1.Huangqi Jianzhongtang Regulates Polarization of Macrophages M1/M2 and Improves Fat Consumption in Cancer Cachexia Mice
Zhiyan FANG ; Haiyan ZHU ; Wenying HUAI ; Cong HUANG ; Ruocong YANG ; Haiyan YU ; Tiane ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):61-69
ObjectiveTo investigate the effects of Huangqi Jianzhongtang (HQJZ) on macrophage polarization and fat consumption in cancer cachexia (CC) mice. MethodsUltra-performance liquid chromatography-quadrupole/electrostatic field Orbitrap high-resolution mass spectrometry (UPLC-Q-Orbitrap HRMS) was used to control the quality of HQJZ. (1) In vitro experiment: HQJZ-containing serum was prepared, and the optimal concentration was determined by cytotoxicity assay. Mouse monocyte-derived macrophages (RAW264.7) were cultured and randomly divided into six groups, including a blank group, a classically activated macrophages (M1) group, an alternatively activated macrophages (M2) group, a HQJZ + blank group, a HQJZ+M1 group, and a HQJZ + M2 group. The relative expression of macrophage marker genes CD86, inducible nitric oxide synthase (iNOS), CD206, and arginase-1 (Arg1) was detected by real-time quantitative polymerase chain reaction (Real-time PCR ). (2) In vivo experiment: Thirty-two BALB/c mice were randomly divided into a control group, a model group, a medroxyprogesterone acetate (MPA) group, and a HQJZ group. Except for the control group, the other mice were injected with CT-26 colon cancer cells to establish a CC model. Mice in the MPA and HQJZ groups were given MPA (0.13 g·kg-1·d-1) or HQJZ (13.13 g·kg-1·d-1) by gavage, respectively, while mice in the control and model groups were given an equal volume of saline by gavage, with interventions continued for 10 d. Real-time PCR was used to detect the expression of macrophage markers (iNOS, Arg1, CD86, CD206) and fat browning-related genes uncoupling protein 1 (UCP1) and peroxisome proliferator-activated receptor γ (PPARγ) in epididymal adipose tissue. Western blot (WB) was used to detect protein expression levels of UCP1 and PPARγ. Micro-computed tomography (micro-CT) was used to measure residual fat volume, and hematoxylin-eosin (HE) staining was used to assess fat browning and calculate pathological scores. ResultsIn vitro, the dominant effective concentration of HQJZ-containing serum was 12.5%. Real-time PCR results showed that, compared with the blank group, Arg1 expression decreased in the HQJZ+blank group (P<0.05), CD206 showed a downward trend without statistical significance, while iNOS and CD86 expression were significantly increased (P<0.05). Compared with the M1 group, Arg1 and CD206 expression decreased in the HQJZ+M1 group (P<0.05). Compared with the M2 group, CD206 expression decreased in the HQJZ+M2 group (P<0.05), CD86 expression increased significantly (P<0.01). In vivo, Real-time PCR results showed that, compared with the control group, CD86 and CD206 expression levels were significantly increased in the model group (P<0.01). Compared with the model group, CD206 expression in the MPA group was significantly decreased (P<0.01). In the HQJZ group, CD206 was significantly decreased (P<0.01). WB results showed that, compared with the model group, protein expression of UCP1 and PPARγ was significantly reduced in the HQJZ group (P<0.05, P<0.01). micro-CT results showed that the total white fat volume in the HQJZ group was greater than that in the model group (P<0.05). HE staining results showed that pathological scores in the HQJZ group were lower than those in the model group (P<0.05). ConclusionHQJZ may inhibit white adipose tissue browning by promoting macrophage M1 polarization and suppressing M2 polarization, thereby delaying fat consumption in CC mice.
