1.Pathogenic Mechanisms of Spleen Deficiency-Phlegm Dampness in Obesity and Traditional Chinese Medicine Prevention and Treatment Strategies:from the Perspective of Immune Inflammation
Yumei LI ; Peng XU ; Xiaowan WANG ; Shudong CHEN ; Le YANG ; Lihua HUANG ; Chuang LI ; Qinchi HE ; Xiangxi ZENG ; Juanjuan WANG ; Wei MAO ; Ruimin TIAN
Journal of Traditional Chinese Medicine 2026;67(1):31-37
Based on spleen deficiency-phlegm dampness as the core pathogenesis of obesity, and integrating recent advances in modern medicine regarding the key role of immune inflammation in obesity, this paper proposes a multidimensional pathogenic network of "obesity-spleen deficiency-phlegm dampness-immune imbalance". Various traditional Chinese medicine (TCM) herbs that strengthen the spleen, regulate qi, and resolve phlegm and dampness can treat obesity by improving spleen-stomach transport and transformation, promoting water-damp metabolism, and regulating immune homeostasis. This highlights immune inflammation as an important entry point to elucidate the TCM concepts of "spleen deficiency-phlegm dampness" and the therapeutic principle of "strengthening the spleen and eliminating dampness to treat obesity". By systematically analyzing the intrinsic connection between "spleen deficiency generating dampness, internal accumulation of phlegm dampness" and immune dysregulation in obesity, this paper aims to provide theoretical support for TCM treatment of obesity based on dampness.
2.In vitro biocompatibility of graded glass infiltrated ultra-translucent zirconia
Qiya ZHANG ; Yixiang TONG ; Shijiao YANG ; Yumeng ZHANG ; Ling DENG ; Wei WU ; Yao XIE ; Jian LIAO ; Ling MAO
Chinese Journal of Tissue Engineering Research 2026;30(2):443-450
BACKGROUND:In previous studies,glass materials were infiltrated into 5Y-PSZ ultra-translucent zirconia by a double sintering method to prepare 5Y-PSZ-YGI graded glass infiltrated ultra-translucent zirconia materials that can maintain high transparency and high flexural strength.OBJECTIVE:To evaluate the in vitro biocompatibility of 5Y-PSZ-YGI graded glass infiltrated ultra-translucent zirconia materials.METHODS:(1)Glass materials were infiltrated into 5Y-PSZ ultra-translucent zirconia by double sintering to prepare 5Y-PSZ-YGI graded glass infiltrated ultra-translucent zirconia.5Y-PSZ-YGI graded glass infiltrated ultra-translucent zirconia(or 5Y-PSZ ultra-translucent zirconia,3Y-TZP transparent zirconia)was placed in DMEM culture medium containing 10%fetal bovine serum for 12,24 and 72 hours,and the surface area ratio of culture medium to sample was 3 mL/cm2,and the 12-,24-and 72-hour material extracts were obtained.(2)After culturing mouse fibroblast L929 for 24 hours,the original culture medium was discarded and divided into 7 groups for culture:the control group was replaced with DMEM culture medium containing 10%fetal bovine serum by volume,and the other 6 groups were replaced with 24-hour extract of 3Y-TZP transparent zirconia,24-hour extract of 5Y-PSZ ultra-translucent zirconia,24-hour extract of 5Y-PSZ-YGI graded glass infiltrated ultra-translucent zirconia,72-hour extract of 3Y-TZP transparent zirconia,72-hour extract of 5Y-PSZ ultra-translucent zirconia,and 72-hour extract of 5Y-PSZ-YGI graded glass infiltrated ultra-translucent zirconia.After 1,3,and 5 days of culture,cell growth was observed under a microscope,and the cell proliferation rate was obtained by CCK-8 assay to determine cytotoxicity.(3)Human anticoagulated blood was mixed with 5Y-PSZ-YGI graded glass infiltrated ultra-translucent zirconia,5Y-PSZ ultra-translucent zirconia,and 3Y-TZP transparent zirconia,and the hemolysis rate was detected after 0.5 hours.Human anticoagulated blood was mixed with 12-hour extract of 3Y-TZP transparent zirconia,12-hour extract of 5Y-PSZ ultra-translucent zirconia,and 12-hour extract of 5Y-PSZ-YGI graded glass infiltrated ultra-translucent zirconia,and the hemolysis rate was detected after 0.5 hours.RESULTS AND CONCLUSION:(1)Under the microscope,it could be seen that the number of cells in each group increased with the extension of culture time,and the cell morphology of each experimental group was basically the same as that of the control group.The cytotoxicity grade of the 24-hour extract of 3Y-TZP transparent zirconia group on the first day of culture was grade 0,and the cytotoxicity grade of the other experimental groups at each time period was grade 1.(2)Neither the material nor the material extract caused obvious hemolytic reaction,and the hemolytic rate was less than 5%.(3)The results showed that 5Y-PSZ-YGI graded glass infiltrated ultra-translucent zirconia had no significant effect on the growth and proliferation of mouse fibroblasts L929,and did not cause hemolytic reaction with human blood,and had good in vitro biocompatibility.
