1.Mechanism of Huayu Jiedu Prescription in Preventing and Treating Cerebral Ischemia Injury by Regulating NETosis After Acute Cerebral Infarction with Blood Stasis and Toxin Syndrome
Wuchaonan LIU ; Dingxiang LI ; Le YANG ; Jing LIU ; Shengping LUO ; Fang LEI ; Hanlin LEI ; Yihui DENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):50-59
ObjectiveTo investigate the material basis of the pathogenesis of cerebral ischemic injury with blood stasis and toxin syndrome and to explore the protective effects of Huayu Jiedu prescription (HYJDP) on neutrophil extracellular trap-related cell death (NETosis) in cerebral ischemic injury following acute cerebral infarction. MethodsSeventy-two Sprague-Dawley (SD) rats were randomly divided into six groups (n=12 per group): sham operation (Sham) group, blood stasis and toxin model (Model) group, low-, medium-, and high-dose HYJDP groups (HYJDP-L, HYJDP-M, and HYJDP-H; 9, 18, and 36 g·kg-1, respectively), and butylphthalide (NBP) group (0.06 g·kg-1). Except for the Sham group, rats in all other groups were subjected to carrageenan/dry yeast combined with a modified intraluminal filament method to establish a focal cerebral ischemia model of the middle cerebral artery with blood stasis and toxin syndrome. Neurological function was evaluated at 24 h after modeling using the Zea-Longa neurological deficit score. Cerebral infarction rate was assessed by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Pathological morphology of brain tissue was observed using hematoxylin-eosin (HE) staining. Enzyme-linked immunosorbent assay (ELISA) was used to determine serum levels of interleukin-8 (IL-8), myeloperoxidase-DNA complexes (MPO-DNA), and citrullinated histone H3 (CitH3). Protein expression of phosphorylated phosphatidylinositol 3-kinase (p-PI3K), protein kinase B (p-Akt), mammalian target of rapamycin (p-mTOR), sequestosome 1 (p62), and CitH3 in brain tissue was detected by Western blot. Immunofluorescence (IF) was used to detect the expression of neutrophil-specific marker Ly6G, CitH3, and neuron-specific nuclear protein (NeuN) in brain tissue. ResultsCompared with the Sham group, neurological deficit scores and cerebral infarction rates in the model group were significantly increased (P<0.01 for both). HE staining showed varying degrees of neuronal degeneration and necrosis, characterized by blurred neuronal structures, nuclear pyknosis and fragmentation, cytoplasmic dissolution into a vacuolated reticular pattern, and mild glial cell proliferation. ELISA results showed that serum levels of IL-8, MPO-DNA, and CitH3 were significantly increased (P<0.01). Western blot analysis demonstrated decreased expression of p-PI3K, p-Akt, p-mTOR, and p62, while CitH3 expression was significantly increased (P<0.01). IF results showed an increased number of NETs+ cells and a significant decrease in NeuN+ cells (P<0.01). Compared with the Model group, neurological deficit scores in the HYJDP-H group were significantly decreased (P<0.05), and cerebral infarction rates in the HYJDP-H and NBP groups were significantly reduced (P<0.01). HE staining showed that brain tissue damage was markedly alleviated in the HYJDP-H group. ELISA results showed that levels of IL-8, MPO-DNA, and CitH3 were significantly decreased in the HYJDP-M, HYJDP-H, and NBP groups (P<0.01). Western blot analysis showed that expression of p-PI3K, p-Akt, p-mTOR, and p62 was significantly increased in the HYJDP-H and NBP groups, while CitH3 expression was significantly reduced in all drug intervention groups (P<0.01). IF results showed that the number of NETs+ cells was significantly decreased and the number of NeuN⁺ cells was significantly increased in all drug intervention groups (P<0.01). ConclusionNETs may be the material basis of the pathogenesis of cerebral ischemic injury characterized by blood stasis and toxin. HYJDP can regulate the PI3K/Akt/mTOR signaling pathway, reduce the release of pro-inflammatory mediators and NETosis-related products, alleviate cerebral ischemic injury caused by autophagy-dependent NETosis, and thereby exert a neuroprotective effect.
