1.Research progress on antibody-drug conjugates in the treatment of triple-negative breast cancer
Danna LIU ; Shuangshuang SONG ; Lu CHEN ; Yongqiang SUN ; Bo SUN ; Hanli ZHOU ; Xiaoli ZHAO ; Tiandong KONG
China Pharmacy 2026;37(1):124-129
Antibody-drug conjugates (ADCs) are a novel class of anti-tumor agents composed of a targeted monoclonal antibody, a cytotoxic drug, and a linker connecting the two. They combine the high specificity of antibodies with the potent cytotoxicity of chemotherapeutic agents. Triple-negative breast cancer (TNBC) is characterized by high aggressiveness, elevated risks of recurrence and metastasis, and poor prognosis, largely due to the lack of effective therapeutic targets. This review summarizes the research progress of ADCs in the treatment of TNBC. It has been found that ADCs targeting human epidermal growth factor receptor 2 (such as trastuzumab deruxtecan), trophoblast cell surface antigen 2 (such as sacituzumab govitecan and datopotamab deruxtecan), zinc transporter LIV-1 (such as ladiratuzumab vedotin), HER-3 (such as patritumab deruxtecan), epidermal growth factor receptor (such as AVID100), and glycoprotein non-metastatic melanoma protein B (such as glembatumumab vedotin) have all demonstrated promising therapeutic effects against TNBC. Despite challenges including acquired resistance and treatment-related toxicities, ADCs are undoubtedly reshaping the therapeutic landscape for TNBC and are expected to occupy a more central position in TNBC treatment in the future.
2.Mechanism of Xiaozheng Zhitong Paste in Alleviating Bone Cancer Pain by Regulating Microglial Pyroptosis Based on PINK1/Parkin/NLRP3 Signaling Pathway
Lingyun WANG ; Guangda ZHENG ; Lu SHANG ; Juanxia REN ; Changlin LI ; Dongtao LI ; Haixiao LIU ; Yaohua CHEN ; Guiping YANG ; Yanju BAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):80-90
ObjectiveThe paper aims to investigate the mechanism by which Xiaozheng Zhitong paste (XZP) alleviates bone cancer pain (BCP) through regulating the PTEN-induced putative kinase 1 (PINK1)/Parkin-mediated mitophagy-NOD-like receptor protein 3 (NLRP3) inflammasome pathway to suppress microglial pyroptosis. MethodsLipopolysaccharide (LPS) and LPS-adenosine triphosphate (ATP) were used to establish an inflammation and pyroptosis model in microglial cells. The cells were randomly divided into the following groups: control group, LPS group, LPS+low-dose XZP group, LPS+high-dose XZP group, LPS-ATP group, LPS-ATP+low-dose XZP group, LPS-ATP+high-dose XZP group, LPS-ATP+XZP group, and LPS-ATP+XZP+CsA group. Techniques including terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining, enzyme-linked immunosorbent assay (ELISA), Western blot, and confocal fluorescence staining were employed to assess the effects of XZP on microglial apoptosis, inflammatory cytokine release, inflammasome activation, pyroptosis, and mitophagy. ResultsIn vitro experiments showed that compared with the blank group, the LPS group exhibited significantly increased levels of microglial apoptosis and pro-inflammatory factors interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α)(P<0.01), along with significantly upregulated protein expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and phosphorylated nuclear factor-κB p65 (p-NF-κB p65) (P<0.01). Compared with the LPS group, the high-dose LPS-XZP group significantly reduced the level of apoptosis (P<0.01) and the content of the aforementioned pro-inflammatory factors (P<0.01). Both the low- and high-dose LPS-XZP groups dose-dependently downregulated the protein expression of iNOS, COX-2, and p-NF-κB p65 (P<0.05, P<0.01). Compared with the blank group, the LPS-ATP group showed significantly upregulated expression of pyroptosis-related proteins, including Caspase-1/pro-Caspase-1, N-terminal fragment of gasdermin D (GSDMD-N)/full-length gasdermin D (GSDMD-F), NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), IL-1β precursor (pro-IL-1β), and mature IL-1β (P<0.01). The levels of pyroptotic factors IL-1β and IL-18 were significantly elevated (P<0.01), and membrane pore formation and intracellular reactive oxygen species (ROS) levels were significantly increased (P<0.01). Compared with the LPS-ATP group, both the low- and high-dose LPS-ATP+XZP groups dose-dependently downregulated the expression of the aforementioned pyroptosis-related proteins (P<0.05, P<0.01). The low-dose LPS-ATP+XZP group reduced IL-1β levels (P<0.01), while the high-dose group reduced both IL-1β and IL-18 levels (P<0.01) Both the low- and high-dose LPS-ATP+XZP groups dose-dependently reduced membrane pore formation and intracellular ROS production (P<0.01). Compared