| Traditional Chinese medicine Lophatheri Herba is the dry leaves and stems of Lophatherum gracile Brongn. in Gramineae. This paper mainly focuses on the protection of Lophatheri Herba to reveal the material basis of myocardial ischemia and its mechanism of action, the research work carried out as follows:1. This study firstly established a rapid high performance liquid chromatography - triple quadrupole ion trap mass spectrometer - multiple reactions monitoring method to measured seven kinds of Lophatheri Herba flavonoids content. Finalized orientin, isoorientin, swertiajaponin, vitexin, isovitexin, afzelin and luteolin-7-O-β-D-glucoside as the standard material for Lophatheri Herba can get repetitive, are composed of carbon glycosides flavonoid and oxygen glycoside flavonoid. Two groups of flavonoids components were isoorientin orientin:swertiajaponin:vitexin:isovitexin:afzelin:luteolin-7-O-β-D-glucoside = 20:15 55:5:5:2:5, as well as isoorientin:orientin:swertiajaponin:vitexin:isovitexin:afzelin: luteolin-7-O-β-D-glucoside = 30:25:30:10:5:2:5.2. Firstly oral administration Lophatheri Herba flavonoids, then establishing the ischemia reperfusion injury model, the results showed that Lophatheri Herba flavonoids can reduce the damage caused by myocardial ischemia through reducing the expression of NF-κB, TNF-α and caspase-3 protein in the myocardial tissue. Therefore, the mechanism of Lophatheri Herba flavonoids protect the ischemic myocardium may scavenge oxygen free radicals, inhibiting oxygen free radicals, inhibiting lipid peroxidation, increased NO levels, inhibited the activity of NF-κB and TNF-a, and lowered the role of caspase-3 protein expression and so on.3. Lophatheri Herba flavonoid monomers and flavonoid components can effectively improve myocardial cell survival, inhibition of apoptosis, and effectively reduce the concentration of free calcium ions. Lophatheri Herba flavonoid components inhibiting the activation of p38MAPK signaling pathway achieve improvement of myocardial ischemia. Lophatheri Herba flavonoid monomers and components may inhibit the expression of p65, up-regulating the expression of p50 achieves suppressing effect of NF-κB activation. Also, Lophatheri Herba flavonoid monomers and components may inducible HSP70, HSP27 transition expression, up-regulating iNOS and COX-2 expression, to achieve the protective effect of myocardial ischemia. It was found different Lophatheri Herba flavonoid monomers at different sites of the role of protein expression in different strength, the second flavonoid compounds effected results show multiple flavonoids act on multiple protein targets and the overall trend showed a complementary.4. Using response surface methodology by total flavonoid content and antioxidant effects of multiple response results optimized the extraction process of Lophatheri Herba flavonoids. The obtained Lophatheri Herba extracts directly reflect its antioxidant effect using pre-column oxidation of HPLC-line detection, combined with standards and mass spectrometry, ultimately revealed the material basis of Lophatheri Herba antioxidant effect were mostly C-glycosides flavonoids and few oxygen glycosides and flavonoid aglycone.5. To further explore the process of Lophatheri Herba flavonoids in vivo, first established flavonoids bind to serum albumin in vitro model. Through the combined location, binding sites numbers, combine forces type, size and other parameters, comprehensive molecular docking simulation experiment, preliminary study the may transport in vivo.6. Established HPLC-MS/MS method and radioactive iodine labeling, determining the Lophatheri Herba flavonoids distribution changes in organ tissue and serum of rats. It was found that Lophatheri Herba carbon glycoside flavonoid absorbed by the gastrointestinal tract into the blood circulation, and into the heart through the blood circulation, the prototype compounds can be found in the serum and the heart. Flavonoid aglycone been detected in heart failure, as oxygen glycoside flavonoid content is very low or no distribution. |
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