3.In vitro biocompatibility of graded glass infiltrated ultra-translucent zirconia
Qiya ZHANG ; Yixiang TONG ; Shijiao YANG ; Yumeng ZHANG ; Ling DENG ; Wei WU ; Yao XIE ; Jian LIAO ; Ling MAO
Chinese Journal of Tissue Engineering Research 2026;30(2):443-450
BACKGROUND:In previous studies,glass materials were infiltrated into 5Y-PSZ ultra-translucent zirconia by a double sintering method to prepare 5Y-PSZ-YGI graded glass infiltrated ultra-translucent zirconia materials that can maintain high transparency and high flexural strength.OBJECTIVE:To evaluate the in vitro biocompatibility of 5Y-PSZ-YGI graded glass infiltrated ultra-translucent zirconia materials.METHODS:(1)Glass materials were infiltrated into 5Y-PSZ ultra-translucent zirconia by double sintering to prepare 5Y-PSZ-YGI graded glass infiltrated ultra-translucent zirconia.5Y-PSZ-YGI graded glass infiltrated ultra-translucent zirconia(or 5Y-PSZ ultra-translucent zirconia,3Y-TZP transparent zirconia)was placed in DMEM culture medium containing 10%fetal bovine serum for 12,24 and 72 hours,and the surface area ratio of culture medium to sample was 3 mL/cm2,and the 12-,24-and 72-hour material extracts were obtained.(2)After culturing mouse fibroblast L929 for 24 hours,the original culture medium was discarded and divided into 7 groups for culture:the control group was replaced with DMEM culture medium containing 10%fetal bovine serum by volume,and the other 6 groups were replaced with 24-hour extract of 3Y-TZP transparent zirconia,24-hour extract of 5Y-PSZ ultra-translucent zirconia,24-hour extract of 5Y-PSZ-YGI graded glass infiltrated ultra-translucent zirconia,72-hour extract of 3Y-TZP transparent zirconia,72-hour extract of 5Y-PSZ ultra-translucent zirconia,and 72-hour extract of 5Y-PSZ-YGI graded glass infiltrated ultra-translucent zirconia.After 1,3,and 5 days of culture,cell growth was observed under a microscope,and the cell proliferation rate was obtained by CCK-8 assay to determine cytotoxicity.(3)Human anticoagulated blood was mixed with 5Y-PSZ-YGI graded glass infiltrated ultra-translucent zirconia,5Y-PSZ ultra-translucent zirconia,and 3Y-TZP transparent zirconia,and the hemolysis rate was detected after 0.5 hours.Human anticoagulated blood was mixed with 12-hour extract of 3Y-TZP transparent zirconia,12-hour extract of 5Y-PSZ ultra-translucent zirconia,and 12-hour extract of 5Y-PSZ-YGI graded glass infiltrated ultra-translucent zirconia,and the hemolysis rate was detected after 0.5 hours.RESULTS AND CONCLUSION:(1)Under the microscope,it could be seen that the number of cells in each group increased with the extension of culture time,and the cell morphology of each experimental group was basically the same as that of the control group.The cytotoxicity grade of the 24-hour extract of 3Y-TZP transparent zirconia group on the first day of culture was grade 0,and the cytotoxicity grade of the other experimental groups at each time period was grade 1.(2)Neither the material nor the material extract caused obvious hemolytic reaction,and the hemolytic rate was less than 5%.(3)The results showed that 5Y-PSZ-YGI graded glass infiltrated ultra-translucent zirconia had no significant effect on the growth and proliferation of mouse fibroblasts L929,and did not cause hemolytic reaction with human blood,and had good in vitro biocompatibility.