with the blank group, the LPS-ATP group showed significantly reduced expression of mitophagy-related proteins PINK1 and Parkin, and a decreased ratio of microtubule-associated protein 1 light chain 3Ⅱ(LC3Ⅱ) to LC3Ⅰ(P<0.01), while p62 expression was significantly increased (P<0.01). Mitochondrial ROS levels were significantly enhanced (P<0.01). Compared with the LPS-ATP group, both the low- and high-dose LPS-ATP+XZP groups dose-dependently reversed the expression of these proteins (P<0.05, P<0.01) and reduced mitochondrial ROS levels (P<0.01). After treatment with the mitophagy inhibitor cyclosporin A (CsA), the beneficial effects of XZP on mitochondrial function and its inhibitory effects on pyroptosis-related protein expression were significantly reversed (P<0.05, P<0.01). ConclusionXZP reduces ROS levels by activating PINK1/Parkin-mediated mitophagy, thereby inhibiting NLRP3 inflammasome activation and microglial pyroptosis, which provides new molecular evidence for the mechanism by which XZP alleviates BCP.
3.Xiaozheng Zhitong Paste Relieves Bone Cancer Pain in Mice by Alleviating Activation of Microglia in Spinal Cord and Damage to Neurons via Blocking PAR2/NF-κB/NLRP3 Pathway
Guangda ZHENG ; Linghan MENG ; Lu SHANG ; Juanxia REN ; Dongtao LI ; Haixiao LIU ; Lingyun WANG ; Changlin LI ; Yaohua CHEN ; Guiping YANG ; Yanju BAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):91-100
ObjectiveTo investigate the effects and underlying mechanisms of Xiaozheng Zhitong Paste (XZP) on bone cancer pain (BCP). MethodsThirty female BALB/c mice were randomly divided into five groups: a Sham group, a BCP group, a BCP+low-dose XZP group, a BCP+high-dose XZP group, and a BCP+high-dose XZP + protease-activated receptor 2 (PAR2) agonist GB-110 group. BCP mice model was constructed by injecting Lewis lung carcinoma cells into the femoral cavity of the right leg, which was followed by being treated with XZP for 21 d. After 21 d, the mice were sacrificed. Nissl staining was used to evaluate the survival of spinal cord neurons. Immunofluorescence staining was conducted to localize ionized calcium-binding adapter molecule 1 (Iba1) and neuronal nuclear antigen (NeuN) in spinal cord tissue, thereby assessing microglial activation and neuronal survival. Enzyme-linked immunosorbent assay (ELISA) was employed to measure the levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), transforming growth factor-β (TGF-β), interleukin-4 (IL-4), and interleukin-10 (IL-10) in spinal cord tissue. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect mRNA expression levels associated with M1/M2 polarization of microglia. Western blot analysis was performed to examine the expression of proteins related to microglial polarization as well as those involved in the PAR2/nuclear factor kappa B (NF-κB)/NOD-like receptor protein 3 (NLRP3) signaling pathway in the spinal cord. ResultsCompared with the Sham group, the spinal cord neurons were damaged, the number of Nissl-positive spinal cord neurons in the spinal cord tissue was significantly reduced (P<0.01), and the rate of NeuN-positive cells was significantly decreased (P<0.01). The spinal cord microglia were activated, the inflammatory level of the spinal cord tissue was enhanced, and Iba1 staining was significantly enhanced (P<0.01). The levels of IL-1β, TNF-α, IL-6, TGF-β, IL-4 and IL-10 were significantly increased (P<0.01). The mRNA expressions of IL-1β, TNF-α and inducible nitric oxide synthase (iNOS) were significantly increased (P<0.01), and the expression of PAR2, NLRP3, ASC and NF-κB p65 proteins in the spinal cord tissue of the BCP mice was significantly enhanced (P<0.01). Compared with the BCP group, high-dose XZP treatment significantly increased the number of Nissl-positive spinal cord neurons in the BCP mice (P<0.01), significantly enhanced the rate of NeuN-positive cells in the spinal cord tissue, and significantly weakened Iba1 staining (P<0.01). In addition, the levels of IL-1β, TNF-α, and IL-6 were significantly decreased, while the levels of TGF-β, IL-4, and IL-10 were significantly increased (P<0.05, P<0.01). The mRNA expression levels of IL-1β, TNF-α, and iNOS were decreased, whereas those of cluster of differentiation 206 (CD206), arginase-1 (Arg-1), and YM1/2 were significantly increased (P<0.05, P<0.01). Low-dose and high-dose XZP treatment significantly decreased the expression of PAR2, NLRP3, ASC, and NF-κB p65 proteins in the spinal cord tissue (P<0.05, P<0.01). These effects could all be significantly eliminated by the PAR2 agonist GB-110. ConclusionXZP can mitigate BCP in mice, which may be achieved through blocking the activated PAR2/NF-κB/NLRP3 pathway.
