1.Pathogenesis and Syndrome Differentiation of "Gaozhuo" of Oxidative Stress in Diabetic Kidney Disease
Yuman YIN ; Yunfeng YU ; Xiangning HUANG ; Jiawang HUANG ; Gang HU ; Juan HUANG ; Rong YU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):226-234
Oxidative stress is a pivotal factor in the onset and progression of diabetic kidney disease (DKD), and it plays an essential role in the prevention and treatment of DKD. The "Gaozhuo" pathogenesis posits that DKD is characterized by the invasion of Gaozhuo and damage to the kidney collaterals, with the underlying cause being the insufficiency of spleen Qi and the internal formation of Gaozhuo, which provides valuable guidance on oxidative stress. The insufficiency of spleen Qi and the internal formation of Gaozhuo represent a dynamic, evolving process. Gaozhuo invades the kidney collaterals, impairs kidney Qi, and progressively leads to the congealing and stagnation of Gaozhuo and blood, ultimately resulting in the failure of both the spleen and kidneys. The damage caused by Gaozhuo to the kidney collaterals and kidney Qi is analogous to the organ and functional damage of the kidneys induced by excessive reactive oxygen species and oxidative stress. Damage to the kidney collaterals means organic injuries to the glomeruli, renal tubules, and renal interstitium, and the depletion of kidney Qi refers to damage to glomerular filtration and renal tubular reabsorption. The congealing and stagnation of Gaozhuo and blood in the kidney collaterals is similar to oxidative stress-induced thickening of the glomerular basement membrane and fibrosis. The interaction between spleen and kidney Qi deficiency and the congealing and stagnation of Gaozhuo and blood creates a vicious cycle that exacerbates the condition, ultimately evolving into the failure of both the spleen and kidneys. The failure of the spleen and kidneys is analogous to renal failure, and its extreme manifestation is end-stage renal disease and uremia. The treatment of oxidative stress in DKD with traditional Chinese medicine (TCM) is based on the principles of strengthening the spleen and tonifying the kidneys, and dispelling turbidity and removing blood stasis. According to the syndrome type, it is recommended to use methods such as strengthening the spleen and tonifying Qi while dispelling dampness and removing turbidity, strengthening the spleen and tonifying the kidneys while dispelling dampness and removing turbidity, strengthening the spleen and tonifying the kidneys while dispelling turbidity and removing blood stasis, or consolidating the spleen and kidneys while clearing away turbidity and blood stasis.
2.Shashen Maidong Tang Enhances Efficacy of Chemotherapy in Mouse Model of Lewis Lung Cancer by Modulating JAK2/STAT3 Signaling Pathway
Lin YU ; Yaoyao WANG ; Limin LIU ; Zuowei HU ; Yanping ZHOU ; Shang WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):1-10
ObjectiveTo predict the mechanism through which Shasheng Maidong Tang enhances the efficacy of chemotherapy for lung cancer via network pharmacology and validate the prediction results in animal experiments. MethodsThe potential mechanism through which Shasheng Maidong Tang enhances the efficacy of chemotherapy for lung cancer was predicted by network pharmacology, liquid chromatography-mass spectrometry (LC-MS), and molecular docking methods. C57/BL6 mice were assigned into normal, model, cisplatin, and Shasheng Maidong Tang+cisplatin groups. In addition to the normal group, the remaining groups were injected subcutaneously with 0.2 mL of 1×107 cells·mL-1 Lewis lung cancer cells to establish the Lewis lung cancer model. The daily gavage dose of Shasheng Maidong Tang was 3.58 g·kg-1, and the concentration of