1.Effect of acupuncture on differentiation and apoptosis of quadriceps muscle satellite cells in knee osteoarthritis model rats and its mechanism
Qu ZHENG ; Baoqiang DONG ; Xingxing LIN ; Yu ZHANG ; Xuefeng GUAN ; Chaojie WANG ; Yiyan HAN
Journal of Jilin University(Medicine Edition) 2025;51(6):1475-1486
Objective:To discuss the effect of acupuncture on the differentiation and apoptosis of quadriceps muscle satellite cells in model rats with knee osteoarthritis(KOA),and to clarify its related mechanism.Methods:A total of 40 SPF-grade rats were selected and randomly divided into control group,model group,celecoxib group,and acupuncture group,with 10 rats in each group.The rats in control group only underwent joint cavity incision followed by suturing,while the rats in model group,celecoxib group,and acupuncture group were used to replicate the KOA models.The maximum circumference of the femoral segment of the affected limb,rat body mass,and quadriceps wet weight of the rats in various groups were measured;the quadriceps wet weight maintenance rate and quadriceps wet weight/body mass ratio of the rats in various groups were calculated.HE staining was used to observe the pathomorphology of articular cartilage and quadriceps muscle tissue of the rats in various groups;terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick end labeling(TUNEL)method was used to detect the apoptosis indexes in articular cartilage and quadriceps muscle tissue of the rats in various groups;immunofluorescence method was used to detect the protein expression levels of interleukin-6(IL-6),Janus kinase(JAK),and signal transducer and activator of transcription 3(STAT3)in quadriceps muscle tissue of the rats in various groups;Western blotting method was used to detect the expression levels of IL-6/JAK/STAT3 signaling pathway proteins,and muscle satellite cells,and apoptosis-related proteins in quadriceps muscle tissue of the rats in various groups.Results:Compared with control group,the affected hind limb circumference,quadriceps wet weight,wet weight maintenance rate,and wet weight/body mass ratio of the rats in model group were significantly decreased(P<0.05);compared with model group,the affected hind limb circumference,quadriceps wet weight,wet weight maintenance rate,and wet weight/body mass ratio of the rats in celecoxib group and acupuncture group were significantly increased(P<0.05);compared with celecoxib group,the affected hind limb circumference,quadriceps wet weight,wet weight maintenance rate,and wet weight/body mass ratio of the rats in acupuncture group were significantly increased(P<0.05).The HE staining results showed that the knee articular cartilage of the rats in control group remained intact,chondrocytes were aggregated and horizontally arranged with smooth edges,and quadriceps muscle cells were long cylindrical,orderly arranged,and regular in shape;in model group,the knee articular cartilage was thinner with rough edges,reduced number of cartilage layers,and disordered arrangement,and the quadriceps muscle fibers were disorganized,with some muscle fiber dissolution and muscle cell membrane damage,accompanied by muscle fiber fragments and a large amount of inflammatory exudate;in celecoxib group,the morphology of knee articular cartilage was generally normal,occasionally with irregular cartilage arrangement and reduced thickness,sporadically visible necrotic chondrocytes,quadriceps muscle fibers and sarcolemma were relatively intact,new muscle fibers appeared,some muscle fiber edges were blurred,accompanied by a small amount of cell debris and mild inflammatory infiltration;in acupuncture group,the knee articular cartilage structure remained intact with smooth edges,occasionally rough edges,and chondrocytes were aggregated and orderly arranged.The TUNEL assay results showed that compared with control group,the apoptosis indexes in articular cartilage and quadriceps muscle tissue of the rats in model group were significantly increased(P<0.05);compared with model group,the apoptosis indexes in articular cartilage and quadriceps muscle tissue of the rats in celecoxib group and acupuncture group were significantly decreased(P<0.05);compared with celecoxib group,the apoptosis index in articular cartilage and