Objective To study expression of NDRG3 in prostatic mesenchyma and effect of exogenous NDRG3 on prostatic stromal cells. Methods Immunohistochemical analysis was used to check expression of NDRG3 in prostate mesnenchy...Objective To study expression of NDRG3 in prostatic mesenchyma and effect of exogenous NDRG3 on prostatic stromal cells. Methods Immunohistochemical analysis was used to check expression of NDRG3 in prostate mesnenchyma. The WPMY-1 prostate immortalized mesenchyma cell line was stably-transfected with a NDRG3 gene expression vector. The NDRG3-stable transfected WPMY-1 sublines were studied along with parental and empty vector transfected WPMY-1 cells as controls. RT-PCR technology was applied to identity downstream gene expression under regulation of NDRG3 expression.Results Expression of DNRG3 was observed in prostate cancer mesenchyma, over-expression of NDRG3 in WPMY-1 cell up-regulated expression of chemotatic factors-CXCL3 and CXCL5. Conclusion Expression of stromal NDRG3 in prostate cancer specimens is significantly higher than that in benign prostatic hypertrophy (BPH) sample. There is a remarkable difference between the two groups of samples, NDRG3 may be related to angiogenesis in prostatic mesenchyma.展开更多
Ovarian mesenchymal cells(oMCs)constitute a distinct microenvironment that supports folliculogenesis under physiological conditions.Supplementation of exogenous non-ovarian mesenchymal-related cells has been reported ...Ovarian mesenchymal cells(oMCs)constitute a distinct microenvironment that supports folliculogenesis under physiological conditions.Supplementation of exogenous non-ovarian mesenchymal-related cells has been reported to be an efficient approach to improve ovarian functions.However,the development and cellular and molecular characteristics of endogenous oMCs remain largely unexplored.In this study,we surveyed the single-cell transcriptomic landscape to dissect the cellular and molecular changes associated with the aging of oMCs in mice.Our results showed that the oMCs were composed of five ovarian differentiatedMC(odMC)populations and one ovarian mesenchymal progenitor(oMP)cell population.These cells could differentiate into various odMCs via an oMP-derived route to construct the ovarian stroma structures.Comparative analysis revealed that ovarian aging was associated with decreased quantity of oMP cells and reduced quality of odMCs.Based on the findings of bioinformatics analysis,we designed different strategies involving supplementation with young oMCs to examine their effects on female fertility and health.Our functional investigations revealed that oMCs supplementation prior to ovarian senescence was the optimal method to improve female fertility and extend the reproductive lifespan of aged females in the longterm.展开更多
目的:构建骨髓间充质干细胞(BMMSC)与多发性骨髓瘤(MM)细胞共培养体系,探讨共培养后BMMSC对M M细胞迁移和归巢的影响。方法:采用贴壁筛选法培养绿色荧光蛋白(GFP)小鼠BM M SC,建立M M细胞株XG-7细胞和BMMSC间接及直接共培养体系;用CD13...目的:构建骨髓间充质干细胞(BMMSC)与多发性骨髓瘤(MM)细胞共培养体系,探讨共培养后BMMSC对M M细胞迁移和归巢的影响。方法:采用贴壁筛选法培养绿色荧光蛋白(GFP)小鼠BM M SC,建立M M细胞株XG-7细胞和BMMSC间接及直接共培养体系;用CD138磁珠法分离直接共培养的XG-7细胞及BMMSC,用台盼蓝染色计数法检测XG-7细胞的增殖水平、Annexin V/PI测定细胞凋亡水平、单丹(磺)酰戊二胺(MDC)法检测细胞自噬泡数量、在共聚焦显微镜下动态观察BMMSC与PE-CD138标记的XG-7细胞共培养过程的分子流向和分布。结果:经共培养后,BMMSC促进XG-7细胞增殖;单独培养后凋亡率为(17.90±1.46)%,直接和间接共培养后凋亡率分别降至(6.23±0.12)%和(6.97±0.03)%(P<0.01);XG-7细胞单独培养及与BMMSC共培养后,荧光显微镜下均可出现自噬,共培养后的XG-7细胞自噬现象相对少见;在共聚焦荧光显微镜下XG-7细胞是以极化的胞膜区同BMMSC接触,加入甲基β环糊精后极化现象消失。结论:BMMSC可促进XG-7细胞的生长,且增强XG-7细胞抗凋亡及抗自噬能力,影响骨髓瘤细胞的迁移和归巢。展开更多