cisplatin intraperitoneally injected on every other day was 2 mg·kg-1. Drugs were administered for 14 d. The changes in the tumor volume and the rate of tumor suppression were monitored, and the tumor histopathological changes were observed by hematoxylin-eosin (HE) staining. Enzyme-linked immunosorbent assay was employed to measure the interleukin (IL)-6 and interferon (IFN)-γ levels in peripheral blood. Real-time PCR was performed to quantify the mRNA levels of Janus kinase 2 (JAK2), signal transducer and activator of transcription 1 (STAT1), and signal transducer and activator of transcription 3 (STAT3) in the tumor tissue of mice. Western blot was employed to determine the protein levels of JAK2, STAT3, B-cell lymphoma-2 (Bcl-2), cysteinyl aspartate-specific proteinase-3 (Caspase-3), and Pim-1 proto1 (PIM1) in the tumor tissue. Immunohistochemistry was employed to detect the expression of Bcl-2 and PIM1 in the tumor tissue. ResultsNetwork pharmacological predictions indicated that Shasheng Maidong Tang might enhance the efficacy of chemotherapy for lung cancer by regulating nitrogen metabolism, AGE-RAGE signaling pathway, cancer pathway, and JAK/STAT signaling pathway. The experimental results demonstrated that tumor volume in the cisplatin group and Shasheng Maidong Tang+cisplatin group was reduced compared with the model group, with statistically distinct differences observed on days 14, 17, 20 post modeling (P<0.05). Notably, the Shasheng Maidong Tang+cisplatin therapy further decreased tumor volume compared with the cisplatin group, showing marked reductions on days 17 and 20 (P<0.05), consistent with trends visualized in tumor volume comparison charts. The Shasheng Maidong Tang+cisplatin group exhibited higher tumor inhibition rate than the cisplatin group (P<0.05). Histopathological analysis via HE staining revealed that the tumors in the model group displayed frequent nuclear mitosis, densely arranged cells, hyperchromatic nuclei, and no necrosis. Cisplatin treatment induced partial necrosis and vacuolization, while the Shasheng Maidong Tang+cisplatin group exhibited extensive necrotic regions, maximal vacuolization, disarranged tumor cells, and minimal mitotic activity. Compared with the model group, the cisplatin group and the Shasheng Maidong Tang+cisplatin group showed elevated level of IFN-γ (P<0.01) and declined level of IL-6 (P<0.01) in the peripheral blood. Compared with the cisplatin group, the Shasheng Maidong Tang+cisplatin group presented elevated level of IFN-γ (P<0.01) and lowered level of IL-6 (P<0.01) in the peripheral blood. Compared with the model group, the cisplatin group and the Shasheng Maidong Tang+cisplatin groups showed down-regulated mRNA levels of JAK2 and STAT3 (P<0.01) and up-regulated mRNA level STAT1 (P<0.01). Compared with the cisplatin group, the Shasheng Maidong Tang+cisplatin group presented down-regulated mRNA levels of JAK2 and STAT3 (P<0.01) and up-regulated mRNA level of STAT1 (P<0.01). Compared with the model group, the cisplatin group and the Shasheng Maidong Tang+cisplatin group showed down-regulated protein levels of JAK2 (P<0.01), Bcl-2 (P<0.01), PIM1 (P<0.01), and STAT3 (P<0.05), and up-regulated protein level of Caspase-3 (P<0.01). Compared with the cisplatin group, Shasheng Maidong Tang+cisplatin group presented down-regulated protein levels of JAK2 (P<0.01), Bcl-2 (P<0.01), PIM1 (P<0.01), STAT3 (P<0.05), and up-regulated protein level of Caspase-3 (P<0.01). The Bcl-2 and PIM1 expression results obtained by immunohistochemistry were consistent with those of Western blot. ConclusionShasheng Maidong Tang may enhance the efficacy of chemotherapy in the mouse model of Lewis lung cancer by regulating the JAK2/STAT3 signaling pathway.
