1.Study on the Interventional Effects of Cornus Officinalis Before and After Wine Preparation in Liu Wei Di Huang Decoction on Postmenopausal Osteoporosis Model Rats Based on Medicinal
Zhi WEN ; Hui YU ; Qiong LONG ; Jiaying TAN ; Li HUANG ; Wangzhong XIAO ; Bing DAI
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(8):2717-2725
Objective To establish a dual wavelength HPLC method to compare the content changes of seven active components:monoglycoside,loganin,swertin,paeoniflorin,gallic acid,5-hydroxymethyl furfural and paeonol in Liuwei Dihuang Decoction(hereinafter referred to as"Liuweidihuang Decoction")before and after the preparation of Cornus wine;To explore the effect and mechanism of Liu(Shan)and Liu(Jiu)on 90 days after ovariectomy in postmenopausal osteoporosis model rats.Methods ①The contents of seven active ingredients in the Cornus wine mixed with Liuwei Dihuang Decoction were determined by dual wavelength HPLC.Chromatographic conditions:hypersil C18 chromatography(4.6 mm×250 mm,5 μm),mobile phase acetonitrile-0.3%phosphoric acid,gradient elution(0-7 min,0%-8%acetonitrile;7-15 min,8%-10%;15-20 min,10%-15%;20-30 min,15%-23%;30-50 min,23%-45%),flow rate 0.6 mL·min-1,detection wavelength:240 nm(monoglycoside,loganin,swertin,paeoniflorin),274 nm(gallic acid,5-hydroxymethyl furfural,paeonol),column temperature 30℃,injection volume 20 μL.②6-8 month old SD rats,divided into sham-operated group,model group,positive drug group,shanshuang flesh with Liuwei Dihuang Decoction group(liu(shan)group)and wine cornelian flesh withLiuwei Dihuang Decoction group(liu(wine)group),dosing started on the 5 d after ovariectomy,weighed weekly,and 5 rats from each group were taken at 90 d after surgery,weighed,anaesthetized,serum taken,bilateral femurs,and organs(uterus,spleen,liver,both kidneys and heart)were weighed.Serum calcium(Ca),phosphorus(P)and estradiol(E2)levels were measured using a fully automated biochemical instrument and chemiluminescent microparticle immunoassay,respectively;bone mineral density(BMD)values were measured using a dual-energy X-ray bone densitometer.HE staining was used to observe the morphological changes of bone tissue in rats.Results ①All the seven potent ingredients mentioned above were contained in the Liuwei Dihuang Decoction before and after the wine preparation of Cornu Cervi Pantotrichum,and the total content of the seven potent ingredients in liu(wine)was higher than that in liu(shan)(P<0.01).Among them,gallic acid and 5-hydroxymethylfurfural were higher in Liu(Jiu)than in Liu(Shan)(P<0.01),while the content of monosidine was lower than in Liu(Shan)(P<0.01).②Comparison with the sham-operated group:90 d after surgery,the serum Ca,P,E2 levels and uterine index of rats in the model group decreased(P<0.01),the BMD value decreased(P<0.01),and the degree of structural damage to bone tissue The degree of damage to the bone tissue structure was deepened,manifested by enlargement of the bone marrow cavity,gradual increase of adipocytes,dilution of the bone trabeculae arrangement,reduction of the number of bone trabeculae or even fracture.③Compared with the model group,90 d after surgery,the serum E2,Ca,P levels and uterine index of the rats in the liu(shan)and liu(wine)groups increased(P<0.05),the bone tissue structure of the liu(shan)and liu(wine)groups improved significantly,and the BMD value increased significantly(The liver and spleen indices of the liu(wine)groups increased(P<0.05).Conclusion The combination of the wine preparation of Cornus officinalis and its use in the formula of LiuWei Di Huang Decoction both contain monosidine,strychnine,sweretin,paeoniflorin,gallic acid,5-hydroxymethyl furfural(5-HMF)and tannin,and can effectively interfere with PMOP.Among them,the anti-osteoporotic effect of Liu Wei Di Huang Tang,which was formulated with wine and dogwood,was better than that of Liu Wei Di Huang Tang,which was formulated with Cornus officinalis,and the former contained more 5-HMF and gallic acid than the latter,which may provide some experimental basis for the selection of wine and dogwood in the 2020 edition of the Pharmacopoeia and the clinical application of Liu Wei Di Huang Tang in the prevention and treatment of PMOP.
2.Study on Optimization of the Nine-time Repeat Steaming and Sun-drying Process Processing Technology of Polygonum multiflorum
Jun’an PU ; Simin ZHANG ; Mengjiao LIU ; Wangzhong XIAO ; Lei YANG
China Pharmacy 2020;31(22):2713-2719
OBJECTIVE:To establish methods for the content determination of stilbene glycoside ,emodin,emodin methyl ether and total polysaccharide in P. multiflorum ,and to optimize the processing technology of traditional nine-time repeat steaming and sun-drying process of P. multiflorum . METHODS :HPLC method was adopted to determine the contents of stilbene glycoside , emodin,and emodin methyl ether. The content of total polysaccharide was determined by UV spectrometry. With steaming time , drying time and drying temperature as independent variables ,the contents of stilbene glycoside ,emodin,emodin methyl ether and total polysaccharide as indexes ,the processing technology of nine-time repeat steaming and sun-drying process of P. multiflorum was optimized by weighted comprehensive score method and central composite design-response surface method ,and the validation tests were conducted. RESULTS :The linear range of sample size of stilbene glycoside ,emodin and emodin methyl ether were 0.27-2.7 µg(r=0.997 1),0.063-0.63 µg(r=0.999 9),0.038-0.38 µg(r=0.990 9),respectively. The linear range of mass concentration of total polysaccharide was 2.07-12.42 µg/mL(r=0.999 6),respectively. RSDs of precision ,stability and reproducibility tests were all lower than 2%. The recoveries were 99.43%-101.06%(RSD=0.63%,n=6),98.74%-120.33% (RSD=1.34%,n=6),98.39%-102.44%(RSD=1.49%,n=6),99.51%-101.98%(RSD=0.87%,n=6),respectively. The optimal processing technology was steaming time for 4.5 h,drying time for 9 h,drying temperatrue for 66 ℃ and repeat processing for 9 times. Results of 3 times of validation tests showed that the comprehensive score of optimal technology was 35.69, relative error of which to predicted value (36.90)was 3.30%. CONCLUSIONS :Established method is simple and reproducible , and the optimal processing technology is predicive ,stable feasible.