目的观察血管紧张素转换酶抑制剂(ACEI)依那普利对单侧输尿管梗阻(UUO)大鼠肾组织胰岛素样生长因子-1(IGF-1)表达及肾小管上皮细胞转分化(EMT)的影响。方法采用UUO大鼠模型,雄性SD大鼠32只随机分为假手术组、模型组和依那普利组。依那...目的观察血管紧张素转换酶抑制剂(ACEI)依那普利对单侧输尿管梗阻(UUO)大鼠肾组织胰岛素样生长因子-1(IGF-1)表达及肾小管上皮细胞转分化(EMT)的影响。方法采用UUO大鼠模型,雄性SD大鼠32只随机分为假手术组、模型组和依那普利组。依那普利组从造模前24 h开始以依那普利10 mg/(kg·d)灌胃,模型组和假手术组以等体积的生理盐水灌胃。于造模后第14天处死大鼠,取梗阻侧肾组织,苏木精-伊红染色(HE)和Masson染色观察肾组织病理改变;实时定量逆转录-聚合酶链反应(RT-q PCR)检测肾组织IGF-1m RNA的表达;Western blot检测肾组织IGF-1、α-血管平滑肌肌动蛋白(α-SMA)和钙黏蛋白(E-cadherin)的表达。结果模型组大鼠肾间质损伤指数逐渐增加,肾间质胶原相对面积逐渐增大,IGF-1、α-SMA蛋白表达增加,IGF-1 m RNA和钙黏蛋白表达减少。经依那普利治疗后,肾间质损伤指数和肾间质胶原相对面积下降,IGF-1、α-SMA蛋白表达减少,IGF-1 m RNA和E-钙黏蛋白表达增加,与模型组比较,差异有统计学意义(P<0.05)。结论依那普利可通过抑制肾组织IGF-1表达抑制EMT,进而防治肾间质纤维化。展开更多
基金Shanghai Municipal Committee of Science and Technology (06ZR14072)
文摘Objective To study expression of NDRG3 in prostatic mesenchyma and effect of exogenous NDRG3 on prostatic stromal cells. Methods Immunohistochemical analysis was used to check expression of NDRG3 in prostate mesnenchyma. The WPMY-1 prostate immortalized mesenchyma cell line was stably-transfected with a NDRG3 gene expression vector. The NDRG3-stable transfected WPMY-1 sublines were studied along with parental and empty vector transfected WPMY-1 cells as controls. RT-PCR technology was applied to identity downstream gene expression under regulation of NDRG3 expression.Results Expression of DNRG3 was observed in prostate cancer mesenchyma, over-expression of NDRG3 in WPMY-1 cell up-regulated expression of chemotatic factors-CXCL3 and CXCL5. Conclusion Expression of stromal NDRG3 in prostate cancer specimens is significantly higher than that in benign prostatic hypertrophy (BPH) sample. There is a remarkable difference between the two groups of samples, NDRG3 may be related to angiogenesis in prostatic mesenchyma.
基金supported by the National Key Research and Development Program of China (2022YFC2703800)the National Natural Science Foundation of China (82230051)the 2115 Talent Development Program of China Agricultural University (1021-00109022)。
文摘Ovarian mesenchymal cells(oMCs)constitute a distinct microenvironment that supports folliculogenesis under physiological conditions.Supplementation of exogenous non-ovarian mesenchymal-related cells has been reported to be an efficient approach to improve ovarian functions.However,the development and cellular and molecular characteristics of endogenous oMCs remain largely unexplored.In this study,we surveyed the single-cell transcriptomic landscape to dissect the cellular and molecular changes associated with the aging of oMCs in mice.Our results showed that the oMCs were composed of five ovarian differentiatedMC(odMC)populations and one ovarian mesenchymal progenitor(oMP)cell population.These cells could differentiate into various odMCs via an oMP-derived route to construct the ovarian stroma structures.Comparative analysis revealed that ovarian aging was associated with decreased quantity of oMP cells and reduced quality of odMCs.Based on the findings of bioinformatics analysis,we designed different strategies involving supplementation with young oMCs to examine their effects on female fertility and health.Our functional investigations revealed that oMCs supplementation prior to ovarian senescence was the optimal method to improve female fertility and extend the reproductive lifespan of aged females in the longterm.
文摘目的观察血管紧张素转换酶抑制剂(ACEI)依那普利对单侧输尿管梗阻(UUO)大鼠肾组织胰岛素样生长因子-1(IGF-1)表达及肾小管上皮细胞转分化(EMT)的影响。方法采用UUO大鼠模型,雄性SD大鼠32只随机分为假手术组、模型组和依那普利组。依那普利组从造模前24 h开始以依那普利10 mg/(kg·d)灌胃,模型组和假手术组以等体积的生理盐水灌胃。于造模后第14天处死大鼠,取梗阻侧肾组织,苏木精-伊红染色(HE)和Masson染色观察肾组织病理改变;实时定量逆转录-聚合酶链反应(RT-q PCR)检测肾组织IGF-1m RNA的表达;Western blot检测肾组织IGF-1、α-血管平滑肌肌动蛋白(α-SMA)和钙黏蛋白(E-cadherin)的表达。结果模型组大鼠肾间质损伤指数逐渐增加,肾间质胶原相对面积逐渐增大,IGF-1、α-SMA蛋白表达增加,IGF-1 m RNA和钙黏蛋白表达减少。经依那普利治疗后,肾间质损伤指数和肾间质胶原相对面积下降,IGF-1、α-SMA蛋白表达减少,IGF-1 m RNA和E-钙黏蛋白表达增加,与模型组比较,差异有统计学意义(P<0.05)。结论依那普利可通过抑制肾组织IGF-1表达抑制EMT,进而防治肾间质纤维化。