RESUMEN
Background@#Focal segmental glomerulosclerosis (FSGS) is a kidney disease that is commonly associated with proteinuria and the progressive loss of renal function, which is characterized by podocyte injury and the depletion and collapse of glomerular capillary segments. The pathogenesis of FSGS has not been completely elucidated; however, recent advances in molecular genetics have provided increasing evidence that podocyte structural and functional disruption is central to FSGS pathogenesis. Here, we identified a patient with FSGS and aimed to characterize the pathogenic gene and verify its mechanism.@*Methods@#Using next-generation sequencing and Sanger sequencing, we screened the causative gene that was linked to FSGS in this study. The patient's total blood RNA was extracted to validate the messenger RNA (mRNA) expression of coenzyme Q monooxygenase 6 (COQ6) and validated it by immunohistochemistry. COQ6 knockdown in podocytes was performed in vitro with small interfering RNA, and then, F-actin was determined using immunofluorescence staining. Cell apoptosis was evaluated by flow cytometry, the expression of active caspase-3 was determined by Western blot, and mitochondrial function was detected by MitoSOX.@*Results@#Using whole-exome sequencing and Sanger sequencing, we screened a new causative gene, COQ6, NM_182480: exon1: c.G41A: p.W14X. The mRNA expression of COQ6 in the proband showed decreased. Moreover, the expression of COQ6, which was validated by immunohistochemistry, also had the same change in the proband. Finally, we focused on the COQ6 gene to clarify the mechanism of podocyte injury. Flow cytometry showed significantly increased in apoptotic podocytes, and Western blotting showed increases in active caspase-3 in si-COQ6 podocytes. Meanwhile, reactive oxygen species (ROS) levels were increased and F-actin immunofluorescence was irregularly distributed in the si-COQ6 group.@*Conclusions@#This study reported a possible mechanism for FSGS and suggested that a new mutation in COQ6, which could cause respiratory chain defect, increase the generation of ROS, destroy the podocyte cytoskeleton, and induce apoptosis. It provides basic theoretical basis for the screening of FSGS in the future.
Asunto(s)
Adolescente , Animales , Femenino , Humanos , Ratones , Apoptosis , Genética , Fisiología , Línea Celular , Citometría de Flujo , Glomeruloesclerosis Focal y Segmentaria , Genética , Inmunohistoquímica , Mutación , Genética , Podocitos , Metabolismo , Patología , ARN Mensajero , Genética , ARN Interferente Pequeño , Genética , Metabolismo , Ubiquinona , Genética , MetabolismoRESUMEN
<p><b>OBJECTIVE</b>To observe the effect of Compound Shenhua Tablet (, SHT) on the sodium-potassium- exchanging adenosinetriphosphatase (Na(+)-K(+)-ATPase) in the renal tubular epithelial cells of rats with acute ischemic reperfusion and to investigate the mechanisms underlying the effects of SHT on renal ischemic reperfusion injury (RIRI).</p><p><b>METHODS</b>Fifty male Wistar rats were randomly divided into the sham surgery group, model group, astragaloside group [150 mg/(kg·d)], SHT low-dose group [1.5 g/(kg·d)] and SHT high-dose group [3.0 g/(kg·d)], with 10 rats in each group. After 1 week of continuous intragastric drug administration, surgery was performed to establish the model. At either 24 or 72 h after the surgery, 5 rats in each group were sacrificed, blood biochemistry, renal pathology, immunoblot and immunohistochemical examinations were performed, and double immunofluorescence staining was observed under a laser confocal microscope.