2.Mediating role of optimism between mindfulness level and subjective well-being in patients after lung transplantation
Liqin SONG ; Ying ZHOU ; Danxia HUANG ; Jing CHENG ; Chunqin LIU ; Qing LUO ; Yimeng LIU ; Xinyang HU ; Wenying TAN
Chinese Journal of Modern Nursing 2025;31(5):603-609
Objective:To explore the relationship between mindfulness, optimism, and subjective well-being in post-lung transplantation patients, so as to provide a basis for nursing staff to improve patients' subjective well-being.Methods:This study was a cross-sectional survey. Convenience sampling was used to select 205 post-lung transplantation patients admitted to the First Affiliated Hospital of Guangzhou Medical University from October 2022 to November 2023 for the study. General Information Questionnaire, World Health Organization-Five Well-Being Scale, Mindful Attention Awareness Scale (MAAS) and Life Orientation Test-R (LOT-R) were used to investigate the study participants. Pearson correlation was used to analyze the correlation between subjective well-being and mindfulness and optimism in patients after lung transplantation. AMOS 24.0 software was used to construct a mediating model to analyze the path relationship between mindfulness, optimism and subjective well-being.Results:A total of 205 questionnaires were distributed and 202 valid questionnaires were recovered, with an effective recovery rate of 98.54% (202/205). In 202 patients after lung transplantation, the WHO-5 Well-Being Scale total score, MAAS total score, and LOT-R score were (16.31±4.73), (56.75±9.44), and (18.49±3.85), respectively. Lung transplantation patients' subjective well-being was positively correlated with mindfulness and optimism ( r=0.570, 0.600, both P<0.01). Optimism partially mediated the relationship between mindfulness and subjective well-being, with an effect value of 0.290 and an effect proportion of 52.35% (0.290/0.554) . Conclusions:Mindfulness and optimism are both positively correlated with subjective well-being in post-lung transplantation patients, and mindfulness could also influence subjective well-being through the mediating effect of optimism. Healthcare professionals should fully explore and cultivate positive psychological resources, such as mindfulness, in lung transplant patients, by increasing optimism as the target of intervention, which in turn improves patients' subjective well-being.
3.Diagnostic value of fetal echocardiography for ventricular septal defect and reasons for missed diagnosis and misdiagnosis
Wenying YUAN ; Qichang ZHOU ; Lin ZHANG ; Xinglan HUANG ; Shuizhen LIU ; Ling HUANG ; Fang LIU
Chinese Journal of Perinatal Medicine 2025;28(7):593-597
Objective:To explore the diagnostic value of fetal echocardiography (ECHO) for ventricular septal defect (VSD) and the causes of missed diagnosis and misdiagnosis by analyzing the standard of ECHO sections and image quality.Methods:A retrospective analysis was conducted on 94 VSD fetuses diagnosed by ECHO at Xiangtan Maternal and Child Health Care Hospital from January 2022 to June 2024. Clinical data including the anatomic subtypes, associated anomalies, chromosomal karyotype results, pregnancy outcomes, and neonatal ECHO results were collected and analyzed. Besides, by reviewing the ultrasound images from the institution's ultrasound workstation, a descriptive analysis was conducted on the ECHO section criteria and image quality in the missed and misdiagnosed cases.Results:Among 10 984 fetuses undergoing ECHO, a total of 94 cases of fetal VSD were detected (0.7%), including 45 (48%) isolated VSD and 49 (52%) non-isolated VSD cases. Anatomic distribution revealed perimembranous type predominance (67, 71%), followed by muscular (24, 26%), subarterial types (2, 2%), and perimembranous+muscular type (1, 1%). Among 29 missed diagnosis cases (fetal ECHO-negative but neonatal ECHO-confirmed VSDs), all were isolated VSD, muscular types accounted for 19 (66%), perimembranous types for 8 (27%), and subarterial types for 2 (7%). The causes of missed diagnosis include: technical limitation in 14 cases (48%), restricted cardiac acoustic window in 11 cases (38%), and operator-dependent factors in four cases (14%). All 18 misdiagnosed cases occurred in isolated VSD, with 15 cases of perimembranous and three cases of muscular types. The causes of misdiagnosis include: nine cases due to non-perpendicular beam alignment to perimembranous septum, six cases due to excessively high color flow gain,and three cases due to the misinterpretation of the right ventricular outflow tract flow above the aortic valve as VSD septal perforation blood flow. The sensitivity and specificity of fetal ECHO for diagnosing VSD were 76.42% and 99.83%. Among the 94 cases of fetal VSD, 38 cases (40%) underwent chromosomal karyotype analysis and chromosomal microarray analysis, which identified chromosomal karyotype abnormalities or pathogenic gene variants in 4/16 non-isolated VSDs (all perimembranous), while no anomalies were detected in 22 isolated VSDs. Pregnancy outcomes showed 31 terminations due to associated structural or chromosomal abnormalities versus 63 live births. In the live births, who underwent echocardiography in the neonatal period, 19 cases (30%) showed no VSD and were considered to have undergone spontaneous in utero closure, and persistent defects were found in 44 (70%).Conclusions:Fetal ECHO demonstrates high sensitivity and specificity in the diagnosis of fetal VSD. Critical quality control measures include individualized parameter adjustment and systematic multiplanar continuous scanning to minimize diagnostic errors. Isolated small and medium-sized muscular and perimembranous VSD usually have a good prognosis.
