根据美国心脏病协会(American Heart Association on Heart Disease and Stroke)的数据统计。全球范围内每年大约有1730万人死于心血管疾病及相关并发症,并且这个数字还在逐年增加。miRNA的发现和研究为诊断和治疗心血管疾病(cardiovasc...根据美国心脏病协会(American Heart Association on Heart Disease and Stroke)的数据统计。全球范围内每年大约有1730万人死于心血管疾病及相关并发症,并且这个数字还在逐年增加。miRNA的发现和研究为诊断和治疗心血管疾病(cardiovascular disease, CVD)提供了新的思路。目前已知的人类miRNA有3000个以上,最初的分析推测其可调控人类至少三分之一的编码基因。也就是说miRNA为多种疾病的诊断、治疗及预后提供了潜在靶点。MicroRNAs (miRNAs)是由RNA聚合酶II (Pol II)转录的初级miRNA (primiRNA)产生的约22个核苷酸的小RNA家族。Mir-30家族是miRNA家族中的一个重要成员,由Mir-30a、Mir-30b、Mir-30c、Mir-30d和Mir-30e组成,Mir-30c又分为Mir-30c-1和Mir-30c-2,由位于人类染色体1、6和8的6个基因编码。展开更多
目的观察威草胶囊对尿酸性肾病大鼠肾组织miR-29a和miR-30c表达的影响,探讨威草胶囊降血尿酸和肾保护作用的具体机制。方法将50只SPF级SD雄性大鼠随机分为正常组、模型组、别嘌呤醇组、威草胶囊小剂量组、威草胶囊大剂量组,每组10只。...目的观察威草胶囊对尿酸性肾病大鼠肾组织miR-29a和miR-30c表达的影响,探讨威草胶囊降血尿酸和肾保护作用的具体机制。方法将50只SPF级SD雄性大鼠随机分为正常组、模型组、别嘌呤醇组、威草胶囊小剂量组、威草胶囊大剂量组,每组10只。正常组给予普通饲料喂养,其余4组均给予高嘌呤饲料喂养,在给予高嘌呤饲料后的第20天停用高嘌呤饲料,均改用普通饲料喂养。正常组和模型组每日给予蒸馏水2 m L灌胃,别嘌呤醇组给予别嘌呤醇0.05 g/(kg·d)灌胃,威草胶囊小剂量组、大剂量组分别给予威草胶囊0.9g/(kg·d)、1.8 g/(kg·d)灌胃,共4周。4周后处死大鼠,检测血清UA、BUN、SCr水平;摘取双侧肾脏,做HE染色、Masson染色,免疫组织化学方法检测CTGF、TGF-β1、ColⅠ、ColⅢ表达情况,实时荧光定量PCR法测定大鼠肾组织中miR-29a和miR-30c表达情况。结果模型组大鼠血清UA、BUN、SCr水平均明显高于正常组(P均<0.05);别嘌呤醇组血清UA水平明显低于模型组(P<0.05),威草胶囊小剂量组、大剂量组血清UA、BUN、SCr水平均明显低于模型组(P均<0.05);威草胶囊大剂量组血清UA、BUN水平均明显低于别嘌呤醇组(P均<0.05),威草胶囊小剂量组血清UA、BUN、SCr水平与威草胶囊大剂量组比较差异均无统计学意义(P均>0.05)。HE染色、Masson染色:正常组少量肾组织纤维化,纤维化程度轻;模型组纤维化区域增多,纤维化程度重;别嘌呤醇组肾组织纤维化改变不明显;威草胶囊小剂量组和大剂量组肾组织间质纤维化程度明显减轻,威草胶囊大剂量组接近正常。模型组大鼠肾组织中CTGF、TGF-β1、ColⅠ、ColⅢ表达量均明显高于正常组(P均<0.05);威草胶囊小剂量组和大剂量组CTGF、ColⅠ、ColⅢ表达量和威草胶囊大剂量组TGF-β1表达量均明显低于模型组和别嘌呤醇组(P均<0.05);威草胶囊小剂量组CTGF、TGF-β1、ColⅠ、ColⅢ表达量与威草胶囊大剂量组比较差异均无统计学意义(P均>0.05)。模型组大鼠肾组织中miR-29a、miR-30c表达水平均明显低于正常组(P均<0.05);威草胶囊小剂量组和大剂量组miR-29a、miR-30c表达水平均明显高于模型组(P均<0.05),别嘌呤醇组与模型组比较差异无统计学意义(P均>0.05);威草胶囊大剂量组miR-29a、miR-30c表达水平均明显高于别嘌呤醇组(P均<0.05);威草胶囊小剂量组miR-29a、miR-30c表达水平与威草胶囊大剂量组比较差异均无统计学意义(P均>0.05)。结论威草胶囊可改善尿酸性肾病大鼠的肾功能,减轻肾组织纤维化程度,与其升高肾组织miR-29a、miR-30c的表达水平,降低TGF-β1、CTGF的表达,减少ColⅠ、ColⅢ的生成有关,别嘌呤醇不具有此作用。展开更多