</p><p><b>RESULTS</b>Compared with the sham surgery group, the serum creatinine (SCr) and blood urea nitrogen (BUN) levels were significantly increased, Na(+)-K(+)-ATPase protein level was decreased, and kidney injury molecule-1 (KIM-1) protein level was increased in the model group after the surgery (P<0.01 or P<0.05). Compared with the model group, the SCr, BUN, pathological scores, Na(+)-K(+)-ATPase, and the KIM-1 protein level of the three treatment groups were significantly improved at 72 h after the surgery (P<0.05 or P<0.01). And the SCr, BUN of the SHT low- and high-dose groups, and the pathological scores of the SHT high-dose group were significantly lower than those of the astragaloside group (P<0.05). The localizations of Na(+)-K(+)-ATPase and megalin of the model group were disrupted, with the distribution areas overlapping with each other and alternately arranged. The severity of the disruption was slightly milder in three treatment groups compared with that of the model group. The results of immunofluorescence staining showed that the SHT high-dose group had a superior effect as compared with the astragaloside group and the SHT low-dose group.</p><p><b>CONCLUSIONS</b>The SHT effectively alleviated RIRI caused by ischemic reperfusion, promoted the recovery of the polarity of renal tubular epithelial cells, and protected the renal tubules. The therapeutic effects of SHT were superior to those of astragaloside as a single agent.</p>
Asunto(s)
Animales , Masculino , Ratas , Enfermedad Aguda , Nitrógeno de la Urea Sanguínea , Moléculas de Adhesión Celular , Metabolismo , Cromatografía Liquida , Creatinina , Sangre , Medicamentos Herbarios Chinos , Farmacología , Usos Terapéuticos , Técnica del Anticuerpo Fluorescente , Immunoblotting , Pruebas de Función Renal , Túbulos Renales , Patología , Proteína 2 Relacionada con Receptor de Lipoproteína de Baja Densidad , Metabolismo , Ratas Wistar , Daño por Reperfusión , Quimioterapia , Patología , Saponinas , ATPasa Intercambiadora de Sodio-Potasio , Metabolismo , Coloración y Etiquetado , ComprimidosRESUMEN
<p><b>OBJECTIVE</b>To investigate the effect of the Uremic Clearance Granule (UCG, ), a Chinese patent medicine, on tubular epithelial-to-mesenchymal transition (EMT) in a unilateral ureteral obstruction (UUO) model in vivo and transforming growth factor (TGF)-β1 induced EMT of HK-2 cells in vitro.</p><p><b>METHODS</b>In vivo study, 50 Sprague Dawley rats were divided into three groups: a sham operation group (n=10), a UUO group (n=20), and a UUO with UCG treatment group (n=20). The UCG was given at a dose of 4.5 g/kg body weight per day by gavage after surgery. In vitro study, HK-2 cells were cultured in 10% fetal bovine serum (FBS), 10% healthy rat serum, 10% FBS and TGF-β1 (10 ng/mL), 10% healthy rat serum and TGF-β1, or 10% rat serum containing the uremic clearance granule and TGF-β1. The expression of the epithelial marker E-cadherin and the mesenchymal markers vimentin and α-smooth muscle actin (α-SMA) in kidney tissues and HK-2 cells were investigated by Western blot analysis and immunofluorescence staining.</p><p><b>RESULTS</b>The rats of the UUO group showed obvious tubulointerstitial fibrosis, compared with the sham operation group rats. Tubulointerstitial fibrosis score was reduced by 17.5%±1.1% at day 7 and by 20.0%±1.2% at day 14 in the UCG-treated group, compared with the UUO group. The UCG could maintained expression of E-cadherin and suppressed expression of vimentin and α-SMA in kidney tissues of UUO rats at days 7 and 14, as determined by Western blot analysis and immunofluorescence staining. Rat serum containing the UCG partially inhibited TGF-β1-induced fibroblast phenotype of HK-2 cells and maintained the epithelial morphology of HK-2 cells in vitro. This occurred partially through a reduction of vimentin expression and an increase of E-cadherin expression.</p><p><b>CONCLUSION</b>These results suggest that the UCG prevents tubular EMT and may be a promising agent for treating tubulointerstitial fibrosis.</p>
Asunto(s)
Animales , Masculino , Ratas , Sangre , Western Blotting , Línea Celular , Medios de Cultivo , Transición Epitelial-Mesenquimal , Técnica del Anticuerpo Fluorescente , Técnicas In Vitro , Túbulos Renales , Patología , Ratas Sprague-Dawley , Uremia , PatologíaRESUMEN