4.Cross sectional and cross lagged network analyses of Internet addiction among university students
GOU Hao, HUANG Wenying, SUN Qunqun, HU Chang, ZHANG Wen, XIANG Luyao, SONG Chao
Chinese Journal of School Health 2025;46(9):1287-1291
Objective:
To understand the dynamic temporal evolution pathways of Internet addiction among university students and to identify the core driving nodes, so as to provide theoretical evidences for the precise implementation of targeted interventions.
Methods:
Using a convenient cluster sampling method, a total of 1 066 full time freshmen and sophomores were recruited from three universities in Guizhou, Jiangxi, and Guangdong Provinces for a follow up survey (T1:January-March 2024; T2:January-March 2025). The Revised Chen Internet Addiction Scale (CIAS-R) was employed to assess the status of Internet addiction among university students, and cross sectional as well as cross lagged panel network models were constructed to analyze Internet addiction and its multidimensional influencing factors.
Results:
The T1 network comprised 19 nodes and 114 non zero edges, while the T2 network comprised 19 nodes and 126 non zero edges. Cross sectional network analysis revealed the strongest association between "insufficient sleep" and "daytime fatigue"; the core nodes were "first thought upon waking for going online" and "feeling low after disconnection" (characteristics of psychological dependence) at T1, while the core nodes shifted to "impaired health" and "excitement when online" (characteristics of functional impairment and addictive psychodynamic features) at T2. Cross lagged network analysis further indicated that "reduced leisure" directly predicted "sleep compression", and a bidirectional relationship was observed between "needing more time to achieve satisfaction" and "academic decline".
Conclusions
Internet addiction among university students exhibits dynamic evolutionary characteristics. Stage specific targeted interventions focusing on core driving nodes are needed, integrating behavioral regulation and academic support to break the vicious cycle and enhancing the ability to cope with real life demands.
5.Effects of acupoint catgut embedding on gut microbiota and fecal short-chain fatty acids in Parkinson's disease patients with constipation.
Xiaomei ZHANG ; Jie JIANG ; Wenying LI ; Juping CHEN ; Yin HUANG ; Wei REN ; Yafang SONG ; Jianhua SUN
Chinese Acupuncture & Moxibustion 2025;45(11):1533-1540
OBJECTIVE:
To observe the effects of acupoint catgut embedding (ACE) on gut microbiota and fecal short-chain fatty acids (SCFAs) levels in patients with Parkinson's disease (PD) with constipation.
METHODS:
A total of 80 PD patients with constipation were randomly divided into an observation group and a control group, 40 cases in each group. Additionally, 40 healthy individuals were recruited as a healthy control group. The control group received conventional Western medical treatment for PD combined with polyethylene glycol (PEG), once daily for eight weeks. The observation group received additional ACE treatment at bilateral Tianshu (ST25), Zusanli (ST36), and Shangjuxu (ST37), once every two weeks for eight weeks. The healthy control group received no intervention. The spontaneous bowel movements (SBMs) per week and patient assessment of constipation quality of life (PAC-QOL) scores were assessed at baseline and after treatment in the two groups. Fecal samples were collected at the end of treatment for the observation and the control groups and at baseline for the healthy control group. Gut microbiota composition and diversity were analyzed using 16S rRNA method, and SCFA levels were measured using high-performance liquid chromatography (HPLC).