3.Effect of summer short-term clinical practice teaching on empathy ability in rehabilitation therapy students
Chinese Journal of Rehabilitation Theory and Practice 2026;32(1):117-124
ObjectiveTo explore an centralized clinical practice teaching model delivered via a summer short-term semester, and to examine its effectiveness on empathy and communication efficacy among undergraduate students majoring in rehabilitation therapy through a mixed-methods design. MethodsFrom June to July, 2025, forty second-year rehabilitation therapy undergraduates from Nanchang Medical College participated in a one-week immersive clinical practice during the summer short-term semester. An action research framework integrating a one-group pre-post experimental design and qualitative research methods were adopted. Quantitative data were collected using the Chinese-Adapted Jefferson Scale of Empathy-Medical Student Version (JSE-HP) to assess changes in empathy. Qualitative data were obtained through semi-structured focus group interviews and structured reflective journals to investigate students' experiences and transformations in empathic cognition, emotional integration and professional identity. ResultsAfter the teaching intervention, students demonstrated significant improvements in the total score of JSE-HP and all subdimensions (perspective taking, compassionate care and standing in the patient's shoes) (t < -3.69, P < 0.01). Qualitative analysis yielded three core themes: reconstruction of clinical reasoning paradigms, emotional-cognitive integration and elevation of professional identity. ConclusionSummer short-term semester clinical practice model, structured around “clinical immersion, narrative reflection, interprofessional collaboration and formative assessment”, effectively facilitates embodied cultivation of empathy. This model not only bridges the gap between theory and clinical practice, but also serves as an educational catalyst for students' transformation from technical performers to humanistic caregivers. It aligns with the core concepts of the World Health Organization rehabilitation competency framework, offering a replicable and scalable approach to advancing systematic reform in medical humanities education.
4.Differences in chemical components and quality analysis of Gardenia jasminoides before and after processing with ginger
Lihua TANG ; Yu WU ; Xuedi HUANG ; Xiaolian HU ; Yi TANG ; Zilong CHEN ; Xiaofan XIAO ; Xide YE
China Pharmacy 2026;37(2):168-173
OBJECTIVE To analyze the differences in chemical components of Gardenia jasminoides before and after processing with ginger, and to evaluate the quality differences among different producing areas. METHODS Ultra-high performance liquid chromatography-tandem time-of-flight mass spectrometry was used to analyze the compositional differences of G. jasminoides before and after processing with ginger. The water content, total ash, and ethanol-soluble extract content of ginger- processed G. jasminoides were determined according to the 2020 edition of Chinese Pharmacopoeia. High performance liquid chromatography was adopted to determine the contents of genipin gentiobioside, geniposide, crocin Ⅰ and crocin Ⅱ in ginger- processed G. jasminoides. RESULTS A total of 49 chemical components were identified from raw G. jasminoides and ginger- processed G. jasminoides, including 14 flavonoids, 15 iridoids, 10 organic acids, 2 alkaloids and 8 other compounds. Among them, 42 components were detected in raw G. jasminoides, 28 in ginger-processed G. jasminoides, and 21 components were common to both. After processing with ginger, raw G. jasminoides lost 21 components (including iridoids, flavonoids, alkaloids, and others), while 7 chemical components were added (including coumarins, organic acids, organic acid esters, and flavonoids). For the 15 batches of ginger-processed G. jasminoides, the water content ranged from 5.64% to 7.11%, total ash from 2.92% to 4.87%, and ethanol-soluble extract from 40.61% to 58.02%. The average contents of genipin gentiobioside, geniposide, crocin Ⅰ and crocin Ⅱ were 0.108 7, 0.542 2, 0.565 0, and 0.012 5 mg/g, respectively. CONCLUSIONS After processing with ginger, G. jasminoides loses 21 components, while 7 new components are added. Differences are observed in the water content, total ash, ethanol-soluble extract, and the contents of genipin gentiobioside, geniposide, crocin Ⅰ, and crocin Ⅱ of ginger-processed G. jasminoides from different producing areas. Notably, samples from Fujian exhibit high contents of genipin gentiobioside and ethanol-soluble extract, while samples from Jiangxi have a high content of crocin Ⅰ.
