ABSTRACT
The direct acting substances of Cuscuta chinensis in vivo were preliminarily identified through the correlation analysis of "metabolites-effect identification" model. The ovariectomized female rats were i.g administered with 95% ethanol extract part, 40% ethanol elution part and n-butanol extract part of Cuscuta chinensis. The serum fingerprints of different parts and times of administration were established by UPLC/Q-TOF-MS. At the same time, serum estradiol (E2), follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels were detected. Bivariate correlation analysis and grey correlation analysis were used to screen estrogenic components. The results showed that nine direct acting substances in vivo highly related to estrogen effect were found in the drug containing serum, which were hyperoside, astragalin, methyl quercetin glucuronide, quercetin-diglucuronide, quercetin, apigenin, isoquercitrin, kaempferol glucuronide and kaempferol. We can preliminarily screen out the direct acting substance of estrogen effect of Cuscuta chinensis in vivo based on the research idea of serum spectrum effect correlation. It provides a reliable basis for revealing the estrogeneffective substances of Cuscuta chinensis and confirming the quality markers. This experiment was approved by Harbin University of Commerce Ethics Committees (Approval No. HSDU2020-065).
ABSTRACT
Objective: To preliminarily screen out the estrogen-like quality markers of small grain Cuscuta chinensis from Heilongjiang Province, so as to provide reference for its subsequent experimental research and quality control. Methods: UPLC- Q-TOFMS/MS was used to qualitatively analyze the extract of C. chinensis and the fingerprints of different polar fractions were established. The estrogenic activity of different polar fractions was evaluated with uterine coefficient, endometrial thickness and serum estrogen level of mice. The bivariate correlation analysis and gray relational analysis were used to construct the composition-activity relationship between chemicals and the effects of estrogen for screening quality markers. And the content determination methods of the five quality markers were established. Results: A total of 10 quality markers related to the estrogenic effect from C. chinensis were found in the positive and negative ion scanning modes. They were hyperoside, astragalin, stigmasterol, neocuscutoside C, apigenin, kaempferol, 6-O-(E)-p-coumaroyl-β-D-fructofuranosyl-(2→1)-α-D-glucopyranoside, quercetin, isorhamnetin, and 2,6-octadecadiynoic acid. The contents of the five quality markers were determined as follows: hyperoside (2.753 ± 0.097) mg/g, quercetin (1.139 ± 0.107) mg/g, apigenin (1.104 ± 0.047) mg/g, kaempferol (1.144 ± 0.079) mg/g and isorhamnetin (0.697 ± 0.074) mg/g. Conclusion: The quality markers of small grain C. chinensis from Heilongjiang Province can be screened out according to the composition-activity relationship, and the method for the detect the concentrations of the quality markers is accurate and stable.
ABSTRACT
The toxic effect of ammonia on rCHO-GS cell decreased obviously due to the transfection of GS system in serum-free culture. The maximum cell density, 15.6 x 10(5) cells/mL was obtained in the culture with 1.42 mmol/L ammonia. The growth of rCHO-GS cell was inhibited with an increased ammonia concentration. However, a cell density of 8.9 x 10(5) cells/mL was obtained when the concentration of ammonia was 12.65mmol/L. The intracellar metabolic pathways were affected due to the decrease of the toxic effect of ammonia on rCHO-GS cell. With the increase of initial ammonia concentration from 0.36mmol/L to 12.65mmol/L, the yield coefficients of cell to glucose and lactate to glucose decreased. The activities of hexokinase (HK), pyruvate kinase (PK), and lactate dehydrogenase (LDH) increased by 43%, 140% and 25%, respectively, indicating that the utilization of glucose increased and the glycolysis pathway was more prone to efficient energy metabolism pathway. An increased activity of glutamate-pyruvate aminotransferase (GPT) showed that the conversation from glutamate to alpha-ketoglutarate was shifted to glutamate-pyruvate transamination pathway. The deamination pathway was inhibited due to a decreased activity of glutamate dehydrogenase. In addition, the number of cells in G0/G1 phase increased and the specific production rate of recombinant protein increased by 2.1-fold with the increase of initial ammonia concentration from 0.36mmol/L to 12.65mmol/L.