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Objective@#A longitudinal study design was used to explore the effect of perceived discrimination on the psychological adaptation of children relocated to alleviate poverty.@*Methods@#Four hundred twenty six children who were relocated to alleviate poverty were measured three times using a perceived discrimination questionnaire, childhood depression inventory, and the Illinois loneliness questionnaire, the data were analyzed using a cross lag model.@*Results@#Depression and loneliness of children relocated to alleviate poverty had an increasing trend during the three surveys( F=18.79, 8.69, P <0.01). Perceived discrimination was positively correlated with depression and loneliness at the time points for the three surveys( r=0.23~0.55, P <0.01). Cross lag analysis showed that perceived discrimination in the pretest (Tn) significantly predicted posttest (Tn+1) depression ( β=0.16, 0.20, P < 0.01 ) and perceived discrimination at time 2 significantly predicted loneliness ( β=0.25, P <0.01) at time 3.@*Conclusion@#Perceived discrimination was shown to be an important factor affecting the psychological adaptation of children relocated to alleviate poverty.
ABSTRACT
OBJECTIVE To explore the protective effect and mechanism of ophiopogonin D (OP-D) on oxidative stress injury of H9c2 cells induced by H2O2. METHODS An oxidative damage model of H9c2 cells was established by H2O2 induction. The cells were divided into control group (cultured in serum-free medium for 28 h), H2O2 injury model group (treated with H2O2400μmol·L-1 for 3 h), OP-D 5, 10 and 20 μmol · L-1 pretreatment groups (treated with H2O2400 μmol · L-1 for 3 h after OP-D pretreat?ment for 24 h), and an inhibitor of CYP2J3, 6-(2-proparglyloxyphenyl) hexanoic acid (PPOH) group (OP-D 20μmol·L-1+PPOH 10μmol·L-1, PPOH was added to the cells 1 h before OP-D treatment). Cell activity was measured by MTT method, levels of dihydroxyeicosatrienoic acid (11,12-DHET and 14,15-DHET respectively) were detected by enzyme-linked immunosorbent assay (ELISA), while levels of malondialdehyde (MDA), nitric oxide (NO) and activity of lactate dehydrogenase (LDH), superoxide dismutase (SOD) were detected by assay kits. Flow cytometry (FCM) was used to detect reactive oxygen species (ROS) and apoptosis. Western blotting was used to detect the expressions of CYP2J3, Akt phos?phorylation (p-Akt) protein and endothelial nitric oxide synthase phosphorylation (p-eNOS) protein in cells, and the possible mechanism by which OP-D reduces oxidative stress was further verified with PPOH. RESULTS H2O2400μmol · L-1 significantly inhibited H9c2 cell viability (P<0.01), and OP-D signifi?cantly increased the cell survival rate after H2O2 injury (P<0.01). Different concentrations of OP-D increased the level of 11,12- DHET and 14,15-DHET (P<0.05, P<0.01). OP-D increased the level of NO in cells after H2O2-induced injury (P<0.05, P<0.01), enhanced the activity of SOD (P<0.05), and decreased the level of MDA and LDH (P<0.05, P<0.01). OP-D significantly reduced oxidative stress and apoptosis after H2O2 injury (P<0.05, P<0.01). OP-D pretreatment increased the protein and mRNA expression of CYP2J3 (P<0.05, P<0.01) and the phosphorylation of PI3K/Akt-eNOS pathway after H2O2 injury (P<0.05, P<0.01). After PPOH was given in advance, the protective effect of OP-D was inhibited (P<0.05, P<0.01). CONCLUSION OP-D can reduce H2O2-induced H9c2 cell damage, which may be related to the activation of PI3K pathway and the phosphorylation of its downstream factors Akt and eNOS by inducing CYP2J3 expression and increasing the contents of 11,12-DHET and 14,15-DHET.
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This paper aimed to study the six chemical components of Polygoni Multiflori Radix (gallic acid, quercetin, luteolin, kaempferol, resveratrol, apigenin). By the established pregnane X receptor (human pregnant X receptor, PXR) CYP3A4 mediated drug induced rapid screening technique, the effect of chemical components on the cell activity was detected by MTS cell method, and the value of IC₅₀ was calculated. The dual luciferase reporter system was used to co-transfect PXR reporter gene expression vector containing transcriptional regulation and CYP3A4 with HepG2 cells, with 10 μmol·L⁻¹ rifampicin (RIF) as a positive control, and 10 μmol·L⁻¹ of ketoconazole (TKZ) as negative control. Gallic acid, quercetin, luteolin, kaempferol, apigenin, resveratrol(5, 10, 20 μmol·L⁻¹) were used to incubate for 24 h, and the luciferase activity was detected. The results showed that when plasmid pcDNA3.1 was co-transfected with pGL4.17-CYP3A4, gallic acid and resveratrol had an inhibitory effect on the regulation of CYP3A4, and quercetin, luteolin, kaempferol had an inductive effect on CYP3A4; when pcDNA3.14-PXR was co-transfected with pGL4.17-CYP3A4, quercetin, luteolin, kaempferol, apigenin, resveratrol had an inductive effect. To sum up, the 6 reported liver injury components had inhibitory or activating effects on CYP3A4. After PXR plasmid was involved, 5 components had an inductive effect on CYP3A4, and the inductive effects of 2 components were significantly different. In this experiment, we found that 2 kinds of potential liver injury components in Polygoni Multiflori Radix had been induced by CYP3A4, which was achieved through PXR regulation. It suggested that attention shall be paid to potential drug interactions when combined with Polygoni Multiflori Radix, so as to improve the safety and efficacy.
