| Object:The object of study to observe the effect of Qingxin Zishen Decoction on skin temperature of the tail of hot flashes in postmenopausal rats,and the effect on ET and NO in blood of hot flashes in postmenopausal rats.To establish an HPLC fingerprint of Qingxin Zishen Decoction and determine the contents of its multiple components,so as to provide a scientific basis for quality control.Methods:1.Seventy adult SD rats were randomly divided into ovariectomized group and control group.The surviving ovariectomized rats were randomly divided into 5 groups(n=10):model group,Qingxin Zishen Decoction low-dose group,moderate-dose group,high-dose group,Estradiol valerate group.Skin temperature of the tail of each group was measured continuously every week.2.Seventy adult SD rats were randomly divided into ovariectomized group and control group.The surviving ovariectomized rats were randomly divided into 5 groups(n=10):model group;low-dose group,moderate-dose group,high-dose group of Qingxin Zishen Decoction;Estradiol valerate group.Abdominal aortic blood of rats in each group was taken and the contents of NO and ET were determined.3.HPLC analysis was performed on a Phenomenex Kinetex C18 column(100 mm×4.60 mm,2.6 μm),gradient eluting with the mobile phase consisting of methanol,acetonitrile and 0.2%formic acid aqueous.The detection wavelength was set at 245 nm and 280 nm,and the column temperature was 40℃.Fingerprints of eight batches of Qingxin Zishen Decoction were determined,and the similarities among fingerprints were evaluated.Attributive analysis and identification of common peaks were performed and the contents of 15 components were determined.Results:1.Compared with the control group,tail temperature of rats in the model group increased significantly after bilateral ovaries were removed(P<0.05).Compared with the model group,the tail temperature of rats in the estradiol valerate group,the moderate-dose group and the high-dose group of Qingxin Zishen Decoction decreased significantly within 1-4 weeks(P<0.05)Within 2-4 weeks after administration,the tail temperature of rats in the low-dose group of Qingxin Zishen Decoction decreased significantly(P<0.05).Within 3-4 weeks of administration,tail temperature of rats in Qingxin Zishen Decoction group and estradiol valerate group decreased significantly,with no statistically significant difference(P>0.05).2.Compared with the control group,the content of NO and NO/ET in the model group decreased significantly(p<0.05),and ET has an upward trend.After administration,the content of NO and NO/ET in the low-dose,moderate-dose group and high-dose groups of Qingxin Zishen Decoction significantly increased(P<0.001),and ET showed a decreasing trend3.The fingerprint similarities of 8 batches of Qingxin Zishen Decoction were all greater than 0.98 compared with the reference fingerprint,and 33 common peaks were identified in the fingerprint.Among them,nine peaks were identified from Salvia Miltiorrhiza,nine peaks were identified from Corni Fructus,two peaks were identified from Ziziphi Spinosae Semen,two peaks were identified from Nelumbinis Plumula,seven peaks were identified from Coptidis Rhizoma,while four peaks cannot be originated and none of the common peaks was identified from Rehmanniae Radix,Uncariae Ramulus Cum Uncis and Triticum aestivum.By comparing with the chemical reference,fifteen components,including gallic acid(P2),5-HMF(P3),danshensu(P4),protocatechuic aldehyde(P5),morroniside(P9),caffeic acid(P10),cornin(P12),loganin(P13),magnoflorine(P14),coptisine(P24),lithospennic acid(P28),berberine(P29),palmatine(P30),salvianolic acid B(P31),and salvianolic acid E(P33),were identified and quantified.The contents of the fifteen components were 159.7-248.2μg/g,244.9-321.3μg/g,63.5-116.2μg/g,28.6-38.6pg/g,804.5-1263.6μg/g,26.6-50.7μg/g,79.4-108.2μg/g,470.4-757.3μg/g,137.1-178.6μg/g,13.2-34.2μg/g,69.4-106.4μg/g,25.9-138.9μg/g,23.0-59.2μg/g,986.2-2244.7μg/g,and 41.9-92.8μg/g,respectively.Conclusion The method established in this study is simple,accurate and highly reproducible,and can provide basis for quality control of Qingxin Zishen DecoctionConclusion:1.Qingxin Zishen Decoction can significantly reduce the skin temperature of the tail of hot flashes in postmenopausal rats2.The content of NO and NO/ET in the serum of postmenopausal rats was increased by Qingxin Zishen Decoction3.The method established in this study is simple,accurate and highly reproducible,and can provide basis for quality control of Qingxin Zishen Decoction. |
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