1.Quality Evaluation of Lycii Cortex and Roasted Lycii Cortex Based on Fingerprint and Content Determination
Yihuan WU ; Wenli ZENG ; Xuemei QIN ; Zongxin SHI ; Chengcheng HUANG ; Yuntao DAI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):165-172
ObjectiveTo establish fingerprint profiles and a quantitative determination method for Lycii Cortex, providing a scientific basis for the formulation of quality standards for Lycii Cortex and its roasted products. MethodsHigh performance liquid chromatography(HPLC) was developed for the quantitative method for determining kukoamine B in Lycii Cortex and its roasted products on an Alphasil XD-C18 CH column(4.6 mm×250 mm, 5 μm). HPLC fingerprint profiles were established for 10 batches of Lycii Cortex and its roasted products, and ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used to identify the common peaks based on reference standards, literature and MS information. Quality evaluation indicators included yield of decoction pieces, appearance properties, content of kukoamine B, and fingerprint profiles. The temperature and time of the roasting process were investigated to select the optimal preparation process, which was then verified. Additionally, chemical pattern recognition was combined to assess the differences in the chemical composition of Lycii Cortex before and after roasting, as well as among samples from different origins. ResultsQuantitative analysis indicated that the contents of kukoamine B in Lycii Cortex and its roasted products were 0.35%-5.51% and 0.24%-4.15%, respectively. The transfer rate of kukoamine B was 58.6%-78.9% after roasting. The fingerprint profile analysis demonstrated that the method established in this study effectively separated kukoamine B from other components in the samples and distinctly differentiated it from its impurity peak, cis-N-caffeoylputrescine. The HPLC fingerprint profiles of Lycii Cortex and its roasted products showed high similarity(all above 0.95), with 7 common peaks identified and five common components, including kukoamine B, cis-N-caffeoylputrescine, N-coumaroyl tyramine, feruloyltyramine, and glucosyringic acid, confirmed. Process optimization confirmed that baking at 110 ℃ for 20 min was a stable and feasible method for roasting Lycii Cortex. Principal component analysis and cluster analysis showed that there was little difference in the chemical composition between raw and roasted Lycii Cortex, but the quality of Lycii Cortex from different origins differed greatly. ConclusionThis study successfully established the fingerprint profiles and a quantitative method for the effective component kukoamine B in Lycii Cortex and roasted Lycii Cortex. The qualitative and quantitative analyses clarified that the impact of the roasting process on the chemical composition of Lycii Cortex was less significant than the variations due to its geographical origin. The findings of this study offer a reference for the development of quality evaluation methods and the establishment of quality standards for Lycii Cortex and its processed products.
2.Effects of Jianpi Yishen Huatan Formula (健脾益肾化痰方)-Medicated Serum on Apoptosis,Migration and the LncRNA ALAL-1/USP4/HDAC2 Pathway in Human Lung Squamous Carcinoma Cells
Yijun FANG ; Xuemei WANG ; Changzhou XIONG ; Liubang LI ; Huimin QIN ; Zhiguang WANG
Journal of Traditional Chinese Medicine 2025;66(14):1481-1488
ObjectiveTo investigate the potential mechanism of the Jianpi Yishen Huatan Formula (健脾益肾化痰方,JPYSHF) in treating squamous cell lung cancer through the LncRNA ALAL-1/USP4/HDAC2 signaling pathway. MethodsForty Sprague-Dawley (SD) rats were randomly divided into a control group and high-, medium-, and low-dose JPYSHF group with 10 rats in each group. Rats in the JPYSHF groups were administered JPYSHF concentrated liquid at doses of 45, 30, and 15 g/(kg·d) via intragastric gavage, respectively, while the control group received 10 ml/(kg·d) of normal saline, once daily for 10 consecutive days before preparation of drug containing serum. Human lung squamous carcinoma SK-MES-1 cells were divided into a