1.In Vitro and in vivo Component Analysis of Total Phenolic Acids from Gei Herba and Its Effect on Promoting Acute Wound Healing and Inhibiting Scar Formation
Xixian KONG ; Guanghuan TIAN ; Tong WU ; Shaowei HU ; Jie ZHAO ; Fuzhu PAN ; Jingtong LIU ; Yong DENG ; Yi OUYANG ; Hongwei WU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):156-167
ObjectiveBased on ultra performance liquid chromatography-quadrupole-electrostatic field orbital trap high-resolution mass spectrometry(UPLC-Q-Orbitrap-MS), to identify the in vivo and in vitro chemical components of total phenolic acids in Gei Herba(TPAGH), and to clarify the pharmacological effects and potential mechanisms of the effective part in promoting acute wound healing and inhibiting scar formation. MethodsUPLC-Q-Orbitrap-MS was used to identify the chemical components of TPAGH and ingredients absorbed in vivo after topical administration. A total of 120 ICR mice were randomly divided into the model group, recombinant human epidermal growth factor(rhEGF) group(4 mg·kg-1), and low, medium, and high dose groups of TPAGH(3.5, 7, 14 mg·kg-1), with 24 mice in each group. A full-thickness skin excision model was constructed, and each administration group was coated with the drug at the wound site, and the model group was treated with an equal volume of normal saline, the treatment was continued for 30 days, during which 8 mice from each group were sacrificed on days 6, 12, and 30. The healing of the wounds in the mice was observed, and histopathological changes in the skin tissues were dynamically observed by hematoxylin-eosin(HE), Masson, and Sirius red staining, and enzyme-linked immunosorbent assay(ELISA) was used to dynamically measure the contents of interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), vascular endothelial growth factor A(VEGFA), matrix metalloproteinase(MMP)-3 and MMP-9 in skin tissues. Network pharmacology was used to predict the targets related to the promotion of acute wound healing and the inhibition of scar formation by TPAGH, and molecular docking of key components and targets was performed. Gene Ontology(GO) biological process analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis were carried out for the related targets, so as to construct a network diagram of herbal material-compound-target-pathway-pharmacological effect-disease for further exploring its potential mechanisms. ResultsA total of 146 compounds were identified in TPAGH, including 28 phenylpropanoids, 31 tannins, 23 triterpenes, 49 flavonoids, and 15 others, and 16 prototype components were found in the serum of mice. Pharmacodynamic results showed that, compared with the model group, the TPAGH groups showed a significant increase in relative wound healing rate and relative scar inhibition rate(P<0.05), and the number of new capillaries, number of fibroblasts, number of new skin appendages, epidermal regeneration rate, collagen deposition ratio, and Ⅲ/Ⅰ collagen ratio in the tissue were significantly improved(P<0.05, 0.01), the levels of IL-6, TNF-α, MMP-3 and MMP-9 in the skin tissues were reduced to different degrees, while the level of VEGFA was increased. Network pharmacology analysis screened 10 core targets, including tumor protein 53(TP53), sarcoma receptor coactivator(SRC), protein kinase B(Akt)1, signal transducer and activator of transcription 3(STAT3), epidermal growth factor receptor(EGFR) and so on, participating in 75 signaling pathways such as advanced glycation end-products(AGE)-receptor for AGE(AGE/RAGE) signaling pathway, phosphatidylinositol 3-kinase(PI3K)/Akt signaling pathway, mitogen-activated protein kinase(MAPK) signaling pathway. Molecular docking confirmed that the key components genistein, geraniin, and casuariin had good binding ability to TP53, SRC, Akt1, STAT3 and EGFR. ConclusionThis study comprehensively reflects the chemical composition of TPAGH and the absorbed components after topical administration through UPLC-Q-Orbitrap-MS. TPAGH significantly regulates key indicators of skin healing and tissue reconstruction, thereby clarifying its role in promoting acute wound healing and inhibiting scar formation. By combining in vitro and in vivo component identification with network pharmacology, the study explores how key components may bind to targets such as TP53, Akt1 and EGFR, exerting therapeutic effects through related pathways such as immune inflammation and vascular regeneration.