2.Protective Effect of Huayu Jiedu Prescription Against PC12 Cell Damage Induced by Combination of NETs and OGD
Wuchaonan LIU ; Ruiyu HUANG ; Qianru ZENG ; Mengge ZHANG ; Yunhao YI ; Tao GAO ; Dingxiang LI ; Yihui DENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(21):120-130
ObjectiveTo investigate the protective effect of Huayu Jiedu prescription against PC12 cell damage induced by a combination of neutrophil extracellular traps (NETs) and oxygen-glucose deprivation (OGD). MethodsFirst, dimethyl sulfoxide (DMSO) was used to induce the differentiation of the human promyelocytic leukemia cell line (HL-60) into neutrophil-like cells (dHL-60), and cell differentiation was assessed through Giemsa staining and flow cytometry. Next, phorbol myristate (PMA) was used to induce the formation of NETs in dHL-60 cells. The formation of NETs was assessed via SYTOX Green staining, and NETs were extracted by low-speed centrifugation. Finally, the NETs prepared through the conditioned medium were combined with OGD to induce a high-differentiation PC12 neuronal cell damage model through the co-culture method. This model was treated with the Huayu Jiedu prescription-containing serum. Cells were randomly allocated into a normal control group, a model (NETs + OGD) group, a traditional Chinese medicine (NETs + OGD + 5% Huayu Jiedu prescription-containing serum) group, and a NETosis inhibitor (100 mg·L-1 DNase I + NETs + OGD) group. The cell counting kit-8 (CCK-8) assay was used to evaluate cell viability and screen the optimal intervention concentrations of NETs and Huayu Jiedu Formula-containing serum, as well as the optimal intervention time for OGD. Enzyme-linked immunosorbent assay (ELISA) was adopted to measure the levels of myeloperoxidase-DNA complexes (MPO-DNA), citrullinated histone H3 (CitH3), and neutrophil elastase (NE). Western blot was employed to quantify the protein levels of CitH3 and cleaved Caspase-3. Terminal-deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL) staining and flow cytometry were employed to assess neuronal apoptosis. ResultsCompared with those in the normal control group, cells in the model group successfully differentiated into dHL60 cells after DMSO induction. Wright-Giemsa staining revealed that the cells in the model group had disappeared nucleoli, distinct reniform and lobulated changes of nuclei, highly condensed chromatin, and the cytoplasm containing pale purple fine granules. Flow cytometry showed that the mean fluorescence intensity (MFI) of CD11b in the model group was enhanced (P<0.01). Compared with those in the normal control group, dHL60 cells in the model group successfully generated NETs after PMA induction. SYTOX Green staining revealed loss of cell membrane integrity, with chromatin released from the cytoplasm to form abundant extracellular, cloud-like fibrillar structures, and a marked increase in fluorescence intensity (P<0.01). Through the CCK-8 assay for assessing the viability of highly differentiated PC12 cells, the optimal concentration for NETs was determined as 1 000 ng·L-1, the optimal concentration of Huayu Jiedu formula-containing serum as 5%, and the optimal duration for OGD as 2 h. Compared with the normal control group, the model group showed elevated levels of MPO-DNA, CitH3, and NE (P<0.01) and upregulated protein levels of CitH3 and cleaved Caspase-3 (P<0.01). Furthermore, TUNEL staining revealed an increased proportion of apoptotic cells (P<0.01) and flow cytometry showed an increased apoptosis rate (P<0.01) in the model group. Compared with the model group, the Huayu Jiedu formula-containing serum group and the NETosis inhibitor group exhibited declined levels of MPO-DNA, CitH3, and NE (P<0.05,P<0.01) and downregulated protein levels of CitH3 and cleaved Caspase-3 (P<0.01). In addition, TUNEL staining showed a decrease in the proportion of apoptotic cells (P<0.01), and flow cytometry revealed a decrease in the apoptosis rate (P<0.01). ConclusionHuayu Jiedu prescription may exert a protective effect against ischemic neuronal damage by inhibiting NET formation or alleviating NET-induced neurotoxicity