control group and low-, medium-, and high-dose JPYSHF-medicated serum groups. The control group was cultured with 10% saline-containing serum, while the JPYSHF groups were cultured with 10% low-, medium-, or high-dose medicated serum. After 48 hours of incubation, flow cytometry was used to detect apoptosis rates, and a cell scratch assay was performed to evaluate migration areas at 0 h and 24 h to calculate migration rate. Additional SK-MES-1 cells were divided into control serum, JPYSHF-medicated serum (low-, medium-, high-) dose, LncRNA-silenced group (transfected with ALAL-1 siRNA), USP4-inhibited group (treated with 35 μmol/L PR-619, a deubiquitinase inhibitor), and HDAC2-inhibited group (treated with 60 μmol/L Vorinostat). After 24 and 48 hours of culture, cell viability was assessed using the CCK-8 assay; LncRNA ALAL-1, USP4, and HDAC2 mRNA levels were quantified by qPCR after 24 hours; USP4 and HDAC2 protein levels were measured by Western Blot after 48 hours. ResultsCompared with the control serum group, the total apoptosis rate of cells in middle- and high-JPYSHF-medicated serum group significantly increased, and the cell migration rate of cells in the low-, middle- and high-JPYSHF-medicated serum group significantly decreased (P<0.05 or P<0.01). The cell migration rate of the low-, medium- and high-JPYSHF-medicated serum groups decreased with the increase of concentration in a concentration-dependent manner (P<0.05 or P<0.01). Compared with the control serum group at the same time, the cell viability at 24 h and 48 h significantly decreased in all groups (P<0.05 or P<0.01). Compared with the low-JPYSHF-medicated serum group at the same time, the cell viability at 24 h and 48 h also decreased in the high-JPYSHF-medicated serum group and the LncRNA silencing group (P<0.05). Compared with the control serum group, the expression of USP4 and HDAC2 mRNA reduced in the low- and medium-dose JPYSHF-medicated serum groups and the USP4 inhibitor group, and the expression of LncRNA ALAL-1, USP4 and HDAC2 mRNA reduced in the high-dose JPYSHF-medicated serum group and LncRNA-silencing group, and HDAC2 mRNA expression reduced in the HDAC2 inhibitor group. USP4 and HDAC2 protein levels were reduced in cells of all groups except for USP4 protein level in HDAC2 inhibitor group (P<0.05 or P<0.01). ConclusionJPYSHF-medicated serum inhibits proliferation and promotes apoptosis of human lung squamous carcinoma cells, and its mechanism of action may be related to its inhibition of the LncRNA ALAL-1/USP4/HDAC2 pathway, with best effect at a high concentration.
3.Riboflavin metabolism involved into comparison of therapeutic action of wild-simulated and transplanted Astragali Radix to treat chronic atrophic gastritis rats based on endogenous-xenobiotics metabolism.
Kaidi FAN ; Xu LIAN ; Weicong ZHANG ; Xuemei QIN ; Yuetao LIU
Chinese Herbal Medicines 2025;17(3):513-528
OBJECTIVE:
Astragali Radix (AR, Huangqi in Chinese) has been utilized generally as a bulk drug for the treatment of chronic atrophic gastritis (CAG) in China. The efficacy of wild-simulated AR (WAR) and transplanted AR (TAR) commercially available is unclear. This study aimed to further clarify the therapeutic action of WAR and TAR to treat CAG rats based on endogenous-xenobiotics metabolism.
METHODS:
Ultra-high performance liquid chromatography coupled with quadrupole-time of flight mass spectrometer (UHPLC-Q-TOF/MS) based metabolomics had been used to analyze the cecal contents metabolic features and metabolic process differences of two ARs in the treatment of CAG.
RESULTS:
Apparent pharmacodynamic indicator examination revealed that the WAR group had a more substantial curative effect. Metabolomics studies revealed that seven endogenous metabolites were significant differences in two ARs. Furthermore, following treatment, 77 and 65 xenobiotics metabolites were identified in the WAR and TAR groups, respectively. Finally, the most critical riboflavin metabolic route in the formation of CAG was chosen for molecular docking with the relevant exogenous components, and WAR scored higher than TAR.