2.Stewed Polygoni Multiflori Radix Treats Androgenic Alopecia in Mice by Activating Wnt/β-catenin Signaling Pathway
Fuzhu PAN ; Mingxia CHEN ; Bin YI ; Yanhua XUE ; Qiuping YU ; Fayun WU ; Enhui JI ; Hongwei WU ; Jing XU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(23):246-253
ObjectiveTo evaluate the therapeutic effect of stewed Polygoni Multiflori Radix on androgenic alopecia (AGA) and study the treatment mechanism. MethodNinety-nine SPF-grade male C57BL/6J mice were randomized into control, model, positive drug (finasteride, 0.65 mg·kg-1), low (0.78 g·kg-1), medium (1.56 g·kg-1), and high (3.12 g·kg-1)-dose stewed Polygoni Multiflori Radix, and Polygoni Multiflori Radix Praeparata groups by the random number table method. The mouse model of AGA was constructed by subcutaneous multi-point injection of testosterone propionate diluent for 60 days, and the mice were administrated with corresponding drugs by gavage from day 11. The therapeutic effects of stewed Polygoni Multiflori Radix and Polygoni Multiflori Radix Praeparata on AGA were evaluated by newly hair area, hair length, hair weight in the hair removal area, and hematoxylin-eosin staining. Enzyme-linked immunosorbent assay was employed to determine the levels of testosterone (T), dihydrotestosterone (DHT), and 5α-reductase (5-AR) in the skin tissue of mice. Western blot was employed to determine the expression levels of key proteins in the Wnt/β-catenin signaling pathway. ResultCompared with the control group, the model group (after 60 days of modeling) showed reductions in the newly hair area, hair length and weight in the back hair removal area, and ratio of hair follicles containing melanin to total hair follicles (P<0.05, P<0.01), elevated levels of T, DHT, and 5-AR, up-regulated expression level of glycogen synthase kinase-3β (GSK-3β) (P<0.05, P<0.01), and down-regulated expression levels of β-catenin, phospho-glycogen synthase kinase-3β (p-GSK-3β), and p-GSK-3β/GSK-3β (P<0.05, P<0.01) in the skin tissue. Compared with the model group, the positive drug, low-, medium-, and high-dose stewed Polygoni Multiflori Radix, and low-, medium-, and high-dose Polygoni Multiflori Radix Praeparata improved the newly hair area and hair length of mice (P<0.01), and stewed Polygoni Multiflori Radix and Polygoni Multiflori Radix Praeparata at low and medium doses improved the weight of newly formed hair in mice (P<0.05, P<0.01). The positive drug, low-, medium-, and high-dose stewed Polygoni Multiflori Radix, and low- and high-dose Polygoni Multiflori Radix Praeparata increased the ratio of hair follicles containing melanin to total hair follicles in the skin tissue (P<0.05, P<0.01). Compared with Polygoni Multiflori Radix Praeparata at the same doses, the medium and high doses of stewed Polygoni Multiflori Radix increased the ratio of melanin-containing hair follicles to total hair follicles (P<0.05). Compared with the model group, stewed Polygoni Multiflori Radix lowered the levels of T and DHT, down-regulated the expression level of GSK-3β (P<0.01), and up-regulated the expression levels of β-catenin, p-GSK-3β, and p-GSK-3β/GSK-3β (P<0.05, P<0.01) in the skin tissue of the mice. ConclusionStewed Polygoni Multiflori Radix can ameliorate androgenic alopecia in mice by reducing the androgen level and promoting Wnt/β-catenin signaling.
