Correlation Based Puerarin Pueraria Estrogen-like And Vasodilatory Effects Of Specific Knockout

The correlation between indicators and its efficacy in the Chinese herbs is the key scientific issues of basic and applied research of Chinese medicine. For most of Chinese medicineherbs, choosingtypical monomer component (representative component) as a starting point in-depth study is one of the main strategies used in the field of Chinese medicine currently,but this approach ignores the unique effect of holistic medicine plays,that is:How representative are the monomer component,if they are associated with traditional Chinese medicineindications?We the need to explore sound, scientific methods and techniques platforms efficacy component analysis to clarify the correlation representative composition and efficacy of traditional Chinese medicine.The small molecule of Chinese herbsspecifitic knockout technology is new technology and methods in studing the correlationefficacy of representative composition and of Chinese herbs. By comparison study, we can reveal the correlation of the representative component and the herbs by the analysis of representative components in addition to efficacy before and after knockout of representative components.Pueraria is an ancient and traditional Chinese herb which is commonly used,Puerarin is a representative isoflavones in Pueraria.The study found that isoflavones as phytoestrogens can protect cardiovascular by regulating NO secretion.Puerarin and other isoflavones in Pueraria have a weak estrogen and vasodilator effect, whichare effective ingredients for the treatment of cardiovascular diseases.In this study, we choose Radix Puerariae which has exact efficacy and basic research results as a template, Puerarin monoclonal antibody as a tool to establish the "specific knock out the Puerarin" technology, and then get the Pueraria water extracts which has specific knock out Puerarin. In order to clarify the correlation between Puerarin and Radix Puerariae in drug efficacy and contribution, we take difference compared method as research methods, use In vitro models and cell models corresponding to the traditional attending effect and the pharmacological effects of the herb, take a combination of efficacy and biological indicators as evaluations criteria through the establishment of the immunoassay method of high sensitivity, specificity, suitable for micro-biological samples. After complete the above-mentioned study and achieve the desired objectives, we can not only clarify the material basis of Radix Puerariae and Puerarine in pharmacodynamics, but also provide the exact basis for index components in the quality control of Pueraria herbs compatibility mechanism, metabolic studies, preparation process and other aspects. We can lay a solid foundation for the relevance of the research methods in Chinese herbal medicines and their efficacy using specific knock out index components method, so that this new technical means can be more widely available.1Investigation of puerarin and Pueraria on estrogen-like activity by knockout methods[OBJECTIVE]Comparative Study of RPE, PRR and PRR-KO-RPE on estrogen receptor-positive human breast cancer cell T47D proliferation, cell cycle effects of and ERa, ERβ protein expression.Explore estrogen-like activity on puerarin andpueraria vasodilatory estrogen-like activity in the efficacy of effect.[METHODS]Take T47D cells as a template, using the MTT method for detecting drugs and IC1182,780intervention changes in cell proliferation, cell cycle was measured by flow cytometry change, using western blot method to detect the protein level of ERa and ERβ protein expression of T47D cells.[RESULTS]1Three drugs have the effect of promoting cell proliferation and showed a dose response relationship. After PRR knockout significantly decreased cell proliferation. Drugs48h, ICI182,780on RPE proliferation inhibition rate strongest knockout PRR inhibition rate.2Three groups of drugs could reduce the proportion of cells in G0/G1phase, increasing the proportion of cells in S phase. RPE and PRR can increase the proportion of cells in different degrees of G2/M phase. After the PRR knockout percentage and proliferation index G2/M phase decreased significantly.3Each group can raise the relative expression levels of ERa, ERβ’s. After the PRR knockout, ERa expression did not change significantly, ERβ expression reduced. IC1182,780for each medication ERa, ERP expression of both antagonism, the PRR knockout, ERa cells expressing the degree of antagonism did not change significantly.