1.Standard deviation method and root mean square method for measuring signal-to-noise ratio of MRI of American College of Radiology phantom based on phased array coils
Songlin SHA ; Qi LIU ; Yanling BAI ; Jin MA ; Hongxia ZHANG ; Yunfeng SUN ; Jianjie LU
Chinese Journal of Medical Imaging Technology 2025;41(1):138-141
Objective To compare the value of standard deviation(SD)method and root mean square(RMS)method for measuring signal-to-noise ratio(SNR)of MRI of American College of Radiology(ACR)phantom based on phased array coils.Methods ACR phantom was scanned with head coil(Head),abdominal coil(Anterior),dual piece flexible coil(Flex L)and abdominal coil+flexible coil(Anterior+Flex L)each for 6 times,respectively,with sequences of axial T 1W-spin echo(SE)and T2W-SE.And the scanning schemes were divided into 8 groups based on 4 types of coils and 2 sequences.SNR of 8 groups were measured with standard deviation method and root mean square method,respectively,and the differences between the above 2 methods were observed.Results For T1W-SE sequence,SNRsD of MRI based on Head,Anterior,Flex L and Anterior+Flex L was 1.25±0.07,0.56±0.02,1.15±0.10 and 1.04±0.11,respectively,while SNRRMs was 1.10±0.07,0.58±0.03,1.01±0.13 and 1.31±0.15,respectively.For T2W-SE sequence,SNRsD was 1.40±0.08,0.48±0.02,1.04±0.07 and 1.08±0.06,respectively,while SNRRMs was 1.29±0.09,0.53±0.03,0.84±0.08 and 1.35±0.12,respectively.Compared pairwise,significant difference of SNRsD was found in 9 pairs(all P<0.05),while of SNRRMs was detected in 10 pairs(all P<0.05).Conclusion Compared with standard deviation method,root mean square method was more suitable for measuring SNR of MRI of ACR phantom based on phased array coils.
2.Effects of heme-binding protein 1 gene knockdown on proliferation,migration,and inflammatory response of microglia BV2 and their mechanisms
Sifan FENG ; Yunfeng LI ; Jiaying WANG ; Fubin MA ; Yan WANG
Journal of Jilin University(Medicine Edition) 2025;51(6):1532-1541
Objective:To explore the effect of heme-binding protein 1(HEBP1)down-regulation on the function of microglia BV2,and to clarify the key role of HEBP1 in the microglia.Methods:Negative control and HEBP1 knockdown small interfering RNA(siRNA)were constructed to knockdown HEBP1 gene in mouse-derived microglial BV2,and the HEBP1 knockdown BV2 cell models were obtained.The BV2 cells were divided into si-NC group,si-HEBP1-1 group,si-HEBP1-2 group,and si-HEBP1-3 group.Real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods were used to detect the expression levels of HEBP1 mRNA and protein in the BV2 cells after knockdown;the siRNA with the most significart knockdown effect was selected for stlbsequent expreriments.The proliferation abilities of the cells in si-NC group and si-HEBP1 group were detected by cell counting kit-8(CCK-8)assay,and the cell migration rates were assessed by scratch assay;the cellular mitochondrial membrane potential and reactive oxygen species(ROS)levels were detected by kits;the cellular mitochondrial respiratory function was detected by mitochondrial respirometer.The BV2 cells were divided into si-NC group,si-NC+lipopolysacch aride(LPS)group,si-HEBP1 group,and si-HEBP1+LPS group.RT-qPCR method was used to detect the expression levels of HEBP1,interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6)mRNA in the BV2 cells in various groups,and Western blotting method was used to detect the expression levels of HEBP1 protein in the BV2 cells in various groups.Results:When the BV2 cells were transfected with siRNA carrying with red fluorescence tag CY3,the transfect effricacy was above 90%;compared with si-NC group,the expression levels of HEBP1 protein in the BV2 cells in si-HEBP1-1 group,si-HEBP1-2 group,and si-HEBP1-3 group were significantly decreased(P<0.05 or P<0.01),especially in si-HEBP1-1 group.Compared with si-NC group,the expression levels of HEBP1 mRNA in the BV2 cells in si-HEBP1-1 group,si-HEBP1-2 group,and si-HEBP1-3 group were significantly decreased(P<0.01),especially