CONCLUSION
In this work, we analyzed the efficacy differences of AR from diverse cultivation forms by combining metabolomics and medicinal chemistry technologies, and it gave a fresh perspective for TCM efficacy evaluation and quality control research.
4.Determination and Mechanism Prediction of Potential Active Ingredients in Erdongtang Based on UHPLC-QqQ-MS and Network Pharmacology
Xueyuan WANG ; Baoxin LI ; Shougang SHI ; Zhengjun HUANG ; Yuntao DAI ; Xuemei QIN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(5):28-34
ObjectiveTo establish an ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry(UHPLC-QqQ-MS) for determination of the active ingredients in Erdongtang, and to predict the targets and pathways of anti-insulin resistance action of this formula. MethodThe analysis was performed on an ACQUITY UPLC BEH C18 column(2.1 mm×100 mm, 1.7 μm) with the mobile phase of 0.1% formic acid aqueous solution(A)-acetonitrile(B) for gradient elution(0-3 min, 90%-87%A; 3-6 min, 87%-86%A; 6-9 min, 86%-83%A; 9-11 min, 83%-75%A; 11-18 min, 75%-70%A; 18-19 min, 70%-52%A; 19-22 min, 52%A; 22-25 min, 52%-5%A; 25-27 min, 5%-90%A; 27-30 min, 90%A). The contents of active ingredients in Erdongtang was detected by electrospray ionization(ESI) and multiple reaction monitoring(MRM) mode under positive and negative ion modes. On this basis, network pharmacology was applied to predict the targets and pathways of Erdongtang exerting anti-insulin resistance effect. ResultThe 20 active ingredients in Erdongtang showed good linear relationships within a certain mass concentration range, and the precision, stability, repeatability and recovery rate were good. The results of determination showed that the ingredients with high content in 15 batches of samples were baicalein(1 259.39-1 635.78 mg·L-1), baicalin(1 078.37-1 411.52 mg·L-1), the ingredients with medium content were mangiferin(148.59-217.04 mg·L-1), timosaponin BⅡ(245.10-604.89 mg·L-1), quercetin-3-O-glucuronide(89.30-423.26 mg·L-1), rutin(46.91-1 553.61 mg·L-1), glycyrrhizic acid(55.97-391.47 mg·L-1), neomangiferin(37.45-127.03 mg·L-1), nuciferine(0.89-63.48 mg·L-1), hyperoside(6.96-136.78 mg·L-1), liquiritin(30.89-122.78 mg·L-1), liquiritigenin(26.64-110.67 mg·L-1), protodioscin(58.57-284.26 mg·L-1), the ingredients with low content were wogonin(7.16-20.74 mg·L-1), pseudoprotodioscin(5.49-22.96 mg·L-1), ginsenoside Rb1(7.31-23.87 mg·L-1), ginsenoside Rg1(10.78-28.33 mg·L-1), ginsenoside Re(7.78-24.76 mg·L-1), ophiopogonin D(2.08-4.29 mg·L-1), methylophiopogonanone A(0.74-1.67 mg·L-1). The results of network pharmacology indicated that the mechanism of anti-insulin resistance exerted by Erdongtang might be related to the phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt) signaling pathway. ConclusionThe established UHPLC-QqQ-MS has the advantages of simple sample processing, strong exclusivity and high sensitivity, and can simultaneously determine the contents of the main ingredients from seven herbs in Erdongtang, which can lay the foundation for the development of Erdongtang compound preparations. The results of the network pharmacology can provide a reference for the mechanism study of Erdongtang in the treatment of type 2 diabetes mellitus.