[CONCLOTION]Puerarin has some relevance in Pueraria efficacy of estrogen-like activity, and the mechanism of cell proliferation and ERβ pathway.2Investigation of puerarin in Pueraria on vasodilatory efficacy by knockout methods[OBJECTIVE]Comparative Studyof radix puerariae extract(RPE), puerarin (PRR) and knockout Puerarin Pueraria Extract (PRR-KO-RPE) impacted on the SD rat thoracic vascular ring tension, explore the role of puerarin in Pueraria vasodilatory efficacy.[METHODS]Isometric tension recordings were applied to evaluate the effects of RPE, PRR and PRR-KO-RPE on resting, NE-Preconsrticting baortic ringswith or without endothelium.[RESULTS]1Cumulative concentration of RPE,PRR and PRR-KO-RPE in endothelial intact and denuded thoracic aortic rings have some relaxing effect and dose-dependent relaxation effects. Puerarin knockout, Kudzu degree of relaxation of vascular rings significantly reduced.2PRR after the last administration over the RP E can quickly reach maximum relaxation effects. RP E vasodilation and overall time than PRR PRR-KO-RPE long.[CONCLOTION]Puerarin has some relevance in endothelium-dependent vasodilatation Pueraria efficacy.3Correlation study on Pueraria and Puerarin on the efficiency of estrogen-like activity and efficacy of endothelium-dependent vasodilatation by knockout methods[OBJECTIVE]Preliminary study of puerarin in human umbilical vein endothelial cells (HUVECs) distribution and metabolic characteristics.Comparative study of RPE, PRR and PRR-KO-RPE on HUVECs proliferation, NO secretion and eNOS, ERa, ERβ protein expression.Investigate the relationship and mechanism of puerarin andpueraria vasodilatory estrogen-like activity and endo the lium-dependent vasodilator efficacy.[METHODS]Take HUVEC cells as a template. Using cell immunohistochemistry and puerarin immune competitive ELISA method, comparative study of puerarin monomer and Pueraria extract puerarin HUVECs metabolic characteristics of the distribution, and metabolic expression of the relationship between PRR and NO secretion; MTT method HUVECs proliferation and secretion of NO, western blot analysis to detect the expression of drugs on HUVECs in ERa, ERβ and eNOS determined under the action of the three drugs and antagonists.[RESULTS]1Immunohistochemical staining cells observed in cytoplasm and nucleus PRR role HUVECs after around a large brown granules.2Take puerarin concentration as standard amount, RPE and PRR role of1.0μg/mL of30min, RPE and PRR role of10.0μg/mL for20min, HUVECs extent of absorption of puerarin monomer is significantly higher than the same concentration of Pueraria Pueraria Su absorption.1.0μg/mL puerarin monomer effect on HUVECs10min, NO secretion levels higher than the same puerarin in Pueraria Extract,60min after Pueraria extract content is higher than that of the cells to secrete NO puerarin.10μg/mL puerarin monomer effect on HUVECs10min, NO secretion was higher than the same puerarin in Pueraria Extract,40min after Pueraria extract content is higher than that of the cells to secrete NO puerarin.3The secretion of NO on RPE was stronger than PRR and PRR-KO-RPE.Knockout PRR NO secretion. L-NAME and ICI182,780inhibited RPE, PRR and PRR-KO-RPE secretion of NO effect on HUVECs. Knockout PRR L-NAME and ICI182,780on the degree of inhibition of NO secretion HUVECs were weakened.4Three drugs could increase the eNOS, ERa and ERβrelative expression levels. PRR group has the highest expression of eNOS and lower expression of ERa than RPE and PRR-KO-RPE. RPE, PRR and PRR-KO-RPE group have no different ERβ expression. L-NAME has the strongest degree of antagonism on PRR relative expression of eNOS, ICI182,780has the strongest degree of antagonism on PRR relative expression of ERa and ER(3[CONCLOTION]Puerarin in heterogeneity and metabolic differences in the distribution of endothelial cells may lead to the foundation of their different biological effects. eNOS-NO pathway and ERβ-NO pathway may be puerarin in Pueraria pharmacodynamic correlation mechanisms to promote endothelial NO secretion.