1.Material Basis of Anti-Inflammatory Efficacy and Mechanism of Action of Bushen Tongdu Prescription Based on UPLC-LTQ-Orbitrap-MS and Network Pharmacology
Yan RONG ; Lulu JING ; Hongping HOU ; Huijun WANG ; Lihua CHEN ; Yunxin CHEN ; Liang LI ; Li LIN ; Xiaoqin LUO ; Haiyu ZHAO ; Xiaolu WEI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):152-161
ObjectiveThis paper aims to investigate the material basis of the anti-inflammatory efficacy and mechanism of action of Bushen Tongdu prescription (BSTDP). MethodsThe chemical components of BSTDP and its blood-absorbed components in vivo were systematically identified by using ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap high-resolution mass spectrometry (UPLC-LIT-Orbitrap-MS). Network pharmacology was employed to screen blood-absorbed bioactive components and potential targets of this formula. A protein-protein interaction (PPI) network of core targets was constructed to conduct enrichment analysis. Molecular docking was further utilized to verify the binding affinity between key components and targets. The inflammatory model was established and verified in vivo by using a transgenic zebrafish Tg (mpx: GFP). At three days post-fertilization (3 dpf), larvae of zebrafish were randomly assigned to blank group, model group, positive drug dexamethasone acetate group (75 μmol·L-1), and BSTDP groups with low, medium, and high doses (500, 1 000, and 2 000 mg·L-1). The distribution and quantity of neutrophils in the yolk sac region were observed under a fluorescence microscope. The mRNA expression levels of key genes in the toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor kappa-B (NF-κB) signaling pathway and inflammatory factors including interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) were detected by Real-time quantitative polymerase chain reaction (Real-time PCR). ResultsA total of 120 chemical components were identified in BSTDP, among which 26 original components were confirmed by using serum pharmacochemical methods. A total of 227 common targets linking rheumatoid arthritis (RA) and the blood-absorbed components were screened by network pharmacology. It is suggested that pseudobrucine, vomicine, sinapine, rehmannioside, cinnamyl alcohol glycoside, and methylephedrine exert anti-inflammatory effects by acting on core targets including protein kinase B1 (Akt1), signal transducer and activator of transcription 3 (STAT3), tumor necrosis factor (TNF), TLR4, mitogen-activated protein kinase 14 (MAPK14), and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit α (PIK3CA), thereby modulating multiple signaling pathways such as TLR4 and NF-κB. In vivo verification in zebrafish demonstrates that the maximum tolerable concentration of Bushen Tongdu Formula is 2 000 mg·L-1. Compared to those in the blank group, zebrafish in the model group showed a significantly higher number of neutrophils in the yolk sac region (P<0.01) and rising mRNA levels of TLR4, MyD88, NF-κB, TNF-α, IL-6, and IL-1β (P<0.01). Compared to that in the model group, the number of neutrophils was significantly reduced in BSTDP groups with medium and high doses, as well as the dexamethasone acetate group (P<0.05, P<0.01). There was no statistically significant difference in the low dose group. The mRNA expression levels of TLR4, MyD88, NF-κB, TNF-α, IL-6, and IL-1β were significantly down-regulated (P<0.05, P<0.01). ConclusionThis paper identifies the material basis of the efficacy of BSTDP, demonstrating that the formula can exert an anti-inflammatory effect through the TLR4/MyD88/NF-κB signaling pathway. The results provide scientific experimental evidence for its further clinical application.