RESULTS:
Compared before treatment, the observation group showed a significant increase in SBMs (P<0.01), and PAC-QOL scores including physical discomfort, psychosocial discomfort, worry and concern, and total score were significantly reduced (P<0.01) after treatment; the control group also showed a reduction in PAC-QOL total score after treatment (P<0.01). After treatment, the observation group had significantly more SBMs (P<0.01), and lower PAC-QOL physical discomfort, psychosocial discomfort, worry and concern scores, and total score (P<0.01), and higher PAC-QOL satisfaction score (P<0.01) than the control group. Compared with the healthy control group, the control group showed decreased Chao1 and Ace indices (P<0.01). Compared with the healthy control group, the relative abundance of Prevotella and Roseburia was increased (P<0.05), while that of Enterobacter and Ruminococcus torques (six species in total) was decreased (P<0.05) in the control group. Compared with the control group, the observation group had increased relative abundance of Dialister, Parabacteroides, and Ruminococcus torques (P<0.05), and decreased relative abundance of Prevotella and Eubacterium ruminantium (P<0.05). Compared with the healthy control group, the control group had increased fecal SCFA levels (P<0.05); compared with the control group, the observation group had reduced fecal SCFA levels (P<0.05). Compared with the healthy control group, acetic acid, propionic acid, and butyric acid levels were elevated in the control group (P<0.05); compared with the control group, acetic acid, propionic acid, and butyric acid levels were decreased in the observation group (P<0.05).
CONCLUSION
ACE could increase spontaneous bowel movements and improve the quality of life in PD patients with constipation, which may be related to the regulation of gut microbiota composition and SCFA levels.
Humans
;
Constipation/metabolism*
;
Male
;
Gastrointestinal Microbiome
;
Acupuncture Points
;
Female
;
Middle Aged
;
Parkinson Disease/complications*
;
Aged
;
Fatty Acids, Volatile/metabolism*
;
Catgut
;
Feces/microbiology*
;
Acupuncture Therapy
;
Quality of Life
;
Adult
6.Mediating role of optimism between mindfulness level and subjective well-being in patients after lung transplantation
Liqin SONG ; Ying ZHOU ; Danxia HUANG ; Jing CHENG ; Chunqin LIU ; Qing LUO ; Yimeng LIU ; Xinyang HU ; Wenying TAN
Chinese Journal of Modern Nursing 2025;31(5):603-609
Objective:To explore the relationship between mindfulness, optimism, and subjective well-being in post-lung transplantation patients, so as to provide a basis for nursing staff to improve patients' subjective well-being.Methods:This study was a cross-sectional survey. Convenience sampling was used to select 205 post-lung transplantation patients admitted to the First Affiliated Hospital of Guangzhou Medical University from October 2022 to November 2023 for the study. General Information Questionnaire, World Health Organization-Five Well-Being Scale, Mindful Attention Awareness Scale (MAAS) and Life Orientation Test-R (LOT-R) were used to investigate the study participants. Pearson correlation was used to analyze the correlation between subjective well-being and mindfulness and optimism in patients after lung transplantation. AMOS 24.0 software was used to construct a mediating model to analyze the path relationship between mindfulness, optimism and subjective well-being.Results:A total of 205 questionnaires were distributed and 202 valid questionnaires were recovered, with an effective recovery rate of 98.54% (202/205). In 202 patients after lung transplantation, the WHO-5 Well-Being Scale total score, MAAS total score, and LOT-R score were (16.31±4.73), (56.75±9.44), and (18.49±3.85), respectively. Lung transplantation patients' subjective well-being was positively correlated with mindfulness and optimism ( r=0.570, 0.600, both P<0.01). Optimism partially mediated the relationship between mindfulness and subjective well-being, with an effect value of 0.290 and an effect proportion of 52.35% (0.290/0.554) . Conclusions:Mindfulness and optimism are both positively correlated with subjective well-being in post-lung transplantation patients, and mindfulness could also influence subjective well-being through the mediating effect of optimism. Healthcare professionals should fully explore and cultivate positive psychological resources, such as mindfulness, in lung transplant patients, by increasing optimism as the target of intervention, which in turn improves patients' subjective well-being.