3.Application of Magnetic Resonance Elastography in the Evaluation of Common Intracranial Tumor Stiffness
Jingyi LIN ; Jing ZHAO ; Tianyu HU ; Manshi HU ; Minfen LIU ; Dingxiang XIE
Journal of Sun Yat-sen University(Medical Sciences) 2026;47(1):143-151
ObjectiveTo explore the clinical potential of magnetic resonance elastography (MRE) for assessing the stiffness of common intracranial tumors. MethodsA retrospective analysis was conducted on 47 patients with brain tumors who underwent surgical treatment at The First Affiliated Hospital of Sun Yat-sen University between April 2023 and September 2024. The cohort included 10 cases of glioma, 12 cases of vestibular schwannoma, and 25 cases of meningioma. All patients underwent multiparameter MRI and MRE preoperatively. After co-registering MRE images with contrast-enhanced 3D T1-weighted images, three regions of interest (ROIs) were placed in the areas of maximum and minimum tumor stiffness respectively to measure the shear stiffness values, and the mean was calculated as the average shear stiffness. Additionally, one ROI was placed in the white matter and one in the grey matter of the posterior horn of the lateral ventricle at the basal ganglia level to obtain normal reference values. A neurosurgeon, blind to the MRE results, graded the intraoperative tumor stiffness based on surgical videos using a 5-point scale (1: soft; 5: hard). Intraclass correlation coefficient (ICC) was used to assess inter-observer consistency. Analysis of variance (ANOVA) and nonparametric tests were employed to compare the differences of shear stiffness values between groups. Spearman correlation analysis was used to examine the correlation between elasticity values and clinicopathological parameters. The discriminatory efficacy of shear stiffness values for tumor types was evaluated with receiver operating characteristic (ROC) curves. ResultsThe ICCs for elasticity values measured by two physicians in the hardest and softest tumor areas were 0.868 and 0.831, respectively (both P< 0.01). The maximum, minimum, and average shear stiffness values of meningiomas[(1.91±0.28) m/s, (1.66±0.25) m/s, (1.78±0.26) m/s, respectively] were significantly higher than those of gliomas[(1.59±0.23) m/s, (1.42±0.23) m/s, (1.50±0.22) m/s, respectively (P<0.05)]. The shear stiffness values of vestibular schwannomas[(1.70±0.30) m/s, (1.51±0.23) m/s, (1.61±0.26) m/s, respectively) were lower than those of meningiomas and higher than those of gliomas, but these differences were not statistically significant (both P>0.05). The shear stiffness values of white matter were significantly higher than that of grey matter[(1.63±0.08) m/s vs. (1.39±0.12) m/s, P<0.05)]. In the overall sample, the maximum, minimum, and average shear stiffness values were significantly positively correlated with the intraoperative hardness grade (rs = 0.591, rs = 0.541, rs = 0.571, respectively; all P<0.001), negatively correlated with WHO grade (rs = -0.458, rs = -0.458, rs = -0.480, respectively; all P<0.01), but showed no correlation with Ki-67 (all P>0.05). ConclusionMRE can be used to assess the stiffness of intracranial tumors and to some extent differentiate between tumor types, which aids in formulating more individualized preoperative surgical plans.
