The Influence Of Pregnancy-related Anxiety On Internalization Behavior Of Offspring

Objective To explore the effect of pregnancy-related anxiety on internalization behavior of children through cohort study,and to explore the mechanism of pregnancy psychological stress inducing internalization behavior in young male mice.Methods Cohort study:A total of 3474 pregnant woman enrolled in Ma’anshan Birth Cohort(MABC)from May 2013 to September 2014 were included in follow-up study subjects.The information about demographic characteristics of pregnant was collected in early,second and third trimester of pregnancy.Use the completed self-made scale of pregnancy-related anxiety to assess pregnancy-related anxiety in each period,and the strength and difficulty questionnaire(SDQ)was used to assess internalization behavior of 4-year-old children.Logistic regression model was conducted to evaluate the associations between pregnancy-related anxiety and internalization behavior of 4-year-old children.Experimental study:Kunming female mice mated with male mice.Female mice with vaginal plugs were raised in single cages and recorded as the first day of pregnancy.The pregnant mice were randomly divided into pregnancy control group and pregnancy psychological stress group.The mice in control group were kept normally,and the mice in pregnancy psychological stress group were subjected to psychological stress.Open field test and elevated plus maze were used to detect the degree of anxiety of offspring male mice at 4-5 weeks after birth.Forced swimming test and sucrose preference test were used to detect the degree of depression of offspring male mice.ELISA kit was used to examine the levels of CRH and GC in mice serum.HE staining and TUNEL staining was used to observe pathological changes in m PFC neurons.The Golgi-Cox method was used to observe the dendrite and dendritic spines in m PFC pyramidal neurons.WB was used to quantitatively detect the expression of m TOR and p-m TOR in m PFC.Results Cohort study:The number of mother-child pairs included in the analysis in the first trimester,middle trimester,and third trimester of pregnancy were 1716,2357,and2323,respectively.The detection rates of pregnancy-related anxiety in early,middle and third trimester of pregnancy were 20.0%,23.3%,and 22.7%,respectively.The detection rates of pregnancy-related anxiety in both the early and middle trimesters was 12.7%,and the detection rates of pregnancy-related anxiety in both the middle and third trimesters was 15.7%,and the detection rates of pregnancy-related anxiety in both the first and third trimesters was 12.0%.The detection rate of pregnancy-related anxiety in all three trimesters of pregnancy was 10.0%,and the overall detection rate of internalization behavior in 4-year-old children was 7.4%.The?~2 test showed that there was a statistical significant difference in the detection rate of internalization behavior between mothers with pregnancy-related anxiety and those without pregnancy-related anxiety in the third trimester(P<0.05).Also,the?~2 test showed that there was a statistical significant difference in the detection rate of internalization behavior between mothers with pregnancy-related anxiety and those without pregnancy-related anxiety in both the middle and the third trimester(P<0.05).Furthermore,analysis of multiple logistic regression showed that after adjusting the two potential confounding factors of family monthly income and induced abortion history,as well as adjusting the age of pregnant women,registered permanent residence,family monthly income,pre-pregnancy smoking history,spontaneous abortion history and induced abortion history,pregnancy-related anxiety in the third trimester of pregnancy increases the risk of children internalization behavior(OR=1.627,95%CI:1.157～2.287;OR=1.603,95%CI:1.136～2.261),and pregnancy-related anxiety of pregnancy increases the risk of children internalization behavior in both the middle and the third trimester(OR=1.546,95%CI:1.051～2.275;OR=1.508,95%CI:1.019～2.232).Experimental study:The open field test and elevated plus maze showed that there was no statistical difference in the degree of anxiety between the pregnancy control group and pregnancy psychological stress group.The forced swimming test showed that compared with the CON group,the offspring male mice in PS group had more motion time(P<0.01).The sucrose preference test showed that compared with the CON group,the offspring male mice in PS group had less sugar preference rate(P<0.05).Compared with the CON group,the level of CRH and GC in mice serum were increased in PS group(P<0.05).HE staining showed that in CON group most pyramidal neurons were clearly stained,neatly arranged,round spheres,round nucleus,and obvious nucleoli in m PFC,Compared with the CON group,the number of neurons in the PS group was reduced(P<0.05),and some pyramidal neurons were disordered,exhibiting loose arrangement and dark-staining,with visible nuclear shrinkage.TUNEL staining showed that compared with the CON group,the number of apoptotic cells and the apoptotic index were increased in the PS group(P<0.01).Moreover,the Golgi-Cox method showed that compared with the CON group,the dendritic complexity and dendritic spine density were decreased in the PS group(P<0.05).Lastly,compared with the CON group,the expression of m TOR and p-m TOR in m PFC were both increased in the PS group(P<0.05).Conclusion Cohort study showed that pregnancy-related anxiety in the third trimester or pregnancy-related anxiety in both the middle and third trimesters are risk factors for internalization behavior in children.Animal experimental study showed that the mechanism of pregnancy psychological stress inducing adverse psychology in offspring male mice may be related to the offspring’s HPA axis dysregulation,and the inhibition of the m TOR signaling pathway which leads to neuron injury in the m PFC of offspring brain.