4.Protective effect and mechanism of genistein on etoposide-induced chondrocyte senescence
Jinhong WANG ; Tianyu CHEN ; Lifang MAO ; Yingjie ZHAO ; Renpeng ZHOU ; Wei HU ; Chao LU
Acta Universitatis Medicinalis Anhui 2026;61(4):636-643
ObjectiveTo investigate the protective effect of genistein (Gen) on etoposide-induced chondrocyte senescence and its underlying mechanism. MethodsThe C28/I2 cell line was treated with different concentrations of Gen and etoposide, and the cell viability was detected by the CCK-8 assay. The senescence model of C28/I2 chondrocytes was induced by etoposide, with Gen intervention. Senescence-associated β-galactosidase (SA-β-gal) staining was performed to detect the senescence-positive rate and staining characteristics of chondrocytes. The expressions of peroxiredoxin 6 (Prdx6), cyclin-dependent kinaseto clarify the functional necessity of Prdx6. ResultsCompared with the etoposide group, the C28/I2 chondrocyte viability significantly increased (P<0.01), the expression ofsenescence-associated proteins p21 and p16 decreased (P<0.01, P<0.05), the expression of senescence-associated genes p21 and p16 reduced (both P<0.01), the fluorescence intensity of senescence-associated proteins p21 and p16 was diminished (P<0.05, P<0.01), and the proportion of SA-β-gal-positive cells decreased (P<0.01) in the Gen+etoposide group. Compared with the Control group, the expression of Prdx6 was downregulated in the etoposide group (P<0.05). Compared with the etoposide group, the expression of Prdx6 was upregulated in the Gen+etoposide group (P<0.01). Compared with the Control group, the GPx activity significantly decreased in the si-Prdx6 group (P<0.01). Furthermore, compared with the si-Prdx6 group, the GPx activity increased in the si-Prdx6+Gen group (P<0.05). Molecular docking results revealed that Gen formed hydrogen bond interactions with the active site of Prdx6. After Prdx6 knockdown, the expression of senescence-associated genes p21 and p16 and the fluorescence intensity of senescence-associated proteins p21 and p16 both increased in the Gen+etoposide+si-Prdx6 group (both P<0.01). ConclusionGen can inhibit etoposide-induced senescence of C28/I2 chondrocytes by upregulating the expression of Prdx6. This study provides potential drug targets and experimental basis for the prevention and treatment of chondrocyte senescence-related diseases.
5.Experimental study of epigallocatechin gallate-loaded hydrogel scaffold for the treatment of periodontitis
Longge LIU ; Jing MAO ; Huimin ZHENG ; Wei LI
Acta Universitatis Medicinalis Anhui 2026;61(6):1013-1020
ObjectiveTo explore the structural characteristics, biocompatibility, anti-inflammatory and antioxidant properties of hyaluronic acid (HA)/silk fibroin (SF) composite hydrogel scaffolds loaded with epigallocatechin gallate (EGCG), and to provide a new material basis for the treatment of periodontitis. MethodsThe composite hydrogel scaffolds were constructed by cross-linking HA and SF and loaded with EGCG. The water absorption rate, degradation property and material characteristics were detected by scanning electron microscopy. The cytocompatibility and anti-inflammatory and antioxidant properties were evaluated by in vitro experiments. ResultsThe EGCG-loaded scaffolds exhibited a porous network structure and desirable swelling properties. In vitro experiments demonstrated that the scaffolds possessed good biocompatibility, along with the ability to inhibit the expression of inflammatory factors and scavenge reactive oxygen species. ConclusionThe HA/SF-EGCG composite scaffolds have excellent biocompatibility and dual functional activities of anti-inflammatory and antioxidant, providing a new strategy for improving the clinical treatment effect of periodontitis.
