| Fructus Corni as the Comaceae plant Cornus(Cornus officinalis Sieb.et Zucc.)is the dried ripe fruit pulp,its flavour acid,acerbity,lukewarm,belongs to the liver and kidney.Fructus Corni has effects on liver and kidney tonic,astringency and stypsis,dizziness,tinnitus and antidiabetes.It mainly produced provinces in China,such as Henan,Shaanxi,Anhui,Zhejiang province and so on.Iridoid glycosides is the main active components of Fructus Corni and exhibits several biological activities,including anti-inflammatory,antioxidant,antithrombotic,reduce blood glucose and immunological regulation effects,and to protect brain and kidney functions.Additionally,it can ameliorated diabetes and its complications.Hence,this studyexplores the iridoid glycoside of Cornus officinals Sieb.et Zucc.(CIG)and it's antidiabetic effect.In this study,CIG extraction process has been optimized using response surface methodology(RSM)and ultrasound assisted microwave extraction(UAME).The D101 macroporous adsorbing resins was used to purify CIG,and offered test samples for its pharmacological studies.Also,contents of CIG of six provinces such as anhui,zhejiang,hubei,henan,shaanxi and 15 different regions were detected,and performed qualitative and quantitative analysis,further confirmed the high yield iridoid glycoside of Cornus officinalis producing area,and through the multiple comparison analysis of the differences of different origins of CIG.Finally,effects of CIG on glucolipid metabolism,liver inflammation,oxidative stress and the key of the skeletal muscle insulin resistence(IR)target gene of mRNA expression in diabetic mice were investigated.This preliminary explored the function of CIG in inhibiting the pathogenesis of diabetes.The main results of this experiment were as follows:(1)The optimal extraction condition of CIG was as follows:microwave power 641W,ultrasonic power 360 W,solvent to sample ratio 21:1 mL/g and extraction for 9 minutes.(2)D101 macroporous resin for CIG static adsorption ratios and desorption ratios was better,AB-8 was worse,and the purity of CIG was up to 36.76%.(3)Detection of CIG contents among different regions showed that the highest the samples was produced in zhejiang ChunAn and up to 32.68 mg/g.Mutiple comparison analysis indicating that chunan county,zhejiang province had significant difference(p<0.05)compared to yangcheng county,shanxi province,yunxian county,anhui province,neixiang county,henna province,and foping county,shaaxi province.Both Chunan county,zhejiang province and foping county,shaaxi province had significant difference(p<0.05)in contrast to yangcheng county,shanxi province,yunxian county,anhui province,and neixiang county,henna province.Meanwhile foping in shaanxi province had significant difference(p<0.05)compared to other regions.(4)RP-HPLC-UV method was eatablished for determination of 4 iridoid glycosides simultaneously.The JADE-PAK ODS-AQ C18 column(250 mm×4.6 mm,5?m)and the ultraviolet detector(UV)were used for separation and detection,respectively.The separation process was carried out using a gradient elution(0-15 min,10%B?20%B;15-19 min,20%B?22%B;19-25 min,22%B?25%B;25-35 min,25%B?27%B;35-50 min,27%B?70%B;50-55 min,70%B?10%B),which was based on mobile phase of the mixture aqueous solution with 0.2%acetic acid(A)-acetonitrile(B)at the flow rate of 1.0 mLˇmin-1.The detection wavelength was 240 nm,the column temperature was 35 ? and the injection volume was 10 uL.The results showed that the chromatographic could be used for the separation and determination of 4 iridoid glycosides,the peak shape was presented well and was separated well.The method was simple and reliable,with high precision and good repeatability,and provided fundamental evidence for theoretical and scientific research of CIG in controlling intrinsic quality and pharmacodynamic evaluation(5)The animal experiments showed that the status among the groups of mice were improved by feeding CIG for 4 weeks,and the weight of mice of the middle of Cornus officinalis iridoid glycoside(CIG-M),high of Corpus officinalis iridoid glycoside(CIG-H),positive compare(PC)groups were raised.The fasting blood glucose(FBG)in each dose groups and PC groups were significantly lower(p<0.01),fasting insulin(FINS),insulin sensitivity index(ISI),high-density lipoprotein cholesterol(HDL-C)increased significantly(p<0.05),while insulin resistance index(IRI),total cholesterol(T-CHO),triglycerides(TG),and low-density lipoprotein cholesterol(LDL-C)were significantly lower(p<0.05).Glucose tolerance(OGTT)and organ index had the degree of improvement,which clearly proved that CIG could improve glucolipid toxicity,stimulate isle beta cells or repair the damaged islet beta cells to release more insulin to reduce blood sugar.Content of tumor necrosis factor-a(TNF-a),interleukin-6(IL-6),C reactive protein(CRP),and malondialdehyde(MDA)was reduced,but superoxide dismutase(SOD)was up-regulated in each does group treated with CIG,indicating that CIG could reduce the content of liver tissue inflammation factors,lower lipid peroxidation,increase the activity of antioxidant enzymes,reducing oxidative stress damage to the organization,and enhancing the antioxidant capacity.The results showed that CIG could improve IR and antidiabetes by regulating multiple targets.Histopathological results indicating that CIG can obviously improve the liver cells and the degree of lesion of pancreatic cells in diabetes mice.CIG-H and PC group were significantly up-regulated the mRNA expression of GLUT-4?INSR?PI3K and PKB gene in mice skeletal muscle(p<0.01),the experimental results indicated that CIG could improve IR and treat diabetes by up-regulating the expression of key target genes in insulin signaling pathway. |
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