ABSTRACT
Objective To establish a method for the determination of 10-hydroxyl carbamazepine (MHD),which is an activity metabolite of oxcarbazepine in human serum. Methods Serum samples were detected by high performance liquid chromatography (HPLC) after being processed by methanol protein deposition.The chromatographic column was Agilent TC-C18 (4.6 mm × 250 mm, 5 μm), with the mobile phase of acetonitrile-10 mmol ? L-1 KH2 PO4 ( 33 : 67) at a flow rate of 1.0 mL?min-1 .The detection wavelength was 230 nm,and phenacetin was used as an internal standard. Results The average recovery range of low,middle and high (1.0,10.0,60.0 μg?mL-1 ) concentrations for MHD was from 100.3％ to 106.0％.The RSD of intra-day and inter-day was ≤5.8％ (n= 5) and ≤7.4％ (n= 5),respectively.The limit detection of analysis method was 0.1 μg?mL-1 .Regression equation was Y = 0.1308X+ 0.0679 ( r = 0.9966,n = 5). Serum samples remained stable at room temperature,freezing and freeze thawing condition. Conclusion This method is sensitive,accurate,simple and quick,and can be used for monitoring the oxcarbazepine metabolites MHD in serum for clinical and pharmacokinetic study.
ABSTRACT
Objective To investigate the effect of TAK-242,an antagonist for Toll-like receptor 4,against myocardial isch?emia/reperfusion injury(I/R)in C57BL/6 mice along with the underlying mechanism. Methods C57BL/6Mice(n=36)were random?ized into three groups:sham group,I(30 min)/R 24 h model group and I/R+TAK-242(3 mg/kg)treatment group. At 24 h after reper?fusion,cardiac function and myocardial infarct size were evaluated with echocardiography and triphenyltetrazolium chloride(TTC), myocardial pathological pattern in mice was detected by HE staining,the mRNA and protein levels of TLR4 were determined by real time PCR and Western blot respectively,and the serum levels of IL-6,TNF-α,IL-10 and HMGB1 were detected by ELISA. Re?sults Our results showed that left ventricular systolic diameters(LVID)were shortened(P<0.01),left ventricular ejection fraction (LVEF)and left ventricular short axis shortening fraction(LVFS)were both decreased significantly(P<0.01)in following I/R mice. Myocardial infarction size was large and myocardial inflammatory cell infiltration was severe in I/R mice. The mRNA and protein levels of TLR4 were elevated(P<0.01),the serum levels of IL-6,TNF-α,IL-10 and HMGB1 in I/R mice were significantly increased com?pared with the sham group(P<0.01). We found that TAK-242 significantly extended LVID(P<0.05),increased LVEF and LVFS (P<0.05),reduced myocardial infarction and improved myocardial inflammatory cell infiltration,inhibited the mRNA and protein levels of TLR4(P<0.05),down-regulated the expression of IL-6 and TNF-α(P<0.01),and up-regulated IL-10 and HMGB1 com?pared with I/R group(P<0. 01). Conclusion Treatment with TAK-242 can significantly reduce myocardial ischemia/reperfusion in?jury partly via regulating inflammation.
ABSTRACT
BACKGROUND: In clinical, propofol can contract cerebral vessels, decrease cerebral blood flow, decrease brain metabolic oxygen consumption,which can decrease pressure in brain. Studies prove that propofol can protect endothelial cell that may be injuried by active oxygen injury and also decrease nerves injury of experimental rats with cerebral ischemia.OBJECTIVE: To investigate the protective effects of propofol on cerebral ischemia-reperfusion injury in rat and its mechanism.DESIGN: Randomized and controlled study.SETTING:Anesthesiological Department of the Second Affiliated Hospital of Xi'an Jiaotong University.PARTICIPANTS: The experiment was conducted at Pharmacological Laboratory of Medical College of Xi' an Jiaotong University in 2004. Totally 40 healthy male SD rats, aged 3-4 months, weighting 200-300 g, were divided randomly into four groups: Model group, control group, nimodipine group and propofol group, with 10 in each group.METHODS: The rats were anesthetized by intraperitoneal methods with ketamine and propofol separately. When righting reflex was abolished, external carotid artery was separated and ligated. A nylon thread was put at the stump site of external carotid artery without ligation. Model group: 10 mL normal saline was injected into intraperitone in 10 minutes before ischemia.Control group: 10 mL normal saline was injected into intraperitone at the end of operation. Nimodipine group: 10 g/L nimodipine (1 mg/kg) was injected into intraperitone in10 minutes before ischemia. Propofol group: 10 g/L propofol (110 mg/Kg) was injected into intraperitone in 10 minutes before ischemia. When ischemia was lasted for 3 hours, nylon thread was with drawed for reperfusion. When reperfusion was lasted for 3 hours, blood samples were obtained from orbit. Skulls were opened and brains were removed.Effect of propofol on cerebral ischemia-reperfusion injury was observed.MAIN OUTCOME MEASURES: Infarction area, cerebral water content,serum lactate dehydrogenase (LDH) and creatine kinase (CK) levels, brain superoxide dismutase (SOD) activity, malondialdehyde (MDA) and Ca2+levels were measured. Ultrastructure of brain tissue was examined under electron microscope.RESULTS: ①Infarct area in propofol group was significantly smaller than that in model group [(10.45±3.65, 19.68±4.03)%, (t=3.493,P ＜ 0.01)]. ② CK level was lower in propofol group than that in model group [(471±200,1 930±917) IU/L, (t=3.493, P ＜ 0.01)]; and LDH level in propofol group [(8 240±2 580) U/L] was significantly different from that in model group [(15 470±2 680) U/L, (t=3.441, P ＜ 0.01)]; And water content in brain tissue was lower in propofol group than that in model group [(78.2±2.4,82.9±2.9)%, (t=3.321, P ＜ 0.01)]. ③ The death rate of rats was 13.6%in propofol group, and 47.6% in model group, the former was decreased obviously as compared with the latter, and the difference was significant (t=6.21,P ＜ 0.05). ④ SOD activity was (1 690±780) U/g in propofol group and (830±110) U/g in model group, the difference was significant (t=3.420, P ＜ 0.01); but MDA content was obviously lower in propofol group than that in model group [(0.058±0.014, 0.115±0.047) μmol/g, (t=3.336, P ＜ 0.01)].CONCLUSION: Propofol has protective effect on cerebral ischemia-reper fusion injury in rats, and the mechanism is related with inhibition of Ca2+overloading and lipid peroxidation.