6.Effect and mechanism of action of Biejiajian Pills on a rat model of hepatic fibrosis induced by carbon tetrachloride
Longda WU ; Yuanqin DU ; Yanfei WEI ; Dewen MAO ; Yong LIN ; Lu LU ; Faming SHU
Journal of Clinical Hepatology 2026;42(8):1866-1877
ObjectiveTo investigate the therapeutic effect of Biejiajian Pills on rats with carbon tetrachloride (CCl4)-induced hepatic fibrosis and its mechanism based on the bile acid (BA)-short-chain fatty acid (SCFA) metabolic axis. MethodsThe method of CCl4 induction was used to establish a rat model of hepatic fibrosis, and 32 male Sprague-Dawley rats were randomly divided into control group, model group, Biejiajian Pills group (2.2 g/kg), and silymarin group (43.19 mg/kg), with 8 rats in each group. All rats except those in the control group were given intraperitoneal injection of CCl4 (diluted with corn oil at a ratio of 4∶6, 2 mL/kg) to induce a model of hepatic fibrosis for 8 consecutive weeks, and the rats in the drug administration groups were given the corresponding drug by gavage. An automatic biochemical analyzer was used to measure the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil), and albumin (Alb); chloramine-T colorimetry was used to measure the content of hydroxyproline (HYP) in liver tissue; HE staining and Masson staining were used to observe liver histopathological changes; UPLC-MS/MS and GC-MS were used to measure the levels of BAs and SCFAs. The Shapiro-Wilk test and the Levene test were performed for all data to determine normality and homogeneity of variance; a one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups; the Kruskal-Wallis H test was used for comparison of Ishak score between groups, and the Dunn’s test was used for further comparison between two groups. The Benjamini-Hochberg method was used for multiple comparison correction in the screening of differentially expressed metabolites in metabolomics data. The variable importance in projection (VIP) values were obtained in combination with the orthogonal partial least squares-discriminant analysis (OPLS-DA) model, and metabolites with VIP>1 and P<0.05 were selected as significantly differentially expressed metabolites between groups. The robustness of the OPLS-DA model was evaluated through 200 permutation tests, and the main parameters of the model were reported, including the interpretation rate of the model for categorical variables (R2Y) and the predictive ability of the model (Q2). A Spearman’s rank correlation analysis was performed. ResultsThe results of staining showed ordered arrangement of hepatocytes in the Biejiajian Pills group, with significant alleviation of inflammatory cell infiltration and collagen fiber deposition. Compared with the control group, the model group had significant increases in the serum levels of TBil and HYP and a significant reduction in the serum level of Alb (all P<0.05). Compared with the control group, the Biejiajian Pills group had significant increases in the serum levels of AST, ALT, TBil, and HYP and a significant reduction in the serum level of Alb (all P<0.05), and the silymarin group had significant increases in the serum levels of ALT, TBil, and HYP and a significant reduction in the serum level of Alb (all P<0.05). Compared with the model group, the Biejiajian Pills group had significant reductions in the serum levels of ALT, AST, TBil, and HYP (all P<0.05), and the silymarin group had significant reductions in the serum levels of ALT, TBil, HYP, and AST and a significant increase in the serum level of Alb (all P<0.05). Mechanism studies showed that compared with the control group, the model group had significant reductions in the levels of taurolithocholic acid, tauro-α-muricholic acid, taurocholic acid, glycodeoxycholic acid, taurohyocholic acid, chenodeoxycholic acid, glycohyodeoxycholic acid, 7,12-diketolithocholic acid, taurochenodeoxycholic acid, and glycochenodeoxycholic acid and significant increases in the levels of hyodeoxycholic acid, norcholic acid, β-muricholic acid, and cholic acid (all P<0.05). Compared with the model group, the Biejiajian Pills group had significant increases in the levels of the protective BAs taurolithocholic acid, chenodeoxycholic acid, taurocholic acid, taurohyocholic