5.Efficacy and Application Characteristics of Cold Chinese Medicines Based on Chinese Pharmacopoeia (2020 Edition)
Lu YUE ; Yilong HU ; Jingying YANG ; Xiangxiang WU ; Mingsan MIAO ; Ming BAI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):241-248
ObjectiveTo provide a reference for the rational clinical use of cold Chinese medicines by sorting and analyzing their properties, flavors, meridian tropism, primary therapeutic indications, methods of administration, dosages, and precautions as recorded in the 2020 edition of Pharmacopoeia of the People's Republic of China (Chinese Pharmacopoeia). MethodsCold Chinese medicines for internal and external use included in the 2020 edition of Chinese Pharmacopoeia were entered one by one, and their efficacy, properties, flavors, meridian tropism, methods of administration, dosages, and usage precautions were statistically classified and summarized to guide clinical medication use. ResultsA total of 259 cold Chinese medicines for internal use were included and categorized into 18 efficacy groups, mainly comprising heat-clearing drugs, water-excreting and dampness-draining drugs, and phlegm-resolving, cough- and asthma-relieving drugs. Their predominant flavors were bitter, sweet, and pungent, and they primarily entered the liver, lung, and stomach meridians. The main methods of administration included decocting first, grinding into powder for oral use, or preparing into pills or powders, with most dosages ranging from 9 to 15 g. A total of 83 cold Chinese medicines for external use were included, involving 16 efficacy categories. Their main flavors were bitter, sweet, and pungent, primarily entering the liver, lung, and large intestine meridians. The main external application methods were grinding into powder for topical use or preparing decoctions for fumigation and washing, with most dosages ranging from 9 to 15 g. Whether for internal or external use, cold Chinese medicines should be used with caution or contraindicated in pregnant women. ConclusionThe cold Chinese medicines included in the 2020 edition of the Chinese Pharmacopoeia are mainly suitable for patients with carbuncles, swellings, and coughs. However, in clinical practice, it is necessary to strictly follow the principles of syndrome differentiation and treatment, pay attention to administration methods and dosages, and use cold medicines rationally and effectively to improve clinical efficacy.
6.Comparison of sleep EEG power spectral density between depressive episode patients and schizophrenia patients with suicidal behavior
Jingwen LIU ; Yunfei ZHOU ; Jingchu HU ; Jiaoyan ZHOU ; Junwei YANG ; Jie LIANG ; Hong XU ; Yu CANG ; Shimeng MA
Sichuan Mental Health 2026;39(1):50-57
BackgroundPatients with depressive episode and schizophrenia have a high risk of suicide. The sleep electroencephalogram power spectral density characteristics of patients with depressive episode accompanied by suicidal behavior and those with schizophrenia may be different, but there is currently a lack of direct comparative studies on these two groups of patients. ObjectiveTo compare the sleep electroencephalogram power spectral density between depressive episode and schizophrenic patients with suicidal behavior, in order to provide references for exploring predictive indicators of suicidal behavior. MethodsFrom June 2018 to December 2020, 20 patients with depressive episode and 20 patients with schizophrenia who had committed suicide within the past month and were treated at the outpatient department of Shenzhen Kangning Hospital were selected. All of them met the diagnostic criteria for depressive episode or schizophrenia as defined in the International Classification of Diseases, tenth edition (ICD-10). Using a random sampling method, 20 volunteers with matching gender and age to the patient groups were selected from the Cuiping community in Shenzhen as the control group. The subjective sleep of the patients was evaluated using the Insomnia Severity Index (ISI), the Dysfunctional Belief and Attitude about Sleep (DBAS), the Disturbing Dreams and Nightmare Severity Index (DDNSI), and the Epworth Somnolence Scale (ESS). The objective sleep of the patients was assessed using polysomnography. The sleep electroencephalogram was filtered and the power spectral density of the brain wave was analyzed and processed for all the subjects. The subjective and objective sleep conditions of the two patient groups were compared, and the sleep electroencephalogram power spectral density of the patient groups and the control group were also compared. ResultsA comparison of subjective and objective sleep conditions between patients with depressive episode accompanied by suicidal behavior and patients with schizophrenia accompanied by suicidal behavior showed no statistically significant differences (P>0.05). Comparisons of sleep electroencephalogram power spectral density in the W stage (average power of α wave, total power of δ wave, average power of δ wave, average power of θ wave), N1 stage (average power of β wave, total power of α wave, total power of δ wave), N2 stage (total power of α wave, average power of α wave, total