Objective Several studies have indicated that miR-15a,miR-15b and miR-16 may be the important regulators of apoptosis.Since attenuate apoptosis could protect myocardium and reduce infarction size,the present study was...Objective Several studies have indicated that miR-15a,miR-15b and miR-16 may be the important regulators of apoptosis.Since attenuate apoptosis could protect myocardium and reduce infarction size,the present study was aimed to find out whether these miRNAs participate in regulating myocardial ischemia reperfusion （I/R） injury.Methods Apoptosis in mice hearts subjected to I/R was detected by TUNEL assay in vivo,while flow cytometry analysis followed by Annexin V/PI double stain in vitro was used to detect apoptosis in cultured cardiomyocytes which were subjected to hypoxia/reoxygenation （H/R）.Taqman real-time quantitative PCR was used to confirm whether miR-15a/15b/16 were involved in the regulation of cardiac I/R and H/R.Results Compared to those of the controls,I/R or H/R induced apoptosis of cardiomyocytes was significantly iucreased both in vivo （24.4％ ± 9.4％ vs.2.2％ ± 1.9％,P ＜ 0.01,n =5） and in vitro （14.12％ ±0.92％ vs.2.22％ ± 0.08％）.The expression of miR-15a and miR-15b,but not miR-16,was increased in the mice I/R model,and the results were consistent in the H/R model.Conclusions Our data indicate miR-15 and miR-15b are up-regulated in response to cardiac I/R injury,therefore,down-regulation of miR- 15a/b may be a promising strategy to reduce myocardial apoptosis induced by cardiac I/R injury.展开更多
目的探讨血清骨膜蛋白(POSTN)、微小核糖核酸-15b(miR-15b)、Clara细胞分泌蛋白16(CC16)与新生儿支气管肺发育不良(BPD)的相关性。方法选取2022年1月至2024年1月保定市妇幼保健院收治的BPD早产儿186例为BPD组,同期非BPD早产儿186例为非...目的探讨血清骨膜蛋白(POSTN)、微小核糖核酸-15b(miR-15b)、Clara细胞分泌蛋白16(CC16)与新生儿支气管肺发育不良(BPD)的相关性。方法选取2022年1月至2024年1月保定市妇幼保健院收治的BPD早产儿186例为BPD组,同期非BPD早产儿186例为非BPD组,比较两组血清POSTN、miR-15b、CC16水平,基线资料,肺功能指标。Spearman分析BPD患儿血清POSTN、miR-15b、CC16与BPD的相关性。多因素Logistic回归分析BPD患儿的影响因素。结果BPD组血清POSTN、miR-15b高于非BPD组,CC16低于非BPD组,差异有统计学意义(P<0.05)。BPD组50%潮气量时呼气流速(50%TEF)、千克体重潮气量(VT/kg)、达峰容积比(VPEF/VE)低于非BPD组,差异有统计学意义(P<0.05)。Spearman相关性分析显示,POSTN、miR-15b与5 min Apgar评分、1,25-(OH)2D3、50%TEF、VT/kg、VPEF/VE呈负相关,POSTN与机械通气时间、吸氧时间、最高氧浓度呈正相关,miR-15b与呼吸窘迫综合征、机械通气时间、吸氧时间、最高氧浓度呈正相关;CC16与机械通气时间、吸氧时间、最高氧浓度呈负相关,与5 min Apgar评分、1,25-(OH)2D3、50%TEF、VT/kg、VPEF/VE呈正相关(P<0.05)。Logistic回归分析结果显示,呼吸窘迫综合征、机械通气时间、吸氧时间、最高氧浓度、POSTN、miR-15b是BPD患儿的危险因素,5 min Apgar评分、1,25-(OH)2D3、50%TEF、VT/kg、VPEF/VE、CC16是BPD患儿的保护因素(P<0.05)。结论BPD患儿血清POSTN、miR-15b升高,CC16降低,其水平变化与BPD发生密切相关。展开更多
AIM To determine whether cell division cycle(Cdc)42 is regulated by microRNA(miR)-15 a in the development of pediatric inflammatory bowel disease(IBD).METHODS We cultured 293 T cells, used plasmids and performed dual-...AIM To determine whether cell division cycle(Cdc)42 is regulated by microRNA(miR)-15 a in the development of pediatric inflammatory bowel disease(IBD).METHODS We cultured 293 T cells, used plasmids and performed dual-luciferase assay to determine whether Cdc42 is a miR-15 a target gene. We cultured Caco-2 cells, and stimulated them with tumor necrosis factor(TNF)-α. We then employed lentiviruses to alter the expression of miR-15 a and Cdc42. We