5.Effect of radix scutellariae microemulsion gel on chronic eczema in mice model
Ke WANG ; Chenbing NIU ; Yinxia CHANG ; Xuemei QIN ; Zhihua LUAN ; Yue ZHANG
Chinese Journal of Clinical Pharmacology and Therapeutics 2024;29(4):447-454
AIM:To prepare radix scutellariae mi-croemulsion gel and investigate its therapeutic ef-fect on chronic eczema based on the previous re-search of radix scutellariae self microemulsion.METHODS:The gel matrix and humectant were op-timized by single factor method and response sur-face method to obtain the formula and preparation technique of the gel.The Franz diffusion cell meth-od was used to evaluate the transdermal proper-ties of microemulsion and microemulsion gel in vi-tro.By establishing a chronic eczema model in the mouse ear,the swelling degree,swelling inhibition rate,pathological changes and tumor necrosis fac-tor a(TNF-a),Interleukin-1β(IL-1β)and interleukin-6(IL-6)of radix scutellariae microemulsion gel were measured,to investigate the therapeutic ef-fect on chronic eczema in mice.RESULTS:The physi-cal and chemical properties of radix scutellariae mi-croemulsion gel were stable.Compared with micro-emulsion,the microemulsion gel had better trans-dermal performance.The cumulative transdermal amount of baicalein and wogonin,the main compo-nents of microemulsion gel,was 1.85 times and 2.77 times of that of microemulsion respectively.Moreover,the steady flow rate and permeability co-efficient of microemulsion gel significantly in-creased,and the lag time significantly shortened.Pharmacodynamic study showed that compared with the model group,the radix scutellariae micro-emulsion gel could significantly reduce the ear swelling of mice(P<0.05),and the serum inflamma-tory factor TNF-a,IL-1β and IL-6 reduced content by over 37%.Compared with the radix scutellaria aqueous extract and aqueous extract gel,the treat-ment of chronic eczema was better.CONCLUSION:The preparation process of radix scutellaria micro-emulsion gel is feasible,with strong transdermal property,and a significant therapeutic effect on chronic eczema.
6.Effect of radix scutellariae microemulsion gel on chronic eczema in mice model
Ke WANG ; Chenbing NIU ; Yinxia CHANG ; Xuemei QIN ; Zhihua LUAN ; Yue ZHANG
Chinese Journal of Clinical Pharmacology and Therapeutics 2024;29(4):447-454
AIM:To prepare radix scutellariae mi-croemulsion gel and investigate its therapeutic ef-fect on chronic eczema based on the previous re-search of radix scutellariae self microemulsion.METHODS:The gel matrix and humectant were op-timized by single factor method and response sur-face method to obtain the formula and preparation technique of the gel.The Franz diffusion cell meth-od was used to evaluate the transdermal proper-ties of microemulsion and microemulsion gel in vi-tro.By establishing a chronic eczema model in the mouse ear,the swelling degree,swelling inhibition rate,pathological changes and tumor necrosis fac-tor a(TNF-a),Interleukin-1β(IL-1β)and interleukin-6(IL-6)of radix scutellariae microemulsion gel were measured,to investigate the therapeutic ef-fect on chronic eczema in mice.RESULTS:The physi-cal and chemical properties of radix scutellariae mi-croemulsion gel were stable.Compared with micro-emulsion,the microemulsion gel had better trans-dermal performance.The cumulative transdermal amount of baicalein and wogonin,the main compo-nents of microemulsion gel,was 1.85 times and 2.77 times of that of microemulsion respectively.Moreover,the steady flow rate and permeability co-efficient of microemulsion gel significantly in-creased,and the lag time significantly shortened.Pharmacodynamic study showed that compared with the model group,the radix scutellariae micro-emulsion gel could significantly reduce the ear swelling of mice(P<0.05),and the serum inflamma-tory factor TNF-a,IL-1β and IL-6 reduced content by over 37%.Compared with the radix scutellaria aqueous extract and aqueous extract gel,the treat-ment of chronic eczema was better.CONCLUSION:The preparation process of radix scutellaria micro-emulsion gel is feasible,with strong transdermal property,and a significant therapeutic effect on chronic eczema.
