problem with a high mortality and case fatality, and multiplies the risk for complications of cardiovascular disease and huge medical costs. Integrated traditional Chinese and Western medicine is effective in preventing and treating CKD with less adverse, however there are a lot of questions that we don't know well. Strategies and approaches of the integrated traditional Chinese and Western medicine in preventing and treating CKD are: (1) enhance the study of optimized scheme for single entity; (2) accelerate the step of new drug exploitation; (3) augment the study of action mechanism of traditional Chinese medicine in treating CKD; (4) strengthen the study of the mechanism of Chinese crude drug which is poisonous to kidney and its prevention and cure; (5) utilize the systems biology to study the essence of kidney; (6) establish a guideline for integrated traditional Chinese and Western medicine in prevention and treatment of CKD; (7) preach up the general knowledge of CKD, pay attention to mass screening and early prevention of CKD. It is expected to improve diagnosis and treatment of CKD with integrated traditional Chinese and Western medicine by carrying out these strategies and methods mentioned above.
Assuntos
Medicina Integrativa , Nefropatias/tratamento farmacológico , Medicina Tradicional Chinesa , Fitoterapia , Doença Crônica , Diagnóstico Diferencial , Medicamentos de Ervas Chinesas/uso terapêutico , Humanos , Falência Renal Crônica/tratamento farmacológico , Biologia de Sistemas/métodosRESUMO
BACKGROUND AND OBJECTIVES: Evidence suggests that dietary fiber benefits patients with chronic kidney disease (CKD); however, this conclusion requires further validation. In this study, we examined the effects of dietary fiber on kidney function, inflammation, indoxyl sulfate, nutritional status, and cardiovascular risk in patients with advanced CKD. METHODS AND STUDY DESIGN: We performed linear regressions to assess the association between dietary fiber intake and CKD parameters. The aforementioned parameters were compared over an 18-month follow- up period. Kaplan-Meier analysis was used to investigate the association between fiber intake and Cardiac vascular disease (CVD). RESULTS: In total, 157 patients were included in this study. Dietary fiber and inflammatory indices were associated (interleukin [IL]-6: ß=-0.024, p=0.035). The differential estimated glomerular filtration rate (ΔeGFR) as well as levels of C-reactive protein, IL-6, indoxyl sulfate, and serum cholesterol in the higher fiber intake (>=25 g/day) group were lower than those in the lower fiber intake (<25 g/day) group (p<0.05). Differences in IL-6 and indoxyl sulfate levels were more significant in patients in the higher protein intake group (p<0.05). Dietary fiber intake may be a protective factor associated with CVD (hazard ratio=0.537 and 0.305- 0.947). The protein nutritional status was not different between the two groups (p>0.05). CONCLUSIONS: Our results suggest that increasing fiber intake can retard the decrease in the eGFR; can reduce the levels of proinflammatory factors, indoxyl sulfate, and serum cholesterol; and is negatively associated with cardiovascular risk, but does not disrupt the nutritional status of patients with CKD.
Assuntos
Doenças Cardiovasculares/etiologia , Fibras na Dieta/administração & dosagem , Fibras na Dieta/farmacologia , Insuficiência Renal Crônica/patologia , Adulto , Estudos de Coortes , Proteínas Alimentares/administração & dosagem , Feminino , Humanos , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Estado NutricionalRESUMO
OBJECTIVE: To investigate the effect of Shenkangwan on secretion of transforming growth factor-beta1 (TGF-beta1) and Fibronectin (FN) and their mRNA expression in rat mesangial cells (MC)cultured by high glucose and explore its molecule mechanism on treating diabetic nephropathy (DN). METHODS: Rat MC were cultured in vivo and divided into six groups: low glucose group, high glucose group, normal rat serum group, Capoten tablets group, Shenkangwan high dose group and low dose group. Seventy two hours later, the secretion of TGF-beta1 and FN in MC were detected by enzyme linked immunosorbent assay (ELISA). The expression of TGF-beta1, and FN mRNA were determined by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: High gluscose could induce secretion of TGF-beta1 and FN in MC and increase expression of TGF-beta1 and FN mRNA. While Shenkangwan could repress these effects. CONCLUSION: Shenkangwan can suppress the secretion of the extracellular matrix in MC via TGF-beta1 signal way.