performed quantitative reverse transcription polymerase chain reaction(qRTPCR) and immunofluorescence to determine whether Cdc42 is regulated by miR-15 a in Caco-2 cells. Finally, we collected ileocecal tissue by endoscopy from patients and performed qRT-PCR to examine the expression of miR-15 a and Cdc42 in pediatric IBD patients.RESULTS Target Scan and dual-luciferase assay revealed thatCdc42 was a miR-15 a target gene. MiR-15 a expression increased(P = 0.0038) and Cdc42 expression decreased(P = 0.0013) in cells stimulated with TNF-α, and the expression of the epithelial junction proteins zona occludens(ZO)-1(P < 0.05) and E-cadherin(P < 0.001) decreased. Cdc42 levels decreased in miR-15 a-mimic cells(P < 0.001) and increased in miR-15 a inhibitor cells(P < 0.05). ZO-1 and E-cadherin decreased in mi R-15 a-mimic cells(P < 0.001) but not in the miR-15 a inhibitor + TNF-α cells. In Lv-Cdc42 + TNF-α cells, ZO-1 and E-cadherin expression increased compared to the Lv-Cdc42-NC + TNF-α(P < 0.05) or miR-15 a-mimic cells(P < 0.05). Fifty-four pediatric IBD patients were included in this study, 21 in the control group, 19 in the Crohn's disease(CD) active(AC) group, seven in the CD remission(RE) group, and seven in the ulcerative colitis(UC) group. MiR-15 a increased and Cdc42 decreased in the CD AC group compared to the control group(P < 0.05). miR-15 a decreased and Cdc42 increased in the CD RE group compared to the CD AC group(P < 0.05). miR-15 a was positively correlated with the Pediatric Crohn's disease Activity Index(PCDAI)(P = 0.006), while Cdc42 was negatively correlated with PCDAI(P = 0.0008). Finally, miR-15 a expression negatively correlated with Cdc42 in pediatric IBD patients(P = 0.0045).CONCLUSION Mi R-15 a negatively regulates epithelial junctions through Cdc42 in Caco-2 cells and pediatric IBD patients.展开更多
文摘Objective Several studies have indicated that miR-15a,miR-15b and miR-16 may be the important regulators of apoptosis.Since attenuate apoptosis could protect myocardium and reduce infarction size,the present study was aimed to find out whether these miRNAs participate in regulating myocardial ischemia reperfusion （I/R） injury.Methods Apoptosis in mice hearts subjected to I/R was detected by TUNEL assay in vivo,while flow cytometry analysis followed by Annexin V/PI double stain in vitro was used to detect apoptosis in cultured cardiomyocytes which were subjected to hypoxia/reoxygenation （H/R）.Taqman real-time quantitative PCR was used to confirm whether miR-15a/15b/16 were involved in the regulation of cardiac I/R and H/R.Results Compared to those of the controls,I/R or H/R induced apoptosis of cardiomyocytes was significantly iucreased both in vivo （24.4％ ± 9.4％ vs.2.2％ ± 1.9％,P ＜ 0.01,n =5） and in vitro （14.12％ ±0.92％ vs.2.22％ ± 0.08％）.The expression of miR-15a and miR-15b,but not miR-16,was increased in the mice I/R model,and the results were consistent in the H/R model.Conclusions Our data indicate miR-15 and miR-15b are up-regulated in response to cardiac I/R injury,therefore,down-regulation of miR- 15a/b may be a promising strategy to reduce myocardial apoptosis induced by cardiac I/R injury.