in si-HEBP1-1 group;indicating that si-HEBP1-1 was the siRNA with best HEBP1 knowdown effect,and the HEBP1 knockdown BV2 cell model was successfully constructed.The CCK-8 resuts showed that compared with si-NC group,the proliferation activities of the BV2 cells in si-HEBP1 group were decreased(P<0.05 or P<0.01);from 90 min,the differences in proliferation activities of the BV2 cells in two groups were obvious.The cell scratch experiment results showed that compared with si-NC group,the cell migration rate in si-HEBP1 group was significantly decreased(P<0.05).The fluorescence microscope results showed that compared with si-NC group,the mitochondrial membrane potential of the BV2 cells in si-HEBP1 group was significantly decreased(P<0.05);compared with si-NC group,the ROS level in the BV2 cells in si-HEBP1 group was significantly increased(P<0.05).The mitochondrial respiration function testing results showed that compared with si-NC group,routine respiration(ROUNTINE)and leak respiration(LEAK)in si-HEBP1 group were significautly decreased(P<0.05 or P<0.01),and electron transfer system capacity(ETS)and residual oxygen consumption(ROX)had no significant differences(P>0.05);the ATP amount was decreased(P<0.05).The RT-qPCR results showed that compared with si-NC group,the expression levels of IL-1β,TNF-α,and IL-6 mRNA in the BV2 cells in si-NC+LPS group were significantly decreased(P<0.01);compared with si-HEBP1 group,the expression levels of IL-1β,TNF-α,and IL-6 mRNA in the BV2 cells in si-HEBP1+LPS group were significantly decreased(P<0.01);compared with si-NC+LPS group,the expression levels of IL-1β,TNF-α,and IL-6 mRNA in the BV2 cells in si-HEBP1+LPS group were significantly increased(P<0.01).Conclusion:Knockdown of HEBP1 gene can decrease the proliferation and migration abilities of the microglia BV2 and increase inflammatory response to LPS stimulus,and their mechanisms may be related to mitochondrial function damage and decreased ATP production of the BV2 cells.
3.Safety and efficacy of early precise lower limb weight-bearing rehabilitation after open reduction and internal fixation of ankle fractures
Jianfeng XUE ; Mingjie TANG ; Lei WANG ; Xu WANG ; Jianhua HUANG ; Yunfeng YANG ; Lei SHEN ; Chao ZHANG ; Fucun LIU ; Yunfeng CHEN ; Xin MA ; Zhongmin SHI
Chinese Journal of Orthopaedic Trauma 2025;27(7):557-564
Objective:To evaluate the safety and efficacy of early precise lower limb weight-bearing functional rehabilitation after open reduction and internal fixation of ankle fractures.Methods:A restropective multi-center study was conducted to enroll the eligible 120 patients with malleolar fracture who received the same surgical treatment from March 2023 to December 2023 at the trauma centers in 7 tertiary hospitals according to the inclusion criteria. They were assigned into a study group ( n=60) for precise lower limb weight-bearing functional rehabilitation with the assistance of intelligent lower limb walking rehabilitation crutches and a control group ( n=60) for conventional weight-bearing functional rehabilitation which was gradually increased according to their own feelings under the protection of a walking boot. All the 60 patients in the study group completed their final follow-ups. They were 25 males and 35 females, with an age of (43.8±16.6) years and a body mass index of (25.3±2.3) kg/m 2. Only 51 patients in the control group completed their final follow-ups. They were 27 males and 24 females, with an age of (45.1±16.4) years and a body mass index of (24.7±2.3) kg/m 2. When their incisions healed and their sutures were removed 2 weeks after operation, the patients were guided to start lower limb weight-bearing functional rehabilitation, and exercises for foot and ankle joint mobility and lower limb muscles. The end point of follow-up was 12 weeks after operation. The 2 groups were compared in terms of the crutch-off rates, thigh circumferences, calf circumferences, dorsiflexions, plantarflexions, ankle swellings, American Orthopaedic