power of δ wave, average power of δ wave), N3 stage (average power of α wave, average power of δ wave), and R stage (total power of α wave, average power of α wave, total power of δ wave, average power of δ wave) between patients with depressive episode accompanied by suicidal behavior, patients with schizophrenia accompanied by suicidal behavior, and the control group showed statistically significant differences (P<0.05 or 0.01). The total power of δ wave in the W stage and the average power of β wave and δ wave in the N1 stage were higher in two patient groups were higher than those of the control group. The total power of α wave and the average power of α wave in the N2 stage were lower than those of the control group, while the average power of δ wave was higher than that of the control group. The average power of α wave in the N3 stage of both patient groups were lower than that of the control group, while the average power of δ wave was higher than that of the control group. The total power and average power of α wave in the R stage were lower than those of the control group, while the total power and average power of δ wave were higher than those of the control group. All the differences were statistically significant. Patients with depressive episode accompanied by suicidal behavior had higher average powers of α wave, δ wave, and θ wave in the W stage compared with the control group, while the total power of α wave in the N1 stage was lower in the former group. All these differences were statistically significant (P<0.05). ConclusionThe depressive episode patients accompanied by suicidal behavior have highly overlapping sleep electroencephalogram abnormal patterns with those of schizophrenia patients, mainly manifested as a general decrease in α wave power (N2, N3, R stage) and a general increase in δ wave power (W, N1, N2, N3, R stage) as well as β wave power in N1 stage. At the same time, patients with depressive episode accompanied by suicidal behavior also show specific changes, including an increase in the average power of α and θ waves during the wakefulness period (W stage), and a decrease in the total power of α wave in N1 stage. [Funded by Guangdong Province High-level Clinical Key Specialty (with supporting funds from Shenzhen City) (number, SZGSP013); Shenzhen Key Medical Discipline (number, SZXK041); Shenzhen Clinical Medicine Research Center Project (number, 20210617155253001)]
7.Improvement effect and mechanism of Dendrobium officinale on skin damage in mice with xeroderma
Penglong YU ; Jianqing DENG ; Shanhong SUN ; Kun GAO ; Jianghua HU
China Pharmacy 2026;37(7):914-919
OBJECTIVE To study the improvement effect and mechanism of Dendrobium officinale on skin damage in mice with xeroderma. METHODS The mice were randomly divided into control group, model group, and D. officinale group, with 5 mice in each group. Except for the control group (which only underwent shaving treatment), the mice in all other groups were induced to develop a xeroderma model using an acetone-ether mixture for five consecutive days. The mice in D. officinale group were treated with 200 μL of D. officinale suspension (0.2 mg/mL) two hours after the first modeling each day. Mice in the control group and the model group were applied with an equal volume of pure water; once a day, until the end of the modeling process. After last medication, skin lesions and pathological morphology of the mice were observed. Immunofluorescence was used to detect the expressions of Filaggrin, Loricrin and Ki67 proteins in skin tissue of the mice. The core pathways through which D. officinale improves skin damage in xeroderma were screened using 16S rRNA sequencing combined with gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses, and subsequent validation was conducted. RESULTS Compared with the control group, the mice in the model group exhibited obvious scratching behavior, with a large amount of scale on the skin, excessive epidermal keratinization, and thickened stratum spinosum. The skin scale score, epidermal thickness, and the expression levels of Ki67, Filaggrin, and Loricrin proteins in the skin tissue were significantly increased/elevated ( P <0.05). Compared with model group, the mice in the D. officinale group exhibited reduced scratching behavior and scaling, along with a mitigated degree of skin keratinization. The aforementioned quantitative indicators were significantly decreased/reduced ( P <0.05). The results of core pathway screening revealed that the KEGG pathways involving differentially expressed genes included signaling pathways such as interleukin-17 (IL-17) and tumor necrosis factor (TNF). Further validation experim ents found that after intervention with D. officinale , mRNA expression of downstream effector molecules CCN1, Hbegf, Tnfrsf12a, and Thbs1 genes in skin tissues were all significantly reduced ( P <0.05). CONCLUSIONS D. officinale can repair skin damage in mice with xeroderma, and its mechanism of action is related to restoring the balance of proliferation and differentiation in keratinocytes and down-regulating the mRNA expressions of CCN1, Hbegf, Tnfrsf12a, and Thbs1.