2.Historical Evolution and Key Information Research on Classic Formula Puji Xiaoduyin
Lianchao ZHU ; Lyuyuan LIANG ; Jing TANG ; Jialei CAO ; Ziming XU ; Huizhen ZHANG ; Zhidan GUO ; Rongze MA ; Zhengshao ZHANG ; Bingqi WEI ; Xiubo DU ; Bingxiang MA
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):238-247
Puji Xiaoduyin, a specialized formula for the swollen-head epidemic, was recorded in the Catalogue of Ancient Classical Formula (the Second Batch)-Han Medicine, published in September 2023. It had been inherited and developed by medical experts of successive generations and passed down to this day. This paper sorted out the historical evolution of this formula using bibliometric methods. It also comprehensively analyzed key information on the formula name, historical origin, drug dosage, herb origin, processing methods, decocting methods, function, and clinical applications. Additionally, this paper analyzed the application of this formula in both modern and ancient times. Results showed that the formula was first recorded as "Puji Xiaodu Yinzi" in LI Dongyuan's Proven Formulas written by LI Gao from the Jin dynasty. The medicinal composition and dosage were: Scutellariae Radix and Coptidis Rhizoma (20.65 g each), Ginseng Radix et Rhizoma 12.39 g, Scrophulariae Radix, Citri Reticulatae Pericarpium, and Glycyrrhizae Radix et Rhizoma (8.26 g each), Forsythiae Fructus, Arctii Fructus, Isatidis Radix, and Lasiosphaera Calvatia (4.13 g each), Bombyx Batryticatus and Cimicifugae Rhizoma (2.891 g each), Bupleuri Radix and Platycodonis Radix (8.26 g each). These medicines were grounded to fine powder. One dose, including 20.65 g of the powder, was mixed with 600 mL of water and decocted to 300 mL. After abandoning slag, the medicine should be taken warm frequently. In the formula, Bombyx Batryticatus is stir-fired. With the effect of dispersing wind and clearing heat, removing stagnation and dissipating mass, the formula is specialized in swollen-head epidemic, pestilence, red and swelling head, face, and neck, dry mouth and tongue, as well as other diseases resulting from toxic heat stagnated in the upper jiao. The formula is widely used in treating diseases involving the respiratory, dermal, ophthalmologic, otolaryngologic, and nervous systems. The formula is most frequently used for respiratory diseases, with a wide range of symptoms including parotitis/mumps (66 times), followed by tonsillitis (28 times). In conclusion, the broadly applied formula has accurate efficacy and great development value.
3.Material Basis of Anti-Inflammatory Efficacy and Mechanism of Action of Bushen Tongdu Prescription Based on UPLC-LTQ-Orbitrap-MS and Network Pharmacology
Yan RONG ; Lulu JING ; Hongping HOU ; Huijun WANG ; Lihua CHEN ; Yunxin CHEN ; Liang LI ; Li LIN ; Xiaoqin LUO ; Haiyu ZHAO ; Xiaolu WEI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):152-161
ObjectiveThis paper aims to investigate the material basis of the anti-inflammatory efficacy and mechanism of action of Bushen Tongdu prescription (BSTDP). MethodsThe chemical components of BSTDP and its blood-absorbed components in vivo were systematically identified by using ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap high-resolution mass spectrometry (UPLC-LIT-Orbitrap-MS). Network pharmacology was employed to screen blood-absorbed bioactive components and potential targets of this formula. A protein-protein interaction (PPI) network of core targets was constructed to conduct enrichment analysis. Molecular docking was further utilized to verify the binding affinity between key components and targets. The inflammatory model was established and verified in vivo by using a transgenic zebrafish Tg (mpx: GFP). At three days post-fertilization (3 dpf), larvae of zebrafish were randomly assigned to blank group, model group, positive drug dexamethasone acetate group (75 μmol·L-1), and BSTDP groups with low, medium, and high doses (500, 1 000, and 2 000 mg·L-1). The distribution and quantity of neutrophils in the yolk sac region were observed under a fluorescence microscope. The mRNA expression levels of key genes in the toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor kappa-B (NF-κB) signaling pathway and inflammatory factors including interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) were detected by Real-time quantitative polymerase chain