ABSTRACT
OBJECTIVE：To stud y the correlatio n between the contents of active ingredients and the color of Morus alba ，to establish fingerprint and conduct cluster analysis of samples from different producing areas ，so as to provide reference for its quality control and evaluation. METHODS ：HPLC and HCl-Mg reaction colorimetry were used to determine the contents of morusin and total flavonoids in M. alba . The color of M. alba was observed by naked eye ，and chromaticity values （L＊，a＊，b＊） were measured by color difference meter and color aberration （E＊ab）were calculated. Pearson correlation of the contents of morusin and total flavonoids with color indicators （L＊，a＊，b＊，E＊ab）were analyzed by SPSS 21.0 software. HPLC method was used to establish the fingerprint of 20 batches of M. alba from 3 different producing areas ，and the similarity analysis was carried out. K-means cluster analysis （based on the contents of morusin and total flavonoids and corlor index ）and hierarchical cluster analysis （based on relative peak area of common peaks in fingerprint ）were performed for 20 batches of samples by SPSS 21.0 software. RESULTS：The average contents of morusin and total flavonoids in M. alba were 0.096 0-0.618 6 mg/g，0.48％-1.51％，which were significantly correlated with each color index （P＜0.01）. The smaller L＊，b＊，E＊ab and the larger a＊were，the higher the content of morusin was ；the higher the value of L＊，b＊，E＊ab and the smaller the value of a＊were，the higher the content of total flavonoids was. The similarity between the fingerprints of 20 batches of samples and the control ranged from 0.883 to 0.983；13 common peaks were demarcated ，and No. 1 peak was identified as chlorogenic acid. K-means cluster analysis showed that 20 batches of samples could be divided into 2 categories. Category Ⅰ were mainly from Anhui province with higher content of morusin，lower content of total flavonoids ，darker and yellowish brown color ；category Ⅱ were mainly from Sichuan province and Guizhou province ，with lower content of morusin ，higher content of total flavonoids ，lighter and yellowish white color. The results of hierarchical cluster analysis were consistent with the results. CONCLUS IONS：The color of M. alba is closely related to the contents of morusin and total flavonoids. The content of morusin in yellow-brown M. albais is higher ，while the content of total flavonoids in yellow-white M. albais is higher.
ABSTRACT
OBJECTIVE:To establish the method for the simultaneous determination of content of 6 active components as neochlorogenic acid,mulberroside A,chlorogenic acid,astragalin,sanggenon C and morusin in Morus alba,and to provide reference for improving quality control standard of M. alba. METHODS:HPLC method was adopted. The determination was performed on Agilent 5 TC-C18with mobile phase consisted of acetonitrile-0.1% formic acid(gradient elution)at the flow rate of 1 mL/min. The detection wavelength of 280 nm. RESULTS:The mass concentration linear range of neochlorogenic acid, mulberroside A,chlorogenic acid,astragalin,sanggenon C and morusin were 0.001 06-0.042 4,0.001 67-0.066 8,0.007 95-0.318, 0.001 65-0.066 0,0.005 00-0.200 and 0.001 24-0.049 6 mg/mL,respectively(all r≥0.999 6);the limits of quantitation were 0.11, 0.14,0.81,0.17,0.45 and 0.12 μg/mL,respectively;the limits of detection were 0.04,0.05,0.41,0.07,0.18 and 0.04 μg/mL, respectively;RSDs of precision test were 0.26%,0.31%,0.24%,0.27%,0.36% and 0.44%(n=6),respectively;RSDs of stability test were 0.68%,0.54%,0.62%,0.53%,0.41% and 0.73%(n=6),respectively;average method recovery rates were 99.1%,98.8%,98.8%,98.4%,98.5% and 99.9%(RSDs were 0.5%-1.5%,n=9),respectively. CONCLUSIONS:The method is simple,accurate,and can be used for simultaneous determination of 6 active components in M.alba.
ABSTRACT
Objective To investigate the effects of Morusin on cell line proliferation, cell cycle and cell apoptosis of colorectal cancer HCT116 cell;To discuss its mechanism. Methods HCT116 cells were treated with different concentrations of Morusin for 72 h. Cell proliferation was detected by CCK-8 assay, and cell growth inhibition rate and IC50 value were calculated. HCT116 cells were treated with 25.4, 50.8 μmol/L Morusin for 24 h, 48 h and 72 h. Cell morphology was observed under the microscope, and cell proliferation was detected by CCK-8 assay. After HCT116 cell line was treated with 25.4μmol/L Morusin for 24 h, cell cycle phase distribution was detected by flow cytometry and the cell apoptosis was detected by TUNEL assay under the fluorescence microscope. Post-intervention protein expressions were detected by Western Blot. Results The inhibitory effects of Morusin on the proliferation of HCT116 cell line was concentration/time dependent and IC50 value at 72 h was 25.4μmol/L;Morusin induced the cell cycle arrest at G0/G1 phase and G2/M phase, but there was no induction of cell apoptosis;Morusin significantly decreased the expression ofβ-catenin and its target c-Myc, and downregulated the expressions of cyclinD1 and cyclinB1, which were involved in cell cycle regulation. Conclusion Morusin can inhibit HCT116 cell cycle and the proliferation of colorectal cancer cells. Its mechanism might be realized by suppressing the activity ofβ-catenin pathway and reducing the protein expressions of cyclinD1 and cyclinB1.