8.Mechanisms of Salvianolic Acid B in Inhibiting Epithelial-mesenchymal Transition in Non-small Cell Lung Cancer by Downregulating PAICS Expression
Bo XU ; Jixian ZHANG ; Linling HU ; Bo JIANG ; Shasha YUAN ; Yiling FAN ; Zhishen RUAN ; Yihan YU ; Qing MIAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):23-33
ObjectiveTo investigate the molecular mechanisms by which salvianolic acid B (SalB) inhibits epithelial-mesenchymal transition (EMT) in non-small cell lung cancer (NSCLC) by downregulating phosphoribosylaminoimidazole carboxylase and phosphoribosylaminoimidazole succinocarboxamide synthetase (PAICS) expression. MethodsNSCLC A549 cells and normal bronchial epithelial cells (bronchial epithelium transformed with Ad12-SV40 2B, BEAS-2B) were used as models. Cell viability was assessed using the cell counting kit-8 (CCK-8) assay after treatment with SalB (0, 50, 100, 200, 300, 400, 500 μmol·L-1 for 24 or 48 h to determine effective and safe intervention concentrations. Cell proliferation, cell cycle distribution, and apoptosis were evaluated by 5-ethynyl-2′-deoxyuridine (EdU) staining and flow cytometry, respectively. Wound healing and Transwell invasion assays were performed to assess cell migration and invasion. RNA sequencing combined with bioinformatic analysis was conducted to identify differentially expressed genes and functional enrichment. Molecular docking was used to predict the binding ability between SalB and PAICS, and the cellular thermal shift assay (CETSA) was performed to evaluate the effect of SalB on the thermal stability of the PAICS protein. Western blot (WB) was used to detect the effects of SalB on PAICS and EMT-related proteins (E-cadherin, N-cadherin, Vimentin, Snail, and Slug). A functional rescue assay was conducted by PAICS overexpression via plasmid transfection. ResultsCompared with the control group, SalB inhibited A549 cell viability in a dose-dependent manner (P<0.05), and the effective concentrations (≤300 μmol·L-1) showed no significant cytotoxicity in BEAS-2B cells. Within this concentration range, SalB significantly inhibited A549 cell proliferation, migration, and invasion, and induced G0/G1 phase arrest and apoptosis (P<0.05). Transcriptomic analysis showed that SalB significantly downregulated PAICS expression, and its functions were enriched in cell proliferation and EMT. Bioinformatic analysis indicated that PAICS is highly expressed in lung adenocarcinoma and is associated with poor prognosis (P<0.01). Molecular docking showed that SalB has strong binding ability to PAICS (binding energy -9.1 kcal·mol-1. CETSA results showed that SalB significantly increased the thermal stability of the PAICS protein (P<0.05). WB results showed that, compared with the control group, SalB dose-dependently downregulated PAICS expression, upregulated E-cadherin, and downregulated N-cadherin, Vimentin, Snail, and Slug (P<0.05). Functional rescue experiments showed that, compared with the empty vector group, PAICS overexpression significantly enhanced A549 cell proliferation, migration, and invasion, promoted cell cycle progression, and inhibited apoptosis (P<0.05). Meanwhile, compared with the empty vector + SalB-H group, PAICS overexpression partially reversed the inhibitory effects of SalB on malignant phenotypes and EMT-related proteins (N-cadherin, Vimentin, Snail, and Slug), and downregulated E-cadherin expression (P<0.05,P<0.01), indicating that PAICS is a key functional target mediating the antitumor effects of SalB. ConclusionSalB effectively inhibits EMT progression and cell cycle progression in A549 cells by downregulating PAICS expression, thereby exerting anti-NSCLC effects. This study not only reveals that PAICS is a key functional target through which SalB regulates EMT, but also provides experimental evidence supporting SalB as a potential candidate drug for inhibiting NSCLC metastasis.
