Study on the anti-mitochondrial apoptosis mechanism of Erigeron breviscapus injection based on UPLC-Q-TOF-MS metabolomics and molecular docking in rats with cerebral ischemia-reperfusion injury.

ETHNOPHARMACOLOGICAL RELEVANCE: Erigeron breviscapus is a common medicine of eight ethnic minorities, including Miao, Naxi, and Yi. As early as the Ming Dynasty (AD 1368-1644), Lanmao's Materia Medica of Southern Yunnan (AD 1436) recorded that the medicine is used for the treatment of "Zuo tan you huan." In modern pharmacological research, Erigeron breviscapus injection is the most commonly used preparation in the treatment of ischemic stroke caused by acute cerebral infarction, but its mechanism of action in the treatment of ischemic stroke is not well understood. AIM OF THE STUDY: In this study, a metabonomics study based on ultraperformance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-QTOF-MS) was used in investigating the effect of a traditional Chinese medicine preparation Erigeron breviscapus injection on the rat model of focal cerebral ischemia-reperfusion and the affinity of its main components with the targets of mitochondrial apoptotic pathways. MATERIALS AND METHODS: This study used molecular docking technology to verify the effective binding ability of main effective components of Erigeron breviscapus injection to target proteins related to mitochondrial apoptosis pathway. This study developed a metabonomics method based on the ultra-performance liquid chromatography combined with quadrupole time-of-flight tandem mass spectrometry (UPLC Q-TOF MS) to evaluate the efficacy and study the mechanism of traditional Chinese medicine preparation. With pattern recognition analysis (principal component analysis and partial least squares-discriminate analysis) of urinary metabolites, a clear separation of focal cerebral ischemia-reperfusion model group and healthy control group was achieved. RESULTS: Erigeron breviscapus injection can significantly reduce the area of cerebral infarction, improve tissue morphological lesion in rats, and can increase the number of Nissl bodies. It may be a promoting factor by inhibiting hippocampal nerve cell apoptosis and Bax protein expression and by exerting effects against ischemia reperfusion after the induction of apoptosis. Thus, it plays a role in brain protection. Moreover, it can considerably promote the recovery of neurological deficiency signs in advance. Meanwhile, Erigeron breviscapus decreased malondialdehyde content and T-NOS activity. Its curative effect from strong to weak order: low dose > high dose > medium dose. The representative components of Erigeron breviscapus have good affinity with the active sites of mitochondrial apoptosis-related proteins. Metabolomics found that the potential biomarkers regulated by breviscapine are kynurequinolinic acid, succinylornithine, and leucine proline. It is speculated that it may participate in TRP-kynurequinolinic acid and succinylornithine-urea cycle-NO metabolic pathways. CONCLUSIONS: This paper revealed the potential biomarkers and metabolic pathways regulated by Erigeron breviscapus. It was speculated that the mechanism is related to its inhibition of mitochondrion-mediated apoptosis. Erigeron breviscapus could restore the metabolic profiles of the model animals to normal animal levels. The mechanism may be related to the potential biomarkers of quinolinic acid, succinylornithine, and leucine proline and the metabolic pathways involved. However, the exact mechanism by which Erigeron breviscapus inhibits mitochondrion-mediated apoptosis remains to be further explored.

[Effect of P and K Fertilization on Four Iridoids in Axial and Lateral Root of Gentiana crassicaulis].

Objective: To study the effect of phosphorus( P) and potassium( K) fertilization on four iridoids in axial and lateral root of Gentiana crassicaulis. Methods: Field experiment was conducted based on phosphorus and potassium,and the content of four iridoids was analyzed in the axial and lateral root of Gentiana crassicaulis. Results: Despite of different proportion of phosphorus and potassium,the content of four iridoids in axial root was always higher than that in the lateral root, and under normal fertilizer condition, the content of axial and lateral root was 12. 36% and 9. 84%, respectively. On the condition of phosphorus and potassium fertilization, the content of gentiopicroside and loganic acid were higher than that of the swertiamarin and sweroside both in the axial and lateral root. And the difference of the two parts could be increased by high potassium with appropriate phosphorus application. Conclusion: The four iridoids were more likely to accumulate in the axial root of Gentiana crassicaulis, and phosphorus and potassium fertilization, only affected the different degrees on the content of ingredients among two parts. Thus, more attention should be paid to the use of phosphorus and potassium fertilization, and the difference of the active ingredient content in the axial and lateral root of Gentiana crassicaulis.

