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Objective:To investigate the role of indoxyl sulfate (IS) in myocardial hypertrophy and fibrosis through organic anion transporter-3 (OAT-3) and oxidative stress in H9C2 cells.Methods:Rat myocardial cells (H9C2) were cultured and divided into four groups: Control group, IS group, siRNA negative control group (siOAT-3), and siOAT-3+ IS group. The control group was cultured routinely without IS stimulation. The IS group was stimulated with 250 μmol IS. The siRNA negative control group was transfected with 20 nmol/L OAT-3 siRNA, and the siOAT-3+ IS group was transfected with OAT-3 siRNA 48 hours later, then stimulated with IS for 24 hours. Real-time polymerase chain reaction (RT-PCR) was used to detect and analyze the mRNA expression levels of OAT-3, NADPH oxidase-4 (Nox-4), antioxidant proteins [nuclear factor E2 related factor 2 (Nrf-2), heme oxygenase 1 (HO-1)], myocardial hypertrophy markers [atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), β-myosin heavy chain gene (β-MHC)], and fibrosis markers [transforming growth factor β1 (TGF-β1), Smad homologous protein 3 (Smad-3), collagen type Ⅰ (Collagen Ⅰ)] in each group. H9C2 cells were further divided into three groups: Control group, IS group, and N-acetylcysteine (NAC) group. The NAC group was pre-treated with the antioxidant NAC before stimulation with 250 μmol IS for 24 hours. RT-PCR was used to detect the mRNA expression levels of the above indicators.Results:The mRNA expression level of OAT-3 in the IS group was significantly higher than that in the control group, while the mRNA expression levels of OAT-3 in the siOAT-3 group and the siOAT-3+ IS group were significantly lower than those in the IS group (all P<0.001). Compared with the Control group, the mRNA relative expression levels of Nox-4, ANP, BNP, β-MHC, TGF-β1, Smad-3, and Collagen Ⅰ in H9C2 cells of the IS group were significantly increased (all P<0.001), while the mRNA expression levels of Nrf-2 and HO-1 were significantly decreased (all P<0.001). Compared with the IS group, the mRNA relative expression levels of Nox-4, ANP, BNP, β-MHC, TGF-β1, Smad-3, and Collagen Ⅰ in H9C2 cells of the siOAT-3 group and the siOAT-3+ IS group were significantly lower (all P<0.001), while the mRNA expression levels of Nrf-2 and HO-1 were significantly increased (all P<0.001). Pre-treatment with NAC significantly inhibited the high expression of Nox-4, ANP, BNP, β-MHC, TGF-β1, Smad-3, and Collagen Ⅰ mRNA induced by IS (all P<0.001), while significantly increasing the mRNA expression levels of Nrf-2 and HO-1 (all P<0.01). Conclusions:IS promotes myocardial hypertrophy and fibrotic factor overexpression in H9C2 cells through OAT-3 and oxidative stress.
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Objective To investigate preoperative myocardial enzymes and realative influencing factors in Stanford B type aortic dissection.Methods From Jan.2004 to Sep.2013,151 consecutive patients with Stanford type B aortic dissection were admitted to hospital,aged from 31 to 76 average:(51.51 ± 10.90)year sold.Ninty-five healthy people with similar age and sex were taken as the control group.Fasting venous blood collected more than 12h was collected,myocardial enzymes indexes such as CK,CKMB,LDH,HBDH were measured by Roche modular automatic biochemical analysis system.Primary entry tear and extent of aortic dissection was measured by Toshiba Aquilion ONE 320 slice CT.Degree of aortic valve insufficiency was measured by Philips Sonos 5500 Color Doppler ultrasonic diagnostic apparatus.Results Compared with control group,the level of myocardial enzymes (LDH,HBDH) of aortic dissection group increased significantly(P < 0.01).part myocardial enzymes indexes(CK,LDH,HBDH) of acute stage group existed difference(P < 0.05).Myocardial enzymes indexes only CK existed difference between acute stage group and subacute stage group and chronic stage group(F =18.72,P =0.000),no difference between subacute stage group and chronic stage group.LDH,HBDH of each sub group of aortic dissection group were higher than that of control group,P < 0.01.Trough correlation analysis,CK negatively correlated with disease course of aortic dissection and patients sex [(r =-0.446 ; P =0.000) ; (r =-0.303 ; P =0.000)],CKMB negatively correlated patients sex [(r=-0.203;P=0.020)],LDH negatively correlated with patients sex [(r =-0.171 ;P =0.049)],positively with left ventricular end-diastolic diameter and left ventricular end-systolic diameter [(r =0.202 ; P =0.022) ; (r =0.271 ; P =0.002)].HBDH positively correlated with left ventricular enddiastolic diameter and left ventricular end-systolic diameter [(r =0.385 ;P =0.002) ; (r =0.515 ; P =0.000)],negatively with degree of aortic insufficiency [(r =-0.528 ;P =0.006)].Conclusions Myocardial enzymes rise in preoperative Stanford B aortic dissection,more representing skeletal muscle injury.Be affected by stage of aortic dissection,lower limb skeletal muscle injury aggravates more seriously in acute stage group persists entering the sub acute stage.