9.Mechanisms of Curcumol in Inhibiting Proliferation and Migration in Non-small Cell Lung Cancer via JAK2/STAT3 Signaling Pathway
Yu QI ; Yihan YU ; Linling HU ; Bo JIANG ; Yilong ZOU ; Cunyu FAN ; Yiling FAN ; Jixian ZHANG ; Bo XU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):34-45
ObjectiveTo investigate the inhibitory effects of curcumol (Cur) on the proliferation and metastasis of non-small cell lung cancer (NSCLC) cells and to explore the underlying mechanisms. MethodsIn vivo, a subcutaneous tumor xenograft model was established to evaluate the antiproliferative effect of Cur. In vitro, the cell counting kit-8 (CCK-8) assay was used to assess the effects of Cur at concentrations of 0, 60, 120, 240, 360, 480, 600, 720, 840, 960 μmol·L-1 on the viability of NCI-A549 and NCI-H23 cells, and to evaluate its inhibitory effect on the proliferation of human bronchial epithelial BEAS-2B cells. Wound healing and Transwell migration assays were conducted to assess changes in cell migratory capacity following Cur treatment. Immunohistochemistry (IHC-P) was used to investigate the regulatory effect of Cur on the Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signaling pathway in tumor tissues. Western blot was performed to determine the protein expression levels of phosphorylated JAK2 (p-JAK2), phosphorylated STAT3 (p-STAT3), proliferating cell nuclear antigen (PCNA), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), and vascular endothelial growth factor A (VEGFA) in tumor tissues and cells. To further verify the role of the JAK2/STAT3 signaling pathway in the pharmacological effects of Cur, rescue experiments were performed using the pathway agonist colivelin. ResultsIn vivo experiments showed that, compared with the model group, the tumor volumes of subcutaneous xenografts in nude mice in both low- and high-dose Cur groups were significantly reduced (P<0.05), and the tumor inhibition rates were significantly increased (P<0.05). The inhibitory effect in the high-dose group was comparable to that of the cisplatin group, and the body weight of mice in the Cur groups remained stable throughout the experiment. In vitro, compared with the control group, Cur at concentrations of 120 and 240 μmol·L-1 inhibited the proliferation of NCI-A549 and NCI-H23 cells in a concentration-dependent manner (P<0.05), with a significant inhibitory effect observed at 360 μmol·L-1 (P<0.01), while no significant effect on the viability of BEAS-2B cells was observed. Migration assays demonstrated that, compared with the control group, Cur treatment significantly reduced the migration rates of both cell lines in a concentration-dependent manner (P<0.05), with an inhibitory effect at 360 μmol·L-1 comparable to that of the cisplatin group. Mechanistic validation showed that, compared with the control group, the protein expression levels of p-JAK2 and p-STAT3 in tumor tissues and cells were significantly downregulated in the Cur groups (P<0.01), and the expression levels of downstream proteins PCNA, MMP-2, MMP-9, and VEGFA were also significantly decreased with increasing Cur concentration (P<0.05). In the rescue experiments, compared with the control group, colivelin pretreatment increased cell proliferation and migration rates (P<0.05) and upregulated the expression of related proteins (P<0.05). Compared with the Cur group, the colivelin+Cur group showed significantly increased proliferation and migration rates (P<0.05), along with significantly upregulated protein expression levels (P<0.05). ConclusionCur can significantly inhibit the proliferation and metastasis of NSCLC both in vivo and in vitro, and its mechanism of action is closely associated with the inhibition of JAK2/STAT3 signaling pathway activation.
