| Background and ObjectiveCytochrome P450s(CYPs)is an important human drug-metabolizing enzyme family,which mainly expressed in the liver,small intestine,and kidney.CYPs is capable of metabolizing approximately 75% of clinical drugs.Changes in the expression and activity of CYPs can lead to alteration in the pharmacokinetics and pharmacodynamics of drugs.In addition to the differences in CYPs activity caused by drug-drug interactions(drug induction or inhibition of CYPs),the basal expression and activity of CYPs exhibit significantly inter-individual differences resulting in individual differences in drug metabolism.Originated in the 1990 s,the developmental origins of health and disease(DOHaD)considered that during sensitive period in development,environmental stress factors may permanently alter the development and function of organization to adapt to the adjustment,which is the root cause of adverse health conditions(eg,chronic diseases)in adulthood.Organism development is highly sensitive to environmental factors such as nutrients,environmental chemicals,drugs,infections,heavy metal and other factors.Parts of the reason for the dramatic increase in the prevalence of many major diseases and dysfunctions,over the past 40 years,seem to be related to nutritional imbalances in developmental stages or exposure to chemicals,inflammation,and other environmental factors.Numerous studies have found that inflammation in most adult animal models causes down-regulation of most drug-metabolizing enzyme genes expression.Some studies have demonstrated that exposure of LPS during mouse pregnancy reduced the Cyp3a11 mRNA and protein level in the mouse placenta and fetus.However,whether LPS exposure during pregnancy affects the expression of Cyp3a11 and other CYPs in offspring mouse remains unknown.Humans may be exposed to inflammation caused by bacterial infections during pregnancy,so inflammatory reactions in the early developmental stage may lead to long-term changes in the drug metabolism.Previous studies are based on short-term effects,and studies on the long-term effects of offspring have not been reported.Pregnane X receptor(PXR)is a key transcriptional regulator in the liver,which regulates the basal expression of hepatic drug-metabolizing enzymes(DMEs)and mediates the induction of CYPs isoenzymes induced by foreign substances.The nuclear factor-?B is an important regulator during the inflammatory response.Inflammatory stimuli can regulate the expression of Pxr by binding the NF-?B p65 subunit to the retinoid X receptor in mice.Previous studies were based on the shortterm effects of inflammation on the expression of metabolic enzymes,whereas the long-term effects on DMEs expression in offspring have not been reported.Therefore,clarifying the long-term effects of early exposure to inflammation is very important for guiding clinical rational drug use.In the current study,we aimed to explore the consequence of LPS exposure during pregnancy on the expression of DMEs genes in offspring mice.This study may provide new insights into the varied expression of drug-metabolizing enzymes expression in adult,and broaden the knowledge of the long-term effects of environmental events during pregnancy on drug metabolism in the offspring.The study mainly carried out research from the following:(1)The impact of exposure to LPS during pregnancy on the expression of hepatic DMEs in offspring at infants(postnatal day 10),adolescents(postnatal day 30),and adulthood(postnatal day 60)age.(2)Effects of LPS exposure during pregnancy on the developmental patterns of hepatic metabolic enzymes in offspring mouse.MethodC57BL/6 mice were used as the animal model,LPS as a model drug and saline as vehicle control.C57BL/6 female mice were randomly divided into 2 groups on the day 10 of pregnancy and intraperitoneally injected with 100 ?g·kg-1 LPS or 10 ?l·kg-1 saline.Offspring mice were sacrificed at postnatal day 10,30 and 60.Liver samples were collected to detect the mRNA and protein expression of selected inflammatory cytokines(NF-?B and IL-6),nuclear receptors(Pxr,Ahr and Car)and DMEs(Cyp3a11,2b10,1a2,Sult1e1 and Ugt1a1).Results 1.Effects of exposure to LPS during pregnancy on the expression of inflammatory cytokines in the liver of offspring at different ages:Compared with the control group of the same age,the expression of NF-?B and IL-6 mRNA in the liver of the 10-day-old female mice in the LPS treatment group was higher than that in the control group,but significantly lower than that in the control group at 60 days of age(n = 4-6/group,P < 0.05);the expression of NF-?B mRNA in the liver of male mice of 30 days was significantly higher than that of the control group,while the 60-day-old group was significantly lower than the control group(n = 5-11/group,P < 0.05).In the LPS treatment group,the expression levels of liver IL-6 mRNA in the male mice of 10 and 30 days old were higher than those in the control group,and 60 days old was lower than the control group,but there was no statistical difference.2.The impact of LPS exposure during pregnancy on hepatic drug-metabolizing enzymes and nuclear receptors expression in 10-day-old offspring mice(neonatal period):mRNA expression level: Compared with the control group,the mRNA expression of nuclear receptors Pxr and Car as well as DMEs Cyp3a11,1a2,2ab10,Ugt1a1 and Sult1e1 was significantly increased(n = 4-8/ group,P < 0.05).Protein expression level: Consistent with the induced mRNA expression,the protein expression of hepatic Cyp3a11 and 1a2,in the offspring mice,was significantly higher in the LPS treatment group in comparison with the control group(n = 4-8/ group,P < 0.05).3.Effects of exposure to LPS during pregnancy on the expression of hepatic nuclear receptors and DMEs in 30-day-old offspring mouse(adolescent period):mRNA expression level: Compared with the control group,offspring mice metabolic enzyme Cyp1a2 and Sult1e1 mRNAexpression level was significantly increased;hepatic nuclear receptor Car and metabolic enzyme Cyp3a11 mRNA expression level significantly decreased(n = 4-11/group,P < 0.05).Ugt1a1 mRNA expression in the liver of female mice was significantly lower than that of the control group(n = 4-11/group,P < 0.01);whereas,there was no change in the mRNA expression of Ugt1a1 in the liver of male mice.