[A comparative study on the absorption kinetics parameters of rhubarb free anthraquinones between normal dogs and dogs with severe acute pancreatitis].

OBJECTIVE: To study the effects of severe acute pancreatitis (SAP) on absorption kinetic parameters of rhubarb free anthraquinones. METHODS: Eleven healthy Beagle dogs were randomly divided into the normal group (n = 6) and the SAP group (n = 5). The SAP animal model was prepared by surgery through portal vein blood channel building to collect blood from normal dogs and dogs with SAP. The free anthraquinones (20 mg/kg) was given by gastrogavage. The concentrations of five anthraquinones (aloe emodin, rhein, emodin, chrysophanol, and physcion) in the blood plasma of the portal vein and the femoral artery were determined using high performance liquid chromatography (HPLC). The kinetic parameters were calculated using MATLAB2007B Software. The half life (t(1/2Ka)), the absorption peak time (Tmax), the peak concentration (Cmax), the area under the curve [AUC(0-infinity)], and the mean residence time (MRT) were calculated using the statistical moment method. The transport velocity of corresponding medicines from the gastrointestinal tract to the blood (ka) was calculated. RESULTS: There was no difference in the chemical composition absorption type of the portal vein and the femoral artery between the two groups. Aloe emodin could be detected in the portal vein of each animal at each time point, and they were in the quantitative range. Rhein could be detected in the portal vein of each animal at each time point, and they were lower than the quantitative limit at few time points. Emodin and chrysophanol could be detected in the portal vein of partial animals at each time point, and most of them were higher than the quantitative limit. Physcion could be detected only in the portal vein of less animals at few time points. Rhein could be detected in the femoral artery at most time points, but the rhein plasma concentration at most time points were lower than the quantitative limit. Lower concentration of aloe emodin, emodin, and chrysophanol could be detected in the femoral artery at only few time points. Physcion was not detected in the femoral artery. The rhein plasma concentration of the femoral artery and the chrysophanol Cmax of the portal vein at 45 min were higher in the SAP group than in the normal group with statistical difference (P<0.05). There was no statistical difference in the rest indices. The AUC of rhein in the two groups were 59.32% and 66.07% of the total free anthraquinones respectively. CONCLUSIONS: SAP could not obviously affect the absorption kinetics parameters of rhubarb free anthraquinones. The intestinal tract and the liver might possibly play important roles in metabolizing or transforming rhubarb free anthraquinones.

[Intestinal absorption of aloe-emodin using single-passintestinal perfusion method in rat].

The intestinal absorption of aloe-emodin was investigated using the single pass intestinal perfusion (SPIP) technique in S/D rats. SPIP was performed in each isolated segment of the intestine (i.e., duodenum, jejunum, ileum and colon) and the different concentrations inhibitor group of P-glycoprotein (P-gp) and multidrug resistance-associated protein (MRP2) with the concentrations of aloe-emodin (0.238 mg x L(-1)) at a flow rate of 0.28 mL x min(-1). The effective absorption rate constant (Ka) and apparent absorption coefficient (Papp) of aloe-emodin for each segment were determined before and after treated with different concentrations of inhibitors of P-gp and MRP2 respectively. Aloe-emodin exhibits a high intestinal permeability except the the ileum, indicative that the compounds are well absorbed. Decreases of Ka and Papp values in the duodenum, jejunum, colon and ileum, furthermore, the duodenum has significant increased compared with the ileum, there are have no significant difference in other isolated region of the intestine. Compared with the group which have no inhibitor of P-gp, the Ka and Papp were significantly increased in inhibitor of P-gp groups. Compared with the group of no inhibitor of MRP2, the Ka and Papp were significantly increased in inhibitor of MRP2 groups with the highest and the middle concentration. The results suggested that the inhibitors of P-gp and MRP2 all can promote the intestinal absorption of aloe-emodin.