目的探讨二氯二苯基三氯乙烷(dichlorodiphenyltrichloroethane,DDT)诱导人结直肠癌增殖、侵袭与微小RNA-129(microRNA-129,miR-129)/细胞周期蛋白依赖性激酶-14(cyclin-dependent kinase-14,CDK-14)表达的关系。方法5×106个/m L...目的探讨二氯二苯基三氯乙烷(dichlorodiphenyltrichloroethane,DDT)诱导人结直肠癌增殖、侵袭与微小RNA-129(microRNA-129,miR-129)/细胞周期蛋白依赖性激酶-14(cyclin-dependent kinase-14,CDK-14)表达的关系。方法5×106个/m L密度的人结直肠癌细胞分别用终浓度为0、100、250和500 nmol/L的DDT处理72 h,运用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,MTT]法及甲基紫染色测定细胞活力的和细胞细胞集落,流式细胞仪测定细胞凋亡水平,Transwell小室(transwell chamber,Transwell)侵袭室及划痕试验测定细胞侵袭迁移水平,实时荧光逆转录(real-time fluorescent reverse transcription,RT-PCR)法及蛋白印迹法测定细胞miR-129、CDK-14、卷曲螺旋结构域-34(coiled-coil domain-34,CCDC34)、半乳糖激酶1(Galactokinase 1,GALK1)水平。结果100、250和500 nmol/L DDT组细胞增殖水平[(70.59±7.72、76.63±8.32、84.25±9.04 vs 62.58±6.83)%]、细胞集落[(99.95±9.58、215.63±19.85、398.96±35.26 vs 62.63±6.52)个]、侵袭数目[(76.25±9.58、154.62±19.51、298.96±38.57 vs 49.65±6.65)个]、迁移水平(75.96±14.58、119.69±20.59、165.63±32.68 vs 11.59±2.28)、CDK-14、CCDC34、GALK1 m RNA蛋白水平明显高于对照组(P<0.01),凋亡率、miR-129水平明显低于对照组(P<0.01);且500 nmol/L DDT组细胞增殖水平、细胞集落、侵袭数目、迁移水平、CDK-14 mRNA蛋白[(1.60±0.28、2.14±0.33、2.99±0.51 vs 0.92±0.16),(0.42±0.07、0.85±0.14、1.20±0.20 vs 0.21±0.03)]、CCDC34(0.48±0.08、0.87±0.15、1.18±0.18 vs 0.28±0.07)、GALK1(0.50±0.09、0.84±0.14、1.22±0.19 vs 0.30±0.08)蛋白水平均高于250 nmol/L DDT组,差异均有统计学意义(均P<0.01),凋亡率(1.70±0.31、1.00±0.16、0.60±0.11 vs 3.00±0.51)、miR-129(1.42±0.24、0.92±0.15、0.64±0.10 vs 1.78±0.28)水平均低于250 nmol/L DDT组,差异均有统计学意义(均P<0.01);250nmol/L DDT组细胞增殖水平、细胞集落�展开更多
BACKGROUND Diabetic intracerebral hemorrhage(ICH)is a serious complication of diabetes.The role and mechanism of bone marrow mesenchymal stem cell(BMSC)-derived exosomes(BMSC-exo)in neuroinflammation post-ICH in patie...BACKGROUND Diabetic intracerebral hemorrhage(ICH)is a serious complication of diabetes.The role and mechanism of bone marrow mesenchymal stem cell(BMSC)-derived exosomes(BMSC-exo)in neuroinflammation post-ICH in patients with diabetes are unknown.In this study,we investigated the regulation of BMSC-exo on hyperglycemia-induced neuroinflammation.AIM To study the mechanism of BMSC-exo on nerve function damage after diabetes complicated with cerebral hemorrhage.METHODS BMSC-exo were isolated from mouse BMSC media.This was followed by transfection with microRNA-129-5p(miR-129-5p).BMSC-exo or miR-129-5poverexpressing BMSC-exo were intravitreally injected into a diabetes mouse model with ICH for in vivo analyses and were cocultured with high glucoseaffected BV2 cells for in vitro analyses.The dual luciferase test and RNA immunoprecipitation test verified the targeted binding relationship between miR-129-5p and high-mobility group box 1(HMGB1).Quantitative polymerase chain reaction,western blotting,and enzyme-linked immunosorbent assay were conducted to assess the levels of some inflammation factors,such as HMGB1,interleukin 6,interleukin 1β,toll-like receptor 4,and tumor necrosis factorα.Brain water content,neural function deficit score,and Evans blue were used to measure the neural function of mice.RESULTS Our findings indicated that BMSC-exo can promote neuroinflammation and functional recovery.MicroRNA chip analysis of BMSC-exo identified miR-129-5p as the specific microRNA with a protective role in neuroinflammation.Overexpression of miR-129-5p in BMSC-exo reduced the inflammatory response and neurological impairment in comorbid diabetes and ICH cases.Furthermore,we found that miR-129-5p had a targeted binding relationship with HMGB1 mRNA.CONCLUSION We demonstrated that BMSC-exo can reduce the inflammatory response after ICH with diabetes,thereby improving the neurological function of the brain.展开更多