Foot & Ankle Society (AOFAS) ankle-hindfoot scores, and visual analog scale (VAS) pain scores at 6 and 12 weeks after operation. The complications were also recorded in the 2 groups. Results:No statistically significant differences were observed in the baseline characteristics between the 2 groups, indicating comparability ( P>0.05). At 6 and 12 weeks after operation, the crutch-off rates [41.7% (25/60) and 100.0% (60/60)], dorsiflexions (69.3%±21.6% and 82.9%±26.3%) and AOFAS ankle-hindfoot scores [(68.5±7.6) points and (96.9±3.7) points] in the study group were significantly better than those in the control group [13.7% (7/51) and 39.2% (20/51), 61.5%±16.5% and 72.0%±14.3%, (61.9±9.3) points and (90.1±7.2) points] ( P<0.05). At 6 weeks after operation, the thigh circumference (97.4%±1.9%), calf circumference (97.3%±1.9%), and plantarflexion (76.6%±19.8%) in the study group were significantly higher than those in the control group (95.9%±2.5%, 94.6%±3.2%, and 63.9%±16.9%) ( P<0.05). There were no significant differences between the 2 groups in ankle swelling at 6 or 12 weeks after operation, or thigh or calf circumference, plantarflexion, or VAS pain score at 12 weeks after operation ( P>0.05). No wound complications, secondary fracture displacement, or loosening of internal fixation occurred in either group during the follow-up period. There were no cases of nonunion or delayed union. Conclusions:Early lower limb weight-bearing functional rehabilitation after open reduction and internal fixation of ankle fractures demonstrates good safety. Precise weight-bearing rehabilitation accelerates functional recovery of the ankle, enabling earlier return to normal daily activities.
4.Engineering of mesenchymal stem cell-derived exosomes and their application progress in the field of oral medicine
ZHAO Yunfeng ; LIU Qian ; LI Meng ; LI Luying ; ZHANG Wei ; HU Xiantong ; MA Chufan
Journal of Prevention and Treatment for Stomatological Diseases 2025;33(9):792-800
In recent years, mesenchymal stem cell-derived exosomes (MSC-EXO) have garnered increasing attention in the field of stomatology and have become an established research area in biomedical research. This article reviews the engineering of exosomes derived from mesenchymal stem cells and their application in the field of stomatology, in order to provide new ideas for the development of stomatology. Exosomes are nanoscale membrane vesicles secreted by cells and contain a variety of proteins, RNAs, lipids, and other biomolecules. They are transported through the circulatory system and can interact with other cells to regulate their biological behavior and participate in a variety of physiological and pathological processes. In the treatment of oral diseases, exosomes have shown great potential due to their natural biological activity and versatility. However, studies have found that relying solely on the function of natural exosomes may not fully meet the complex clinical requirements. Therefore, the concept of engineered exosomes has emerged. Engineered exosomes can be modified by bioengineering technology to enhance their targeting, allowing them to reach the lesion site more accurately. At the same time, engineered exosomes can also be surface modified or loaded internally to carry specific therapeutic molecules, such as drugs, gene editing tools or signaling molecules to improve the therapeutic effect. In addition, this engineered treatment can also confer greater stability to exosomes, making them better able to resist clearance by the immune system when circulating in the body, extending their half-life, and improving the effectiveness of treatment. Although engineered exosomes have attracted extensive attention in the fields of stomatology and other fields, their application is still mainly in the stage of basic research. To promote the clinical application of engineered exosomes, it is necessary to provide more sufficient evidence of biocompatibility and clarify their therapeutic effect and mechanism.
