The Mechanism Of Fear Of Kidney Injury During Pregnancy Affecting Offspring Learning And Memory Ability Based On Reelin Protein

Objective: According to the traditional Chinese medicine theory of "kidney dominates fear" and "kidney generates marrow and brain",we used prenatal compound fear stimulation to act on pregnant rats to establish a model of fear of injury to the kidney during pregnancy,and to observe the behavior of offspring rats induced by fear stimulation during pregnancy To change the behavioral characteristics of Zuogui Pill with Bushen Bujing Fang,and to explore the possible mechanism of fear of kidney injury during pregnancy affecting the brain function of offspring rats.Methods: Healthy adult SD rats were selected,and the pregnant rats were randomly divided into experimental group（fear of stimulation during pregnancy,fear of kidney injury group）and control group（normal feeding）.Starting from the 7th day of gestation,the pregnant mice in the group of pregnant mice in the kidney injury group were randomly stimulated with plum-blossom needles,electric shocks on the feet,simulated earthquake platform,and high decibel noise until they gave birth.The control group was fed completely normally and guaranteed to be undisturbed.The first day of birth was recorded on the day of the birth of the pregnant rats,and they were weaned on the 21 st day after birth,and then separated,and the offspring male rats of the prenatal "fear of injury to the kidney" group at 42 days postpartum were randomly divided into prenatal "fear of injury to the kidney" stimulation + Zuogui pill group（n=10）,prenatal "fear of hurting the kidney" stimulation + normal saline group（n=10）.And blank control + normal saline group,gavage,once a day for 4 consecutive weeks（n=10）.On the 16 th day of pregnancy,the sugar water preference test and open field test were used to evaluate the rat model of fear of kidney injury during pregnancy.After 21 days of weaning,the offspring were separated and fed,and42 days after birth,they were given gavage for four consecutive weeks.The Morris water maze test was performed to evaluate their learning and memory ability.The brain weight of the offspring in each group was weighed,real-time polymerase chain reaction（RT-PCR）and Western blot（WB）were used to detect the m RNAs of Reelin（Reelin）,LIMK and Cofilin in the hippocampus of the offspring rats and protein,as well as the protein expression changes of synaptic structural protein synaptophysin（SYN）and postsynaptic densification protein9（PSD-95）.Results:（1）The comparison of sugar water preference test and open field experiment of pregnant rats in the control group and the fear-injured-kidney group showed that the pregnancy-injured-kidney rats showed obvious stress state,indicating that the modeling was successful.（2）In the positioning navigation experiment,compared with the control group,the offspring of kidneys may be damaged.The escape latency was significantly prolonged（P<0.01）,indicating that the learning ability of the offspring might be damaged during pregnancy.（3）In the space exploration experiment,the number of offspring crossing the platform with fear of kidney injury was significantly reduced,and the percentage of staying in the third target quadrant was significantly shortened（P<0.05）,indicating that the scaring stimulus during pregnancy reduced the offspring’s memory ability.The Zuogui Pill intervention could improve the learning and memory decline of the offspring of fear of kidney injury（P<0.01）.（4）The expression of SYN and PSD-95 proteins in the hippocampus of the offspring of fear of kidney injury was significantly decreased,indicating that the synaptic structure of the offspring of fear of kidney injury during pregnancy was injured.（5）In the follow-up experiments,by comparing the m RNA and protein expressions of Reelin,LIMK,and Cofilin in the hippocampus of the offspring rats of the control group and the two groups,it was found that the m RNA and protein of Reelin and Cofilin in the offspring induced by scaring stimulation during pregnancy were significantly higher.It was down-regulated（P<0.01）,and this change could be reversed by Zuoguiwan.Conclusion: Fear-injured kidney during pregnancy can induce abnormal changes in offspring rats with decreased learning and memory ability,and postpartum intervention with the kidney-invigorating formula Zuogui Pill can improve the learning and memory decline of the fear-injured kidney rat to a certain extent,further demonstrating the fear of injury.The related theory that the kidney causes fetal damage and the kidney is the innate foundation.Fear injury during pregnancy may affect the expression of Reelin in the hippocampus of offspring,reduce the phosphorylation of LIMK,inhibit the phosphorylation of Cofilin,and then affect the stability of the actin skeleton,resulting in the reduction of synaptic structure and the decline of learning and memory ability.