ABSTRACT OBJECTIVE To establish the HPLC fingerprint of Paeoniae Radix Alba and investigate the spectrum-effect relationship between the fingerprint and the protection against vascular injury.METHODS The HPLC method was used to establish the fingerprint of Paeoniae Radix Alba. Lipopolysaccharide-induced human umbilical vein endothelial cells (HUVEC) and Tg(fli1:egfp) zebrafish models were used to evaluate in vivo and in vitro protective effects of 17 batches of Paeoniae Radix Alba extracts. Grey correlation degree and partial least squares method were used to analyze the spectrum-effect relationship between thefingerprint of Paeoniae Radix Alba and the protection effects.RESULTS The HPLC fingerprint of Paeoniae Radix Alba was constructed, and 11 common peaks were obtained, and the correlation was high, indicating that multiple components of Paeoniae Radix Alba may had synergistic effects on the protection of vascular endothelial cell activity. The correlation sequence with in vivo activitywasP6>P7>P10>P11>P9>P5>P3>P2>P8>P1>P4, and in vitro activity was P6>P4>P2>P3>P5>P1>P10>P9>P7>P11>P8, respectively. Peak 6 was identified as paeoniflorin. The regression coefficient of the partial least squares standard equation was positive, positively correlated with the protection of vascular endothelial cell activity and the vascular development in Tg(fli1:egfp) zebrafish, with VIP>1.0. Furthermore, the verification experiments of active ingredient showed that paeoniflorin could improve cell viability, migration, and tube formation, as well as promote vascular development in Tg(fli1:egfp) zebrafish in a dose-dependent manner. The above results indicated that paeoniflorin might be the major active component of Paeoniae Radix Alba for the protection effect on vascular endothelial cell.CONCLUSION The main active substance of Paeoniae Radix Alba in protecting against vascular inflammation and damage is paeoniflorin, which can provide reference for the quality control of medicinal materials.

Parthenogenetic embryonic stem (pES) cells isolated from parthenogenetic activation of oocytes and embryos, also called parthenogenetically induced pluripotent stem cells, exhibit pluripotency evidenced by both in vitro and in vivo differentiation potential. Differential proteomic analysis was performed using differential in-gel electrophoresis and isotope-coded affinity tag-based quantitative proteomics to investigate the molecular mechanisms underlying the developmental pluripotency of pES cells and to compare the protein expression of pES cells generated from either the in vivo-matured ovulated (IVO) oocytes or from the in vitro-matured (IVM) oocytes with that of fertilized embryonic stem (fES) cells derived from fertilized embryos. A total of 76 proteins were upregulated and 16 proteins were downregulated in the IVM pES cells, whereas 91 proteins were upregulated and 9 were downregulated in the IVO pES cells based on a minimal 1.5-fold change as the cutoff value. No distinct pathways were found in the differentially expressed proteins except for those involved in metabolism and physiological processes. Notably, no differences were found in the protein expression of imprinted genes between the pES and fES cells, suggesting that genomic imprinting can be corrected in the pES cells at least at the early passages. The germline competent IVM pES cells may be applicable for germ cell renewal in aging ovaries if oocytes are retrieved at a younger age.
Animals ; Cell Line ; Electrophoresis, Gel, Two-Dimensional ; Mice ; Parthenogenesis ; physiology ; Pluripotent Stem Cells ; metabolism ; Proteomics ; methods

OBJECTIVE To investigate the components and metabolites of leaves of Cyclocarya paliurus in rats plasma. METHODS UHPLC-Q/TOF-MS method was applied to identify the components and metabolites of rat plasma. The prototype components and metabolites were analyzed in rats plasma of leaves of Cyclocarya paliurus water extract after intragastric administration by comparing the chromatograms of Cyclocarya paliurus, blank serum and drug-containing serum. RESULTS A total of 15 compounds including 6 prototype components and 9 metabolites were identified. The main prototype components in blood were caffeylquinic acid, flavonoids and saponins, and the metabolic pathways were methylation and hydroxylation. CONCLUSION This experiment showes that the prototype components absorbed into blood may be the potential bioactive components in leaves of Cyclocarya paliurus, providing a scientific basis for clarifying its material basis in pharmacodynamics.

Abstract Diabetic osteoporosis is a systemic metabolic bone disease caused by long-term hyperglycemia and has become one of the major complications of diabetes. It is characterized by decreased bone mass, destroyed bone microstructure, and increased bone fragility. Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex herb pair is a classic drug combination, and has been widely used in many anti-diabetic osteoporosis prescriptions. The pharmacological studies have shown that the effects of this herb pair on promoting formation and inhibiting bone resorption are closely related to the Nrf2, MAPK, Wnt and RANK/RANKL signaling pathways as well as autophagy. The compatibility of this herb pair changes the dissolution and pharmacokinetics of the active substances, which have synergistic effects. This article summarized the effective substances of Anemarrhenae Rhizoma-Phellodendri Chinensis Cortex and its mechanism of action against diabetes and osteoporosis, providing a reference for further research and development of the drug pair.