Objective:To explore the effect of Hupo powder(HP)on autophagy in menstrual blood-derived stem cells(MenSCs)with endometriosis(EMT).Methods:EMT MenSCs(E-MenSCs)and healthy MenSCs(H-MenSCs)were isolated from the menstr...Objective:To explore the effect of Hupo powder(HP)on autophagy in menstrual blood-derived stem cells(MenSCs)with endometriosis(EMT).Methods:EMT MenSCs(E-MenSCs)and healthy MenSCs(H-MenSCs)were isolated from the menstrual blood of patients with EMT and healthy female participants,respectively.We identified their stem cells’characteristics via adipogenic and osteogenic differentiation.Twelve male SpragueeDawley rats received 0.9% NaCl and HP-dispensing granules by gastric irrigation to prepare blank serum and medicated serum,respectively.We used serum concentrations of 5%,10%,and 20%,each at administered times of 12,24,and 48 h to select the best condition.These cells were divided into three groups:blank serum of the control group,blank serum of the model group,and medicated serum of the HP group.H-MenSCs were used in the control group,while E-MenSCs were used in the model and HP groups.We analyzed cell viability using a cell counting kit-8 assay,observed cell morphology,evaluated the amounts of auto-phagosomes and autolysosomes by transmission electron microscopy,and detected the protein expression of autophagy markers(LC3-II and Beclin1)by Western blot.Results:E-MenSCs and H-MenSCs became long fusiform with a diffuse radial pattern,forming lipid droplets and calcium nodules after adipogenic and osteogenic differentiation.We then used the best conditiond 20% serum and 48 hdfor the subsequent experiments.In contrast to the model group,the HP group exhibited lower cell viability(=0.007),larger amounts of autophagosomes and autolysosomes(P<0.001 and P=0.001,respectively),and higher expression of LC3-II and Beclin1(P=0.021 and P=0.019,respectively).Conclusion:Hupo powder can promote autophagy in E-MenSCs,which might be one of the mechanisms underlying its therapeutic effects.展开更多
基金This study was supported by the National Natural Science Foundation of China(81973895)the Key Research Projects of Beijing University of Chinese Medicine(2020-JYB-ZDGG-143-3).
文摘Objective:To explore the effect of Hupo powder(HP)on autophagy in menstrual blood-derived stem cells(MenSCs)with endometriosis(EMT).Methods:EMT MenSCs(E-MenSCs)and healthy MenSCs(H-MenSCs)were isolated from the menstrual blood of patients with EMT and healthy female participants,respectively.We identified their stem cells’characteristics via adipogenic and osteogenic differentiation.Twelve male SpragueeDawley rats received 0.9% NaCl and HP-dispensing granules by gastric irrigation to prepare blank serum and medicated serum,respectively.We used serum concentrations of 5%,10%,and 20%,each at administered times of 12,24,and 48 h to select the best condition.These cells were divided into three groups:blank serum of the control group,blank serum of the model group,and medicated serum of the HP group.H-MenSCs were used in the control group,while E-MenSCs were used in the model and HP groups.We analyzed cell viability using a cell counting kit-8 assay,observed cell morphology,evaluated the amounts of auto-phagosomes and autolysosomes by transmission electron microscopy,and detected the protein expression of autophagy markers(LC3-II and Beclin1)by Western blot.Results:E-MenSCs and H-MenSCs became long fusiform with a diffuse radial pattern,forming lipid droplets and calcium nodules after adipogenic and osteogenic differentiation.We then used the best conditiond 20% serum and 48 hdfor the subsequent experiments.In contrast to the model group,the HP group exhibited lower cell viability(=0.007),larger amounts of autophagosomes and autolysosomes(P<0.001 and P=0.001,respectively),and higher expression of LC3-II and Beclin1(P=0.021 and P=0.019,respectively).Conclusion:Hupo powder can promote autophagy in E-MenSCs,which might be one of the mechanisms underlying its therapeutic effects.
文摘目的研究琥珀丸对绝经后骨质疏松大鼠骨密度、骨强度及骨代谢的影响。方法采用双侧卵巢摘除法建立绝经后骨质疏松症(postmenopausal osteoporosis,PMOP)大鼠模型,随机分为假手术组、模型组、琥珀丸组、阿仑膦酸钠组,各组分别予以灌胃给药。给药6周后处死取材,双能X线检测大鼠骨密度;ELISA检测各组大鼠血清I型原胶原分子N端前肽(PINP)、骨钙素(BGP)及钙磷的变化;骨生物力学生物仪进行三点弯曲的生物力学测定试验,测定最大载荷、最大应力、弹性模量。结果与假手术组相比较,模型组血清中PINP、BGP、核因子κB受体活化因子配体(RANKL)、血清抗酒石酸酸性磷酸酶(TRACP5b)、血清碱性磷酸酶(alkaline phosphatase,ALP)和白细胞介素-6(interleukin-6,IL-6)水平均显著增高,血清钙、磷、骨保护素(OPG)含量明显降低(P<0.05)。与模型组相比较,琥珀丸组和阿仑膦酸钠组血清中PINP、BGP、RANKL、TRACP5b、ALP和IL-6水平均显著降低,血清钙、磷、OPG含量明显增高(P<0.05)。与假手术组相比,模型组大鼠股骨骨密度(bone mineral density,BMD)值明显降低(P<0.05)。与模型组相比,琥珀丸组与阿仑膦酸钠组大鼠股骨BMD明显增高(P<0.05)。与假手术组相比,模型组大鼠股骨最大载荷、最大应力、弹性模量明显降低(P<0.05)。与模型组相比,琥珀丸组与阿仑膦酸钠组大鼠股骨最大载荷、最大应力、弹性模量明显升高(P<0.05)。结论琥珀丸能够提高PMOP大鼠的骨密度,改善骨强度,其作用机制可能与PINP、BGP、血清钙、磷水平相关,从而治疗绝经后骨质疏松症。