7.Diagnostic value of fetal echocardiography for ventricular septal defect and reasons for missed diagnosis and misdiagnosis
Wenying YUAN ; Qichang ZHOU ; Lin ZHANG ; Xinglan HUANG ; Shuizhen LIU ; Ling HUANG ; Fang LIU
Chinese Journal of Perinatal Medicine 2025;28(7):593-597
Objective:To explore the diagnostic value of fetal echocardiography (ECHO) for ventricular septal defect (VSD) and the causes of missed diagnosis and misdiagnosis by analyzing the standard of ECHO sections and image quality.Methods:A retrospective analysis was conducted on 94 VSD fetuses diagnosed by ECHO at Xiangtan Maternal and Child Health Care Hospital from January 2022 to June 2024. Clinical data including the anatomic subtypes, associated anomalies, chromosomal karyotype results, pregnancy outcomes, and neonatal ECHO results were collected and analyzed. Besides, by reviewing the ultrasound images from the institution's ultrasound workstation, a descriptive analysis was conducted on the ECHO section criteria and image quality in the missed and misdiagnosed cases.Results:Among 10 984 fetuses undergoing ECHO, a total of 94 cases of fetal VSD were detected (0.7%), including 45 (48%) isolated VSD and 49 (52%) non-isolated VSD cases. Anatomic distribution revealed perimembranous type predominance (67, 71%), followed by muscular (24, 26%), subarterial types (2, 2%), and perimembranous+muscular type (1, 1%). Among 29 missed diagnosis cases (fetal ECHO-negative but neonatal ECHO-confirmed VSDs), all were isolated VSD, muscular types accounted for 19 (66%), perimembranous types for 8 (27%), and subarterial types for 2 (7%). The causes of missed diagnosis include: technical limitation in 14 cases (48%), restricted cardiac acoustic window in 11 cases (38%), and operator-dependent factors in four cases (14%). All 18 misdiagnosed cases occurred in isolated VSD, with 15 cases of perimembranous and three cases of muscular types. The causes of misdiagnosis include: nine cases due to non-perpendicular beam alignment to perimembranous septum, six cases due to excessively high color flow gain,and three cases due to the misinterpretation of the right ventricular outflow tract flow above the aortic valve as VSD septal perforation blood flow. The sensitivity and specificity of fetal ECHO for diagnosing VSD were 76.42% and 99.83%. Among the 94 cases of fetal VSD, 38 cases (40%) underwent chromosomal karyotype analysis and chromosomal microarray analysis, which identified chromosomal karyotype abnormalities or pathogenic gene variants in 4/16 non-isolated VSDs (all perimembranous), while no anomalies were detected in 22 isolated VSDs. Pregnancy outcomes showed 31 terminations due to associated structural or chromosomal abnormalities versus 63 live births. In the live births, who underwent echocardiography in the neonatal period, 19 cases (30%) showed no VSD and were considered to have undergone spontaneous in utero closure, and persistent defects were found in 44 (70%).Conclusions:Fetal ECHO demonstrates high sensitivity and specificity in the diagnosis of fetal VSD. Critical quality control measures include individualized parameter adjustment and systematic multiplanar continuous scanning to minimize diagnostic errors. Isolated small and medium-sized muscular and perimembranous VSD usually have a good prognosis.