Assuntos
Nefropatias Diabéticas/patologia , Medicamentos de Ervas Chinesas/farmacologia , Fibronectinas/biossíntese , Células Mesangiais/efeitos dos fármacos , Fator de Crescimento Transformador beta/biossíntese , Animais , Células Cultivadas , Meios de Cultura , Nefropatias Diabéticas/prevenção & controle , Medicamentos de Ervas Chinesas/administração & dosagem , Matriz Extracelular/efeitos dos fármacos , Matriz Extracelular/metabolismo , Fibronectinas/genética , Glucose/farmacologia , Masculino , Células Mesangiais/metabolismo , Plantas Medicinais/química , RNA Mensageiro/biossíntese , Ratos , Ratos Wistar , Fator de Crescimento Transformador beta/genéticaRESUMO
OBJECTIVE: To investigate the effects of Shenkang pill on renal function and extracellular matrix secretion on the diabetic rats. METHOD: The diabetic rat models were induced by intraperitoneal injection of streptozotocin (STZ) and randomly divided into 3 groups' model control group; Capoten group and Shenkangwan group. Some normal other rats were used as normal control group. All rats were treated with corresponding drugs for 8 weeks. During and after the treatment, the general state, blood and urine glucose levels, excretion rate of the 24 hour urine protein and albumin, serum creatinine and blood urea nitrogen contents, kidney weight and relative kidney weight were measured. The mRNA of fibronectin(FN) in the kidney also detected by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR). RESULT: Diabetes mellitus and renal lesions occurred in the three model groups. The expression of FN mRNA of the kidney in diabetic rats increased obviously. Shenkang pill could improve the general state and renal function of the diabetic rats, decrease the blood glucose levels and the excretion rate of the 24 hour urine protein and albumin, reduce the expression of FN mRNA in kidney. CONCLUSION: Shenkang pill has a certain protective effect on the diabetic kidney.
Assuntos
Nefropatias Diabéticas/metabolismo , Medicamentos de Ervas Chinesas/farmacologia , Fibronectinas/biossíntese , Plantas Medicinais , Animais , Glicemia/metabolismo , Nitrogênio da Ureia Sanguínea , Nefropatias Diabéticas/induzido quimicamente , Nefropatias Diabéticas/patologia , Combinação de Medicamentos , Medicamentos de Ervas Chinesas/isolamento & purificação , Fibronectinas/genética , Hemoglobinas Glicadas/metabolismo , Rim/metabolismo , Masculino , Plantas Medicinais/química , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Distribuição Aleatória , Ratos , Ratos Wistar , EstreptozocinaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Existing evidences suggest that Radix Astragali and its polysaccharides composition (APS) can improve muscle mass, but the mechanisms need more research. AIM OF THE STUDY: In this study, we aimed to examine the effects of APS on muscle wasting at molecular level in 5/6 nephrectomised rats. MATERIALS AND METHODS: We performed 5/6 nephrectomy or sham operation in 160 6-week-old Sprague-Dawley rats, and feed animals with or without 2% APS for 155 days. After treatment, we compared the change of weight, muscle fibre, protein metabolism, pro-inflammatory factors (TNF-α, IL-15, CRP) and oxidative factors (MDA, SOD) among each group. In addition, we detected the Akt/mTOR, ubiquitin proteasome, autophagy signalling and AA transporters in vivo and in vitro. RESULTS: Data in vivo show 2% APS could alleviate weight loss and improve protein metabolism in nephrectomised rats. The levels of serum pro-inflammatory factors and oxidative factors were restored by APS treatment. In molecular levels, APS restored Akt/mTOR, MAFbx, MuRF1, Atg7, LC3B-II/LC3B-I and SLC38A2 which changed in nephrectomised rats. Data in vitro show the optimal dose of APS is 0.2mg/mL, and SLC38A2 siRNA attenuated the effects of 0.2mg/mL APS on atrophy and autophagy. CONCLUSIONS: Our results suggested APS could improve muscle wasting through Akt/mTOR, ubiquitin proteasome and autophagy signalling, and SLC38A2 may be one of potential targets.