acid, and taurodeoxycholic acid and significant reductions in the levels of tauro-α-muricholic acid, tauro-β-muricholic acid, taurohyodeoxycholic acid+tauroursodeoxycholic acid, α-muricholic acid, glycoursodeoxycholic acid, and norcholic acid (alll P<0.05). As for SCFAs, compared with the model group, the Biejiajian Pills group had significant increases in the levels of acetic acid, propionic acid, isobutyric acid, butyric acid, isopentanoic acid, pentanoic acid, and hexanoic acid (all P<0.05). Compared with the model group, the silymarin group had significant reductions in the levels of taurolithocholic acid, tauro-α-muricholic acid, norcholic acid, glycocholic acid, deoxycholic acid, tauro-β-muricholic acid, taurodeoxycholic acid, taurohyodeoxycholic acid+tauroursodeoxycholic acid, and α-muricholic acid and significant increases in the levels of taurocholic acid and chenodeoxycholic acid, as well as significant increases in the levels of the SCFAs acetic acid, propionic acid, pentanoic acid, and hexanoic acid (all P<0.05). The correlation analysis showed that the metabolism of BAs and SCFAs was correlated with the changes in serum indicators. ConclusionBiejiajian Pills can alleviate CCl4-induced hepatic fibrosis in rats by regulating the BA/SCFA metabolic axis.
7.Establishment and Evaluation of Different Disease-Syndrome Combination Mice Models of Knee Osteoarthritis with Cold-Dampness Stagnation Syndrome
Enrui HU ; Yibao WEI ; Deren LIU ; Aobuliaisan MAIMAITITUXUN ; Licheng ZHANG ; Jiachen SONG ; Jun MAO ; Taiyang LIAO ; Peimin WANG
Journal of Traditional Chinese Medicine 2026;67(19):2120-2129
ObjectiveTo establish and evaluate different disease-syndrome combination mice models of knee osteoarthritis (KOA) with cold-dampness obstruction syndrome, and to identify the optimal modeling method. MethodsEighty mice were randomly divided into a control group, a treadmill plus climate chamber group, an anterior cruciate ligament transection (ACLT) plus climate chamber group, an ACLT group, a destabilization of the medial meniscus (DMM) plus climate chamber group, a DMM group, an intra-articular injection plus climate chamber group, and an intra-articular injection group, with 10 mice in each group. Mice in the ACLT group underwent surgical transection of the knee anterior cruciate ligament to establish the model, while those in the DMM underwent surgical transection of the medial meniscal tibial ligament of the knee, and those in the intra-articular injection group were modeled by an intra-articular injection of 50 μl of normal saline containing 1mg of monosodium iodoacetate (MIA) into the medial side of the patellar ligament. Mice in the ACLT plus climate chamber, DMM plus climate chamber, and intra-articular injection plus climate chamber groups were placed in an artificial climate chamber (relative humidity 60%, temperature 4 ℃, wind speed 6 m/s) simultaneously with aforementioned interventions, once daily for 2 h each time, for 14 consecutive days. Mice in the control group underwent sham surgery, involving only the incision of the skin and subcutaneous tissue at the knee joint to expose the articular cavity, followed by layer-by-layer suturing. Mice in the treadmill plus climate chamber group were placed on an animal treadmill starting on the 4th day after the sham operation, undergoing high-intensity treadmill exercise at a speed of 20 m/min, an inclination of 5°, for 30min per session, 5 times per week for 28 consecutive days, while being housed in the artificial climate chamber. Cold pain thresholds of the mice in each group were compared using the cold plate test and the acetone low-temperature test before and after modeling. Following modeling, knee joint pathological changes were observed using HE and Safranin O-Fast Green staining, and quantified using Mankin and Osteoarthritis Research Society International (OARSI) scores. Serum levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were measured by ELISA. Protein and mRNA expression levels of pain-related markers, including calcitonin gene-related peptide (CGRP), protein gene product 9.5 (PGP9.5), and cellular proto-oncogene Fos (c-Fos), and the cold hyperalgesia-related markers including transient receptor potential