During vascular remodeling after intimal injury,circulating platelets are activated by exposed collagen and release platelet-derived microvesicles(PMVs),which may contribute to the injury-induced apoptosis of vascular...During vascular remodeling after intimal injury,circulating platelets are activated by exposed collagen and release platelet-derived microvesicles(PMVs),which may contribute to the injury-induced apoptosis of vascular smooth muscle cells(VSMCs).However,the mechanisms in this process are still unclear.Using a rat balloon injury model,platelet adhesion at the injured site was detected,and promoted medial VSMC apoptosis was revealed with dT-mediated dUTP nick-end labeling(TUNEL)in vivo.VSMC apoptosis was promoted by coincubation with PMVs in vitro.Transcriptomics analysis indicated the abound expression of the microRNA-30(miR-30)family in platelets,and the expression of the miR-30 family in platelets and PMVs was confirmed with qPCR.Ingenuity Pathway Analysis(IPA)software identified transcription factor 2(RUNX2)as a pivotal molecule linking miR-30 and cellular apoptosis.In vitro,PMVs delivered miR-30e,an important member of the miR-30 family,into VSMCs and increased cell apoptosis.A dual-luciferase assay was adopted to verify the interaction of miR-30e with RUNX2,and fluorescence in situ hybridization(FISH)proved the cytoplasmic localization of miR-30e in VSMCs.The apoptotic effect on VSMCs was further confirmed by using miR-30e mimics and RUNX2 siRNA.Furthermore,RUNX2 siRNA altered the expression of its downstream genes,Tnfrsf11b,Smad6 and Mmp13,which may participate in apoptosis,as suggested by IPA.Our results indicated that PMVs play important roles in the interactions between circulating elements and VSMCs by delivering miR-30e to VSMCs and then promoting apoptosis.PMVs and potential intercellular messages may be novel therapeutic targets for preventing pathological vascular remodeling after intimal injury.展开更多
文摘根据美国心脏病协会(American Heart Association on Heart Disease and Stroke)的数据统计。全球范围内每年大约有1730万人死于心血管疾病及相关并发症,并且这个数字还在逐年增加。miRNA的发现和研究为诊断和治疗心血管疾病(cardiovascular disease, CVD)提供了新的思路。目前已知的人类miRNA有3000个以上,最初的分析推测其可调控人类至少三分之一的编码基因。也就是说miRNA为多种疾病的诊断、治疗及预后提供了潜在靶点。MicroRNAs (miRNAs)是由RNA聚合酶II (Pol II)转录的初级miRNA (primiRNA)产生的约22个核苷酸的小RNA家族。Mir-30家族是miRNA家族中的一个重要成员,由Mir-30a、Mir-30b、Mir-30c、Mir-30d和Mir-30e组成,Mir-30c又分为Mir-30c-1和Mir-30c-2,由位于人类染色体1、6和8的6个基因编码。
