ABSTRACT
Objective:To study the protective effect and mechanism of Ranae Oviductus protein hydrolysate （ROPH） on the expression of pathway-related proteins in ethanol-induced L-02 cell injury. Method:The ROPH was prepared by compound enzymatic hydrolysis. L-02 cell injury model was induced with 400 mmol·L<sup>-1 </sup>ethanol. Cell viability was detected by cell counting kit-8 （CCK-8） assay. Cell cycle and apoptosis were examined by flow cytometry. JC-1/Hochest staining was employed for qualitative investigation. The expression of related proteins in apoptosis， mitogen-activated protein kinase （MAPK） signaling pathway， and pyroptosis in L-02 cells was detected by Western blot. Result:The results of the CCK-8 assay showed that 400 mmol·L<sup>-1 </sup>ethanol could induce L-02 cell injury within 12 hours. Compared with the blank group， the model group showed decreased viability of L-02 cells （<italic>P</italic><0.01）， elevated percentage of the cell cycle in the G<sub>0</sub>/G<sub>1</sub> phase （<italic>P</italic><0.01）， increased total cell apoptosis rate （<italic>P</italic><0.01）， reduced mitochondrial membrane potential （<italic>P</italic><0.01）， up-regulated expression of apoptosis-related proteins ［B-cell lymphoma-2 （Bcl-2）-associated X protein （Bax）， Cytochrome C （Cyt C）， and cysteine-dependent aspartate specific protease-3 （Caspase-3）］ （<italic>P</italic><0.05， <italic>P</italic><0.01） and MAPK signaling pathway-related proteins ［C-Jun amino-terminal kinase （JNK） and p38 MAPK］ （<italic>P</italic><0.05， <italic>P</italic><0.01）， and potentiated expression of pyrolysis-related proteins Caspase-1 and interleukin-1<italic>β </italic>（IL-1<italic>β</italic>） （<italic>P</italic><0.05）. Compared with the model group， the ROPH treatment group exhibited improved cell cycle arrest （<italic>P</italic><0.05， <italic>P</italic><0.01）， diminished total cell apoptosis rate （<italic>P</italic><0.01）， elevated mitochondrial membrane potential in a dose-dependent manner， down-regulated expression of Bax， Cyt C， and Caspase-3 proteins （<italic>P</italic><0.05， <italic>P</italic><0.01）， up-regulated expression of Bcl-2 protein （<italic>P</italic><0.05， <italic>P</italic><0.01）， and a downward trend in expression of proteins related to MAPK signaling pathway and pyrolysis （<italic>P</italic><0.05， <italic>P</italic><0.01）. Conclusion:ROPH could inhibit oxidative stress-triggered liver injury in ethanol-induced cells by improving mitochondrial membrane potential， reducing the expression of proteins in the mitochondria-mediated apoptosis pathway， and inhibiting the expression of proteins related to the MAPK signaling pathway and pyrolysis pathway to reduce the mitochondrial dysfunction and inflammatory response in ethanol-induced L-02 liver cells and inhibit oxidative stress， thereby exerting a therapeutic role in alcoholic liver injury.
ABSTRACT
A rapid Liquid chromatography-tandem mass spectrometry method was developed for the simultaneous determination of six neurotransmitters in rat plasma,adrenal gland and hypothalamus,including γ-aminobutyric acid (GABA),serotonin(5-HT), tyrosine(L-Tyr), dopamine(DA), norepinephrine(NE) and epinephrine(E). This method was used for the neurotransmitters detection in rat adrenal, blood and hypothalamus. The samples were pre-column derivatized with dansyl chloride,and 5-HTP and CA were used as internal standards. A Thermo C18column (50 mm×3 mm,2.7 μm) was used for sample separation with 0.1% aqueous formic acid solution as phase A and methanol as phase B at a flow rate of 0.2 mL/min, and the injection volume was set at 2 μL. The detection was carried out with multi-reaction monitoring(MRM) in electron spray ionization (ESI) positive mode. The linear ranges for detection of GABA,5-HT, L-Tyr, DA, NE and E were 0.26-620.80 μmol/L,0.03-11.20 μmol/L,1.20-88.00 μmol/L,0.03-41.02 μmol/L, 0.01-47.20 μmol/L, 0. 01-90. 24 μmol/L, respectively, with recoveries varying from 91. 16% to 116.20%. This method can detect the neurotransmitters rapidly and accurately, providing a platform for the determination of neurotransmitters in rat plasma, adrenal gland and hypothalamus in pharmacological experiments.
ABSTRACT
<p><b>OBJECTIVE</b>To determine serum carnitine levels in patients with liver diseases and to investigate their significance.</p><p><b>METHODS</b>25 patients with acute viral hepatitis, 34 with chronic viral hepatitis, 22 with post hepatitis cirrhosis with normal renal function, 9 with post hepatitis cirrhosis but with renal disfunction, and 40 healthy subjects (serving as controls) were enrolled in this study. An enzymatic cycling method was used to determine the serum free carnitine levels.</p><p><b>RESULTS</b>The serum free carnitine level was (48.3+/-10.2)micromol/L in the healthy control group. It was (35.2+/-13.2)micromol/L in the acute viral hepatitis group, (36.5+/-9.9)micromol/L in the chronic viral hepatitis group, (45.0+/-11.0)micromol/L in the post hepatitis cirrhosis with normal renal function group, and (83.6+/-50.4)micromol/L in the post hepatitis cirrhosis with renal dysfunction group. Serum free carnitine levels in the acute viral hepatitis and chronic viral hepatitis groups were significantly lower than those in the healthy controls. There were no significant differences in serum free carnitine levels of the post hepatitis cirrhosis group and the normal control group.</p><p><b>CONCLUSIONS</b>Patients with liver diseases can have carnitine metabolism errors. One of the secondary carnitine lack causes is liver disease.</p>
Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Carnitine , Blood , Chronic Disease , Hepatitis, Viral, Human , Blood , Liver Cirrhosis , BloodABSTRACT
Objective To investigate plasma levels of total carnitine (TC) and free camitine (FC) in patients undergoing hemodialysis and peritoneal dialysis.Methods 200 cases of normal group came from physical examination in this hospital,all testing cases were the in-hospital patients in the department of nephropathy.TC and FC were determined by use of an enzymatic cycle assay on Hitachi 7170 automatic biochemical analyzer.Results In 200 cases of normal group,TC level was (56.52?9.61) ?mol/L,and FC was (46.60?8.23) ?mol/L.In 37 hemodialysis patients,TC and FC levels were (41.47?13.22) ?mol/L and (24.58?8.91)?mol/L before dialysis,a statistic difference was observed against the control group (P0.05).Conclusions Carnitine deficiency was seen in most patients undergoing hemodialysis and peritoneal dialysis.Furthermore,the deficiency status got worse along with the dialysis course in hemodialysis patients.Carnitine infusion can effectively improve the status of these patients.