10.Mechanisms of Oxyresveratrol in Inhibiting Epithelial-mesenchymal Transition in Non-small Cell Lung Cancer via PI3K/Akt Signaling Pathway
Linling HU ; Bo JIANG ; Yu QI ; Yilong ZOU ; Cunyu FAN ; Yiling FAN ; Yihan YU ; Bo XU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):46-57
ObjectiveTo investigate the mechanisms by which oxyresveratrol (OXY) inhibits epithelial-mesenchymal transition (EMT) in non-small cell lung cancer (NSCLC) through the phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway. MethodsCell counting kit-8 (CCK-8) assays were used to determine the survival rates of A549 and H1299 cells treated with different concentrations of OXY, and appropriate concentrations (0, 30, 60, 90 μmol·L-1) were selected. The effects of OXY on the proliferation of A549 and H1299 cells were evaluated using 5-ethynyl-2′-deoxyuridine (EdU) assays and colony formation assays. Wound healing assays and Transwell invasion assays were performed to assess the effects of OXY on cell migration and invasion. Western blot (WB) was used to detect the expression levels of Snail, E-cadherin, N-cadherin, and Vimentin in A549 and H1299 cells. Network pharmacology and molecular docking were applied to predict the mechanism of action of OXY, and WB was used to evaluate the effects of OXY on proteins in the PI3K/Akt signaling pathway. Rescue experiments were conducted using the PI3K/Akt signaling pathway agonist 740Y-P. Under activation of the PI3K/Akt pathway, the effect of OXY on proliferation, migration, and invasion phenotypes, as well as on the expression levels of PI3K/Akt pathway-related proteins and EMT markers (Snail, E-cadherin, N-cadherin, and Vimentin), were examined. ResultsIn the forward experiments, CCK-8 assay results showed that, compared with the control group, the survival rates of NSCLC cells in the OXY-treated groups (20-120 μmol·L-1) were significantly decreased (P<0.05). The half-maximal inhibitory concentration (IC50) values of A549 and H1299 cells after 48 h of OXY treatment were 113.6 μmol·L-1 and 92.53 μmol·L-1, respectively. Therefore, concentrations of 0, 30, 60, 90 μmol·L-1 were selected as the gradient for subsequent phenotypic and mechanistic studies. Compared with the control group, the proliferation rate, colony number, migration rate, and invasion number of NSCLC cells in the OXY groups (30, 60, and 90 μmol·L-1) were significantly decreased (P<0.01, P<0.05). WB results showed that, compared with the control group, the protein expression levels of Snail, N-cadherin, and Vimentin in NSCLC cells of the OXY groups were significantly decreased (P<0.05), whereas E-cadherin expression was significantly increased (P<0.01). Network pharmacology and molecular docking results indicated that OXY could act on the PI3K/Akt signaling pathway and exhibited good binding affinity with PI3K and Akt proteins. Further WB results showed that, compared with the control group, there were no statistically significant differences in the expression levels of PI3K and Akt proteins in NSCLC cells of the OXY groups, whereas the expression levels of phosphorylated PI3K (p-PI3K) and phosphorylated Akt (p-Akt) were significantly decreased (P<0.05). In the rescue experiments, compared with the control group, the proliferation rate, colony number, migration rate, and invasion number of NSCLC cells in the 740Y-P group (15 μmol·L-1) were significantly increased (P<0.01). Compared with the control + OXY group (90 μmol·L-1), these indices in the 740Y-P + OXY group (15 μmol·L-1 + 90 μmol·L-1) were also significantly increased (P<0.01). WB results showed that, compared with the control group, there were no statistically significant differences in the expression levels of PI3K and Akt proteins in the 740Y-P group. However, the expression levels of p-PI3K, p-Akt, Snail, N-cadherin, and Vimentin were significantly increased (P<0.05), while E-cadherin expression was significantly decreased (P<0.01). Compared with the control + OXY group, there were no statistically significant differences in PI3K and Akt protein expression in the 740Y-P + OXY group. However, the expression levels of p-PI3K, p-Akt, Snail, N-cadherin, and Vimentin were significantly increased (P<0.05), while E-cadherin expression was significantly decreased (P<0.05). ConclusionOXY inhibits the PI3K/Akt signaling pathway and suppresses the EMT process, thereby exerting anti-metastatic effects in NSCLC.

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