ankyrin 1 (TRPA1) and transient receptor potential melastatin 8 (TRPM8) in the dorsal root ganglion (DRG) tissues were detected by Western Blot and qPCR. TRPA1 and TRPM8 fluorescent intensity in the DRG was detected by immunofluorescence assay. ResultsCompared to the control group, the remaining groups showed decreased cold pain thresholds in the cold plate tese, increased cold pain thresholds in the acetone low-temperature test, elevated Mankin and OARSI scores, and increased serum TNF-α and IL-1β levels, upregulated protein and mRNA expression levels of CGRP, PGP9.5, c-Fos, TRPA1, and TRPM8 in the DRG, and increased immunofluorescence intensities of TRPA1 and TRPM8 in the DRG tissue (P<0.05 or P<0.01), along with severe pathological damage and cartilage degeneration in the knee joints. Compared to the ACLT, DMM, and intra-articular injection groups, the ACLT plus climate chamber, DMM plus climate chamber, and intra-articular injection plus climate chamber groups exhibited more significant changes in all the aforementioned indicators (P<0.01). Furthermore, changes in the aforementioned indicators were more pronounced in the ACLT plus climate chamber group compared to the Treadmill plus climate chamber group, DMM plus climate chamber and intra-articular injection plus climate chamber group (P<0.01), exhibiting the most severe degree of pathological damage and cartilage degeneration. ConclusionThe combination of an artificial climate chamber can successfully establish a disease-syndrome combined animal model of KOA with cold-damp obstruction syndrome, with the ACLT combined with the artificial climate chamber being the optimal modeling approach.
8.Establishment and Evaluation of Different Disease-Syndrome Combination Mice Models of Knee Osteoarthritis with Cold-Dampness Stagnation Syndrome
Enrui HU ; Yibao WEI ; Deren LIU ; Aobuliaisan MAIMAITITUXUN ; Licheng ZHANG ; Jiachen SONG ; Jun MAO ; Taiyang LIAO ; Peimin WANG
Journal of Traditional Chinese Medicine 2026;67(19):2120-2129
ObjectiveTo establish and evaluate different disease-syndrome combination mice models of knee osteoarthritis (KOA) with cold-dampness obstruction syndrome, and to identify the optimal modeling method. MethodsEighty mice were randomly divided into a control group, a treadmill plus climate chamber group, an anterior cruciate ligament transection (ACLT) plus climate chamber group, an ACLT group, a destabilization of the medial meniscus (DMM) plus climate chamber group, a DMM group, an intra-articular injection plus climate chamber group, and an intra-articular injection group, with 10 mice in each group. Mice in the ACLT group underwent surgical transection of the knee anterior cruciate ligament to establish the model, while those in the DMM underwent surgical transection of the medial meniscal tibial ligament of the knee, and those in the intra-articular injection group were modeled by an intra-articular injection of 50 μl of normal saline containing 1mg of monosodium iodoacetate (MIA) into the medial side of the patellar ligament. Mice in the ACLT plus climate chamber, DMM plus climate chamber, and intra-articular injection plus climate chamber groups were placed in an artificial climate chamber (relative humidity 60%, temperature 4 ℃, wind speed 6 m/s) simultaneously with aforementioned interventions, once daily for 2 h each time, for 14 consecutive days. Mice in the control group underwent sham surgery, involving only the incision of the skin and subcutaneous tissue at the knee joint to expose the articular cavity, followed by layer-by-layer suturing. Mice in the treadmill plus climate chamber group were placed on an animal treadmill starting on the 4th day after the sham operation, undergoing high-intensity treadmill exercise at a speed of 20 m/min, an inclination of 5°, for 30min per session, 5 times per week for 28 consecutive days, while being housed in the artificial climate chamber. Cold pain thresholds of the mice in each group were compared using the cold plate test and the acetone low-temperature test before and after modeling. Following modeling, knee joint pathological changes were observed using HE and Safranin O-Fast Green staining, and quantified using Mankin and Osteoarthritis Research