quadriceps muscle tissue of the rats in acupuncture group were significantly decreased(P<0.05).The immunofluorescence assay results showed that compared with control group,the expression levels of IL-6,JAK,and STAT3 proteins in quadriceps muscle tissue of the rats in model group were significantly decreased(P<0.05);compared with model group,the expression levels of IL-6,JAK,and STAT3 proteins in quadriceps muscle tissue of the rats in celecoxib group and acupuncture group were significantly increased(P<0.05);compared with celecoxib group,the expression levels of IL-6,JAK,and STAT3 proteins in quadriceps muscle tissue of the rats in acupuncture group were significantly increased(P<0.05).The Western blotting results showed that compared with control group,the expression levels of IL-6,JAK,STAT3,paired box transcription factor 7(Pax7),Desmin,Myosin,and Myogenin proteins in quadriceps muscle tissue of the rats in model group were significantly decreased(P<0.05);compared with model group,the expression levels of IL-6,JAK,STAT3,Pax7,Desmin,Myosin,and Myogenin proteins in quadriceps muscle tissue of the rats in celecoxib group and acupuncture group were significantly increased(P<0.05);compared with celecoxib group,the expression levels of IL-6,JAK,STAT3,Pax7,Desmin,Myosin,and Myogenin proteins in quadriceps muscle tissue of the rats in acupuncture group were significantly increased(P<0.05).Compared with control group,the expression levels of B-cell lymphoma 2(Bcl-2),B-cell lymphoma-xl(Bcl-xl),and myeloid cell leukemia 1(MCL1)proteins in quadriceps muscle tissue in model group were significantly decreased(P<0.05),and the expression levels of Bcl-2-associated X protein(Bax)and cysteinyl aspartate specific proteinase-3(Caspase-3)proteins were significantly increased(P<0.05);compared with model group,the expression levels of Bcl-2,Bcl-xl,and MCL1 proteins in quadriceps muscle tissue of the rats in celecoxib group and acupuncture group were significantly increased(P<0.05),and the expression levels of Bax and Caspase-3 proteins were significantly decreased(P<0.05);compared with celecoxib group,the expression levels of Bcl-2,Bcl-xl,and MCL1 proteins in quadriceps muscle tissue of the rats in acupuncture group were significantly increased(P<0.05),and the expression levels of Bax and Caspase-3 proteins were significantly decreased(P<0.05).Conclusion:Acupuncture can promote the differentiation of quadriceps muscle satellite cells and inhibit muscle cell apoptosis in the model rats with KOA,and the mechanism may be related to the up-regulation of expressions of IL-6,JAK,and STAT3 proteins in the quadriceps muscle tissue.
2.Correlation between lower limb alignment and risk factors of patellofemoral pain syndrome in young men
Bing HAN ; Hongbin LIU ; Hehong WANG ; Hanqing ZHAO ; Riguang ZHAO ; Yiyan SUN ; Yu ZHANG
Chinese Journal of Tissue Engineering Research 2024;28(8):1211-1216
BACKGROUND:The risk factors for patellofemoral pain syndrome are still unclear and research on risk factors is gradually becoming a hot topic in order to improve the therapeutic outcome of this disease. OBJECTIVE:To investigate the relationship between lower limb alignment and patellofemoral pain syndrome in young male runners. METHODS:In the same running population,21 patients with bilateral knee pain,29 patients with single knee pain and 20 normal runners were collected from January 2021 to July 2022.Full-length X-ray examination of both lower limbs in standing position was performed in all subjects.The lateral distal femoral angle,medial tibial proximal angle,joint line congruence angle,hip-knee-ankle angle,and knee physical valgus angle were measured and statistically analyzed. RESULTS AND CONCLUSION:There was no significant difference in the parameters between the left and right knees in each group(P>0.05).Compared with the normal group,no significant changes in lateral distal femoral angle,medial tibial proximal angle,joint line congruence angle,and hip-knee-ankle angle were observed in the single knee pain group and double knee pain group,while the knee physical valgus angle was significantly increased in these two groups(P<0.05).To conclude,increased knee physical valgus angle may be a risk factor for patellofemoral pain syndrome.