文摘目的探讨二氯二苯基三氯乙烷(dichlorodiphenyltrichloroethane,DDT)诱导人结直肠癌增殖、侵袭与微小RNA-129(microRNA-129,miR-129)/细胞周期蛋白依赖性激酶-14(cyclin-dependent kinase-14,CDK-14)表达的关系。方法5×106个/m L密度的人结直肠癌细胞分别用终浓度为0、100、250和500 nmol/L的DDT处理72 h,运用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐[3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide,MTT]法及甲基紫染色测定细胞活力的和细胞细胞集落,流式细胞仪测定细胞凋亡水平,Transwell小室(transwell chamber,Transwell)侵袭室及划痕试验测定细胞侵袭迁移水平,实时荧光逆转录(real-time fluorescent reverse transcription,RT-PCR)法及蛋白印迹法测定细胞miR-129、CDK-14、卷曲螺旋结构域-34(coiled-coil domain-34,CCDC34)、半乳糖激酶1(Galactokinase 1,GALK1)水平。结果100、250和500 nmol/L DDT组细胞增殖水平[(70.59±7.72、76.63±8.32、84.25±9.04 vs 62.58±6.83)%]、细胞集落[(99.95±9.58、215.63±19.85、398.96±35.26 vs 62.63±6.52)个]、侵袭数目[(76.25±9.58、154.62±19.51、298.96±38.57 vs 49.65±6.65)个]、迁移水平(75.96±14.58、119.69±20.59、165.63±32.68 vs 11.59±2.28)、CDK-14、CCDC34、GALK1 m RNA蛋白水平明显高于对照组(P<0.01),凋亡率、miR-129水平明显低于对照组(P<0.01);且500 nmol/L DDT组细胞增殖水平、细胞集落、侵袭数目、迁移水平、CDK-14 mRNA蛋白[(1.60±0.28、2.14±0.33、2.99±0.51 vs 0.92±0.16),(0.42±0.07、0.85±0.14、1.20±0.20 vs 0.21±0.03)]、CCDC34(0.48±0.08、0.87±0.15、1.18±0.18 vs 0.28±0.07)、GALK1(0.50±0.09、0.84±0.14、1.22±0.19 vs 0.30±0.08)蛋白水平均高于250 nmol/L DDT组,差异均有统计学意义(均P<0.01),凋亡率(1.70±0.31、1.00±0.16、0.60±0.11 vs 3.00±0.51)、miR-129(1.42±0.24、0.92±0.15、0.64±0.10 vs 1.78±0.28)水平均低于250 nmol/L DDT组,差异均有统计学意义(均P<0.01);250nmol/L DDT组细胞增殖水平、细胞集落�
基金Supported by the National Natural Science Foundation of China,No.81900743Heilongjiang Province Outstanding Young Medical Talents Training Grant Project,China,No.HYD2020YQ0007.
文摘BACKGROUND Diabetic intracerebral hemorrhage(ICH)is a serious complication of diabetes.The role and mechanism of bone marrow mesenchymal stem cell(BMSC)-derived exosomes(BMSC-exo)in neuroinflammation post-ICH in patients with diabetes are unknown.In this study,we investigated the regulation of BMSC-exo on hyperglycemia-induced neuroinflammation.AIM To study the mechanism of BMSC-exo on nerve function damage after diabetes complicated with cerebral hemorrhage.METHODS BMSC-exo were isolated from mouse BMSC media.This was followed by transfection with microRNA-129-5p(miR-129-5p).BMSC-exo or miR-129-5poverexpressing BMSC-exo were intravitreally injected into a diabetes mouse model with ICH for in vivo analyses and were cocultured with high glucoseaffected BV2 cells for in vitro analyses.The dual luciferase test and RNA immunoprecipitation test verified the targeted binding relationship between miR-129-5p and high-mobility group box 1(HMGB1).Quantitative polymerase chain reaction,western blotting,and enzyme-linked immunosorbent assay were conducted to assess the levels of some inflammation factors,such as HMGB1,interleukin 6,interleukin 1β,toll-like receptor 4,and tumor necrosis factorα.Brain water content,neural function deficit score,and Evans blue were used to measure the neural function of mice.RESULTS Our findings indicated that BMSC-exo can promote neuroinflammation and functional recovery.MicroRNA chip analysis of BMSC-exo identified miR-129-5p as the specific microRNA with a protective role in neuroinflammation.Overexpression of miR-129-5p in BMSC-exo reduced the inflammatory response and neurological impairment in comorbid diabetes and ICH cases.Furthermore,we found that miR-129-5p had a targeted binding relationship with HMGB1 mRNA.CONCLUSION We demonstrated that BMSC-exo can reduce the inflammatory response after ICH with diabetes,thereby improving the neurological function of the brain.