Society International (OARSI) scores. Serum levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were measured by ELISA. Protein and mRNA expression levels of pain-related markers, including calcitonin gene-related peptide (CGRP), protein gene product 9.5 (PGP9.5), and cellular proto-oncogene Fos (c-Fos), and the cold hyperalgesia-related markers including transient receptor potential ankyrin 1 (TRPA1) and transient receptor potential melastatin 8 (TRPM8) in the dorsal root ganglion (DRG) tissues were detected by Western Blot and qPCR. TRPA1 and TRPM8 fluorescent intensity in the DRG was detected by immunofluorescence assay. ResultsCompared to the control group, the remaining groups showed decreased cold pain thresholds in the cold plate tese, increased cold pain thresholds in the acetone low-temperature test, elevated Mankin and OARSI scores, and increased serum TNF-α and IL-1β levels, upregulated protein and mRNA expression levels of CGRP, PGP9.5, c-Fos, TRPA1, and TRPM8 in the DRG, and increased immunofluorescence intensities of TRPA1 and TRPM8 in the DRG tissue (P<0.05 or P<0.01), along with severe pathological damage and cartilage degeneration in the knee joints. Compared to the ACLT, DMM, and intra-articular injection groups, the ACLT plus climate chamber, DMM plus climate chamber, and intra-articular injection plus climate chamber groups exhibited more significant changes in all the aforementioned indicators (P<0.01). Furthermore, changes in the aforementioned indicators were more pronounced in the ACLT plus climate chamber group compared to the Treadmill plus climate chamber group, DMM plus climate chamber and intra-articular injection plus climate chamber group (P<0.01), exhibiting the most severe degree of pathological damage and cartilage degeneration. ConclusionThe combination of an artificial climate chamber can successfully establish a disease-syndrome combined animal model of KOA with cold-damp obstruction syndrome, with the ACLT combined with the artificial climate chamber being the optimal modeling approach.
9.Pulchinenoside A downregulates LRP1 to inhibit the proliferation, migration, and invasion of synovial inflammatory cells
Lifang MAO ; Tianyu CHEN ; Jinhong WANG ; Hao WANG ; Yingjie ZHAO ; Renpeng ZHOU ; Wei HU ; Chao LU
Acta Universitatis Medicinalis Anhui 2026;61(9):1584-1593
ObjectiveTo investigate the effects and mechanisms of pulchinenoside A (PSA) on tumor necrosis factor-α (TNF-α) -induced abnormal proliferation, migration, and invasion of synovial fibroblasts. MethodsA synovitis model was established by stimulating SW982 synovial cells with TNF-α. The low-density lipoprotein receptor-related protein 1 (LRP1) was knocked down using LRP1-specific siRNA or inhibited by the receptor-associated protein (RAP). Cells were treated with various concentrations of PSA, and cell viability was assessed by CCK-8 assay. Protein levels of LRP1, matrix metalloproteinase-3 (MMP3), and N-cadherin were measured by Western blotting. Proliferation was evaluated using an EdU incorporation assay. Migration and invasion capacities were examined by Transwell and wound-healing assays. mRNA expression of LRP1, MMP3, N-cadherin, IL-1β, IL-6, and TNF-α was detected by reverse transcription quantitative real-time PCR (RT-qPCR). Immunofluorescence staining was used to visualize the expression of MMP3 and N-cadherin. The involvement of LRP1 was further verified with the inhibitor RAP. ResultsCompared with the TNF-α-treated model group, PSA treatment significantly reduced SW982 cell viability (P<0.05), downregulated MMP3 and N-cadherin expression (both P<0.01), and decreased the proportion of EdU-positive cells (P<0.05). Moreover, TNF-α stimulation upregulated LRP1 expression compared with normal controls (P<0.01), whereas PSA treatment reversed this increase (P<0.05). LRP1 knockdown or inhibition markedly decreased MMP3 and N-cadherin levels compared with the model group (both P<0.01). Similarly, the combination of PSA and RAP led to significant reductions in MMP3 and N-cadherin expression compared with the model group (both P<0.01). ConclusionPSA inhibits TNF-α-induced abnormal proliferation, migration, and invasion of synovial, at least in part, by downregulating LRP1 expression.