3.Celoside I enhances mitophagy of retinal ganglion cells in optic nerve in-jury model by regulating ROS-mediated JNK/c-Jun signaling pathway
Yiyan HAN ; Qu ZHENG ; Lei ZHAO ; Zhihao NING ; Baoqiang DONG ; Tao ZUO
Chinese Journal of Pathophysiology 2024;40(10):1898-1905
AIM:This study aimed to investigate the mechanism by which celoside I enhances mitophagy in a model of optic nerve injury through regulation of reactive oxygen species(ROS)-mediated c-Jun N-terminal kinase(JNK)/c-Jun signaling pathway.METHODS:Twenty-four New Zealand white rabbits were randomly divided into four groups:sham surgery,model,mecobalamin,and experimental group.Optic nerve injury was induced in the model,mecobala-min,and experimental groups,while the sham surgery group underwent a sham procedure.The mecobalamin group re-ceived mecobalamin,the experimental group received celoside I,and the sham surgery and model groups received saline.Interventions were administered daily for 28 d.Various techniques including endoscopy,hematoxylin-eosin(HE)stain-ing,TUNEL method,immunofluorescence staining and Western blot were used to assess fundus condition,retinal mor-phology,apoptosis,ROS expression,and protein levels in the retina.RESULTS:Fundus examination revealed im-proved blood flow in the mecobalamin and experimental groups compared to the model group.Retinal morphology showed enhanced retinal ganglion cells(RGCs)in the mecobalamin and experimental groups.Apoptosis index was lower in the mecobalamin group compared to the experimental group.Immunofluorescence staining indicated reduced ROS and P62 ex-pression and increased parkin and microtubule-associated protein light chain 3(LC3)expression in the experimental group compared to the mecobalamin group.Protein analysis showed decreased JNK,c-Jun,and P62 levels,and increased parkin and LC3 levels in the mecobalamin and experimental groups compared to the model group.CONCLUSION:Celo-side I reduces ROS expression,inhibits the JNK/c-Jun pathway,enhances mitophagy,reduces apoptosis,and protects RGCs in optic nerve injury models.
4.Study on protective effects and mechanism of ziyuglycoside Ⅰ on acute lung injury in sepsis rats
Daiqin BAO ; Yiyan LIU ; Zisen ZHANG ; Han SHE ; Lei TAN ; Tao LI ; Qingxiang MAO ; Liangming LIU
China Pharmacy 2023;34(5):537-543
OBJECTIVE To investigate the protective effects and mechanism of ziyuglycoside Ⅰ on acute lung injury in sepsis rats based on network pharmacology, and conduct experimental verification. METHODS The network pharmacology was used to predict the potential target of ziyuglycoside Ⅰ in the treatment of acute lung injury following sepsis. The rat model of sepsis was reproduced by cecum ligation and puncture for experimental verification. Totally 192 SD rats were randomly divided into the sham operation group (Sham group), sepsis group (Sep group), conventional therapy group (CT group) and ziyuglycoside Ⅰ group (Zg Ⅰ group), respectively. Sham group and Sep group were given sterile normal saline, and CT group and ZgⅠ group were given relevant volume of Ringer’s solution and ziyuglycoside Ⅰ. The arterial blood gas, serum inflammatory factors, lung wet/dry mass ratio, pathological changes of lung tissue, pulmonary vascular permeability, the expressions of pulmonary vein tight junction protein 1 (ZO-1) and vascular endothelial cadherin (VE-cadherin) protein and 72-hour survival were observed in each group. RESULTS Results of network pharmacology showed that there were 47 potential targets of ziyuglycoside Ⅰ in the treatment of sepsis. The results of gene ontology function enrichment analysis and Kyoto encyclopedia of genes and genomes pathway enrichment analysis showed that the mechanism could 598486924@qq.com be correlated with biological processes such as positive regulation of reactive oxygen species metabolism, wound healing, regulation of endothelial cell proliferation, cell activation, blood vessel development, response to oxidative stress, etc., and with signaling pathway such as apoptosis, tight junction, HIF-1 signaling pathway, etc. The results of experimental verification showed that compared with Sham group, pH value and the level of partial arterial oxygen pressure were decreased significantly in Sep group (P<0.05), while the level of partial pressure of carbon dioxide, serum levels of tumor necrosis factor α, interleukin 6 were increased significantly (P<0.05); the ratio of lung wet/dry mass was increased significantly (P<0.05); the protein expressions of ZO-1 and VE-cadherin were decreased significantly (P<0.05); 72 h survival rate decreased,the survival time was significantly shortened (P<0.05); the results of pathological observation of lung tissue showed that the rats’ alveoli were extensively ruptured, the alveolar wall was thickened and accompanied with edema, and there was obvious inflammatory cell infiltration; the results of pulmonary vascular permeability observation showed that the lung surface of rats was dark, with a large amount of Evans blue exudation, and the left lower lung was obviously dark blue. Compared with Sep group, the levels of above indexes almost were reversed significantly in CT group and ZgⅠ group (P<0.05); the lung histopathology and pulmonary vascular permeability were significantly improved, and the recovery degree of ZgⅠ group was greater than that of CT group, which was close to the results of Sham group. CONCLUSIONS Ziyuglycoside Ⅰ can significantly reduce inflammatory reaction and acute lung injury in septic rats, which is related to vascular function and tight junction signal pathway.