Assuntos
Astrágalo/química , Nefrectomia , Polissacarídeos/farmacologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Serina-Treonina Quinases TOR/metabolismo , Ubiquitina/metabolismo , Animais , Autofagia/fisiologia , Linhagem Celular , Regulação da Expressão Gênica/efeitos dos fármacos , Inativação Gênica , Masculino , Músculo Esquelético/patologia , Mioblastos/efeitos dos fármacos , Polissacarídeos/química , Complexo de Endopeptidases do Proteassoma/genética , Complexo de Endopeptidases do Proteassoma/metabolismo , Proteínas Proto-Oncogênicas c-akt/genética , Ratos , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/fisiologia , Serina-Treonina Quinases TOR/genética , Ubiquitina/genéticaRESUMO
This paper studied the chronic fatigue induced by excessive exercise and the restoration effects of Astragalus polysaccharides (APS) on mitochondria. In vivo, we found that excessive exercise could cause oxidative stress statue which led to morphological and functional changes of mitochondria. The changes, including imbalance between mitochondria fusion-fission processes, activation of mitophagy, and decrease of PGC-1α expression, could be restored by APS. We further confirmed in vitro, and what is more, we found that APS may ameliorate mitochondrial dysfunction through Sirt1 pathway. Based on the results, we may figure out part of the molecular mechanism of mitochondrial amelioration by APS.
Assuntos
Astrágalo/química , Dinâmica Mitocondrial/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Polissacarídeos/farmacologia , Animais , Autofagia/efeitos dos fármacos , Autofagia/genética , Suplementos Nutricionais , Regulação da Expressão Gênica/efeitos dos fármacos , Masculino , Camundongos Endogâmicos BALB C , Microscopia de Fluorescência , Estresse Oxidativo/genética , Condicionamento Físico Animal , Resistência Física , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Proteína Supressora de Tumor p53/metabolismoRESUMO
OBJECTIVE: To investigate the effects of the Yifuning soft gelatin capsules(YFN)on estrogen receptor (ER) and nitric oxide (NO) levels in ovariectomized rats. METHOD: Fifty female mature sprague-dawley rats were randomly divided into 5 groups: normal control; model control; diethylstilbestrol tablets (DT); YFN (high dose and low dose). After 4 weeks' treatment, the serum E2 levels were detected by radioimmunoassay. The estrogen receptor (ER) levels of uterus and artery were detected with method of radioligand binding assay of receptor (RBAR). The uterus pathologic changes were investigated with light microscope. NO and NOS levels of the artery and the uterus index were detected too. RESULT: YFN can obviously improve serum E2, increase index and ER of uterus (P < 0.01 or P < 0.05), and ameliorate the uterus' pathologic changes in OVX rats. It also can increase the artery' ER, NO and NOS levels. CONCLUSION: YFN can cure the climacteric syndrome and prevent the cardiovascular disease after post-menopause.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Materia Medica/farmacologia , Óxido Nítrico/metabolismo , Receptores de Estrogênio/metabolismo , Útero/patologia , Animais , Aorta/metabolismo , Cápsulas , Curcuma/química , Combinação de Medicamentos , Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/isolamento & purificação , Estradiol/sangue , Feminino , Materia Medica/administração & dosagem , Materia Medica/isolamento & purificação , Óxido Nítrico Sintase/metabolismo , Ovariectomia , Oviductos/química , Plantas Medicinais/química , Rana temporaria , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Útero/metabolismoRESUMO
The aim is to study the effects and underlying mechanisms of astragalus polysaccharide (APS) on the peroxide-induced injury in C2C12 myoblasts in vitro. Cell viability in the presence or absence of APS was detected by the methyl thiazolyl tetrazolium colorimetric assay. The autophagosomes were observed by electron microscopy to examine the influence of APS on autophagy caused by H2O2 in C2C12 cells, and the percentage of apoptosis cells was measured by flow cytometry. To further confirm the effect of H2O2 on C2C12 cells, the protein expression of LC3 and RARP, which are the markers of autophagy and apoptosis, respectively, was analyzed by Western blot, as well as the expression levels of p-p70S6K, p70S6K, Bcl-2, Bax, cyto-C, and Caspase-3, to reveal the underlying mechanisms. We observed multiple effects of APS on C2C12 functionality. APS treatment of C2C12 cells at 1 mg/mL reduced cell viability to less than 70 %, and analysis by electron microscopy revealed that APS also reduced the number of H2O2-induced autophagosome formation. Similarly, APS abated the H2O2-mediated increase in cell apoptosis, which was accompanied by the inhibition of LC3 II and RARP that are normally upregulated by H2O2. The expression of p-p70S6K and p70S6K, however, remained unchanged in C2C12 cells in the Control, H2O2 and H2O2 + APS groups. In addition, APS promoted the expression of protein Bcl-2 in H2O2-treated C2C12 cells, but did not change Bax, thus reducing the Bax/Bcl-2 ratio that in turn prevented the release of cytochrome c and the activation of caspase-3. APS inhibits the autophagy and apoptosis induced by peroxide injury in C2C12 myoblasts through two independent signaling pathways: the mTOR-independent pathway for the inhibition of autophagy, and the caspase-3-dependent pathway for the suppression of apoptosis.
