Investigation of 6-O-methyl-scutellarein metabolites in rats by ultra-flow liquid chromatography/quadrupole-time-of-flight mass spectrometry.

Context Scutellarin (1) has been widely used in China to treat acute cerebral infarction and paralysis induced by cerebrovascular diseases. However, scutellarin (1) has unstable metabolic characteristics. Objective The metabolic profile of 6-O-scutellarein was studied to determine its metabolic stability in vivo. Materials and methods In this study, a method of UFLC/Q-TOF MS was used to study the 6-O-methyl-scutellarein metabolites in rat plasma, urine, bile and faeces after oral administration of 6-O-methyl-scutellarein (3). One hour after oral administration of 6-O-methyl-scutellarein (3) (34 mg/kg), approximately 1 mL blood samples were collected in EP tubes from all groups. Bile, urine and faeces samples were collected from eight SD rats during 0-24 h after oral administration. The mass defect filtering, dynamic background subtraction and information dependent acquisition techniques were also used to identify the 6-O-methyl-scutellarein metabolites. Results The parent compound 6-O-methyl-scutellarein (3) was found in rat urine, plasma, bile and faeces. The glucuronide conjugate of 6-O-methyl-scutellarein (M1, M2), diglucuronide conjugate of 6-O-methyl-scutellarein (M3), sulphate conjugate of 6-O-methyl-scutellarein (M4), glucuronide and sulphate conjugate of 6-O-methyl-scutellarein (M5), methylated conjugate of 6-O-methyl-scutellarein (M6) were detected in rat urine. M1, M2 and M3 were detected in rat bile. M1 was found in rat plasma and M7 was detected in faeces. Discussion and conclusion Because the parent compound 6-O-methyl-scutellarein (3) was found in rat urine, plasma, bile and faeces, we speculate that 6-O-methyl-scutellarein (3) had good metabolic stability in vivo. This warrants further study to develop it as a promising candidate for the treatment of ischemic cerebrovascular disease.

Design, Synthesis, and Biological Evaluation of Scutellarein Derivatives Based on Scutellarin Metabolic Mechanism In Vivo.

Three series of scutellarein derivatives have been designed and synthesized based on metabolic mechanism of scutellarin (1) in vivo. Their thrombin inhibition activities were tested through the analyzation of prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), and fibrinogen (FIB). The antioxidant activities of these target products were assessed by 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) assay and the ability to protect PC12 cells against H2 O2 -induced cytotoxicity, and their solubilities were evaluated by ultraviolet (UV) spectrophotometer. The results showed that the two isopropyl groups substituted derivative (18c) demonstrated stronger anticoagulant activity, better water solubility, and good antioxidant activity compared with scutellarein (2), which warrants further development of 18c as a promising agent for ischemic cerebrovascular disease treatment.

Protective mechanism of cerebrospinal fluid containing qingxin kaiqiao recipe on PC12 cell injury induced by glutamate / 中国中药杂志

<p><b>OBJECTIVE</b>To study the protective effect of cerebrospinal fluid containing Qingxin Kaiqiao recipe on PC12 cell injury induced by glutamate (Glu), in order to provide basis for the conical application of the recipe.</p><p><b>METHOD</b>SD rats were orally administered with decoction of Qingxin Kaiqiao recipe (7.9 g x kg(-1)) for three and a half days, 2 times a day, in order to prepare cerebrospinal fluid containing Qingxin Kaiqiao recipe. PC cells were divided into the normal group, the model group, the nimodipine group, the 10% normal CSF group, the 10% medicated CSF group, the 20% normal CSF group, the 20% medicated CSF group. Except for the normal group, other groups were cultured with PC12 cells and Glu with the final concentration of 20 mmol x L(-1) to establish the nerve cell injury model. Apart from the model group and the normal group, other groups were intervened with nimodipine, normal cerebrospinal fluid, and 10% and 20% medicated CSF. RT-PCR was used to detect the expression level of Bax mRNA, Bcl-2 mRNA and Caspase-3 mRNA, and MTT method was used to detect the activity of PC12 cells.</p><p><b>RESULT</b>The activity of PC12 cells of all of medicated CSF groups was higher than that of the model group, with the decrease in the expression of Bax mRNA and Caspase-3 mRNA and the increase in the expression of Bcl-2 mRNA. They showed a significant different with the model group (P < 0.01). The 20% medicated CSF group was superior than the 10% medicated CSF group (P < 0.01).</p><p><b>CONCLUSION</b>Qingxin Kaiqiao recipe shows an apparent protective effect on PC12 cells injured by Glu.</p>