4.Protective Effect of Huayu Jiedu Prescription Against PC12 Cell Damage Induced by Combination of NETs and OGD
Wuchaonan LIU ; Ruiyu HUANG ; Qianru ZENG ; Mengge ZHANG ; Yunhao YI ; Tao GAO ; Dingxiang LI ; Yihui DENG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(21):120-130
ObjectiveTo investigate the protective effect of Huayu Jiedu prescription against PC12 cell damage induced by a combination of neutrophil extracellular traps (NETs) and oxygen-glucose deprivation (OGD). MethodsFirst, dimethyl sulfoxide (DMSO) was used to induce the differentiation of the human promyelocytic leukemia cell line (HL-60) into neutrophil-like cells (dHL-60), and cell differentiation was assessed through Giemsa staining and flow cytometry. Next, phorbol myristate (PMA) was used to induce the formation of NETs in dHL-60 cells. The formation of NETs was assessed via SYTOX Green staining, and NETs were extracted by low-speed centrifugation. Finally, the NETs prepared through the conditioned medium were combined with OGD to induce a high-differentiation PC12 neuronal cell damage model through the co-culture method. This model was treated with the Huayu Jiedu prescription-containing serum. Cells were randomly allocated into a normal control group, a model (NETs + OGD) group, a traditional Chinese medicine (NETs + OGD + 5% Huayu Jiedu prescription-containing serum) group, and a NETosis inhibitor (100 mg·L-1 DNase I + NETs + OGD) group. The cell counting kit-8 (CCK-8) assay was used to evaluate cell viability and screen the optimal intervention concentrations of NETs and Huayu Jiedu Formula-containing serum, as well as the optimal intervention time for OGD. Enzyme-linked immunosorbent assay (ELISA) was adopted to measure the levels of myeloperoxidase-DNA complexes (MPO-DNA), citrullinated histone H3 (CitH3), and neutrophil elastase (NE). Western blot was employed to quantify the protein levels of CitH3 and cleaved Caspase-3. Terminal-deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL) staining and flow cytometry were employed to assess neuronal apoptosis. ResultsCompared with those in the normal control group, cells in the model group successfully differentiated into dHL60 cells after DMSO induction. Wright-Giemsa staining revealed that the cells in the model group had disappeared nucleoli, distinct reniform and lobulated changes of nuclei, highly condensed chromatin, and the cytoplasm containing pale purple fine granules. Flow cytometry showed that the mean fluorescence intensity (MFI) of CD11b in the model group was enhanced (P<0.01). Compared with those in the normal control group, dHL60 cells in the model group successfully generated NETs after PMA induction. SYTOX Green staining revealed loss of cell membrane integrity, with chromatin released from the cytoplasm to form abundant extracellular, cloud-like fibrillar structures, and a marked increase in fluorescence intensity (P<0.01). Through the CCK-8 assay for assessing the viability of highly differentiated PC12 cells, the optimal concentration for NETs was determined as 1 000 ng·L-1, the optimal concentration of Huayu Jiedu formula-containing serum as 5%, and the optimal duration for OGD as 2 h. Compared with the normal control group, the model group showed elevated levels of MPO-DNA, CitH3, and NE (P<0.01) and upregulated protein levels of CitH3 and cleaved Caspase-3 (P<0.01). Furthermore, TUNEL staining revealed an increased proportion of apoptotic cells (P<0.01) and flow cytometry showed an increased apoptosis rate (P<0.01) in the model group. Compared with the model group, the Huayu Jiedu formula-containing serum group and the NETosis inhibitor group exhibited declined levels of MPO-DNA, CitH3, and NE (P<0.05,P<0.01) and downregulated protein levels of CitH3 and cleaved Caspase-3 (P<0.01). In addition, TUNEL staining showed a decrease in the proportion of apoptotic cells (P<0.01), and flow cytometry revealed a decrease in the apoptosis rate (P<0.01). ConclusionHuayu Jiedu prescription may exert a protective effect against ischemic neuronal damage by inhibiting NET formation or alleviating NET-induced neurotoxicity