reaction (Real-time PCR). ResultsA total of 120 chemical components were identified in BSTDP, among which 26 original components were confirmed by using serum pharmacochemical methods. A total of 227 common targets linking rheumatoid arthritis (RA) and the blood-absorbed components were screened by network pharmacology. It is suggested that pseudobrucine, vomicine, sinapine, rehmannioside, cinnamyl alcohol glycoside, and methylephedrine exert anti-inflammatory effects by acting on core targets including protein kinase B1 (Akt1), signal transducer and activator of transcription 3 (STAT3), tumor necrosis factor (TNF), TLR4, mitogen-activated protein kinase 14 (MAPK14), and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit α (PIK3CA), thereby modulating multiple signaling pathways such as TLR4 and NF-κB. In vivo verification in zebrafish demonstrates that the maximum tolerable concentration of Bushen Tongdu Formula is 2 000 mg·L-1. Compared to those in the blank group, zebrafish in the model group showed a significantly higher number of neutrophils in the yolk sac region (P<0.01) and rising mRNA levels of TLR4, MyD88, NF-κB, TNF-α, IL-6, and IL-1β (P<0.01). Compared to that in the model group, the number of neutrophils was significantly reduced in BSTDP groups with medium and high doses, as well as the dexamethasone acetate group (P<0.05, P<0.01). There was no statistically significant difference in the low dose group. The mRNA expression levels of TLR4, MyD88, NF-κB, TNF-α, IL-6, and IL-1β were significantly down-regulated (P<0.05, P<0.01). ConclusionThis paper identifies the material basis of the efficacy of BSTDP, demonstrating that the formula can exert an anti-inflammatory effect through the TLR4/MyD88/NF-κB signaling pathway. The results provide scientific experimental evidence for its further clinical application.
4.Historical Evolution and Key Information Research on Classic Formula Puji Xiaoduyin
Lianchao ZHU ; Lyuyuan LIANG ; Jing TANG ; Jialei CAO ; Ziming XU ; Huizhen ZHANG ; Zhidan GUO ; Rongze MA ; Zhengshao ZHANG ; Bingqi WEI ; Xiubo DU ; Bingxiang MA
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):238-247
Puji Xiaoduyin, a specialized formula for the swollen-head epidemic, was recorded in the Catalogue of Ancient Classical Formula (the Second Batch)-Han Medicine, published in September 2023. It had been inherited and developed by medical experts of successive generations and passed down to this day. This paper sorted out the historical evolution of this formula using bibliometric methods. It also comprehensively analyzed key information on the formula name, historical origin, drug dosage, herb origin, processing methods, decocting methods, function, and clinical applications. Additionally, this paper analyzed the application of this formula in both modern and ancient times. Results showed that the formula was first recorded as "Puji Xiaodu Yinzi" in LI Dongyuan's Proven Formulas written by LI Gao from the Jin dynasty. The medicinal composition and dosage were: Scutellariae Radix and Coptidis Rhizoma (20.65 g each), Ginseng Radix et Rhizoma 12.39 g, Scrophulariae Radix, Citri Reticulatae Pericarpium, and Glycyrrhizae Radix et Rhizoma (8.26 g each), Forsythiae Fructus, Arctii Fructus, Isatidis Radix, and Lasiosphaera Calvatia (4.13 g each), Bombyx Batryticatus and Cimicifugae Rhizoma (2.891 g each), Bupleuri Radix and Platycodonis Radix (8.26 g each). These medicines were grounded to fine powder. One dose, including 20.65 g of the powder, was mixed with 600 mL of water and decocted to 300 mL. After abandoning slag, the medicine should be taken warm frequently. In the formula, Bombyx Batryticatus is stir-fired. With the effect of dispersing wind and clearing heat, removing stagnation and dissipating mass, the formula is specialized in swollen-head epidemic, pestilence, red and swelling head, face, and neck, dry mouth and tongue, as well as other diseases resulting from toxic heat stagnated in the upper jiao. The formula is widely used in treating diseases involving the respiratory, dermal, ophthalmologic, otolaryngologic, and nervous systems. The formula is most frequently used for respiratory diseases, with a wide range of symptoms including parotitis/mumps (66 times), followed by tonsillitis (28 times). In conclusion, the broadly applied formula has accurate efficacy and great development value.