<p><b>BACKGROUND</b>It has been suggested that glycated hemoglobin (HbA1c) underestimate the actual glycemic control levels in maintenance hemodialysis (MHD) patients, because of anemia and the using of erythropoietin (EPO); it was recommended that glycated albumin (GA) should be an alternative marker. Therefore, the assessment performances of glycemic control were compared between GA and HbA1c in this research by referring to mean plasma glucose (MPG) in diabetes mellitus (DM) patients undergoing MHD or not.</p><p><b>METHODS</b>MPG was calculated according to the data registered at enrollment and follow-up 2 months later and corresponding HbA1c, albumin (ALB), GA, etc. were measured in 280 cases. A case-control study for comparing GA and HbA1c was done among the groups of MHD patients with DM (n=88) and without DM (NDM; n=90), and non-MHD ones with DM (n=102) using MPG for an actual glycemic control standard.</p><p><b>RESULTS</b>In these 3 groups, only for DM patients' (whether undergoing MHD or not), GA and HbA1c correlated with MPG significantly (P < 0.01). Through linear regression analysis, it could be found that the regression curves of GA almost coincided in MHD and non-MHD patients with DM, because the intercepts (2.418 vs. 2.329) and slopes (0.053 vs. 0.057) were very close to each other. On the contrary, regression curves of HbA1c did not coincide in the two groups, because variance of the slopes (0.036 vs. 0.052) were relatively large. Through comparing receiver operating characteristic (ROC) areas under the curve (AUC), it could be understood that the assessment performances of GA and HbA1c in MHD patients were lower than those in non-MHD ones, and assessment performance of HbA1c in MHD patients was better than GA (P < 0.05). In addition, the effects of Hb and EPO dose on HbA1c, or that of ALB on GA were unobvious in our study.</p><p><b>CONCLUSIONS</b>Actual glycemic control level in MHD patients with DM may be underestimated by HbA1c, and it could be avoided by GA; however, glycemic evaluating performance of HbA1c may be still better than that of GA. Therefore, HbA1c should not be replaced completely although GA can be used as a choice to monitor glycemic level.</p>
Asunto(s)
Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Biomarcadores , Metabolismo , Estudios de Casos y Controles , Diabetes Mellitus Tipo 2 , Metabolismo , Terapéutica , Hemoglobina Glucada , Metabolismo , Diálisis Renal , Albúmina Sérica , MetabolismoRESUMEN
The aim of this study was to investigate the immunological function regulated by Fufang Hongjingtian capsule (HJT) in mice. The mice were given ig HJT 25, 250 and 750 mg/kg, once daily, for 30 - 38 d, respectively. The mice in control group were given ig corresponding solvent. After the last time of administration, the immunological parameters of the mice were measured. The results showed that compared with negative control group, the delayed type hypersensitivity, spleen lymphocyte proliferation and number of spleen IgM antibody forming cells increased in HJT groups. In conclusion the HJT has the effect to improve the immunological functions of mice.
Asunto(s)
Animales , Femenino , Ratones , Medicamentos Herbarios Chinos , Farmacología , Inmunoglobulina M , Alergia e Inmunología , Recuento de Linfocitos , Linfocitos , Biología Celular , Ratones Endogámicos , Rhodiola , Bazo , Biología Celular , Alergia e InmunologíaRESUMEN
The aim of this study was to investigate the protective effects of compound tianpupian (TPP) and its compositions against oxidative damage in mouse erythrocytes. The protective effect of TPP and its compositions against the red cell hemolysis induced by (2)O(2) or auto-oxidation were observed by scanning electron microscopy and spectrophotomety. The result indicated that compound TPP and all of its four components including extract of Rhodiola sachalinensis, Grape Seed Extract proanthocyanidins, Acanthopanax senticosus extract, and tea polyphenols had significant inhibitory activities for the oxidative damage of mouse erythrocytes, out of which the Grape seed extract proanthocyanidins showed the maximal protective effect. It is concluded that compound TPP can protect erythrocytes against oxidative stress and can be used as a valuable Chinese traditional medicine.