5.Lupeol Alleviates Chondrocytes Senescence in Osteoarthritis by Regulating Autophagy via the Sirtuin 3/Mechanistic Target of Rapamycin Kinase Pathway
Yunfeng MA ; Yujing CAO ; Xiaofei HAN
Journal of Sichuan University (Medical Sciences) 2025;56(1):83-93
Objective To investigate the role of lupeol in mitigating chondrocyte senescence in osteoarthritis(OA)by regulating autophagy through the sirtuin 3(SIRT3)/mechanistic target of rapamycin kinase(mTOR)pathway.Methods Knee articular chondrocytes from primary-generation mice were isolated and divided into different groups,including a control group,a lupeol group(given 2.5,5,10,20,and 40 μmol/L lupeol),a tert-butyl hydrogen peroxide(TBHP)group(receiving 50 μmol/L TBHP),TBHP+lupeol group,TBHP+lupeol+chloroquine(CQ)group(receiving 20 μmol/L CQ,an autophagy inhibitor),TBHP+lupeol+si-NC group,and TBHP+lupeol+si-SIRT3 group.Cell proliferation,reactive oxygen species(ROS)levels,and apoptosis were determined by CCK-8,DCFH-DA probe,and flow cytometry.Cell senescence was evaluated by β-gal staining.Western blot was used to determine the expressions of SIRT3,mTOR,senescence marker proteins(p21 and p16),extracellular matrix(ECM)degradation-related proteins(aggrecan,collagen Ⅱ,ADAMTS5,and MMP13),and autophagy-related proteins(LC3B Ⅰ,LC3BⅡ,and P62).RT-qPCR was used to determine the mRNA levels of senescence-associated secretory phenotypes(SASP),including IL-6,Cxcl10,MCP1,and MMP3.The expression of LC3 was detected by immunofluorescence.Autophagosomes were observed by transmission electron microscopy.A total of 30 male wild-type C57BL/6 mice were divided into different groups(n=10),including a Sham group,an OA group,and an OA+lupeol group receiving 50 mg/(kg·d)lupeol via gastric gavage.Cartilage damage was evaluated by safranin O-fast green staining.Results Based on the results of cell viability assay,20 μmol/L lupeol treatment for 24 h was identified as the optimal intervention concentration and duration.Compared with that in the TBHP group,cell viability was elevated in the TBHP+lupeol group(P<0.05);ROS production,the proportion of β-gal-positive cells,the protein expression levels of p21 and p16,and the mRNA levels of SASP were decreased(P<0.05);the protein levels of aggrecan and collagen Ⅱ were elevated and the protein levels of ADAMTS5 and MMP13 were decreased(P<0.05);apoptosis was reduced(P<0.05);P62 protein levels were reduced and the LC3B Ⅱ/LC3B Ⅰ ratio,the intensity of LC3B fluorescence spots,and the number of autophagosomes were increased(P<0.05);the expression level of SIRT3 was elevated and the level of mTOR phosphorylation was reduced(P<0.05)in the TBHP+Lupeol group.CQ treatment effectively abolished the promotion effects of lupeol on cell viability and autophagy,and the inhibitory effects of lupeol on ROS level,cell senescence,ECM degradation,and apoptosis(P<0.05).Silencing of SIRT3 reversed the inhibitory effect of lupeol on mTOR phosphorylation level and the promotion effect of lupeol on autophagy(P<0.05).In the in vivo experiment,compared with the OA group,the OA+lupeol group showed reduced cartilage degeneration and lower scores for the Osteoarthritis Research Society International grading system(P<0.05).The OA+lupeol group also showed up-regulated SIRT3 expression,reduced mTOR phosphorylation level,increased LC3B Ⅱ/LC3B Ⅰ ratio,reduced MMP13 protein level,and reduced mRNA level of SASP(P<0.05).Conclusion Lupeol alleviates chondrocyte senescence in osteoarthritis by regulating autophagy through the SIRT3/mTOR pathway.
6.Significance of precise classification of sacral meningeal cysts by multiple dimensions radiographic reconstruction MRI in guiding operative strategy and rehabilitation.
Jianjun SUN ; Qianquan MA ; Xiaoliang YIN ; Chenlong YANG ; Jia ZHANG ; Suhua CHEN ; Chao WU ; Jingcheng XIE ; Yunfeng HAN ; Guozhong LIN ; Yu SI ; Jun YANG ; Haibo WU ; Qiang ZHAO
Journal of Peking University(Health Sciences) 2025;57(2):303-308
OBJECTIVE:
To precise classify sacral meningeal cysts, effective guide minimally invasive neurosurgery and postoperative personalized rehabilitation by multiple dimensions radiographic reconstruction MRI.
METHODS:
From March to December 2021, based on the original 3D-fast imaging employing steadystate acquisition (FIESTA) scanning sequence, 92 patients with sacral meningeal cysts were pre-operatively evaluated by multiple dimensional reconstruction MRI. The shape of nerve root and the leakage of cyst were reconstructed according to the direction of nerve root or leakage track showed on original MRI scans. Sacral canal cysts were accurately classified as including nerve root and without nerve root, so as to accurately design the incision of skin and formulate corresponding open range of the posterior wall of the sacral canal. Under the microscope intraoperation, the shape of the nerve roots inside cysts or leakage track of the cysts without nerve roots were verified and explored. After the reinforcement and shaping operation, several reexaminations of multiple dimensional reconstruction MRI were performed to understand the deformation of the nerve root and hydrops in the operation cavity, so as to formulate a persona-lized rehabilitation plan for the patients.