5.A prediction model for sarcopenia in postmenopausal women:information analysis based on the China Health and Retirement Longitudinal Study database
Guangzheng LI ; Wei LI ; Bochun ZHANG ; Haoqin DING ; Zhongqi ZHOU ; Gang LI ; Xuezhen LIANG
Chinese Journal of Tissue Engineering Research 2026;30(4):849-857
BACKGROUND:Sarcopenia is an age-related systemic skeletal muscle disease,which is associated with a variety of adverse outcomes such as falls,functional decline,frailty,and death.Postmenopausal women are one of the high-risk groups for sarcopenia.OBJECTIVE:To develop a predictive model for assessing the risk of sarcopenia in Chinese postmenopausal women based on high-quality database.METHODS:Data for this study were derived from 2 370 postmenopausal women from the China Health and Retirement Longitudinal Study(CHARLS),and sarcopenia was assessed using the Asian Working Group on Sarcopenia 2019(AWGS2019)recommended metrics.The study cohort was randomized into a training set(70%)and a validation set(30%).Risk factors for sarcopenia in postmenopausal women were screened using the least absolute shrinkage and selection operator,ten-fold cross-validation,and logistic regression.Nomogram predicting the risk of sarcopenia in postmenopausal women was constructed based on the risk factors,and the model efficacy was evaluated by the receiver operating characteristic curve and area under the curve(AUC),calibration curve,and decision curve analysis.RESULTS AND CONCLUSION:The prevalence of sarcopenia in this study was 23.50%and age,place of residence,sleep quality,cognitive function,depression,and the number of chronic diseases were selected as predictors of sarcopenia in postmenopausal women.The nomogram model showed good discrimination between the training and validation sets,with an AUC value of 0.751(95%confidence interval=0.724-0.778,P<0.001),a specificity of 72.2%,and a sensitivity of 63.2%in the training set,and an AUC value of 0.763(95%confidence interval=0.721-0.805,P<0.001),with a specificity of 69.6%and a sensitivity of 70.8%.The calibration curve showed a relatively significant agreement between the nomogram model and the actual observations,and the decision curve analysis demonstrated broad and good clinical utility.To conclude,the nomogram to assess the risk of sarcopenia constructed based on age,place of residence,sleep quality,cognitive function,depression,and number of chronic diseases,provides an effective tool for identifying and eliminating risk factors for sarcopenia in Chinese postmenopausal women,and helps to reduce the incidence of sarcopenia.
6.Ancient and Modern Documentation of Classic Formula Sangjuyin
Xiaofang WANG ; Lyuyuan LIANG ; Jialei CAO ; Ziming XU ; Wangju ZHOU ; Yiping WANG ; Yujie CHANG ; Ruiting SU ; Yihan LI ; Jingwen LI ; Bingqi WEI ; Bingxiang MA ; Wenli SHI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):200-207
Sangjuyin, as a pungent and cooling agent with precise therapeutic effect, is a classic pungent formula for cooling relief of the epidermis, which is highly respected by medical practitioners. This formula is from the Wenbing Tiaobian written by WU Jutong in the Qing dynasty, on the basis of which subsequent medical practitioners have made additions and subtractions to apply it. The authors used the bibliometric method to systematically organize the medical books from the Qing dynasty and the Republic of China and modern literature to analyze the composition, concoction, decoction, efficacy, and previous and modern application of Sangjuyin. After examination, the drug base of this formula is basically clear. Armeniacae Semen Amarum is the dried mature seeds of Armeniaca vulgaris, family Rosaceae. Forsythiae Fructus is the dried fruit of Forsythia suspensa, family Mulleinaceae. Menthae Haplocalycis Herba is the dried above-ground part of Mentha haplocalyx, family Labiatae. Mori Folium is the dried leaves of Morus alba, family Moraceae. Chrysanthemi Flos is the dried head of Chrysanthemum morifolium, family Asteraceae. Platycodonis Radix is the dried root of Eryngium