1.LC-MS Analysis on Material Basis of Famous Classical Formula Xuefu Zhuyutang
Shujing ZHONG ; Zhaoyue LIU ; Mengge WANG ; Lan MIAO ; Changying REN ; Mingqian SUN ; Li LIN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(6):188-196
ObjectiveTo systematically identify the chemical constituents of Xuefu Zhuyutang(XFZY) and quantitatively determine its main components, aiming to elucidate its pharmacodynamic material basis and provide a scientific foundation for improving its quality control standards. MethodsUltra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) was employed for qualitative analysis of XFZY, and the identification of compounds was accomplished by comparing their retention times, secondary MS fragment ion information, 52 reference standards and relevant databases, followed by attribution of their herbal sources. A total of 22 representative compounds were screened out, and UPLC-quadrupole-linear ion trap mass spectrometry(UPLC-Q-TRAP-MS/MS) was applied for quantitative analysis of the compounds in the formula. ResultsA total of 77 compounds were identified in XFZY, including 31 flavonoids mainly derived from Aurantii Fructus, Glycyrrhizae Radix et Rhizoma, Persicae Semen, Carthami Flos, Bupleuri Radix, Paeoniae Radix Rubra and Achyranthis Bidentatae Radix, 24 terpenoids mainly derived from Platycodonis Radix, Glycyrrhizae Radix et Rhizoma, Paeoniae Radix Rubra and Rehmanniae Radix, 9 phenylpropanoids and their derivatives mainly derived from Chuanxiong Rhizoma, Angelicae Sinensis Radix and Rehmanniae Radix, 4 phenolic acids mainly derived from Chuanxiong Rhizoma, Angelicae Sinensis Radix and Paeoniae Radix Rubra, 3 saccharides mainly derived from Rehmanniae Radix and Achyranthis Bidentatae Radix, and 6 other compounds mainly derived from Persicae Semen, Rehmanniae Radix and Angelicae Sinensis Radix. The results of quantitative analysis showed that the contents of protocatechuic acid, hydroxypaeoniflorin, amygdalin, vanillic acid, paeoniflorin, liquiritin apioside, liquiritin, isoquercitrin, naringin, cosmosiin, hesperidin, neohesperidin, isoliquiritin, liquiritigenin, naringenin, benzoylpaeoniflorin, hesperetin, isoliquiritigenin, formononetin, glycyrrhizic acid, nobiletin and ligustilide in XFZY were determined to be 0.12, 1.57, 54.53, 0.29, 36.17, 4.29, 4.84, 0.09, 46.67, 0.04, 3.44, 31.95, 0.82, 0.10, 0.11, 0.43, 0.07, 0.03, 0.01, 8.24, 0.13, 1.81 mg·g-1. ConclusionThe qualitative method established in this study enables rapid and sensitive analysis of the chemical constituents in XFZY. Among the identified compounds, 52 are confirmed by reference standards, ensuring the accuracy of identification. The quantitative analysis of 22 key components provides a reliable experimental basis for the pharmacodynamic material basis research and quality control standard improvement of XFZY.
2.Decoction for soothing liver and removing stasis and toxicity inhibits he-patocellular carcinoma proliferation in nude mice by inducing ferropto-sis via p53 pathway
Jing LI ; Xiaojun CAI ; Renyi YANG ; Zhibin WANG ; Shujing ZHU ; Ying QU ; Chong ZHONG
Chinese Journal of Pathophysiology 2023;39(12):2176-2184
AIM:To investigate the inhibitory effect of the"decoction for soothing liver and removing stasis and toxicity(SGQYJDF)"on hepatocellular carcinoma(HCC)proliferation in nude mice by inducing ferroptosis via the tumor protein 53(p53)/solute carrier family 7 member 11(SLC7A11/xCT)/glutathione peroxidase 4(GPX4)pathway.METHODS:An ectopic subcutaneous tumor model was established by injecting SK-Hep-1 cells subcutaneously into the right axilla of nude mice.Upon formation of tumor,the mice were randomly divided into five groups(i.e.,control group,low-,medium-and high-dose SGQYJDF groups and medium-dose SGQYJDF plus Sorafenib group).Each group of mice was orally administered with the corresponding therapy for 14 consecutive days,during which the tumor size was observed regularly.At the end of treatment,the tumor growth inhibition rate was calculated based on tumor mass,and histopatho-logical changes were observed by HE staining.Then,the levels of malondialdehyde(MDA),glutathione(GSH)and fer-rous ions(Fe2+)were detected by colorimetric assays.The expression of the proliferation markers Ki-67 and GPX4 was de-tected by immunohistochemistry(IHC).The expression of p53 and xCT was detected by Immunofluorescence(IF).And the expression of p53,xCT and GPX4 was determined by Western blot.RESULTS:(1)SGQYJDF was found to dose-de-pendently decrease tumor volume(P<0.01)and inhibit tumor mass growth(P<0.01),and meanwhile,reduce the per-centage of Ki-67-positive cells(P<0.01)and their proliferation ability in tumor tissues,as compared to the control group.(2)In terms of Ferroptosis-related indicators,SGQYJDF was found to dose-dependently increase the levels of Fe2+ and MDA but decrease the level of GSH in tumor tissues(P<0.01),as compared to the control group.(3)In terms of protein expression,SGQYJDF was found to dose-dependently upregulate the expression of p53(P<0.05)but inhibit the expres-sion of xCT(P<0.05)and GPX4(P<0.01).Notably,medium-dose SGQYJDF plus sorafenib showed a stronger effect than high-dose SGQYJDF.CONCLUSION:Our findings suggest that SGQYJDF can induce Ferroptosis to inhibit the proliferation of subcutaneously transplanted tumor tissues in nude mice by upregulating the expression of p53,and inhibit-ing the expression of xCT and GPX4.

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