RESULTS:
Among the 92 patients with sacral mengingeal cyst, 58 (63.0%) cysts with nerve root cyst, 29 (31.5%) cysts without nerve root cyst, and 5 (5.4%) cysts with mixed sacral canal cyst. In 58 patients with nerve root cysts, the accuracy of preoperative clinical classification on MRI image reached 96.6% (56/58) through confirmation by operating microscope. Only 2 cases of large single cyst with nerve root on the head of cyst were mistaken for without nerve root type. In 29 patients with sacral cyst without nerve root, the accuracy of preoperative image reached 100% through confirmation by operating microscope. The accuracy of judging the internal nerve root and leakage of 12 cases with recurrent sacral cyst was also 100%. Two cases of delayed postoperative hydrops were found one month after operation. After rehabilitation treatment by moxibustion and bathing, the hydrops disappeared 4-6 months after operation.
CONCLUSION
Multiple dimensional reconstruction MRI can precisely make clinical classification of sacral meningeal cysts before operation, guide minimally invasive neurosurgery effectively, and improve the rehabilitation effect.
Humans
;
Magnetic Resonance Imaging/methods*
;
Male
;
Female
;
Sacrum/surgery*
;
Adult
;
Middle Aged
;
Imaging, Three-Dimensional/methods*
;
Cysts/rehabilitation*
;
Aged
;
Adolescent
;
Young Adult
;
Spinal Nerve Roots/diagnostic imaging*
;
Minimally Invasive Surgical Procedures
;
Neurosurgical Procedures/methods*
7.Standard deviation method and root mean square method for measuring signal-to-noise ratio of MRI of American College of Radiology phantom based on phased array coils
Songlin SHA ; Qi LIU ; Yanling BAI ; Jin MA ; Hongxia ZHANG ; Yunfeng SUN ; Jianjie LU
Chinese Journal of Medical Imaging Technology 2025;41(1):138-141
Objective To compare the value of standard deviation(SD)method and root mean square(RMS)method for measuring signal-to-noise ratio(SNR)of MRI of American College of Radiology(ACR)phantom based on phased array coils.Methods ACR phantom was scanned with head coil(Head),abdominal coil(Anterior),dual piece flexible coil(Flex L)and abdominal coil+flexible coil(Anterior+Flex L)each for 6 times,respectively,with sequences of axial T 1W-spin echo(SE)and T2W-SE.And the scanning schemes were divided into 8 groups based on 4 types of coils and 2 sequences.SNR of 8 groups were measured with standard deviation method and root mean square method,respectively,and the differences between the above 2 methods were observed.Results For T1W-SE sequence,SNRsD of MRI based on Head,Anterior,Flex L and Anterior+Flex L was 1.25±0.07,0.56±0.02,1.15±0.10 and 1.04±0.11,respectively,while SNRRMs was 1.10±0.07,0.58±0.03,1.01±0.13 and 1.31±0.15,respectively.For T2W-SE sequence,SNRsD was 1.40±0.08,0.48±0.02,1.04±0.07 and 1.08±0.06,respectively,while SNRRMs was 1.29±0.09,0.53±0.03,0.84±0.08 and 1.35±0.12,respectively.Compared pairwise,significant difference of SNRsD was found in 9 pairs(all P<0.05),while of SNRRMs was detected in 10 pairs(all P<0.05).Conclusion Compared with standard deviation method,root mean square method was more suitable for measuring SNR of MRI of ACR phantom based on phased array coils.