4.Traditional Chinese Medicine for Cancer Pain Management: A Review
Lingyun WANG ; Guangda ZHENG ; Lu SHANG ; Juanxia REN ; Changlin LI ; Dongtao LI ; Haixiao LIU ; Yaohua CHEN ; Guiping YANG ; Yanju BAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):114-123
Cancer pain is one of the most common complications in patients with malignant tumors, severely affecting their quality of life. Its pathogenesis involves complex interactions among the tumor microenvironment, peripheral sensitization, and central sensitization. The tumor microenvironment initiates peripheral pain sensitization by secreting algogenic mediators, activating ion channels and related receptor signaling pathways, driving abnormal osteoclast activation, and mediating neuro-immune crosstalk. Persistent nociceptive input further triggers increased excitability of central neurons, activation of glial cells, and neuroinflammatory cascade reactions, ultimately leading to central pain sensitization. Although traditional opioid drugs can alleviate pain to some extent, they still have many limitations, such as incomplete analgesia, drug tolerance, and adverse reactions. In recent years, traditional Chinese medicine (TCM) compounds have made continuous progress in the treatment of cancer pain. Studies have shown that they can not only effectively relieve cancer pain and reduce the dosage of opioids but also significantly improve patients' quality of life. TCM treatment of cancer pain follows the principle of syndrome differentiation and treatment. Based on this, targeted therapeutic principles have been proposed, including promoting blood circulation, removing stasis, regulating Qi, and unblocking collaterals; tonifying the kidney, replenishing essence, warming Yang, and dispersing cold, activating blood, resolving phlegm, detoxifying, and dispersing nodules, as well as strengthening the body, replenishing deficiency, and harmonizing Qi and blood. Modern research indicates that TCM compounds can exert synergistic effects through multiple pathways, inhibiting inflammatory responses, regulating nerve conduction, intervening in bone metabolism and related gene expression, thereby producing anti-inflammatory and bone-protective effects to achieve the goal of alleviating cancer pain. This article systematically elaborates on the pathogenesis of cancer pain, the clinical application of TCM in treating cancer pain, and its related mechanisms of action, aiming to provide a theoretical basis and new strategies for the integration of TCM into comprehensive cancer pain management.
5.Empirical study of input, output, outcome and impact of community-based rehabilitation stations
Xiayao CHEN ; Ying DONG ; Xue DONG ; Zhongxiang MI ; Jun CHENG ; Aimin ZHANG ; Didi LU ; Jun WANG ; Jude LIU ; Qianmo AN ; Hui GUO ; Xiaochen LIU ; Zefeng YU
Chinese Journal of Rehabilitation Theory and Practice 2026;32(1):83-89
ObjectiveTo investigate the present situation of input, output, outcome and impact of all registered community-based rehabilitation stations in Inner Mongolia in China, and analyze how the input predict the output, outcome and impact. MethodsFrom March 1st to April 30th, 2025, a questionnaire survey was conducted on all registered community-based rehabilitation stations in Inner Mongolia, covering four dimensions: input, output, outcome and impact. A total of 1 365 questionnaires were distributed. The input included four items: laws and policies, human resources, equipment and facilities, and rehabilitation information management. The output included two items: technical paths and benefits/effectiveness. The outcome included three items: coverage rates, rehabilitation interventions and functional results. The impact included two items: health and sustainability. Each item contained several questions, all of which were described in a positive way. Each question was scored from one to five. A lower score indicated that the situation of the community-based rehabilitation station was more in line with the content described in the question. Regression analysis was performed using the total score of each item of input dimension as independent variables, and the total scores of the output, outcome and impact dimensions as dependent variables. ResultsA total of 1 262 valid questionnaires were collected. The mean values of input, output, outcome and impact of community-based rehabilitation stations were 1.827 to 1.904, with coefficient of variation of 45.892% to 49.239%. The regression analysis showed that, rehabilitation information management, human resources, and laws and policies significantly predicted the output dimension (R² = 0.910, P < 0.001). Meanwhile, all four items in the input dimension predicted both the outcome (R² = 0.850, P < 0.001) and impact dimensions (R² = 0.833, P < 0.001). ConclusionInput, output, outcome and impact of the community-based rehabilitation stations in Inner Mongolia were generally in line with the content of the questions, although some imbalances were observed. Additionally, the input of community-based rehabilitation stations could significantly predict their output, outcome and impact.