7.Effect of radix scutellariae microemulsion gel on chronic eczema in mice model
Ke WANG ; Chenbing NIU ; Yinxia CHANG ; Xuemei QIN ; Zhihua LUAN ; Yue ZHANG
Chinese Journal of Clinical Pharmacology and Therapeutics 2024;29(4):447-454
AIM:To prepare radix scutellariae mi-croemulsion gel and investigate its therapeutic ef-fect on chronic eczema based on the previous re-search of radix scutellariae self microemulsion.METHODS:The gel matrix and humectant were op-timized by single factor method and response sur-face method to obtain the formula and preparation technique of the gel.The Franz diffusion cell meth-od was used to evaluate the transdermal proper-ties of microemulsion and microemulsion gel in vi-tro.By establishing a chronic eczema model in the mouse ear,the swelling degree,swelling inhibition rate,pathological changes and tumor necrosis fac-tor a(TNF-a),Interleukin-1β(IL-1β)and interleukin-6(IL-6)of radix scutellariae microemulsion gel were measured,to investigate the therapeutic ef-fect on chronic eczema in mice.RESULTS:The physi-cal and chemical properties of radix scutellariae mi-croemulsion gel were stable.Compared with micro-emulsion,the microemulsion gel had better trans-dermal performance.The cumulative transdermal amount of baicalein and wogonin,the main compo-nents of microemulsion gel,was 1.85 times and 2.77 times of that of microemulsion respectively.Moreover,the steady flow rate and permeability co-efficient of microemulsion gel significantly in-creased,and the lag time significantly shortened.Pharmacodynamic study showed that compared with the model group,the radix scutellariae micro-emulsion gel could significantly reduce the ear swelling of mice(P<0.05),and the serum inflamma-tory factor TNF-a,IL-1β and IL-6 reduced content by over 37%.Compared with the radix scutellaria aqueous extract and aqueous extract gel,the treat-ment of chronic eczema was better.CONCLUSION:The preparation process of radix scutellaria micro-emulsion gel is feasible,with strong transdermal property,and a significant therapeutic effect on chronic eczema.
8.Effect of radix scutellariae microemulsion gel on chronic eczema in mice model
Ke WANG ; Chenbing NIU ; Yinxia CHANG ; Xuemei QIN ; Zhihua LUAN ; Yue ZHANG
Chinese Journal of Clinical Pharmacology and Therapeutics 2024;29(4):447-454
AIM:To prepare radix scutellariae mi-croemulsion gel and investigate its therapeutic ef-fect on chronic eczema based on the previous re-search of radix scutellariae self microemulsion.METHODS:The gel matrix and humectant were op-timized by single factor method and response sur-face method to obtain the formula and preparation technique of the gel.The Franz diffusion cell meth-od was used to evaluate the transdermal proper-ties of microemulsion and microemulsion gel in vi-tro.By establishing a chronic eczema model in the mouse ear,the swelling degree,swelling inhibition rate,pathological changes and tumor necrosis fac-tor a(TNF-a),Interleukin-1β(IL-1β)and interleukin-6(IL-6)of radix scutellariae microemulsion gel were measured,to investigate the therapeutic ef-fect on chronic eczema in mice.RESULTS:The physi-cal and chemical properties of radix scutellariae mi-croemulsion gel were stable.Compared with micro-emulsion,the microemulsion gel had better trans-dermal performance.The cumulative transdermal amount of baicalein and wogonin,the main compo-nents of microemulsion gel,was 1.85 times and 2.77 times of that of microemulsion respectively.Moreover,the steady flow rate and permeability co-efficient of microemulsion gel significantly in-creased,and the lag time significantly shortened.Pharmacodynamic study showed that compared with the model group,the radix scutellariae micro-emulsion gel could significantly reduce the ear swelling of mice(P<0.05),and the serum inflamma-tory factor TNF-a,IL-1β and IL-6 reduced content by over 37%.Compared with the radix scutellaria aqueous extract and aqueous extract gel,the treat-ment of chronic eczema was better.CONCLUSION:The preparation process of radix scutellaria micro-emulsion gel is feasible,with strong transdermal property,and a significant therapeutic effect on chronic eczema.