8.Safety and efficacy of early precise lower limb weight-bearing rehabilitation after open reduction and internal fixation of ankle fractures
Jianfeng XUE ; Mingjie TANG ; Lei WANG ; Xu WANG ; Jianhua HUANG ; Yunfeng YANG ; Lei SHEN ; Chao ZHANG ; Fucun LIU ; Yunfeng CHEN ; Xin MA ; Zhongmin SHI
Chinese Journal of Orthopaedic Trauma 2025;27(7):557-564
Objective:To evaluate the safety and efficacy of early precise lower limb weight-bearing functional rehabilitation after open reduction and internal fixation of ankle fractures.Methods:A restropective multi-center study was conducted to enroll the eligible 120 patients with malleolar fracture who received the same surgical treatment from March 2023 to December 2023 at the trauma centers in 7 tertiary hospitals according to the inclusion criteria. They were assigned into a study group ( n=60) for precise lower limb weight-bearing functional rehabilitation with the assistance of intelligent lower limb walking rehabilitation crutches and a control group ( n=60) for conventional weight-bearing functional rehabilitation which was gradually increased according to their own feelings under the protection of a walking boot. All the 60 patients in the study group completed their final follow-ups. They were 25 males and 35 females, with an age of (43.8±16.6) years and a body mass index of (25.3±2.3) kg/m 2. Only 51 patients in the control group completed their final follow-ups. They were 27 males and 24 females, with an age of (45.1±16.4) years and a body mass index of (24.7±2.3) kg/m 2. When their incisions healed and their sutures were removed 2 weeks after operation, the patients were guided to start lower limb weight-bearing functional rehabilitation, and exercises for foot and ankle joint mobility and lower limb muscles. The end point of follow-up was 12 weeks after operation. The 2 groups were compared in terms of the crutch-off rates, thigh circumferences, calf circumferences, dorsiflexions, plantarflexions, ankle swellings, American Orthopaedic Foot & Ankle Society (AOFAS) ankle-hindfoot scores, and visual analog scale (VAS) pain scores at 6 and 12 weeks after operation. The complications were also recorded in the 2 groups. Results:No statistically significant differences were observed in the baseline characteristics between the 2 groups, indicating comparability ( P>0.05). At 6 and 12 weeks after operation, the crutch-off rates [41.7% (25/60) and 100.0% (60/60)], dorsiflexions (69.3%±21.6% and 82.9%±26.3%) and AOFAS ankle-hindfoot scores [(68.5±7.6) points and (96.9±3.7) points] in the study group were significantly better than those in the control group [13.7% (7/51) and 39.2% (20/51), 61.5%±16.5% and 72.0%±14.3%, (61.9±9.3) points and (90.1±7.2) points] ( P<0.05). At 6 weeks after operation, the thigh circumference (97.4%±1.9%), calf circumference (97.3%±1.9%), and plantarflexion (76.6%±19.8%) in the study group were significantly higher than those in the control group (95.9%±2.5%, 94.6%±3.2%, and 63.9%±16.9%) ( P<0.05). There were no significant differences between the 2 groups in ankle swelling at 6 or 12 weeks after operation, or thigh or calf circumference, plantarflexion, or VAS pain score at 12 weeks after operation ( P>0.05). No wound complications, secondary fracture displacement, or loosening of internal fixation occurred in either group during the follow-up period. There were no cases of nonunion or delayed union. Conclusions:Early lower limb weight-bearing functional rehabilitation after open reduction and internal fixation of ankle fractures demonstrates good safety. Precise weight-bearing rehabilitation accelerates functional recovery of the ankle, enabling earlier return to normal daily activities.