6.Effect and mechanism of Biejiajian Pill on subcutaneous xenograft tumor model of hepatocellular carcinoma Huh7 cells
Lu LU ; Huanling CHEN ; Jian XU ; Yuanqin DU ; Xiaoli LIU ; Yingsheng WU ; Chengting WU ; Wei BAN ; Jingjing HUANG ; Hongna HUANG
Journal of Clinical Hepatology 2026;42(1):125-133
ObjectiveTo investigate the inhibitory effect of Biejiajian Pills (BJJW) on the growth of liver cancer, as well as its potential mechanism in mediating the AMP-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) pathway through mitochondrial energy metabolism. MethodsHuman hepatoma Huh7 cells were used to establish a nude mouse model of subcutaneous xenograft tumor. A total of 18 tumor-bearing nude mice were randomly divided into model group, BJJW group (2.2 g/kg), and metformin group (250 mg/kg), and the corresponding drug was given by gavage for 14 consecutive days. Tumor volume and weight were monitored during the experiment; HE staining was used to observe histopathological changes; the levels of reactive oxygen species (ROS) and adenosine triphosphate (ATP) in tumor tissue were measured; immunohistochemistry and Western blotting were used to measure the expression levels of proteins associated with the AMPK/mTOR pathway. A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups, and the Tukey’s test was used for further comparison between two groups; the Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups, and the Dunn’s test was used for further comparison between two groups. ResultsCompared with the model group, the BJJW group had a tumor inhibition rate of 45.73%, with significant reductions in both tumor volume and weight (P<0.01). Pathological examination showed that compared with the model group, the BJJW group had a significant reduction in the number of tumor cells and the presence of extensive necrosis. Mechanistic studies showed that compared with the model group, the BJJW group had a significant increase in ROS level (P<0.001) and a significant reduction in ATP level (P<0.001), as well as significant increases in p-AMPK/AMPK ratio (0.81±0.20 vs 0.13±0.04, P<0.01) and p-ULK1/ULK1 ratio (0.69±0.17 vs 0.18±0.13, P<0.01) and a significant reduction in p-mTOR/mTOR ratio (1.34±0.16 vs 3.20±0.62, P<0.01). ConclusionBJJW may inhibit the growth of liver cancer by inducing mitochondrial energy metabolism dysfunction, increasing the level of ROS, reducing the level of ATP, and activating the AMPK/mTOR signaling pathway.