(n = 4-11/group,P > 0.05).Cyp2ab10 mRNA expression level significantly higher only in male mice(n = 4-11/group,P < 0.01),but no statistical significance in female mice.The expression level of hepatic nuclear receptor Ahr was only significantly decreased in male mice(n = 5-11/group,P < 0.01).Protein expression level: Compared with the control group,hepatic Cyp3a11 protein expression in the offspring mice was significantly decreased in the LPS treated groups,while Cyp1a2 protein expression was significantly increased in male offspring(n = 4-8/group,P < 0.05)in comparison with the control groups.4.Effects of exposure to LPS during pregnancy on the expression of hepatic drugmetabolizing enzymes and nuclear receptors expre in 60-day-old offspring mice(adult period):mRNA expression level: For the 60-day offspring,the mRNA expression of Cyp1a2,Ugt1a1 and Cyp2ab10 was significantly decreased in the LPS treated groups for both gender(n = 5-6/group,P < 0.05).In addition,a higher mRNA expression of Pxr,Cyp3a11,and Sult1e1 was seen in female not male offspring delivered by LPStreated mice(n = 5-6 /group,P < 0.01).For the mRNA expression of Car,an increased expression was found in the female offspring(n = 5-6/group,P < 0.001),while a decreased expression was seen in the male offspring in LPS treatment groups(n = 4-6/group,P < 0.05).Protein expression level: Compared with the control group,the protein expression level of the liver metabolizing enzyme Cyp3a11 in the female mice was significantly increased,and the protein expression level of Cyp1a2 was significantly decreased(n = 4-8/group,P < 0.05);Cyp3a11 protein level in male mouse liver was significantly reduced,while Cyp1a2 protein level significantly increased(n = 5-7/group,P < 0.05).5.Effects of exposure to LPS during pregnancy on the ontogenic expression patterns of hepatic nuclear receptors and DMEs in offspring:Female offspring: In the control female offspring,the nuclear receptors Pxr,Ahr,Car and the metabolic enzymes Cyp3a11,Ugt1a1,and Sult1e1 were enriched in adolescence;the metabolic enzymes Cyp1a2,2b10 were enriched in adulthood.For the female mice in the treatment group,the nuclear receptor Ahr mRNA expression level was enriched in the neonatal period;the nuclear receptor Pxr and the metabolic enzymes Cyp1a2,Ugt1a1,and Sult1e1 were enriched in adolescence;nuclear receptor Car and metabolic enzymes Cyp3a11,2b10 Enriched in adulthood.Male offspring: In the control group of male rats,the nuclear receptors Pxr,Ahr,Car and the metabolic enzymes Cyp3a11,2b10,Ugt1a1,and Sult1e1 were enriched in adolescence;the metabolic enzyme Cyp1a2 was enriched in adulthood.For the female mice of the treatment group,the nuclear receptor Ahr mRNA expression level was enriched in the neonatal period;the nuclear receptor Pxr and the metabolic enzymes Cyp1a2,2b10,Ugt1a1,and Sult1e1 were enriched in adolescence;the metabolic enzyme Cyp3a11 was in adolescence and adulthood.The nuclear receptor Car is enriched in the neonatal and adolescent stages.6.Correlation of expression levels liver between NF-?B and metabolic enzymerelated genes in offspring mouseIn the liver tissues of the control group,the mRNA expression level of NF-?B was weakly correlated with the expression levels of Pxr,Car,Cyp2b10,Ugt1a1 and Sult1e1,however,it was not statistically significant;In the liver tissues of the treatment group mice,In mRNA expression levels,there was a moderate positive correlation between NF-?B and Pxr(r = 0.47,P < 0.001);weak negative correlation with Car(r =-0.31,P < 0.05);moderate positive correlation with Cyp2b10,(r = 0.43,P < 0.01);there is a weak positive correlation with Ugt1a1(r = 0.33,P < 0.01).7.Correlation of mRNA expression levels between liver Pxr and metabolic enzymes in offspring mouseIn the liver of the control group,there was a strong positive correlation between Pxr and Cyp3a11 in mRNA expression(r = 0.74,P < 0.001);no correlation with Cyp1a2(r = 0.09,P > 0.05);and Sult1e1 There was a moderate positive correlation(r = 0.55,P < 0.01);there was a moderate positive correlation with Ugt1a1(r = 0.59,P < 0.01).There was a weak positive correlation between Pxr and Cyp3a11 in mRNA expression levels in the progeny mice(r = 0.4,P < 0.01);there was a strong positive correlation with Cyp1a2(r =-0.79,P < 0.001);There was a weak positive correlation between Sult1e1(r = 0.28,P > 0.05);there was a strong positive correlation with Ugt1a1(r = 0.72,P < 0.001).ConclusionExposure to LPS during pregnancy significantly affected the expression levels of inflammation-related genes(NF-?B,IL-6),nuclear receptors(Pxr,etc.)and metabolic enzymes(Cyp3a11,etc.)in the liver of offspring mouse and gender differences.1.Exposure to LPS during pregnancy has different degrees of up-regulation or down-regulation of hepatic nuclear receptors and metabolic enzymes in offspring of the same age.2.Exposure to LPS during pregnancy has different degrees of up-regulation or down-regulation of the same metabolic enzyme genes in different ages,and interferes with the developmental expression pattern of metabolic enzymes during liver development.3.There was a correlation between NF-?B,Pxr,Car,Cyp2b10 and Ugt1a1 mRNA expression in the offspring liver of LPS treated group. |
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