ABSTRACT
The protective effect of different polar fractions of Carbonized Rubiae Radix et Rhizoma (cRRR) against ox-LDL-induced damage to human umbilical vein endothelial cells (HUVECs) was investigated by MTT assay, and the components were identified by using UPLC-Q-TOF-MS. According to the study, ethyl acetate extract and n-butanol extract could increase cell viability (P<0.01), while petroleum ether extract had no influence, and water extract could even inhibit the cell viability to some degree. Moreover, 32 compounds in four polar fractions were analyzed, including 31 quinones and their glycosides, and one rubiprasins C. Petroleum ether extract, ethyl acetate extract, n-butanol extract and water extract contained 23, 32, 26, 15 compounds, respectively. According to cell experiments in vitro, active fractions were ethyl acetate extract and n-butanol extract. The results could provide scientific references for further studies on effective material basic of cRRR, and lay a foundation for studies on the relationship between efficacies and materials.
ABSTRACT
In order to explore the effect on chemical constituents after carbonized, the changes of chemical constituents in raw and carbonized Rubiae Radix et Rhizoma were analyzed by UPLC-Q-TOF-MS. The research also used principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) for data statistics to find out the main differences on components before and after carbonized. The accurate m/z values of Q-TOF-MS and Q-TOF-MS-MS fragments were applied to identify the structures. The results showed that 6 more discrepant constituents were existed between raw and carbonized Rubiae Radix et Rhizoma. Three constituents were selected as the main discrepant components according to the peak area (276 nm) and identified, as lucidin, xanthopurpurin and 1,3,6-trihydroxy-2-methylanthraquinone. After carbonized, contents of xanthopurpurin and 1,3,6-trihydroxy-2-methylanthraquinone were observably increasing, while lucidin was obviously decreasing. They could be used as the chemical markers for the differentiation between raw and carbonized Rubiae Radix et Rhizoma. The results of this experiment played an important role in the study of processing principle of carbonized Rubiae Radix et Rhizoma. It also provided important evidences for the interpretation of effective material based on carbonized Rubiae Radix et Rhizoma.
ABSTRACT
Objective: To observe the effect of different extracts from carbonized Rubiae Radix et Rhizoma (RRR) on acute blood stasis model of rats, in order to screen the optimal extraction method for its hemostatic effect by removing blood stasis. Methods: The acute blood stasis models of rats were established with sc injection of high-dose adrenaline hydrochloride after continuous administration for 8 d and being socked in ice-water, using Yunnan Baiyao as positive drug. The effect of the alcohol extract, compound (alcohol + aqueous) extract, aqueous extract, and aqueous extract after alcohol extracting on hemorheology, in vitro thrombus, blood plotelets system, and fibrinolytic system in acute blood stasis model of rats were observed. Results: Compared with the control group, the whole blood viscosity in each shear rate (P < 0.01) and the wet and dry weight of in vitro thrombus could be increased (P < 0.05), the coagulation time was prolonged (P < 0.05, 0.01), the FIB content and platelet aggregation rate induced by ADP increased (P < 0.01), the PAI-1 and TXB2 contents increased, and the 6-keto-PGF1α content decreased in the rats of model group (P < 0.05, 0.01). Compared with the model group, both the alcohol extract and compound extract could significantly improve the whole blood viscosity, decrease the wet and dry weight of external thrombus, shorten the coagulation time, increase the contents of PAI-1 and 6-keto-PGF1α, and decrease the TXB2 content (P < 0.05, 0.01). Both of them could make platelet aggregation rate increased, but have no significant influence on the content of FIB. The alcohol extract significantly decreased the content of t-PA (P < 0.05), while the compound extract significantly increased t-PA content (P < 0.01). The compound extract has influenced the hemorheology and the wet and dry weight of in vitro thrombus greater than the alcohol extract. While on the coagulation time, the effect of alcohol extract is greater. Conclusion: The hemostatic effects by removing blood stasis of alcohol extract and compound extract are stronger than those of others, especially the compound extract. In other words, the optimal extraction method of its hemostatic effect by removing blood stasis is the compound extract.