The Quality Standard Study Of Shenguien Mixture And The Effects Of MEK1 Of ERK1/2 Signaling Pathways In The Brain Tissue Of The Postpartum Depression Rats

Object At present,the number of women with postpartum depression is increasing,more and more scholars in China and abroad.Because of the complex pathogenesis of postpartum depression,a large number of studies have confirmed that the rapid change of hormone levels in the body is the biological basis of postpartum depression.However,there is no report on the relationship between postpartum depression and estrogen and nerve signal transduction pathway."The shenguiren mixture"for clinical experience of Affiliated Hospital of Chengdu University of Traditional Chinese Medicine,composed of ginseng,angelica,Suanzaoren,with Replenishing Qi and blood,nourishing qi,the efficacy of ningxinanshen.The topic for postpartum depression is"Qi deficiency blood stasis syndrome�,she put forward the"treatment of postpartum depression Buxu Huayu method",and further study the shenguiren mixture on the mechanism,the effect of MEK1 in brain tissue of rats with postpartum depression in ERK1/2 pathway.Whether to improve depressive symptoms using MEK1 expression levels of estrogen and neuronal signal transduction pathways,which is the subject of a major innovation,but also provide a scientific basis for Chinese medicine treatment of postpartum depression,it lays the foundation for the development of new drugs.Methods On the basis of the preparation technology,TLC was used to analyze qualitatively the contents of ginsenoside,angelica and sour jujubes,and the contents of ginsenoside Rg1,ginsenoside Re and ginsenoside Rb1were determined by high performance liquid chromatography?HPLC?,to reseash the development of the quality standard of Shenghui mixture.The PPD rats were subjected to ovariectomy and the behavioral changes were observed.It is detected the protein expression of P-MEK1?phospho-mitogen-activatedextracellularregulatedkinase1??MEK1?mitogen-activated extracellular regulated kinase1?of the ERK1/2?extracellular signal-regulated kinase1/2?pathway of the brain tessiue of the rats by Western-blot.1.Study on Preparation Technology of Shengui MixtureThe?34?orthogonal test was carried out in the early stage of the decoction of ginseng and angelica mixture,and the optimum extraction process was determined by L9.Results showed that the extraction process of ginseng alcohol:8 times amount of70%ethanol extract three times,each time 1 h;Angelica distilling with water extraction process:12 times the amount of water for 12 h,a total of 10 h distilled water,collecting volatile oil;extraction technology of jujube kernel water:10 times of water extracting three times,each time 1 h.Suanzaoren aqueous extract by precipitation,less the amount of the loss of Jujuboside A.The experimental results show that the preparation process is stable and feasible.2.Study on quality standard of the Shenguiren mixtureThe TLC identification of ginseng,angelica and wild jujube kernel,the separation effect is good,the negative control without interference.The content of ginsenoside Rg1,ginsenoside Re and ginsenoside Rb1 were determined by HPLC.Chromatographic conditions:Dimma Diamosil C18?250 mm x 4.6 mm,5 um?column,acetonitrile water two-phase,gradient elution,column temperature:25degrees centigrade,flow rate:1 mL�min^-1,detection wavelength:203 nm,sample volume:10 uL,the number of theoretical plates must be more than 6000 calculated by ginsenoside R_g1peak.3.The effect of the Shenguirenmixture of MEK1 of ERK1/2 pathway of postpartum depression on rats.?1?150 female rats were randomly divided into blank group?n=16?and model group?n=121?.The model group were subjected to bilateral ovariectomy on the vaginal smears were 2-5 days after operation.The rats were injected with estradiol benzoate and progesterone for 16 days continuously,and the pregnant state was simulated.The model rats were randomly divided into 6 groups,each group of 16 rats.That is,the blank group,PPD?Postpartum Depression,postpartum depression?model group,Seculin group,estradiol group,the high dose,middle dose and low dose group.Each group according to the corresponding drug?distilled water,distilled water,Seculin,estradiol,shenguiren?on the gastric perfusion,and set up 1,2 and 4 weeks respectively for all observation points,group behavior tests were carried out,including sugar consumption test,open field test and forced swimming test,observe the changes of.?2?After 30 days of drug intervention,They were taken out and stored at-80?about the hypothalamus,hippocampus and the frontal lobe in the brain tissue of rats of each group.The P-MEK1 and MEK1 in the frontal lobe,hippocampus and hypothalamus were detected by Western-blot.Gray-scale of P-MEK1 was detected by ImageJ software.The gray value of P-MEK1 was higher.The data were analyzed by GraphPad Prism 5 software.Results?1?Ginseng,Angelica and Semen Ziziphi spinosae were identified by thin-layer chromatography.The specificity and specificity of the ginsenoside Rb1,Re,Rg1 were 0.424⁴.240 mg�ml^-1,.04312⁴.312 mg�ml^-1,0.412⁴.120mg�ml^-1?R=0.9999?.The reseach of the methodological RSD were all up to the standard.?2?Compared with the blank control group,the body weight of the PPD model group increased slowly.The level of exercise in the open field,the vertical movement score,the central lattice residence time and the consumption of sugar water decreased significantly?P<0.001?.Compared with the PPD model group,there were significant differences between the high,middle and low groups and the positive control group?P<0.001 or P<0.01?.?3?Western blotting showed that the protein expressions of P-MEK1 and MEK1 on the frontal lobe,hippocampus and hypothalamus of depressive model group were significantly decreased,and the protein expression of P-MEK1 in the frontal lobe,hippocampus and hypothalamus were increased significantly,especially in the middle and low groups.Conclusion?1?TLC is the best method for analyzing qualitatively the contents of ginsenoside,angelica and sour jujubes.The determination of the Ginsenoside Rg1,GinsenosideReandGinsenosideRb1isHPLC.Themethodis simple,feasible,accurate and reproducible.And it can be used quality Control of the Ginsenoside Rg1,Ginsenoside Re and Ginsenoside Rb1.?2?The postpartum depression rats showed depression-like behavior,with antidepressant effect,Shen Gui mixture is improved its behavioral indicators.?3?The protein expression of P-MEK1 and MEK1of ERK1/2 signal pathway was up-regulated in a certain extent by Shengui Mixture,and the effect of the middle and low groups was significant.Indicating that the mechanism of this drug is regulated the ERK1/2 pathway and improved the depressive symptoms.