| Objective:In this study,lipid metabolism in plasma of ovariectomized rats and normal rats was observed by Lipidomics analysis,and different lipid metabolites related to menopausal rats were screened.Through the change of relative content of lipid metabolites,we analyzed the status of lipid metabolism in menopausal rats and its relationship with senile diseases.We studied the regulation of QXZST on lipid metabolism in menopause from the change of metabolites content and metabolic pathway.we hope to reveal the regulation mechanism of QXZST on menopausal syndrome from the level of Lipidomics,and lay a foundation for further development and utilization of QXZST.Methods:Four-month-old female SD rats were randomly divided into blank group and ovariectomized group.In the blank group,only the pelvic cavity was opened and the ovaries were not removed.The ovariectomized group underwent bilateral ovariectomy,and the results of modeling were observed by smears of exfoliated vaginal cells in rats.After operation,ovariectomized rats were randomly divided into five groups:model group(group M),QXZST low-dose group(group D),QXZST moderate-dose group(group Z),QXZST high-dose group(group G),Remifemin group(group Y),and blank group(group K),totally 6 groups.The rats were anesthetized with 10%chloral hydrate at a weight of 0.35 ml per kilogram after fasting for 24 hours.Carotid artery blood was taken from the carotid artery to the blood vessels.The centrifuge was centrifuged at a speed of 3000 rpm for 20 minutes.Serum was taken for reserve.UPLC/LTQ-Orbitrap-MS technology was used to detect plasma in each group.The differential lipid metabolites between model group and blank group were found by principal component analysis(PCA),orthogonal partial least squares analysis(OPLS-DA)and differential multiple analysis(FC).The chemspider database was searched based on the precise molecular mass and mass ratio information detected,and the fragment ion information.The metabolites were identified and the differential metabolites were input into KEGG database for checking.Find metabolic pathways.Using multivariate statistical analysis to compare the relative content of these different lipid metabolites in rats of each dose group of QXZST and Remifemin group,we speculate the therapeutic mechanism of QXZST on ovariectomized rats.Results:1.PCA method was used to model the rat samples of model group and blank group.Most of the samples were concentrated in the elliptic 95%confidence region(95%confidence region)of the scatter plot,showing a certain separation trend,but there was still overlap.The OPLS-DA model was further established.The parameters were plasma(positive ion model):R2Y=0.996,Q2=0.789;plasma(negative ion model):R2Y=0.999,Q2=0.694.R2Y represents the interpretation rate of the model and Q2 represents the prediction rate of the model.The above parameters show that the model established in this experiment has high reliability and OVX rat model has been successfully established.2.Lipidomics analysis was performed on the plasma of the group M and the group K.The lipids of OPLS-DA model were analyzed by VIP>1 and univariate statistical analysis P<0.05,and the coefficient of variation(FC)>1.2 were calculated as significant difference lipids.The experiment revealed the differential metabolites related to menopause syndrome.There were 9 types of plasma lipids with significant difference and 76 kinds of lipid compounds.12 ceramides(Cers),29 sphingomyelins(SMs),5 triglycerides(TGs),5 Lysophosphatidylcholines(LPCs),2 fatty acids(FAs),and 1 platelet activating factor(PAF),their relative content increased compared with the blank group(p<0.05).4 phosphatidylglycerols(PGs),4 phosphatidylserines(PS s),14 diacylglycerols(DGs),their relative content decreased compared with the blank group(p<0.05).3.After searching the metabolic pathways of different lipid metabolites,six metabolic pathways were found to be associated with menopausal syndrome,including Glycerophospholipid metabolism,sphingolipid metabolism,glyceride metabolism,cholesterol metabolism,ether lipid metabolism and unsaturated fatty acid synthesis.4.By comparing the changes of relative contents of lipid metabolites in rats before and after administration of QXZST,it was found that QXZST can regulate several disorders of lipid differential metabolites related to menopausal syndrome.After the intervention of QXZST on the plasma metabolism of ovariectomized rats,5 types of differential lipids were identified,which could up-regulate 2 indexes and down-regulate 3 indexes.The difference in relative content of the above indexes was statistically significant compared with that of the model group(p<0.05).QXZST plays a therapeutic role by regulating the metabolism of glycerolphospholipid metabolism,sphingolipid metabolism and glycerol ester metabolism pathway.Conclusion:l.UPLC/LTQ-Orbitrap-MS metabonomics can preliminarily distinguish two different lipid metabolism states between normal and menopausal rats.The results showed that 76 different lipid compounds were related to lipid metabolic disorders in menopausal syndrome,and the changes of these metabolites could be used as predictive markers of menopausal-related degenerative diseases in the elderly.2.Ovariectomized rats have disorders in glycerophospholipid metabolism,sphingolipid metabolism,glycerol ester metabolism,cholesterol metabolism,ether lipid metabolism and unsaturated fatty acid synthesis.3.QXZST can regulate the lipid disorder caused by menopausal syndrome to a certain extent,mainly through regulating glycerol phospholipid metabolism,sphingolipid metabolism and glycerol ester metabolism.4.Through the changes of relative contents of these metabolites and the regulation of metabolic pathways after administration of QXZST,it is revealed that an important therapeutic mechanism of QXZST is to prevent and delay the occurrence of postmenopausal cardiovascular and cerebrovascular diseases,Alzheimer’s disease,diabetes mellitus and metabolic diseases.To provide ideas for further pharmacological research. |
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