Extracellular cysteine/cystine redox potential affects mitochondrial func-tion in NAFLD LO2 cells / 中国病理生理杂志

AIM:To investigate the effects of extracellular cysteine/cystine redox potential (EhCys/CySS) on the mitochondrial function of nonalcoholic fatty liver disease ( NAFLD) hepatocytes.METHODS:LO2 cells were incuba-ted with EhCys/CySS of the oxidized (0 mV), the normal (-80 mV), or the reduced (-150 mV) status medium, then treated with oleic acid to establish NAFLD model in vitro.DCFH-DA and MitoSOX were used as the fluorescent probes for determining reactive oxygen species (ROS).Apocynin (NADPH oxidase inhibitor), MitoQ10 (mitochondria-targeted an-tioxidant), rotenone (mitochondrial respiratory chain complex I inhibitor) and antimycin A (mitochondrial respiratory chain complex III inhibitor) were used to investigate the sources of ROS.RESULTS:An increase in ROS in LO2 cells by oleic acid was aggravated by the oxidized extracellular EhCys/CySS (0 mV), which was removed by the reduced EhCys/CySS (-150 mV) .ROS generation by 0 mV was significantly eliminated by MitoQ10 .ROS levels were dependent on ex-tracellular Eh Cys/CySS in rotenone treated LO2 cells.A decline of mitochondrial membrane potential in the cells with NAFLD was aggravated by 0 mV and reversed by -150 mV.CONCLUSION:The oxidized extracellular Eh Cys/CySS via inhibitiing of complex I intensifies ROS generation and reducing the mitochondrial membrane potential in the NAFLD hepa-tocytes, which were reversed by reduced Eh Cys/CySS.

The effect of fasudil via Rho/ROCK signaling pathway on the inflammation and fibrosis in human mesangial cells in high glucose medium / 中华内科杂志

Objective To study the effect of fasudil on inhibiting the Rho/ROCK signaling pathway under high glucose in human mesangial cells (HMCs) inflammation and fibrosis. Methods Synchronized HMCs were divided into following groups: (1) Normal glucose control group ( NG, 5. 5 mmol/L glucose) ;(2) High glucose group (HG, 30 mmol/L glucose) ; (3) Mannitol group (Man, 5.5 mmol/L glucose + 24. 5 mmol/L mannitol) ; (4) High glucose + fasudil group ( HG + F, the concentrations of fasudil were 25 ,50 and 100 μmol/L, respectively). Collect the supernatant and cells at 0, 12, 24, 36, 48 and 72 h respectively, and determine the concentration changes of the RhoA, ROCK- Ⅰ, connective tissue growth factor (CTGF)mRNA with real-time PCR method in the cells, then used the ELISA method to check the protein content of the fibronectin ( FN) , CTGF, TNFα in the supernatant. Results ( 1) RhoA, ROCK- Ⅰand CTGF mRNA of the HMCs cultured under the high glucose expressed significantly higher than those in the normal group, and there was certain time-dependence. Besides, there was no statistic significance by comparing Man and NG. (2) Under the high glucose situation, after the fasudil pretreatment with different concentrations and 24 h or 48 h culture with high glucose, RhoA, ROCK- Ⅰ , CTGF mRNA expression was significantly decreased in HG + F, compared with HG, and there was certain concentration-dependence. (3) High glucose increased the FN, CTGF, TNFα protein secretion of HMCs in a time-dependent manner, but normal glucose and mannitol had no such effect. (4) After the fasudil pretreatment with different concentrations and culture with high glucose for 12, 24, 36, 48, 72 h, the FN, CTGF, TNFα protein secretion was significantly reduced compared with HG. Conclusion Fasudil can reduce the secretion of downstream inflammatory factors and cytokines by inhibiting high glucose-activated HMCs Rho/ROCK signaling pathway, and reduce the inflammation and fibrosis of HMCs. This provides a new basis for the therapeutic target in the treatment of diabetic nephropathy.

Clinical significance of serum cardiac troponin T in patients with congestive heart failure / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To determine whether the level of serum cardiac troponin T (cTnT) was increased in patients with congestive heart failure (CHF).</p><p><b>METHODS</b>This study consisted of 265 patients with CHF and 75 healthy people. Serum cTnT was measured by electrochemiluminescence immunoassay using an Elecsys 1010 automatic analyzer.</p><p><b>RESULTS</b>cTnT concentration was 0.181 +/- 0.536 ng/mL in CHF patients and 0.003 +/- 0.001 ng/mL in controls (P < 0.001). Patients were categorized according to the levels of heart function and left ventricular ejection fraction (LVEF). In the first group consisting of 105 patients with LVEF </= 35%, cTnT was 0.311 +/- 0.221 ng/mL. In the second group of 106 patients with LVEF > 35%, cTnT was 0.07 +/- 0.0 5 ng/mL (P < 0.01). In patients with NYHA class I, II, III and IV, cTnT values were 0.062 +/- 0.022 ng/mL, 0.113 +/- 0.121 mg/mL, 0.191 +/- 0.231 mg/ml and 0.384 +/- 0.211 mg/mL, respectively (class I vs class II P > 0.05, class II vs class III P < 0.01, class III vs class IV P < 0.01). A negative correlation was observed between serum cTnT concentration and LVEF in 265 patients with CHF (r = -0.493, P < 0.001).</p><p><b>CONCLUSIONS</b>This study shows that the level of serum cTnT is increased in patients with CHF and that the increased level indicates the severity of CHF.</p>