5.Mechanism of Modified Erxian Decoction Regulating Perimenopausal Syndrome via SIRT1/Kisspeptin/GnRH Signaling Pathway
Ruiyu HUANG ; Fang LEI ; Wuchaonan LIU ; Jingjing YANG ; Qianru ZENG ; Shengping LUO ; Yanling CHEN ; Mengge ZHANG ; Fanshun SHEN ; Yihui DENG ; Dingxiang LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(22):51-62
ObjectiveTo explore the regulation of hypothalamic-pituitary-gonadal (HPG) axis by modified Erxian decoction in rats with perimenopausal syndrome (PMS) and to further analyze the expression of proteins related to the silent information regulator 1 (SIRT1)/hypothalamic kisspeptin (Kisspeptin)/gonadotropin-releasing hormone (GnRH) signaling pathway in the arcuate nucleus region (ARC) of the hypothalamus, so as to reveal the potential target of action and molecular biological mechanism of modified Erxian decoction for the treatment of perimenopausal syndrome. MethodsAn animal model was established via the incomplete castration method, with successful modeling confirmed by the exfoliated cervical cell smear method. The 48 rats were divided into six groups based on the randomization principle after successful modeling, including a sham operation group, a model group, an estradiol valerate group (0.09 mg∙kg-1∙d-1), high-, medium-, and low-dose modified Erxian decoction groups (7.614, 3.807,1.903 5 g∙kg-1∙d-1), with 8 rats in each group. The estradiol valerate group and the high-, medium- and low-dose modified Erxian decoction groups were continuously administered by gavage for 28 days, and the indicators were detected 24 hours after the last administration. Body weights and uterine indices were measured. The pathological changes of the uterus were observed by hematoxylin-eosin (HE) staining. Enzyme-linked immunosorbent assay (ELISA) was performed to measure the levels of estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and gonadotropin-releasing hormone (GnRH). Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to determine the expression levels of SIRT1, Kisspeptin, kisspeptin receptor (GPR54), and GnRH in the ARC region of the hypothalamus and gonadotropin-releasing hormone receptor (GnRH-R) in pituitary. ResultsCompared with the sham operation group, rats in the model group had a significantly increased body weight (P0.01), reduced wet weight and index of uterus (P0.01), endometrial thinning or atrophy, glandular atrophy, and a decreasing number of glands. Additionally, serum levels of E2 and the expression of SIRT1 in the ARC region of the hypothalamus significantly decreased (P0.01). Serum levels of FSH, LH, and GnRH, the expression of Kisspeptin, GPR54, and GnRH in the ARC region of the hypothalamus, and GnRH-R in pituitary significantly increased (P0.01). Compared with the model group, the estradiol valerate group and the high-, medium-dose modified Erxian decoction groups had significantly reduced body weight, serum levels of FSH, LH, and GnRH, and expression of Kisspeptin, GPR54, and GnRH in the ARC region of the hypothalamus and GnRH-R in pituitary (P0.05, P0.01) and significantly increased wet weight and index of uterus, serum level of E2, and expression of SIRT1 in the ARC region of the hypothalamus (P0.05, P0.01). In addition, they showed thickened endometrium, increased number of endometrial glands, and improved glandular atrophy. ConclusionModified Erxian decoction regulates the function of the HPG axis through multi-targets, and its mechanism of action may be related to the up-regulation of the expression of SIRT1 in the ARC region of the hypothalamus, the inhibition of the over-activation of the Kisspeptin/GnRH signaling pathway, the regulation of the expression of GnRH-R in the pituitary, the restoration of secretion balance of gonadotropins, and the elevation of the estrogen level. This study provides an experimental basis for the interpretation of the scientific connotation of modified Erxian decoction in the treatment of perimenopausal syndrome and a theoretical reference for the development of a novel therapeutic strategy based on the SIRT1/Kisspeptin/GnRH pathway.