5.Clinicopathological features and prognosis of immunoglobulin A nephropathy after renal transplantation
Tianjing ZHANG ; Pingfan LU ; Yuanjun DENG ; Yang CAI ; Lele LIU ; Chunjiang ZHANG ; Yiyan GUO ; Qian LI ; Na ZHU ; Beichen TIAN ; Min HAN
Chinese Journal of Organ Transplantation 2020;41(2):84-88
Objective:To summarize the relationship between the clinicopathological features and prognosis of immunoglobulin A nephropathy (IgAN) after renal transplantation.Methods:A total of 34 patients with IgAN after renal transplantation confirmed by renal biopsy were enrolled. And another 34 patients with primary IgAN confirmed by initial renal biopsy were adopted as controls. Clinical and pathological features of two groups were compared to explore the relationship between clinicopathological features and prognosis of allograft IgAN.Results:As compared with primary IgAN group, renal function in allograft IgAN group included serum creatinine [(158.5±75.9) vs (84.8±26.8) umol/L], urea nitrogen [(9.7±6.1) vs (5.2±1.4) mmol/L], uric acid [(406.7±87.8) vs (359.0±92.6) umol/L], estimated glomerular filtration rate {(57.4±25.4) vs (91.2±28.6) [ml/(min·1.73m 2)]}. All were statistically significantly higher ( P<0.05) while other parameters showed no differences. Pathologically, the proportion of T1 type (50.0% vs 17.6%) of renal tubular atrophy/interstitial fibrosis was significantly higher in allograft IgAN group than control group ( P<0.05). Furthermore, univariate and multivariate Logistic regression analyses were performed between various pathological parameters and prognosis in allograft IgAN patients. It indicated that the degree of mesangial hyperplasia of patients with transplanted IgAN had a significantly negative impact on the prognosis. Conclusions:The clinicopathological features of patients with allograft IgAN show no difference from those of patients with primary IgAN. And among patients with allograft IgAN, those with severe mesangial hyperplasia often have a worse prognosis.
6.Expression and significance of Mnk2 and eIF4E in esophageal squamous cell carcinoma
Bo ZENG ; Yanfen FENG ; Qitao HUANG ; Jingfu CHEN ; Xin ZHANG ; Xiangqian HAN ; Shuishen ZHANG ; Jianyong ZOU ; Chunhua SU ; Zhenguang CHEN ; Honghe LUO ; Yiyan LEI
Chinese Journal of Pathophysiology 2017;33(2):349-352
AIM: To investigate the expression and significance of MAPK-interacting kinase-2 ( Mnk2 ) and eukaryotic initiation factor 4E ( eIF4E) in the patients with resected esophageal squamous cell carcinoma ( ESCC ). METHODS:The protein expression of Mnk2 and eIF4E in ESCC tissues (98 cases) and normal esophageal tissues (20 cases) were assessed by immunohistochemistry (IHC), and their correlations with clinicopathological features were statisti-cally analyzed.RESULTS:The over-expression rate of Mnk2 and eIF4E was 68.4%(67/98) and 61.2%(60/98), re-spectively.The expression of Mnk2 had a positive correlation with eIF4E (P<0.05).Clinicopathologic analysis showed that Mnk2 expression was significantly correlated with T classification ( P<0.05 ) and clinical stage ( P<0.05 ) .CON-CLUSION:The over-expression of Mnk2 was significantly related to the tumor invasive depth , TNM stages and expression of eIF4E in ESCC.Expression of Mnk2 and eIF4E may have a cooperative formation mechanism in the development of ESCC.

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