7.Preliminary application of sacral neuromodulation in patients with benign prostatic hyperplasia complicated with underactive bladder after transurethral resection of the prostate
Ning LIU ; Yan ZHANG ; Tao LI ; Qiang HU ; Kai LU ; Lei ZHANG ; Jianping WU ; Shuqiu CHEN ; Bin XU ; Ming CHEN
Journal of Modern Urology 2025;30(1):39-42
[Objective] To evaluate the efficacy and safety of sacral neuromodulation (SNM) in the treatment of patients with benign prostatic hyperplasia (BPH) complicated with underactive bladder (UAB) who respond poorly to transurethral resection of the prostate (TURP). [Methods] A retrospective analysis was performed on 10 patients with BPH and UAB treated with TURP by the same surgeon in Zhongda Hospital Southeast University during Jan.2018 and Jan.2023.The residual urine volume was not significantly relieved after operation, and the maximum urine flow rate and urine volume per discharge were not significantly improved.All patients underwent phase I SNM, and urinary diaries were recorded before and after surgery to observe the average daily frequency of urination, volume per urination, maximum urine flow rate, and residual urine volume. [Results] The operation time was (97.6±11.2) min.During the postoperative test of 2-4 weeks, if the residual urine volume reduction by more than 50% was deemed as effective, SNM was effective in 6 patients (60.0%). Compared with preoperative results, the daily frequency of urination [(20.2±3.8) times vs. (13.2±3.2) times], volume per urination [(119.2±56.7) mL vs. (246.5±59.2) mL], maximum urine flow rate [(8.7±1.5) mL/s vs. (16.5±2.6) mL/s], and residual urine volume [(222.5±55.0) mL vs. (80.8±16.0) mL] were significantly improved, with statistical significance (P<0.05). There were no complications such as bleeding, infection, fever or pain.The 6 patients who had effective outcomes successfully completed phase II surgery, and the fistula was removed.During the follow-up of 1 year, the curative effect was stable, and there were no complications such as electrode displacement, incision infection, or pain in the irritation sites.The residual urine volume of the other 4 unsuccessful patients did not improve significantly, and the electrodes were removed and the vesicostomy tube was retained. [Conclusion] SNM is safe and effective in the treatment of BPH with UAB patients with poor curative effects after TURP.
8.Shaoyaotang Containing Serum Mediates Fas/FasL Pathway to Inhibit Lipopolysaccharide Induced Inflammation and Apoptosis of Caco-2 Cells
Yuting YANG ; Dongsheng WU ; Hui CAO ; Yu ZHANG ; Nianjia XIE ; Bo ZOU ; Daguang CHEN ; Erle LIU ; Yi LU ; Zhaowen LYU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):62-69
ObjectiveTo investigate the effects of different concentrations of Shaoyaotang-containing serum on lipopolysaccharide (LPS)-induced inflammation of human colorectal adenocarcinoma (Caco-2) cells by inhibiting apoptosis via activating the tumor necrosis factor (TNF) receptor superfamily member 6 (Fas)/Fas ligand (FasL) pathway. MethodsCaco-2 cells were allocated into blank, model (LPS, 10 mg·L-1), Shaoyaotang-containing serum (5%, 10%, 15%, 20%), and Fas inhibitor (KR-33493, 20 mmol·L-1) groups. Except the blank group, the other groups were stimulated with 10 mg·L-1 LPS for 24 h for the modeling of inflammation. After successful modeling, the blank, Fas inhibitor, and model groups were treated with blank serum, and the Shaoyaotang-containing serum groups were treated with the serum samples at corresponding concentrations for 24 h. The Fas inhibitor group was subjected to KR-33493 pretreatment for 1 h. Cell proliferation and viability were examined by the cell-counting kit-8 (CCK-8) method. The levels of interleukin (IL)-6, IL-1β, and TNF-α were measured by enzyme-linked immunosorbent assay. Apoptosis was detected by flow cytometry. The protein and mRNA levels of Fas, FasL, cysteinyl aspartate-specific proteinase (Caspase)-3, Caspase-9, B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax) were determined by Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), respectively. ResultsCompared with the blank group, the model group presented a decrease in cell survival rate (P<0.01). Compared with that in the model group, the cell survival rate showed no significant change in the 5% Shaoyaotang-containing serum group but increased in the 10%, 15%, and 20% Shaoyaotang-containing serum groups (P<0.01). Since there was no statistical difference between the 5% Shaoyaotang-containing serum group and the model group, 10%, 15%, and 20% Shaoyaotang-containing sera were selected for the follow-up study. Compared with the blank group, the model group showed risen levels of IL-6, IL-1β, and TNF-α (P<0.01), an increased apoptosis rate (P<0.01), up-regulated protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.01), and down-regulated protein and mRNA levels of Bcl-2 (P<0.01). Compared with the model group, the Fas inhibitor group and the 10%, 15%, and 20% Shaoyaotang-containing serum groups showed declined levels of IL-6, IL-1β, and TNF-α (P<0.01), decreased apoptosis rates (P<0.01), down-regulated protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.05, P<0.01), and up-regulated protein and mRNA levels of Bcl-2 (P<0.05, P<0.01). In addition, the 15% and 20% Shaoyaotang-containing serum groups had lower levels of IL-6, IL-1β, and TNF-α (P<0.05, P<0.01), lower apoptosis rates (P<0.05, P<0.01), lower protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.05, P<0.01), and higher protein and mRNA levels of Bcl-2 (P<0.05, P<0.01) than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can reduce the content of inflammatory factors in Caco-2 cells, down-regulate the protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax, and up-regulate the protein and mRNA levels of Bcl-2 under the intervention of LPS by regulating the Fas/FasL pathway and inhibiting the apoptosis of intestinal epithelial cells in ulcerative colitis.