6.Effects and Mechanism of Cyclocaryae Paliuri Folium in Sodium Palmitate-Induced Lipid Deposition in HepG2 Cells
Yixuan LIU ; Haoyang WU ; Jingjing YANG ; Yuzhe CAI ; Zheng LUO ; Dingxiang LI ; Yihui DENG
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(4):72-78
Objective To investigate the ameliorative effect and potential mechanism of Cyclocaryae Paliuri Folium in sodium palmitate-induced lipid deposition in HepG2 cells.Methods The effect of sodium palmitate and lyophilized powder of Cyclocaryae Paliuri Folium on the viability of HepG2 cells was determined by the CCK-8 method to determine the subsequent dosage administered.The HepG2 cells were divided into blank group,model group,pioglitazone group and Cyclocaryae Paliuri Folium low-,medium-and high-dosage group,the lipid deposition model of HepG2 cells was established using 350 μmol/L sodium palmitate,the medication group were given pioglitazone and low-,medium-and high-dosage of Cyclocaryae Paliuri Folium(100,250,500 μg/mL)for 12 h respectively.The intracellular lipid deposition was observed by Nile red staining and BODIPY493/503 fluorescent probe staining,the content of TNF-α and IL-6 in supernatant of cell culture medium were detected by ELISA,Western blot was used to detect PI3K,Akt,sterol-regulatory element binding protein-1(SREBP-1),fatty acid synthase(FAS),Bcl-2,Bax protein expression,qPCR was used to detect the mRNA expression of adipose triglyceride lipase(ATGL)and CD36.Results Compared with the blank group,the lipid deposition of HepG2 cells in the model group increased,TNF-α and IL-6 contents in the supernatant of cell culture medium significantly increased(P<0.01),the protein expressions of p-PI3K,p-Akt,SREBP-1,FAS and Bax in cells significantly increased,while the protein expression of Bcl-2 significantly decreased(P<0.01),the mRNA expression of ATGL significantly decreased,and the mRNA expression of CD36 significantly increased(P<0.01).Compared with the model group,the intracellular lipid deposition of the pioglitazone group and Cyclocaryae Paliuri Folium groups improved to varying degrees,the contents of TNF-α and IL-6 in the cell supernatant decreased,the expressions of p-PI3K,p-Akt,SREBP-1,FAS and Bax proteins decreased,the expression of Bcl-2 protein increased,the expression of ATGL mRNA increased,and the expression of CD36 mRNA decreased,with statistical significance in pioglitazone group and Cyclocaryae Paliuri Folium high-dosage group(P<0.05,P<0.01).Conclusion Cyclocaryae Paliuri Folium may ameliorate sodium palmitate-induced lipid deposition in HepG2 model cells by modulating the PI3K/Akt/SREBP-1/FAS signaling pathway and affecting triacylglycerol metabolism.
7.Specialty training demands among plastic surgery nurses: a survey
Xuefeng CHEN ; Na LIU ; Dingxiang LIU ; Yuanbo LIU
Chinese Journal of Medical Aesthetics and Cosmetology 2025;31(3):298-303
Objective:To investigate the specialized training needs of plastic surgery nurses.Methods:A convenience sampling method was employed. A self-designed questionnaire was developed to survey registered nurses engaged in plastic surgery nationwide, covering general information, attitudes toward specialized nurse training, willingness and needs for training, recognition of training evaluation methods, and interest levels in training content.Results:A total of 177 valid questionnaires were collected, yielding a 100% valid response rate. Participants included 15 males and 162 females, aged 22-54 (37.6±8.9) years, with 1-38 years of work experience in plastic surgery. Among them, 19.2% (34/177) were certified plastic surgery specialists, 92.1% (163/177) agreed on the necessity of establishing specialized plastic surgery nurse positions, 91.0% (161/177) supported the creation of specialized training bases, and 84.7% (150/177) expressed participation needs. Clinical practice and theoretical knowledge training needs were both highly demanded.Conclusion:Plastic surgery nurses demonstrate significant demand for specialized training, the establishment of specialized plastic surgery nurse positions and dedicated training bases should be prioritized.
