1.Quality Evaluation of Gegen Qinlian Tablets Based on HPLC Multi-component Quantification Combined with Chemical Pattern Recognition and TOPSIS Analysis
Ping QIN ; Yingying LU ; Wenming ZHANG ; Zifang FENG ; Lihong GU ; Chenjie XIA ; Minmin HU ; Xiaowei CHEN ; Zhenhua BIAN ; Xiwan LU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(6):217-224
ObjectiveTo establish a high-performance liquid chromatography(HPLC) for the quantitative analysis of multiple components in Gegen Qinlian tablets, and to comprehensively evaluate the quality of samples from different manufacturers by integrating chemical pattern recognition and technique for order preference by similarity to ideal solution(TOPSIS), in order to provide a reference basis for quality evaluation and control of Gegen Qinlian tablets. MethodsHPLC was employed to determine the contents of 10 components in 28 batches of Gegen Qinlian tablets collected from 6 manufacturers, and taking the detection results as variables, SIMCA 14.1 and SPSS 26.0 were employed for cluster analysis(CA), principal component analysis(PCA), and orthogonal partial least squares-discriminant analysis(OPLS-DA) to identify key components affecting the quality. Then, TOPSIS analysis was employed to rank the quality of Gegen Qinlian tablets from the 6 manufacturers and establish a comprehensive quality evaluation method. ResultsA quantitative method for Gegen Qinlian tablets was established. After methodological validation, the method was found to be stable and reliable, and could be used for the quantitative analysis of this preparation. The contents of 3′-hydroxy puerarin, puerarin, 3′-methoxy puerarin, daidzein, coptisine hydrochloride, epiberberine, jatrorrhizine hydrochloride, berberine hydrochloride, palmatine hydrochloride and baicalin in 28 batches of samples were 3.58-7.35, 24.88-42.32, 4.20-9.36, 4.33-7.60, 2.52-6.44, 0.93-4.10, 0.58-3.05, 10.68-22.92, 0.82-4.82, 11.73-60.16 mg·g-1, respectively. Among them, puerarin, berberine hydrochloride and baicalin all met the limit requirements for this preparation specified in the 2025 edition of the Pharmacopoeia of the People's Republic of China. CA and PCA clustered the 28 batches of samples into 5 categories, PCA extracted 2 principal components with a cumulative variance contribution rate of 90.588%, and OPLS-DA screened out 4 differential markers with variable importance in the projection(VIP) values>1.0, namely baicalin, 3′-hydroxy puerarin, coptisine hydrochloride and palmatine hydrochloride, which might be the main components affecting the quality of Gegen Qinlian tablets. TOPSIS analysis showed that the comprehensive score of each evaluation index(Ci) values of different manufacturers were different. Among them, the Ci of manufacturer B was ranked higher, indicating potentially superior quality, while the Ci of manufacturer A was ranked lower, suggesting potentially inferior quality. ConclusionThis study establishes a quantitative method for Gegen Qinlian tablets, and the content uniformity of the same manufacturer is good, while there are differences in the contents of active components among different manufacturers. Through the chemical pattern recognition analysis, it is found that the content differences of Gegen Qinlian tablets may be related to baicalin, 3′-hydroxy puerarin, coptisine hydrochloride and palmatine hydrochloride.
2.Polysaccharide of atractylodes macrocephala inhibits the growth of mice in-situ colon cancer HT-29 cell xenograft via activating immune cells
FENG Zifang ; TANG Shihua ; GUO Lijia ; HE Ling ; YANG Ruibin
Chinese Journal of Cancer Biotherapy 2019;26(11):1209-1213
Objective: To investigate the effect and mechanism of polysaccharide of atractylodes macrocephala (PAM) on the growth of colon cancer cells in mice bearing in-situ colon cancer transplantation tumor. Methods: 1×107 colon cancer HT-29 cells labeled with luciferase were injected into colon serosa of the mice to establish the in-situ colon cancer transplantation tumor model. When the tumor volume reached 230 mm3, the mice were given 30 mg/kg PAM (PAM group) or equal volume of normal saline (Control group) by gavage for 10 consecutive days. The effect of PAM on the growth of colon cancer cells in mice was tested by in vivo tumor imaging technology. The expressions of MHCII and IL-12 in granulocytes, dendritic cells and macrophages, the activation of lymphocytes, and IFN-γ expression in CD4+ and CD8+ cells of tumor tissues were detected by Flow cytometry. Results: PAM significantly inhibited the growth of colon cancer cells in mice bearing in-situ colon cancer transplantation tumor (P<0.01). PAM activated immune cells though increasing the expression levels of MHCII and IL-12 in dendritic cells and macrophages (both P<0.01). PAM significantly increased the frequency of CD8+ cells, NK cells, CD44+/NK cells and CD44+/CD4+ cells in tumor tissues and the number of CD8+ cells and NK cells per unit volume (all P<0.01). PAM significantly increased the IFN-γ secretion of CD4+ and CD8+ cells (both P<0.01), too. Conclusion: PAM inhibits the growth of colon cancer by activating immune cells in tumor tissues of mice bearing in-situ colon cancer transplantation tumor.
3.Immunomodulatory effects of polysaccharopeptide on EAC tumor-bearing mice through MyD88-dependent signaling pathway
Zifang FENG ; Zhixue WANG ; Lijing ZHOU ; Yixi BAO
Chinese Journal of Immunology 2015;(4):480-484,489
Objective:To detect the effects of polysaccharopeptide(PSP) on MyD88-dependent signaling pathway in EAC tumor-bearing mice,and explore the immunomodulatory mechanism of PSP.Methods: Ehrlich′s ascites carcinoma(EAC) C57BL/6 mice were used to establish the animal model for solid tumor.Mice were divided into two groups:WT group and MyD88-/-group.After 22 days of treatment,quantitative real-time PCR( Q-PCR) ,Western blot and were used to detect the related gene and protein expression of TLR4 pathway in spleens,ELISA were used to detect the terminal effect factors secretion eyeball blood from each group.Results:Related genes and proteins of TLR4 pathway in spleen were up-regulated significantly from two groups.Compared with WT group, related genes and proteins in MyD88-dependent pathway(MyD88,TRAF6,NF-κB,AP-1)was down-regulated in MyD88-/-group(P<0.05).Meanwhile,There was no significant change of the related genes and proteins of MyD88-independent pathway(TRAM,TRIF)in MyD88-/-group( P>0.05 ) .The terminal effect factors secretion of IL-2, IL-6, IL-10, TNF-αand IFN-γin MyD88-/-group were decreased significantly compared with WT group(P<0.05).Conclusion:The immunoregulatory effect of PSP on EAC tumor-bearing mice may be implement through the regulation of MyD88-dependent signaling pathway.

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