grandiflorum, family Eryngium. Glycyrrhizae Radix et Rhizoma is the dried root and rhizome of Glycyrrhiza uralensis of the Leguminosae family, and Phragmitis Rhizoma is the fresh or dried rhizome of Phragmites communis of the Gramineae family. It is recommended that the eight drugs be used in raw form as medicine. The dosage and method of decoction were converted into a modern single dosage of 7.46 g Armeniacae Semen Amarum, 5.60 g Forsythiae Fructus, 2.98 g Menthae Haplocalycis Herba, 9.33 g Mori Folium, 3.73 g Chrysanthemi Flos, 7.46 g Platycodonis Radix, 2.98 g Glycyrrhizae Radix et Rhizoma, and 11.19 g Phragmitis Rhizoma, with 400 mL water added, and the solution was boiled to obtain 200 mL, taken twice a day. Sangjuyin has the efficacy of dispersing wind and clearing heat, promoting lung and relieving cough, and it is used for treating the initial onset of wind-warmth and the evidence of evil spirits in the lungs and collaterals. Modern research has shown that Sangjuyin is often used in the treatment of cough, pneumonia, rhinitis, and other respiratory diseases, and the results of this study provide a reference for the later development of Sangjuyin.
7.Improvement effects and mechanism of Achyranthes bidentata total saponins extract on vascular endothelial dysfunction in spontaneously hypertensive rats
Ruifeng LIANG ; Wenjing GE ; Xiaobo KOU ; Ping TIAN ; Hongzhi AN ; Zheng WEI ; Mingli ZHANG
China Pharmacy 2026;37(3):331-337
OBJECTIVE To investigate the improvement effects and mechanism of Achyranthes bidentata total saponins (ABS) extract on vascular endothelial dysfunction in spontaneously hypertensive rat (SHR) based on cytochrome P450 4A (CYP4A)/20-hydroxyeicosatetetraenoic acid (20-HETE)/G protein-coupled receptor 75 (GPR75) axis. METHODS Ten Wistar- Kyoto rats were taken as the normal control group. Forty SHR were first stratified by systolic blood pressure and then, within each stratum, randomly assigned using a random-number table to the model group (MOD group), captopril positive control group (CAP group, 10 mg/kg), ABS low- and high-dose extract groups (ABS-L group, ABS-H group, 60 and 120 mg/kg), with 10 rats in each group. Animals in each group were given the corresponding drug or equal volume of pure water by gavage, once a day, for 28 consecutive days. After the last administration, systolic blood pressure of rats was measured. The levels of vasoactive substances, inflammatory factors and oxidative stress indicators in serum were measured. The pathological changes of rat thoracic aorta were observed. The level of reactive oxygen species (ROS) in aortic tissue was analyzed. The expressions of endothelial nitric oxide synthase (eNOS), CYP4A, GPR75, nuclear factor-κB p65 (NF-κB p65), phosphorylated NF-κB p65, p22phox, and reduced nicotinamide adenine dinucleotide phosphate oxidase 4(NOX4) in thoracic aorta tissue were detected. RESULTS After 28 d of treatment, compared with MOD group, the systolic blood pressure of rats in the ABS-L and ABS-H groups decreased significantly. The levels of 20-HETE, angiotensin Ⅱ, interleukin-1β, interleukin-6, tumor necrosis factor-α, intercellular cell adhesion molecule-1 and malondialdehyde in serum were significantly reduced (P<0.05 or P<0.01), while the levels of nitric oxide, superoxide dismutase, glutathione peroxidase and catalase were significantly increased (P<0.05 or P<0.01). Intimal damage of thoracic aorta was reduced, and endothelial cell morphology was improved. The expressions of ROS, CYP4A, GPR75, p22phox, NOX4 and the phosphorylation level of NF-κB p65 protein in thoracic aorta were down-regulated or reduced (P<0.05 or P<0.01), while the expression of eNOS was up-regulated (P<0.05 or P<0.01). CONCLUSIONS ABS extract may alleviate the inflammatory response and oxidative stress in SHR effectively by down-regulating the expression of CYP4A, reducing the production of 20-HETE, inhibiting the activation of GPR75, and subsequently suppressing the activation of downstream NF-κB and NOX4, thereby improving hypertension-related vascular endothelial dysfunction.