10.Effect of interleukin-8 regulation on monocyte chemotactic protein-1 secretion and expression through the NF-kappaB/p65 signaling pathway on the migration of residual epithelial cells in the lens capsule
Wei SI ; Su XU ; Yuhang ZHANG ; Yi MAO ; Keyu GUO ; Yanzhong HU ; Fengyan ZHANG
International Eye Science 2025;25(4):537-543
AIM: To investigate the effect of interleukin-8(IL-8)on the regulation of monocyte chemotactic protein-1(MCP-1)secreted by lens epithelial cells(LEC)during cell migration in the development of posterior capsule opacification(PCO).METHODS: A rat lens capsule model was established and cultured in medium supplemented with 10% fetal bovine serum. Upon migration of LEC to 30%-50% of the posterior capsule, serum was removed. The capsule was subsequently divided into two groups: a control group and an IL-8(15 ng/mL)treatment group. LEC migration was captured at multiple time points. The secretion and mRNA expression of MCP-1 were quantified using ELISA and RT-qPCR, respectively. Immunofluorescence was used to assess MCP-1 expression in the different experimental groups. SRA01/04 cells were divided into three groups: control, IL-8(15 ng/mL), and IL-8(15 ng/mL)+200 μmol/L Bindarit(BND)groups, with migration measured by the Transwell assay. Additionally, SRA01/04 cells were divided into negative control(NC), NC+15 ng/mL IL-8, and 15 ng/mL IL-8+p65 siRNA groups, and MCP-1 secretion and mRNA expression were further analyzed by ELISA and RT-qPCR.RESULTS:LEC migration in the rat lens capsule cultured in vitro showed that the cells migration of the 15 ng/mL IL-8 group significantly increased at 48, 72 and 96 h(all P<0.05). ELISA results revealed that MCP-1 levels in SRA01/04 cells from the 15 ng/mL IL-8-treated group were markedly higher than those in the control group at both 12 and 24 h(all P<0.05). RT-qPCR analysis also demonstrated a significant increase in MCP-1 mRNA expression in the 15 ng/mL IL-8 group at both time points(all P<0.05). Immunofluorescence staining indicated greater MCP-1 expression in capsular epithelial cells of the 15 ng/mL IL-8 group at 24 h(P=0.007). Transwell assays further confirmed increased cell migration in the 15 ng/mL IL-8 group compared to the control group(P=0.001), while the migration reduced in the 15 ng/mL IL-8+200 μmol/L BND group compared to the 15 ng/mL IL-8 group(P=0.003). Moreover, ELISA and RT-qPCR results demonstrated a significant increase in MCP-1 secretion and mRNA expression in the NC+15 ng/mL IL-8 group at both 12 and 24 h compared to the NC group(all P<0.01). In contrast, MCP-1 secretion and mRNA expression were reduced in the 15 ng/mL IL-8+p65 siRNA group compared to the NC+15 ng/mL IL-8 group at both time points(all P<0.01).CONCLUSION: IL-8 promotes the migration of residual epithelial cells and regulates the secretion and expression of MCP-1 in LEC. The mechanism underlying IL-8's effects appears to be mediated through the activation of the NF-κB/p65 signaling pathway.

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