8.Effects of Zuogui Jiangtang Tongmai Prescription on Short-chain Fatty Acids and G Protein-coupled Receptor 109A in Diabetic Atherosclerotic Mice
Jing CHEN ; Qin WU ; Yanan ZHANG ; Jingjing YANG ; Yanling CHEN ; Wuchaonan LIU ; Fang LEI ; Dingxiang LI ; Yihui DENG
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(3):76-82
Objective To investigate the effects of Zuogui Jiangtang Tongmai Prescription on short-chain fatty acids(SCFAs)and G protein-coupled receptor 109A(GPR109A)in diabetic atherosclerotic mice;To explore its mechanism of improving diabetic atherosclerosis.Methods ApoE-/-mice were fed with high sugar and high fat diet and intraperitoneal injection of streptozotocin to establish atherosclerosis model of diabetes.After modeling,the mice were randomly divided into the model group,Western medicine group(metformin hydrochloride+atorvastatin),and Zuogui Jiangtang Tongmai Prescription low-,medium-and high-dosage groups(19,38,76 g/kg);and the other C57BL/6J mice were set as the control group,with 6 mice in each group.Each group was given solution for gavage,once a day for 4 weeks.Blood glucose of the mice were detected,HE staining was used to observe the morphology of the aorta,TG,TC,LDL-C and HDL-C contents were detected by fully automated biochemistry analyzers,ELISA was used to detect the contents of serum HbA1c,fasting insulin(FINS),interleukin(IL)-1β,IL-6 and tumor necrosis factor(TNF)-α,gas chromatography was used to detect the content of intestinal SCFAs,and RT qPCR and Western blot were used to detect the mRNA and protein expressions of GPR109A in ileal tissue and nuclear factor(NF)-κB p65 in aortic tissue,respectively.Results Compared with the control group,obvious plaques and inflammatory cells infiltration were seen in the aorta of the model group,the blood glucose and serum HbA1c,TG,TC,LDL-C,IL-1β,IL-6 and TNF-α contents increased(P<0.01),FINS and HDL-C content decreased(P<0.01),intestinal acetate,propionate,butyric acid,isobutyric acid and isovaleric acid contents decreased(P<0.01),GPR109A mRNA and protein expression in ileal tissue decreased,NF-κB p65 mRNA and protein expression in aortic tissue increased(P<0.01).Compared with the model group,the aortic plaque area and inflammatory cells infiltration were significantly improved in each drug intervention group,the blood glucose and serum HbA1c,TG,TC,LDL-C,IL-1β,IL-6 and TNF-α contents decreased(P<0.05,P<0.01);the FINS and HDL-C content increased(P<0.05,P<0.01),intestinal acetic,propionic,butyric acid,isobutyric acid and isovaleric acid contents increased(P<0.05,P<0.01),GPR109A mRNA and protein expression in ileal tissue increased and NF-κB p65 mRNA and protein expression in ileal tissue decreased decreased(P<0.05,P<0.01).Conclusion Zuogui Jiangtang Tongmai Prescription can improve glucolipid metabolism and inflammatory response in diabetic atherosclerotic mice,which may be related to regulating SCFAs/GPR109A pathway.
9.Effects and Mechanism of Cyclocaryae Paliuri Folium in Sodium Palmitate-Induced Lipid Deposition in HepG2 Cells
Yixuan LIU ; Haoyang WU ; Jingjing YANG ; Yuzhe CAI ; Zheng LUO ; Dingxiang LI ; Yihui DENG
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(4):72-78
Objective To investigate the ameliorative effect and potential mechanism of Cyclocaryae Paliuri Folium in sodium palmitate-induced lipid deposition in HepG2 cells.Methods The effect of sodium palmitate and lyophilized powder of Cyclocaryae Paliuri Folium on the viability of HepG2 cells was determined by the CCK-8 method to determine the subsequent dosage administered.The HepG2 cells were divided into blank group,model group,pioglitazone group and Cyclocaryae Paliuri Folium low-,medium-and high-dosage group,the lipid deposition model of HepG2 cells was established using 350 μmol/L sodium palmitate,the medication group were given pioglitazone and low-,medium-and high-dosage of Cyclocaryae Paliuri Folium(100,250,500 μg/mL)for 12 h respectively.The intracellular lipid deposition was observed by Nile red staining and BODIPY493/503 fluorescent probe staining,the content of TNF-α and IL-6 in supernatant of cell culture medium were detected by ELISA,Western blot was used to detect PI3K,Akt,sterol-regulatory element binding protein-1(SREBP-1),fatty acid synthase(FAS),Bcl-2,Bax protein expression,qPCR was used to detect the mRNA expression of adipose triglyceride lipase(ATGL)and CD36.Results Compared with the blank group,the lipid deposition of HepG2 cells in the model group increased,TNF-α and IL-6 contents in the supernatant of cell culture medium significantly increased(P<0.01),the protein expressions of p-PI3K,p-Akt,SREBP-1,FAS and Bax in cells significantly increased,while the protein expression of Bcl-2 significantly decreased(P<0.01),the mRNA expression of ATGL significantly decreased,and the mRNA expression of CD36 significantly increased(P<0.01).Compared with the model group,the intracellular lipid deposition of the pioglitazone group and Cyclocaryae Paliuri Folium groups improved to varying degrees,the contents of TNF-α and IL-6 in the cell supernatant decreased,the expressions of p-PI3K,p-Akt,SREBP-1,FAS and Bax proteins decreased,the expression of Bcl-2 protein increased,the expression of ATGL mRNA increased,and the expression of CD36 mRNA decreased,with statistical significance in pioglitazone group and Cyclocaryae Paliuri Folium high-dosage group(P<0.05,P<0.01).Conclusion Cyclocaryae Paliuri Folium may ameliorate sodium palmitate-induced lipid deposition in HepG2 model cells by modulating the PI3K/Akt/SREBP-1/FAS signaling pathway and affecting triacylglycerol metabolism.