Assuntos
Apoptose/efeitos dos fármacos , Astrágalo/metabolismo , Autofagia/efeitos dos fármacos , Peróxido de Hidrogênio/toxicidade , Polissacarídeos/farmacologia , Animais , Western Blotting , Caspase 3/metabolismo , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Citocromos c/metabolismo , Camundongos , Microscopia Eletrônica , Proteínas Associadas aos Microtúbulos/metabolismo , Mioblastos/citologia , Mioblastos/efeitos dos fármacos , Mioblastos/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Proteínas Quinases S6 Ribossômicas 70-kDa/metabolismo , Transdução de Sinais/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismo , Proteína X Associada a bcl-2/metabolismoRESUMO
OBJECTIVE: To investigate the therapeutic effect of Shenshuai Yangzhen capsule, a preparation of traditional Chinese medicine, on lipid metabolism disorder in rats with chronic renal failure (CRF) and explore its mechanism. METHODS: Fifty male SD rats received 5/6 nephrectomy for preparation of CRF models and were randomized into CRF group, gemfibrozil group, high-, moderate- and low-dose Shenshuai Yangzhen groups, and normal control group. After 4-week treatment as indicated, myocardial lipoprotein lipase messenger RNA (LPL mRNA) level were measured by RT-PCR in rats with surgically induced renal failure (two-stage subtotal nephrectomy). The blood lipid parameters in CRF rats were also determined. RESULTS: Compared with those in the CRF group, the plasma total cholesterol, triglyceride, low-density lipoprotein cholesterol and very-low-density lipoprotein cholesterol concentrations in the treatment groups were significantly lower with substantially elevated plasma high-density lipoprotein cholesterol and LPL gene expression. No significant differences were noted between different dose groups of Shenshuai Yangzhen. CONCLUSION: Shenshuai Yangzhen capsule can regulate blood lipid levels in rats with renal insufficiency possibly by enhancing LPL gene expression.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Falência Renal Crônica/metabolismo , Lipídeos/sangue , Animais , Cápsulas , Lipase Lipoproteica/genética , Masculino , Miocárdio/enzimologia , RNA Mensageiro/análise , Ratos , Ratos Sprague-DawleyRESUMO
This article reviews the current status of progress in the research of renal interstitial fibrosis therapy using traditional Chinese herbal medicine, which exerts its therapeutic effect through inhibiting cytokine expression and fibroblast proliferation, inducing apoptosis of the renal myofibroblasts and other mechanisms.
Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Glomerulosclerose Segmentar e Focal/tratamento farmacológico , Apoptose , Divisão Celular , Fibroblastos/citologia , Fibrose , Expressão Gênica/efeitos dos fármacos , Glomerulosclerose Segmentar e Focal/metabolismo , Glomerulosclerose Segmentar e Focal/fisiopatologia , Medicina Tradicional Chinesa , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta1RESUMO
Muscle atrophy poses a serious concern to patients inflicted with inflammatory diseases. There is now increasing evidence which suggests a vital role for tumor necrosis factor alpha (TNF-α) in muscle pathology associated with impairment of differentiation and muscle wasting. Resveratrol has been an ascribed inhibitory effect on glucocorticoid-induced muscle atrophy in vitro, but the influence of resveratrol on the growth of C2C12 myotubes exposed to TNF-α remains unclear. The present study aimed to investigate the involvement of TNF-α in the regulation of skeletal muscle hypertrophy and atrophy, and the possibility to interfere with such modulations by means of resveratrol supplementation. For this purpose, C2C12 myotubes were treated with TNF-α in the presence or absence of resveratrol. Myotube treatment with TNF-α contributes to both hyperexpression of the muscle-specific ubiquitin ligase MAFbx and MuRF1, and these alterations are linked to a decrease of anabolic targets (Akt, mTOR, p70S6k and 4E-BP1) and an increase of catabolic targets (FoxO1, FoxO3a, MAFbx and MuRF1). Resveratrol supplementation effectively counteracts TNF-α induced muscle protein loss and reverses declining expression of Akt, mTOR, p70S6K, 4E-BP1and FoxO1, but exerts no influence of FoxO3a expression. Our study demonstrates that resveratrol can reverse the muscle cell atrophy caused by TNF-α through regulation of the Akt/mTOR/FoxO1 signaling pathways, followed by inhibition of the atrophy-related ubiquitin ligase. Our findings suggested that resveratrol could represent a possible strategy to improve muscle mass.