9.Establishment and Evaluation of Rat Model of Myocardial Ischemia-reperfusion Injury with Phlegm and Blood Stasis Blocking Collaterals Syndrome Based on Metabolomics
Longxiao HU ; Jiabei GAO ; Weihao MA ; Jieming LU ; Yunxiao GAO ; Yue YUAN ; Qiuyan ZHANG ; Xiaoxiao CHEN ; Yali SHI ; Jianxun LIU ; Junguo REN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(12):41-51
ObjectiveTo explore the feasibility, evaluation methods and metabolic differences of high-fat diet(HFD) combined with myocardial ischemia-reperfusion injury(MIRI) to establish a rat model of myocardial ischemia-reperfusion with phlegm and blood stasis blocking collaterals syndrome(PBSBCS). MethodsThirty-two SD rats were randomly divided into the sham operation, HFD, MIRI, and MIRI+HFD groups. Rats in the sham operation and MIRI groups were fed a standard diet(regular chow), while the HFD and MIRI+HFD groups received a HFD for 10 weeks. Rats in the MIRI and MIRI+HFD groups underwent myocardial ischemia-reperfusion surgery, while the sham operation group underwent only thread placement without ligation. Cardiac function was assessed via small-animal echocardiography, including left ventricular ejection fraction(EF), left ventricular fractional shortening(FS), cardiac output(CO), and stroke volume(SV). Serum levels of creatine kinase(CK), CK-MB, triglyceride(TG), total cholesterol(TC), high-density lipoprotein cholesterol(HDL-C), low-density lipoprotein cholesterol(LDL-C), lactate dehydrogenase(LDH), endothelin-1(ET-1), endothelial nitric oxide synthase(eNOS), tumor necrosis factor-α(TNF-α), interleukin-18(IL-18), oxidized LDL(ox-LDL), and cardiac troponin T(cTnT) were measured by biochemical assays and enzyme-linked immunosorbent assay(ELISA). Myocardial histopathology was evaluated via hematoxylin-eosin(HE) staining, while myocardial infarction and no-reflow area were assessed using 2,3,5-triphenyltetrazolium chloride(TTC), Evans blue, and thioflavin staining. Changes in syndrome characteristics[body weight, tongue surface red-green-blue [RGB] values, and pulse amplitude] of PBSBCS were recorded. Serum differential metabolites were analyzed by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS). ResultsCompared with the sham operation group, the HFD and MIRI+HFD groups showed significant increases in body weight(P<0.01), RGB values and pulse amplitude decreased in the HFD, MIRI and MIRI+HFD groups, TC, TG, LDL-C and ox-LDL levels increased in the HFD and MIRI+HFD groups, while HDL-C decreased. Blood perfusion peak time and myocardial no-reflow area increased, serum eNOS level decreased, and CK-MB, LDH, and cTnT activities increased in the HFD, MIRI and MIRI+HFD groups(P<0.05, P<0.01). Whole blood viscosity was increased in the HFD group at medium shear rate, and in the MIRI and MIRI+HFD groups at low, medium and high shear rates(P<0.05, P<0.01). Platelet aggregation rate increased in the MIRI and MIRI+HFD groups, accompanied by elevated ET-1, TNF-α, and IL-18 levels, reduced cardiac function indices, expanded myocardial no-reflow and infarction areas, and increased serum CK, CK-MB, LDH, and cTnT activities(P<0.05, P<0.01). Compared with the MIRI group, the HFD and MIRI+HFD groups showed significant increase in body weight, TC, TG, LDL-C and ox-LDL levels, and significant decrease in HDL-C content(P<0.01). The MIRI+HFD group showed decrease in RGB values and pulse amplitude, and an increase in whole blood viscosity, platelet aggregation, blood perfusion peak time, myocardial no-reflow and infarction areas, elevated ET-1, TNF-α and IL-18 levels, decreased eNOS content, EF and SV, increased serum CK, CK-MB and cTnT activities, and worsened myocardial pathology(P<0.05). Compared with the HFD group, the MIRI+HFD group showed similar aggravated trends(P<0.05, P<0.01). Metabolomics results showed that 34 potential biomarkers involving 13 common metabolic pathways were identified in the MIRI+HFD group compared with the sham operation group. ConclusionThe MIRI group resembles blood stasis syndrome in hemodynamics and myocardial injury, and the HFD group mirrors phlegm-turbidity syndrome in lipid profiles and tongue characteristics. While the MIRI+HFD group aligns with PBSBCS in comprehensive indices, effectively simulating clinical features of coronary heart disease(CHD), which can be used for the evaluation of the pathological mechanism and pharmacodynamics of CHD with PBSBCS.