Subject(s)
Humans , Cytochrome P-450 CYP3A , Metabolism , Drugs, Chinese Herbal , Pharmacology , Hep G2 Cells , Liver , Phytochemicals , Pharmacology , Plant Roots , Chemistry , Polygonum , Chemistry , Pregnane X Receptor , MetabolismABSTRACT
OBJECTIVE This study investigated transcriptional regulation of the main chemical con-stituents of Polygonum multiflorum Thunb. including Stilbene Glucoside (THSG) and anthraquinone constituents (Emodin, Rhein, Aloeemodin, Chrysophanol and Physcion) and six potential liver injury constituents(gallic acid,quercetin,luteolin,kaempferol,resveratrol)on mediated by PXR CYP3A4.Early establishment of pregnane X receptor mediated CYP3A4 drug induced rapid screening technique was used to determine the effects of these constituents. METHODS First,effect of constituents on the cell activity was detected by MTS cell viability assay. IC50was calculated. Second, the expression vector and reporter vector were co-transfected into Hep G2 cells,10 μmol·L-1Rifampicin as a positive control, 10 μmol·L-1Ketoconazole as a negative control. After treated with different concentrations of (the an-thraquinone constituents concentrations were 2.5,5 and 10 μmol·L-1;the concentrations of Gallic Acid, Quercetin,Luteolin,Kaempferol,Apigenin,Resveratrol concentrations were 5,10 and 20 μmol·L-1)for 24 h,the cells were tested for dual luciferase activity.RESULTS The results show that the inhibitory ef-fect of THSG,Chrysophanol,Emodin,Rhein and Aloeemodin on CYP3A4 was inhibited by co-transfec-tion of pcDNA3.1 and pGL4.17-CYP3A4. The expression of pcDNA3.14-PXR and pGL4.17-CYP3A4 was induced by the four constituents. Besides, Emodin has a directly inducing effect. Four anthraqui-none constituentscan induce the effect of CYP3A4 by PXR, but Emodin can directly induce CYP3A4. THSG can inhibit CYP3A4,but in the presence of PXR plasmid can induce CYP3A4.For the six poten-tial liver injury constituents, results show that the plasmid pcDNA3.1 was cotransfected with pGL4.17-CYP3A4 regulation of Gallic Acid and Resveratrol on CYP3A4 inhibitory effects of Quercetin,Luteolin, Kaempferol have an induce effect; after pcDNA3.14-PXR and pGL4.17-CYP3A4 cotransfected, Quercetin, Luteolin, Kaempferol, Apigenin, Resveratrol have induced effect, three constituents'induc-tion effect had significant difference.CONCLUSION 12 kinds of Polygonum multiflorum Thunb.constit-uents have inhibitory or activating effects on CYP3A4, after the participation of PXR, 9 components have induced effects on CYP3A4, and the induction effect of 6 components has significant difference. The results suggested that we should pay attention to potential drug interactions when combined with Polygonum multiflorum Thunb.,and improve safety and efficacy.
ABSTRACT
The rapid screening technology was used to investigate the transcriptional regulation effect of main chemical constituents in tubers of Polygonum multiflorum, including 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucopyranoside(THSG) and anthraquinones (such as rhein, chrysophanol, aloe-emodin, emodin) on CYP3A4 drug inducers induced by human pregnancy X receptor (PXR).The effect of chemical composition on the cell activity was detected by MTS cell viability assay. IC₅₀ was calculated. The expression vector and the reporter vector were co-transfected into HepG2 cells, with 10 μmol•L⁻¹ rifampicin (RIF) as a positive control, and 10 μmol•L⁻¹ ketoconazole (TKZ) as a negative control. After treated with different concentrations of anthraquinones (2.5, 5, 10 μmol•L⁻¹) for 24 h, the cells were tested for dual luciferase activity. The results show that the inhibitory effect of THSG, chrysophanol, emodin, rhein and aloe-emodin on CYP3A4 was inhibited by co-transfection of pcDNA3.1 and pGL4.17-CYP3A4. The expressions of pcDNA3.14-PXR and pGL4.17-CYP3A4 were induced by the four compounds. Besides, emodin had a direct inducing effect. In conclusion, the four anthraquinone compounds have an inducing effect on CYP3A4 by PXR, but emodin can directly induce CYP3A4. THSG can inhibit CYP3A4, but plasmid can induce CYP3A4 after intervened with PXR.These results suggest that we should pay attention to the liver function and avoid liver damage in the combined administration of drugs.