10.Effects of Zuogui Jiangtang Tongmai Prescription on Short-chain Fatty Acids and G Protein-coupled Receptor 109A in Diabetic Atherosclerotic Mice
Jing CHEN ; Qin WU ; Yanan ZHANG ; Jingjing YANG ; Yanling CHEN ; Wuchaonan LIU ; Fang LEI ; Dingxiang LI ; Yihui DENG
Chinese Journal of Information on Traditional Chinese Medicine 2025;32(3):76-82
Objective To investigate the effects of Zuogui Jiangtang Tongmai Prescription on short-chain fatty acids(SCFAs)and G protein-coupled receptor 109A(GPR109A)in diabetic atherosclerotic mice;To explore its mechanism of improving diabetic atherosclerosis.Methods ApoE-/-mice were fed with high sugar and high fat diet and intraperitoneal injection of streptozotocin to establish atherosclerosis model of diabetes.After modeling,the mice were randomly divided into the model group,Western medicine group(metformin hydrochloride+atorvastatin),and Zuogui Jiangtang Tongmai Prescription low-,medium-and high-dosage groups(19,38,76 g/kg);and the other C57BL/6J mice were set as the control group,with 6 mice in each group.Each group was given solution for gavage,once a day for 4 weeks.Blood glucose of the mice were detected,HE staining was used to observe the morphology of the aorta,TG,TC,LDL-C and HDL-C contents were detected by fully automated biochemistry analyzers,ELISA was used to detect the contents of serum HbA1c,fasting insulin(FINS),interleukin(IL)-1β,IL-6 and tumor necrosis factor(TNF)-α,gas chromatography was used to detect the content of intestinal SCFAs,and RT qPCR and Western blot were used to detect the mRNA and protein expressions of GPR109A in ileal tissue and nuclear factor(NF)-κB p65 in aortic tissue,respectively.Results Compared with the control group,obvious plaques and inflammatory cells infiltration were seen in the aorta of the model group,the blood glucose and serum HbA1c,TG,TC,LDL-C,IL-1β,IL-6 and TNF-α contents increased(P<0.01),FINS and HDL-C content decreased(P<0.01),intestinal acetate,propionate,butyric acid,isobutyric acid and isovaleric acid contents decreased(P<0.01),GPR109A mRNA and protein expression in ileal tissue decreased,NF-κB p65 mRNA and protein expression in aortic tissue increased(P<0.01).Compared with the model group,the aortic plaque area and inflammatory cells infiltration were significantly improved in each drug intervention group,the blood glucose and serum HbA1c,TG,TC,LDL-C,IL-1β,IL-6 and TNF-α contents decreased(P<0.05,P<0.01);the FINS and HDL-C content increased(P<0.05,P<0.01),intestinal acetic,propionic,butyric acid,isobutyric acid and isovaleric acid contents increased(P<0.05,P<0.01),GPR109A mRNA and protein expression in ileal tissue increased and NF-κB p65 mRNA and protein expression in ileal tissue decreased decreased(P<0.05,P<0.01).Conclusion Zuogui Jiangtang Tongmai Prescription can improve glucolipid metabolism and inflammatory response in diabetic atherosclerotic mice,which may be related to regulating SCFAs/GPR109A pathway.

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