10.Shaoyaotang Containing Serum Mediates Fas/FasL Pathway to Inhibit Lipopolysaccharide Induced Inflammation and Apoptosis of Caco-2 Cells
Yuting YANG ; Dongsheng WU ; Hui CAO ; Yu ZHANG ; Nianjia XIE ; Bo ZOU ; Daguang CHEN ; Erle LIU ; Yi LU ; Zhaowen LYU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(13):62-69
ObjectiveTo investigate the effects of different concentrations of Shaoyaotang-containing serum on lipopolysaccharide (LPS)-induced inflammation of human colorectal adenocarcinoma (Caco-2) cells by inhibiting apoptosis via activating the tumor necrosis factor (TNF) receptor superfamily member 6 (Fas)/Fas ligand (FasL) pathway. MethodsCaco-2 cells were allocated into blank, model (LPS, 10 mg·L-1), Shaoyaotang-containing serum (5%, 10%, 15%, 20%), and Fas inhibitor (KR-33493, 20 mmol·L-1) groups. Except the blank group, the other groups were stimulated with 10 mg·L-1 LPS for 24 h for the modeling of inflammation. After successful modeling, the blank, Fas inhibitor, and model groups were treated with blank serum, and the Shaoyaotang-containing serum groups were treated with the serum samples at corresponding concentrations for 24 h. The Fas inhibitor group was subjected to KR-33493 pretreatment for 1 h. Cell proliferation and viability were examined by the cell-counting kit-8 (CCK-8) method. The levels of interleukin (IL)-6, IL-1β, and TNF-α were measured by enzyme-linked immunosorbent assay. Apoptosis was detected by flow cytometry. The protein and mRNA levels of Fas, FasL, cysteinyl aspartate-specific proteinase (Caspase)-3, Caspase-9, B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax) were determined by Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), respectively. ResultsCompared with the blank group, the model group presented a decrease in cell survival rate (P<0.01). Compared with that in the model group, the cell survival rate showed no significant change in the 5% Shaoyaotang-containing serum group but increased in the 10%, 15%, and 20% Shaoyaotang-containing serum groups (P<0.01). Since there was no statistical difference between the 5% Shaoyaotang-containing serum group and the model group, 10%, 15%, and 20% Shaoyaotang-containing sera were selected for the follow-up study. Compared with the blank group, the model group showed risen levels of IL-6, IL-1β, and TNF-α (P<0.01), an increased apoptosis rate (P<0.01), up-regulated protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.01), and down-regulated protein and mRNA levels of Bcl-2 (P<0.01). Compared with the model group, the Fas inhibitor group and the 10%, 15%, and 20% Shaoyaotang-containing serum groups showed declined levels of IL-6, IL-1β, and TNF-α (P<0.01), decreased apoptosis rates (P<0.01), down-regulated protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.05, P<0.01), and up-regulated protein and mRNA levels of Bcl-2 (P<0.05, P<0.01). In addition, the 15% and 20% Shaoyaotang-containing serum groups had lower levels of IL-6, IL-1β, and TNF-α (P<0.05, P<0.01), lower apoptosis rates (P<0.05, P<0.01), lower protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax (P<0.05, P<0.01), and higher protein and mRNA levels of Bcl-2 (P<0.05, P<0.01) than the 10% Shaoyaotang-containing serum group. ConclusionThe Shaoyaotang-containing serum can reduce the content of inflammatory factors in Caco-2 cells, down-regulate the protein and mRNA levels of Fas, FasL, Caspase-3, Caspase-9, and Bax, and up-regulate the protein and mRNA levels of Bcl-2 under the intervention of LPS by regulating the Fas/FasL pathway and inhibiting the apoptosis of intestinal epithelial cells in ulcerative colitis.

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