文摘目的探讨血清骨膜蛋白(POSTN)、微小核糖核酸-15b(miR-15b)、Clara细胞分泌蛋白16(CC16)与新生儿支气管肺发育不良(BPD)的相关性。方法选取2022年1月至2024年1月保定市妇幼保健院收治的BPD早产儿186例为BPD组,同期非BPD早产儿186例为非BPD组,比较两组血清POSTN、miR-15b、CC16水平,基线资料,肺功能指标。Spearman分析BPD患儿血清POSTN、miR-15b、CC16与BPD的相关性。多因素Logistic回归分析BPD患儿的影响因素。结果BPD组血清POSTN、miR-15b高于非BPD组,CC16低于非BPD组,差异有统计学意义(P<0.05)。BPD组50%潮气量时呼气流速(50%TEF)、千克体重潮气量(VT/kg)、达峰容积比(VPEF/VE)低于非BPD组,差异有统计学意义(P<0.05)。Spearman相关性分析显示,POSTN、miR-15b与5 min Apgar评分、1,25-(OH)2D3、50%TEF、VT/kg、VPEF/VE呈负相关,POSTN与机械通气时间、吸氧时间、最高氧浓度呈正相关,miR-15b与呼吸窘迫综合征、机械通气时间、吸氧时间、最高氧浓度呈正相关;CC16与机械通气时间、吸氧时间、最高氧浓度呈负相关,与5 min Apgar评分、1,25-(OH)2D3、50%TEF、VT/kg、VPEF/VE呈正相关(P<0.05)。Logistic回归分析结果显示,呼吸窘迫综合征、机械通气时间、吸氧时间、最高氧浓度、POSTN、miR-15b是BPD患儿的危险因素,5 min Apgar评分、1,25-(OH)2D3、50%TEF、VT/kg、VPEF/VE、CC16是BPD患儿的保护因素(P<0.05)。结论BPD患儿血清POSTN、miR-15b升高,CC16降低,其水平变化与BPD发生密切相关。
基金Supported by the National Natural Science Foundation of Shanghai,No.201540068
文摘AIM To determine whether cell division cycle(Cdc)42 is regulated by microRNA(miR)-15 a in the development of pediatric inflammatory bowel disease(IBD).METHODS We cultured 293 T cells, used plasmids and performed dual-luciferase assay to determine whether Cdc42 is a miR-15 a target gene. We cultured Caco-2 cells, and stimulated them with tumor necrosis factor(TNF)-α. We then employed lentiviruses to alter the expression of miR-15 a and Cdc42. We performed quantitative reverse transcription polymerase chain reaction(qRTPCR) and immunofluorescence to determine whether Cdc42 is regulated by miR-15 a in Caco-2 cells. Finally, we collected ileocecal tissue by endoscopy from patients and performed qRT-PCR to examine the expression of miR-15 a and Cdc42 in pediatric IBD patients.RESULTS Target Scan and dual-luciferase assay revealed thatCdc42 was a miR-15 a target gene. MiR-15 a expression increased(P = 0.0038) and Cdc42 expression decreased(P = 0.0013) in cells stimulated with TNF-α, and the expression of the epithelial junction proteins zona occludens(ZO)-1(P < 0.05) and E-cadherin(P < 0.001) decreased. Cdc42 levels decreased in miR-15 a-mimic cells(P < 0.001) and increased in miR-15 a inhibitor cells(P < 0.05). ZO-1 and E-cadherin decreased in mi R-15 a-mimic cells(P < 0.001) but not in the miR-15 a inhibitor + TNF-α cells. In Lv-Cdc42 + TNF-α cells, ZO-1 and E-cadherin expression increased compared to the Lv-Cdc42-NC + TNF-α(P < 0.05) or miR-15 a-mimic cells(P < 0.05). Fifty-four pediatric IBD patients were included in this study, 21 in the control group, 19 in the Crohn's disease(CD) active(AC) group, seven in the CD remission(RE) group, and seven in the ulcerative colitis(UC) group. MiR-15 a increased and Cdc42 decreased in the CD AC group compared to the control group(P < 0.05). miR-15 a decreased and Cdc42 increased in the CD RE group compared to the CD AC group(P < 0.05). miR-15 a was positively correlated with the Pediatric Crohn's disease Activity Index(PCDAI)(P = 0.006), while Cdc42 was negatively correlated with PCDAI(P = 0.0008). Finally, miR-15 a expression negatively correlated with Cdc42 in pediatric IBD patients(P = 0.0045).CONCLUSION Mi R-15 a negatively regulates epithelial junctions through Cdc42 in Caco-2 cells and pediatric IBD patients.