9.Eriodictyol accelerates glucocorticoid-induced apoptosis by promoting osteoblast autophagy
Chinese Journal of Tissue Engineering Research 2024;28(28):4498-4504
BACKGROUND:Glucocorticoid-induced osteoporosis is a common complication of systemic glucocorticoid therapy,which is mainly characterized by its inhibitory effect on osteoblasts.Eriodictyol inhibits osteoclast differentiation and osteoporosis-induced by ovariectomy.However,it is unclear whether eriodictyol regulates glucocorticoid-induced osteoblasts. OBJECTIVE:To explore whether eriodictyol plays a role in glucocorticoid-induced osteoblast apoptosis and its potential regulatory mechanisms. METHODS:Dexamethasone-pretreated osteoblasts MC3T3-E1 were treated with the different concentrations(0,0.5,1,2.5,5,10 μmol/L)of eriodictyol or 5 μmol/L 3-methyladenine,an autophagy inhibitor,and then transfected with heme oxygenase 1 overexpression vector(pcDNA-HMOX1)and empty vector(pcDNA vector).Cell proliferation and apoptosis were assessed by using cell counting kit-8 assay and flow cytometry,respectively.The activity of caspase-3 was detected with ELISA.Western blot assay was used to detect the protein expression of autophagy-related proteins LC3-Ⅱ/LC3-Ⅰ,p62,Atg5 and Atg12,the expression of apoptotic related proteins Bax and Bcl-2,as well as the protein expression of AMPK and p-AMPK. RESULTS AND CONCLUSION:Low concentrations of eriodictyol were non-toxic to MC3T3-E1 cells and promoted cell proliferation,as well as increased the expression of autophagy related proteins LC3-Ⅱ/LC3-Ⅰ,p62,Atg5 and Atg12,decreased caspase-3 enzyme activity,inhibited Bax protein expression,promoted Bcl-2 protein expression and reduced dexamethasone-induced apoptosis in MC3T3-E1 cells in a dose-dependent manner.Moreover,eriodictyol significantly promoted heme oxygenase 1 expression in osteoblasts,whereas overexpression of heme oxygenase 1 promoted AMPK phosphorylation,activated autophagy,and inhibited dexamethasone-induced osteoblast apoptosis.While 3-methyladenine treatment counteracted the effects of heme oxygenase 1 overexpression on MC3T3-E1 cells.To conclude,low concentration of Eriodictyol is non-toxic to osteoblasts and activates AMPK signaling pathway by upregulating the expression of heme oxygenase 1,thereby promoting autophagy and inhibiting dexamethasone-induced osteoblast apoptosis.Eriodictyol has great potential for the treatment of glucocorticoid-induced osteoporosis.
10.Hypidone hydrochloride(YL-0919)improves ischemic stroke in rats by inhibiting ferroptosis
Xiaojuan HOU ; Yue ZHANG ; Yafan BAI ; Hailong LI ; Yixin YANG ; Yunfeng LI ; Hui MA ; Henglin WANG
Chinese Journal of Pharmacology and Toxicology 2024;38(11):807-815
OBJECTIVE To study the way in which hypidone hydrochloride(code:YL-0919)improves motor function after ischemic stroke(IS)and explore the related mechanism.METHODS Adult male SD rats were used to establish a middle cerebral artery occlusion(MCAO)model that simulated acute IS.All animals were randomly divided into four groups:sham group,MCAO group,MCAO+YL-0919 group,and MCAO+YL-0919+erastin(Era,ferroptosis inducer)group.The drug administration groups received the first ip injection 6 h after operation,followed by continuous ip injection once per day.After 7-10 d of drug administration,the effect of YL-0919 on motor function after IS were evaluated via neu-rological function test,adhesive-removal test,rotarod test,balance beam test and open field test.After 7 d of drug administration,TTC staining was used to detect the cerebral infarction area while the colo-rimetry method was used to measure the contents of glutathione(GSH),malondialdehyde(MDA),and ferrous ions(Fe2+)in the penumbra of the cerebral cortex.Western blotting was used to detect the expression levels of glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(xCT),acyl-CoA synthetase long-chain family member 4(ACSL4),and transferrin receptor 1(TFR1)in the cortical penumbra.RESULTS Compared with the sham group,the MCAO group showed higher neurological function scores(P<0.01),with notably prolonged time for tape removal and first contact with the right forepaw(P<0.01),spent significantly more time crossing the balance beam(P<0.01)but endured a notably shorter duration on the rotarod(P<0.01),reduced the movement distance in the open field(P<0.01),had a remarkably increased infarct area(P<0.01)but significantly level of GSH in the cortical penumbra region decreased(P<0.01),while MDA and Fe2+levels were markedly increased(P<0.01).Protein expression levels of GPX4 and xCT were reduced(P<0.05),while those of ACSL4 and TFR1 were elevated(P<0.05).Compared with the MCAO group,these changes were significantly reversed after YL-0919 administration.However,when Era and YL-0919 were administered simultaneously,the reversal effect of YL-0919 was significantly weakened.CONCLUSION YL-0919 can improve motor function impairment and reduce cerebral infarction areas in rats after IS,and the mechanism may be related to the inhibition of ferroptosis.


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