3.Intervention effects of Naoxintong capsules on psychological and cardiac status in depressed rats after heart failure
Jie ZHAO ; Yi OUYANG ; Shaowei HU ; Guanghuan TIAN ; Xixian KONG ; Fuzhu PAN ; Hongwei WU ; Liying TANG ; Hongjun YANG
Science of Traditional Chinese Medicine 2024;2(4):319-326
Background: Depression is a common clinical phenomenon in the patients with heart failure (HF). In traditional Chinese medicine (TCM), diseases in the brain and heart are thought to be correlated and interact. Naoxintong capsules (NXT) has been used for treating cardio-cerebrovascular diseases, while its therapeutic effect on depression after HF remains unclear. Objective: The aim of the study is to evaluate the intervention effect of NXT on depression after HF. Methods: Sprague-Dawley rats were assigned into the following 5 groups: sham, model, NXT (250, 1000 mg/kg), and valsartan (8 mg/kg). Coronary artery occlusion was performed to induce HF and subsequent depression in rats. The cardiac function was evaluated by echocardiography, hematoxylin-eosin staining, and Masson trichrome staining. The sucrose preference test and Morris water maze test were carried out to assess the depressive behaviors in rats. The ultrastructure of hippocampal CA1 neurons was observed and the levels of corticotropin-releasing hormone in the hypothalamus, brain-derived neurotrophic factor in the cortex, and adrenocorticotropic hormone (ACTH) in the plasma were determined by enzyme-linked immunosorbent assay. The levels of dopamine, 5-hydroxytryptamine, norepinephrine, and γ-aminobutyric acid in the hippocampus were measured by UPLC-QQQ-MS. Results: NXT reduced myocardial injury and pathological changes in the cardiac tissue and increased the left ventricular ejection fraction, left ventricular fractional shortening, and cardiac output. NXT increased the sugar preference rate and number of crossings and shortened the escape latency. Furthermore, the NXT treatment restored the levels of corticotropin-releasing hormone, adrenocorticotropic hormone, brain-derived neurotrophic factor, dopamine, and γ-aminobutyric acid to the baseline values. Conclusions: NXT not only demonstrates cardioprotective effect but also attenuates depression in the rats after HF. It may exert the antidepressant effect by inhibiting the hyperactivity of the hypothalamic-pituitary-adrenal axis and recovering the levels of neurotrophic factors and neurotransmitters.
4.Herbal Textual Research on Gei Herba and Its Pharmacological Effect on Cardiovascular and Cerebrovascular Diseases
Guanghuan TIAN ; Tong WU ; Fuzhu PAN ; Xixian KONG ; Huaqing LAI ; Haiyun ZHENG ; Bing LI ; Jianyong ZHANG ; Hongwei WU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(21):274-282
Gei Herba is a traditional folk herbal medicine with a variety of functions such as replenishing Qi and invigorating spleen, tonifying blood and nourishing Yin, moistening lung and resolving phlegm, activating blood and alleviating edema, moving Qi, and activating blood. The reports about the pharmacological effects of this herbal medicine have been increasing in recent years. By reviewing the ancient and modern literature about Gei Herba, we systematically organized the name, original plants, nature, taste, and functions of this herbal medicine, and summarized the modern pharmacological studies and clinical applications of Gei Herba in cardiovascular and cerebrovascular diseases. Gei Herba was first recorded in the name of "Dijiao" in the Geng Xin Yu Ce(《庚辛玉册》) written in the Ming Dynasty. It is derived from Geum japonicum var. chinense (Rosaceae) and sometimes confused with Adina rubella (Rubiaceae). This medicine had numerous synonyms in the local materia medica books. Gei Herba is widely distributed and harvested in summer and autumn, with the dried whole grass used as medicine. The historical records of the nature, taste, meridian tropism, main functions, and indications of Gei Herba are not consistent. It is generally believed that Gei Herba is pungent, bitter, sweet, cool, and has tropism to the liver, spleen, and lung meridians. Based on the effects of tonifying Qi, activating blood, and nourishing Yin, modern pharmacological studies have reported that the extracts of Gei Herba and the tannin phenolic acid compounds and triterpenoids isolated from Gei Herba have therapeutic effects on cardiovascular and cerebrovascular diseases such as hypertension, myocardial ischemia, cerebral ischemia, and vascular dementia. This study provides a reference for discovering the clinical advantages of Gei Herba and developing new drugs.

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