8.Ancient and Modern Application and Key Information Analysis of Classic Formula Erchentang
Qing TANG ; Lyuyuan LIANG ; Jialei CAO ; Lan LIU ; Hejia WAN ; Chengxin LUO ; Bingqi WEI ; Yamin KONG ; Bingxiang MA ; Wenli SHI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(1):262-270
Erchentang is a classic formula widely used by medical practitioners throughout history. In this paper,ancient and modern literature of Erchentang were collected, and bibliometrics was employed to analyze its historic evolution,prescription meaning,herbs origin, processing method,preparation methods, and clinical application. A total of 84 pieces of data were collected, and 58 pieces of data involving 53 ancient medical Chinese books were screened, sorted, and processed. Combined with research of modern scholars,the research has found that the Erchentang originated from the Taiping Huimin Huiye Shijie Fang compiled by the Imperial Medical Bureau of the Song Dynasty. The basic information about the origin of the drugs is quite clear. Pinelliae rhizoma in the formula is the dried tuber of Pinellia ternata. Citri exocarpium rubrum is the dried mature peel of Citrus reticulata and its cultivated varieties, with the inner white membrane removed. Poria is the whitest dry sclerotia of Poria cocos; Glycyrrhizae radix et rhizoma is the dried root and rhizome of the Glycyrrhiza uralensis. The dosage is 5.70 g Pinelliae rhizome and Citri exocarpium rubrum, 3.43 g Poria, and 1.69 g Glycyrrhizae radix et rhizoma praeparata cum melle. During the decoction process, the above-mentioned herbs should be chopped, with 300 mL water, 7 g ginger in thick slices, and 2 g Mume fructus added, and it was then simmered together to 180 mL. After removing the medicinal residue, it can be taken warmly. Erchentang has the effect of drying dampness and resolving phlegm, regulating Qi and harmonizing the middle. It can be used in treating the syndrome of phlegm and dampness,as well as symptoms such as frequent cough,white phlegm,fullness in chest and diaphragm,nausea and vomiting,limb drowsiness,anorexia,dizziness,palpitations,white and greasy tongue coating, and slippery pulse. The above results provide reference for future research and development of Erchentang.
9.The effect of body mass index and inferior pulmonary ligament division on the residual lung expansion after right upper lobectomy: A retrospective cohort study in a single center
Guang MU ; Wenhao ZHANG ; Hongchang WANG ; Yan GU ; Chenghao FU ; Wentao XUE ; Shiyuan XIE ; Tong WANG ; Ke WEI ; Yang XIA ; Liang CHEN ; Jun WANG
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(02):261-266
Objective To analyze the effect of releasing the lower pulmonary ligament on right residual lung expansion after right upper lobe resection under different body mass index (BMI) levels. Methods The clinical data of patients who underwent thoracoscopic right upper lobe resection in the First Affiliated Hospital with Nanjing Medical University from 2021 to 2022 were retrospectively analyzed. Patients were divided into a group A (17 kg/m2<BMI≤23 kg/m2), a group B (23 kg/m2<BMI≤29 kg/m2) and a group C (BMI>29 kg/m2) according to BMI. The presence of residual cavity was judged by chest X-ray at 7-10 days after operation, the degree of compensation change of the right main bronchus angle was measured, and the changes in lung volume were determined by CT three-dimensional reconstruction. Results A total of 157 patients who underwent thoracoscopic right upper lobe resection were included, including 71 males and 86 females, with an average age of (59.7±11.2) years. There were 50 patients in the group A, 75 patients in the group B, and 32 patients in the group C. In the group A, compared with those without releasing the lower pulmonary ligament, patients with releasing had a lower incidence of postoperative residual cavity (P=0.016), greater changes in bronchus angle (P<0.001), and smaller changes in lung volume (P<0.001). In the group B and C, there was no significant effect of releasing the lower pulmonary ligament on postoperative residual cavity, bronchus angle, and lung volume changes (P>0.05). Conclusion For patients with thin and long body shape and low BMI, releasing the lower pulmonary ligament is helpful to promote the expansion of the residual lung after right upper lobe resection and reduce the occurrence of postoperative residual cavity in patients.