9.Effect of radix scutellariae microemulsion gel on chronic eczema in mice model
Ke WANG ; Chenbing NIU ; Yinxia CHANG ; Xuemei QIN ; Zhihua LUAN ; Yue ZHANG
Chinese Journal of Clinical Pharmacology and Therapeutics 2024;29(4):447-454
AIM:To prepare radix scutellariae mi-croemulsion gel and investigate its therapeutic ef-fect on chronic eczema based on the previous re-search of radix scutellariae self microemulsion.METHODS:The gel matrix and humectant were op-timized by single factor method and response sur-face method to obtain the formula and preparation technique of the gel.The Franz diffusion cell meth-od was used to evaluate the transdermal proper-ties of microemulsion and microemulsion gel in vi-tro.By establishing a chronic eczema model in the mouse ear,the swelling degree,swelling inhibition rate,pathological changes and tumor necrosis fac-tor a(TNF-a),Interleukin-1β(IL-1β)and interleukin-6(IL-6)of radix scutellariae microemulsion gel were measured,to investigate the therapeutic ef-fect on chronic eczema in mice.RESULTS:The physi-cal and chemical properties of radix scutellariae mi-croemulsion gel were stable.Compared with micro-emulsion,the microemulsion gel had better trans-dermal performance.The cumulative transdermal amount of baicalein and wogonin,the main compo-nents of microemulsion gel,was 1.85 times and 2.77 times of that of microemulsion respectively.Moreover,the steady flow rate and permeability co-efficient of microemulsion gel significantly in-creased,and the lag time significantly shortened.Pharmacodynamic study showed that compared with the model group,the radix scutellariae micro-emulsion gel could significantly reduce the ear swelling of mice(P<0.05),and the serum inflamma-tory factor TNF-a,IL-1β and IL-6 reduced content by over 37%.Compared with the radix scutellaria aqueous extract and aqueous extract gel,the treat-ment of chronic eczema was better.CONCLUSION:The preparation process of radix scutellaria micro-emulsion gel is feasible,with strong transdermal property,and a significant therapeutic effect on chronic eczema.
10.Effect of radix scutellariae microemulsion gel on chronic eczema in mice model
Ke WANG ; Chenbing NIU ; Yinxia CHANG ; Xuemei QIN ; Zhihua LUAN ; Yue ZHANG
Chinese Journal of Clinical Pharmacology and Therapeutics 2024;29(4):447-454
AIM:To prepare radix scutellariae mi-croemulsion gel and investigate its therapeutic ef-fect on chronic eczema based on the previous re-search of radix scutellariae self microemulsion.METHODS:The gel matrix and humectant were op-timized by single factor method and response sur-face method to obtain the formula and preparation technique of the gel.The Franz diffusion cell meth-od was used to evaluate the transdermal proper-ties of microemulsion and microemulsion gel in vi-tro.By establishing a chronic eczema model in the mouse ear,the swelling degree,swelling inhibition rate,pathological changes and tumor necrosis fac-tor a(TNF-a),Interleukin-1β(IL-1β)and interleukin-6(IL-6)of radix scutellariae microemulsion gel were measured,to investigate the therapeutic ef-fect on chronic eczema in mice.RESULTS:The physi-cal and chemical properties of radix scutellariae mi-croemulsion gel were stable.Compared with micro-emulsion,the microemulsion gel had better trans-dermal performance.The cumulative transdermal amount of baicalein and wogonin,the main compo-nents of microemulsion gel,was 1.85 times and 2.77 times of that of microemulsion respectively.Moreover,the steady flow rate and permeability co-efficient of microemulsion gel significantly in-creased,and the lag time significantly shortened.Pharmacodynamic study showed that compared with the model group,the radix scutellariae micro-emulsion gel could significantly reduce the ear swelling of mice(P<0.05),and the serum inflamma-tory factor TNF-a,IL-1β and IL-6 reduced content by over 37%.Compared with the radix scutellaria aqueous extract and aqueous extract gel,the treat-ment of chronic eczema was better.CONCLUSION:The preparation process of radix scutellaria micro-emulsion gel is feasible,with strong transdermal property,and a significant therapeutic effect on chronic eczema.

Result Analysis
Print
Save
E-mail