| In recent years,the incidence of diabetes has shown a continuous increase and has become a global health problem.At the same time,there will be a decline in cognitive function in patients with T2DM.Therefore,neurological diseases caused by diabetes are getting more and more attention.There is currently no effective drug that can cure T2DM and its associated cognitive dysfunction,and its pathogenesis has not yet been fully explained.As a shrub-like aromatic plant,rosemary has pharmacological effects such as calming and soothing,anti-oxidation,antibacterial,anti-tumor,anti-inflammatory,and anti-depression.At present,it has been reported in the literature that rosemary has a certain regulatory effect on the blood sugar and blood lipids of diabetic rats,but there are few reports on the effect of rosemary on improving diabetic cognitive impairment and the specific mechanism of action.Rosemary is not included in the current edition of"Chinese Pharmacopoeia",and there is no evaluation standard for its medicinal use.This article takes rosemary as the object of research to study its improvement of diabetic cognitive dysfunction.At the same time,it is based on high performance liquid chromatography,quadrupole time-of-flight mass spectrometer and triple quadrupole mass spectrometer.Qualitative and quantitative studies were carried out on the ingredients in Rosemary.The pharmacokinetic characteristics of the 4ingredients in rosemary in rats were also studied,which provided a certain scientific basis for the clinical application of rosemary.The content of the thesis is divided into four chapters,and the research is carried out from the following aspects:1.The research of rosemary on improving diabetic cognitive impairment.Taking T2DM db/db mice as a model,the mice were administered by intragastric administration,and the metabolism,inflammation and cognition and other related indicators of the mice were detected after 8 weeks.Experimental results show that rosemary can reduce the weight of mice,significantly reduce the fasting blood glucose and triglyceride content of mice,reduce free fatty acids and insulin,and at the same time increase insulin sensitivity and increase high-density lipoprotein blood lipids.Pathological results show that rosemary can significantly reduce fat deposition and tissue inflammation.Morris water maze test results show that rosemary can significantly improve the learning and cognitive ability of db/db mice.The q PCR results showed that rosemary can reduce the expression of TNF--inflammatory cytokine in the hippocampus of db/db mice,and help to increase the expression of IL-4 and ARG1 anti-inflammatory cytokines.The results of ELISA kit and Western Blot analysis showed that rosemary increased the content of BDNF in the hippocampus of db/db mice and decreased the level of MDA,thereby enhancing the plasticity of hippocampal nerves,and at the same time increasing the expression of NF200 and PSD95 in the hippocampus of db/db mice.,Make the level of GFAP significantly reduce,indicating that rosemary can reduce the activation of astrocytes in db/db mice while inhibiting the expression of astrocytes,thereby reducing neuroinflammation..These results confirm that rosemary has a certain therapeutic effect on DCI at the animal level.2.Research on the chemical constituents of rosemary based on UPLC-Q-TOF/MS.An Agilent Zorbax-C18(100 mm×2.1 mm,1.8μm)chromatographic column,acetonitrile-0.1%formic acid aqueous solution as the mobile phase,ESI ion source,and positive and negative ion modes were used for collection.A total of 17 chemical components were identified in the experiment,which can be divided into flavonoids and phenolic acids.The method is accurate and rapid,and provides scientific basis for the qualitative,quantitative and medicinal material basis of the chemical components of rosemary decoction.3.Study on HPLC fingerprint of rosemary and its multi-component quantitative analysis.The Yuexu Ultimate XB-C18 analytical column(250 mm×4.6 mm,5μm)was used on the chromatographic column with acetonitrile-0.1%formic acid aqueous solution as the mobile phase,and the detection ultraviolet wavelength was 284nm.Establish fingerprints of 24 batches of rosemary with similarity between 0.885 and0.967.Systematic clustering analysis is divided into three categories.Through principal component analysis and orthogonal partial least squares discriminant analysis,6 large differences are found.Markers.After comparing with the control,5chromatographic peaks were identified,namely,Danshensu,chlorogenic acid,caffeic acid,psyllium,and peak rosmarinic acid.Simultaneously determine the content of these five components in rosemary,and investigate its methodology.The regression equation has a good linear relationship(r2=0.9996～0.9999);the method has good repeatability(RSD≤2.54%,n=6);the sample recovery rate is 84.5%～116.88%(RSD≤2.84%,n=3);The samples are kept at room temperature for 24h(RSD≤2.97%),and the stability is good.The method has good specificity,high accuracy,quickness and simplicity,and provides a basis for the quality standard of rosemary.4.The pharmacokinetic study of the four ingredients in rosemary.The LC-MS/MS method was established to determine the plasma concentrations of Danshensu,chlorogenic acid,plantagoside and rosmarinic acid in rat plasma.An Agilent Zorbax-C18(100 mm×2.1 mm,1.8μm)chromatographic column was used for gradient elution with 0.1%formic acid-acetonitrile solution as the mobile phase.The plasma concentration was determined and the pharmacokinetic parameters were calculated.The results show that each component can achieve baseline separation within 8 minutes of detection time,the linear relationship of the regression equation is relatively good(r~2≥0.9978),the RSD of the relative standard deviation of the inter-day and intra-day precision is less than 6.24%,and the average extraction recovery rate is Between 86.2%and 115.47%.The method was verified by methodology and successfully applied to rats,and the pharmacokinetic study of four components in plasma after gavage rosemary perfume decoction. |
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