10.Experimental study on interferon-stimulated gene myxovirus resistance protein 2-mediated restriction of tumor cell sensitivity to reovirus oncolysis
LIANG Dan1,2 ; YANG Zailing1,2 ; YU Jiani1,2 ; LI Xinlan2,3 ; SHEN Tao1,2 ; SUN Yongshun2,4 ; WEI Yongzhu2,4 ; ZHAO Xing1,2
Chinese Journal of Cancer Biotherapy 2026;33(2):132-139
[摘 要] 目的:探讨干扰素刺激基因家族成员黏病毒抵抗蛋白2(MX2)在调控肿瘤细胞对呼肠孤病毒(Reo)溶瘤敏感性中的作用及其机制。方法:选取4株具有不同耐药特征的人源肿瘤细胞,通过CCK-8法评估其对Reo的溶瘤敏感性;通过转录组测序筛选出差异表达基因MX2,qPCR法及WB法验证MX2在4株人源肿瘤细胞中的表达;使用siRNA敲低溶瘤低敏感的COC1/DDP细胞中的MX2基因。在细胞感染Reo病毒后,通过CCK-8法检测细胞存活率;qPCR法检测细胞中Reo病毒S1基因表达;免疫荧光法检测细胞内Reo病毒蛋白的积累;半数组织培养感染剂量(TCID50)法测定病毒滴度;流式细胞术分别检测细胞内Reo病毒dsRNA、活性氧(ROS)水平以及细胞凋亡率;透射电镜观察细胞内质网形态变化并采用WB法检测内质网应激相关蛋白(JNK、p-JNK、eIF2α、p-eIF2α、CHOP、PERK)的表达。结果:在4株肿瘤细胞中,SKOV3细胞对Reo溶瘤作用高度敏感,而COC1/DDP、HuH-7SRB及SNU-398细胞均为溶瘤低敏感性。转录组测序结果显示,MX2在溶瘤低敏感肿瘤细胞中的表达水平显著高于溶瘤高敏感细胞(P < 0.01);在溶瘤低敏感性的COC1/DDP细胞中,敲低MX2显著促进Reo病毒复制、诱导细胞凋亡增加,并升高细胞内活性氧水平(均P < 0.001)。透射电镜观察显示,敲低MX2的COC1/DDP细胞感染Reo病毒后出现内质网肿胀、扩张及断裂等典型内质网应激超微结构改变。WB结果显示,内质网应激关键标志物eIF2α/p-eIF2α、PERK、CHOP及凋亡相关调节蛋白JNK/p-JNK的表达均显著上调(P < 0.05或P < 0.01)。结论:肿瘤细胞对Reo的溶瘤敏感性与其细胞内MX2表达水平密切相关。敲低MX2可显著增强Reo在细胞内的复制,进而促进ROS积累,触发内质网应激并促进凋亡。病毒复制增加与细胞凋亡激活的双重作用,最终协同增强Reo的溶瘤作用。

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