8.Epidemiological and clinical characteristics of 101 patients with brucellosis
Yiling HUANG ; Fei JIN ; Fang NI ; Wenying XIA ; Chunliang ZHU
Chinese Journal of Endemiology 2024;43(5):393-397
Objective:To analyze the epidemiological and clinical characteristics of patients with brucellosis.Methods:Medical records of confirmed brucellosis patients ( n = 101) were collected from January 2015 to December 2022 at the First Affiliated Hospital of Nanjing Medical University. Patients were divided into acute phase group (≤3 months, n = 89) and non acute phase group (> 3 months, n = 12) according to the course of the disease. The epidemiological characteristics, clinical symptoms, laboratory indicators, treatment plan and curative effect of the patients were analyzed retrospectively. Results:Data of a total of 101 cases of brucellosis were collected, including 72 males and 29 females. The disease occurred throughout the year, with summer (June to August) being the peak period (43.56%, 44/101); 72.28% (73/101) cases had a clear history of contact with animal. Ninety-two point zero eight percent (93/101) of the patients visited the Department of Infectious Diseases for the first time. Clinical manifestations included fever, accounting for 82.18% (83/101), chills accounting for 36.63% (37/101), backache accounting for 33.66% (34/101), night sweats accounting for 22.77% (23/101), and arthralgia accounting for 20.79% (21/101). The symptoms of backache (75.00%, 9/12) and arthralgia (41.67%, 5/12) were more common in patients in the non acute phase group than those of the acute phase group [28.09% (25/89), 17.98% (16/89), P < 0.05]. The most common laboratory test abnormal items were elevated C-reactive protein (CRP, 68.32%, 69/101), erythrocyte sedimentation rate (ESR, 61.39%, 62/101), aspartate aminotransferase (AST, 58.42%, 59/101), and alanine aminotransferase (ALT, 48.51%, 49/101). Elevated ESR (66.29%, 59/89) was more common in patients in the acute phase group than that of the non acute phase group [25.00 (3/12), χ 2 = 7.48, P = 0.006]. All patients were treated with a combination therapy, with a recovery rate of 100.00% (101/101). Conclusions:Brucellosis patients are more common in males, with a higher incidence in summer and often accompanied by a history of contact with animal. The clinical manifestations are diverse and non-specific.
9.Effect of esketamine on cerebral ischemia-reperfusion injury and association with mitochondrial stress in mice
Xia WANG ; Peilong LI ; Yaru HUANG ; Wenying CHI ; Gongming WANG ; Fanjun MENG
Chinese Journal of Anesthesiology 2024;44(2):176-181
Objective:To evaluate the effect of esketamine on cerebral ischemia-reperfusion (I/R) injury and the association with mitochondrial stress in mice.Methods:The experiment was performed in two parts. Part Ⅰ Eighteen SPF male C57BL/6 mice, aged 8-12 weeks, with body mass index of 28-30 g, were divided into 3 groups ( n=6 each) by a random number table method: sham operation group (S group), cerebral I/R group (IR group), and esketamine plus cerebral I/R group (E+ IR group). Cerebral I/R was produced by occlusion of middle cerebral artery for 1 h followed by 24-h reperfusion in anesthetized mice.Esketamine 10 mg/kg was intraperitoneally injected at 20 min before developing the model in E group. Neurological function was evaluated using the Zea Longa score and balance beam test (Feeney score). The cerebral infarct size was determined by TTC staining. Part Ⅱ Primary cortical neurons were isolated and cultured and then divided into 3 groups ( n=42 each) using a random number table method: control group (group C), oxygen-glucose deprivation-reoxygenation (OGD/R) group, and esketamine plus OGD/R group (group E+ OGD/R). Cells were subjected to O 2-glucose deprivation for 1 h followed by restoration of O 2-glucose supply for 24 h. The cells were treated with 25 μmol/L esketamine for 40 min before preparing the model in E+ OGD/R group. The neuronal viability was measured by the CCK-8 assay. The ultrastructure of neurons was observed with a transmission electron microscope. The levels of reactive oxygen species (ROS), glutathione peroxidase (GSH-px) and malondialdehyde (MDA) were determined, and the mitochondrial membrane potential was determined by JC-1 kit. The neuronal apoptosis was detected by TUNEL staining, and the apoptosis rate of neurons was calculated. The expression of Bax, cytochrome C (CytC), cleaved-caspase-9, caspase-3 and cleaved-caspase-3 was detected by Western blot. Results:Part Ⅰ Compared with S group, the Zea Longa score, Feeney score and cerebral infarct size were significantly increased in IR group ( P<0.01). Compared with IR group, the Zea Longa score, Feeney score and cerebral infarct size were significantly decreased in E+ IR group ( P<0.01). Part Ⅱ Compared with C group, the cell viability and activity of GSH-px were significantly decreased, the apoptosis rate of neurons, levels of ROS and MDA, mitochondrial membrane potential, and cleaved-caspase-3/caspase-3 ratio were increased, and the expression of Bax, Cyt C and cleaved-caspase-9 was up-regulated in OGD/R group ( P<0.01). Compared with OGD/R group, the cell viability and activity of GSH-px were significantly increased, the apoptosis rate of neurons, levels of ROS and MDA, mitochondrial membrane potential, and cleaved-caspase-3/caspase-3 ratio were decreased, and the expression of Bax, Cyt C and cleaved-caspase-9 was down-regulated in E+ OGD/R group ( P<0.01). Conclusions:Esketamine can alleviate cerebral I/R injury in mice, and the mechanism may be related to inhibition of mitochondrial stress in neurons, improvement in mitochondrial function, and inhibition of mitochondria-dependent apoptosis in neurons.
10.Effects of Siglec-E on parthenolide inhibiting microglia M1 polarization and targeting MAPK/NF-κB pathway
Wenying FU ; Pengbo WANG ; Xiangyu TANG ; Liyuan FENG ; Yuting HUANG ; Peng LI
Journal of Army Medical University 2024;46(7):651-660
Objective To explore the impact of the sialic acid binding lectin-E(Siglec-E)on the inhibitory properties of parthenolide(PTL)against lipopolysaccharide(LPS)-induced M1 polarization of microglia(BV2).Methods ①Single cell sequencing data of Siglece related mouse brain tissue was obtained from Gene Expression Omnibus(GEO)database and divided into the WT group(n=3)and the Siglece-/-group(n=4).The microglia cells were screened,and the enrichment analysis was performed to analyze related differential genes and pathways.BV2 cells were constructed by the shRNA interference technique and were divided into NC-shRNA and Siglece-shRNA to detect the expression level of Siglec-E(Siglece).② NC-shRNA and Siglece-shRNA cells were respectively divided into the Control group,LPS group,PTL group and PTL+LPS group(n=3).The mRNA levels of markers of M1 polarization in microglia,iNOS,IL-1 β and IL-6,were detected by RT-qPCR.Siglecefl/fl and Cx3cr1cre mice were mated to obtain microglia-specific Siglece deletion(Siglecefl/fl×Cx3cr1cre)mice,and LPS-induced neuroinflammation model was established.③ Nine WT and Siglecefl/fl×Cx3cr1cre male mice were assigned to the Control group,LPS group and PTL+LPS group(n=3).RT-qPCR,immunofluorescence assay and Western blotting were used to verify the knock-out effect and polarization-related pathways,and to investigate the mechanism of Siglec-E affecting PTL inhibition of M1 polarization of microglia.Results Compared with the NC-shRNA group,the expression of Siglec-E in the Siglece-shRNA group was significantly decreased(P<0.01),indicating that the Siglec-E knock-down cell model was successfully established.With the stimulation of LPS,mRNA levels ofiNOS,IL-1 β and IL-6 were significantly up-regulated compared with the Control group both in shRNA cells and Siglece-shRNA cells(P<0.01).With the influence of PTL and LPS,the markers of M1 polarization in NC-shRNA cells mentioned before were significantly decreased(P<0.05),while for Siglice-shRNA cells,there were no significant changes in the markers of M1 polarization.PTL inhibited the phosphorylation of JNK and IκB protein(P<0.01)and the nuclear translocation of NF-κB in BV2 cells,down-regulated Siglec-E,and weakened the inhibitory effect.Compared with mice in the WT group,the expression of Siglec-E in microglia of Siglecefl/fl×Cx3cr1cre mice was decreased significantly(P<0.01),and the inhibitory effect of PTL on the phosphorylation of NF-κB in microglia of Siglecefl/fl×Cx3cr1cre mice was also decreased.Conclusion The absence of Siglec-E in microglia attenuates the inhibition of M1 polarization by the MAPK/NF-κB pathway targeted by PTL.


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