文摘目的观察威草胶囊对尿酸性肾病大鼠肾组织miR-29a和miR-30c表达的影响,探讨威草胶囊降血尿酸和肾保护作用的具体机制。方法将50只SPF级SD雄性大鼠随机分为正常组、模型组、别嘌呤醇组、威草胶囊小剂量组、威草胶囊大剂量组,每组10只。正常组给予普通饲料喂养,其余4组均给予高嘌呤饲料喂养,在给予高嘌呤饲料后的第20天停用高嘌呤饲料,均改用普通饲料喂养。正常组和模型组每日给予蒸馏水2 m L灌胃,别嘌呤醇组给予别嘌呤醇0.05 g/(kg·d)灌胃,威草胶囊小剂量组、大剂量组分别给予威草胶囊0.9g/(kg·d)、1.8 g/(kg·d)灌胃,共4周。4周后处死大鼠,检测血清UA、BUN、SCr水平;摘取双侧肾脏,做HE染色、Masson染色,免疫组织化学方法检测CTGF、TGF-β1、ColⅠ、ColⅢ表达情况,实时荧光定量PCR法测定大鼠肾组织中miR-29a和miR-30c表达情况。结果模型组大鼠血清UA、BUN、SCr水平均明显高于正常组(P均<0.05);别嘌呤醇组血清UA水平明显低于模型组(P<0.05),威草胶囊小剂量组、大剂量组血清UA、BUN、SCr水平均明显低于模型组(P均<0.05);威草胶囊大剂量组血清UA、BUN水平均明显低于别嘌呤醇组(P均<0.05),威草胶囊小剂量组血清UA、BUN、SCr水平与威草胶囊大剂量组比较差异均无统计学意义(P均>0.05)。HE染色、Masson染色:正常组少量肾组织纤维化,纤维化程度轻;模型组纤维化区域增多,纤维化程度重;别嘌呤醇组肾组织纤维化改变不明显;威草胶囊小剂量组和大剂量组肾组织间质纤维化程度明显减轻,威草胶囊大剂量组接近正常。模型组大鼠肾组织中CTGF、TGF-β1、ColⅠ、ColⅢ表达量均明显高于正常组(P均<0.05);威草胶囊小剂量组和大剂量组CTGF、ColⅠ、ColⅢ表达量和威草胶囊大剂量组TGF-β1表达量均明显低于模型组和别嘌呤醇组(P均<0.05);威草胶囊小剂量组CTGF、TGF-β1、ColⅠ、ColⅢ表达量与威草胶囊大剂量组比较差异均无统计学意义(P均>0.05)。模型组大鼠肾组织中miR-29a、miR-30c表达水平均明显低于正常组(P均<0.05);威草胶囊小剂量组和大剂量组miR-29a、miR-30c表达水平均明显高于模型组(P均<0.05),别嘌呤醇组与模型组比较差异无统计学意义(P均>0.05);威草胶囊大剂量组miR-29a、miR-30c表达水平均明显高于别嘌呤醇组(P均<0.05);威草胶囊小剂量组miR-29a、miR-30c表达水平与威草胶囊大剂量组比较差异均无统计学意义(P均>0.05)。结论威草胶囊可改善尿酸性肾病大鼠的肾功能,减轻肾组织纤维化程度,与其升高肾组织miR-29a、miR-30c的表达水平,降低TGF-β1、CTGF的表达,减少ColⅠ、ColⅢ的生成有关,别嘌呤醇不具有此作用。
基金This research was supported by grants from the National Natural Science Foundation of China,No.11625209.
文摘During vascular remodeling after intimal injury,circulating platelets are activated by exposed collagen and release platelet-derived microvesicles(PMVs),which may contribute to the injury-induced apoptosis of vascular smooth muscle cells(VSMCs).However,the mechanisms in this process are still unclear.Using a rat balloon injury model,platelet adhesion at the injured site was detected,and promoted medial VSMC apoptosis was revealed with dT-mediated dUTP nick-end labeling(TUNEL)in vivo.VSMC apoptosis was promoted by coincubation with PMVs in vitro.Transcriptomics analysis indicated the abound expression of the microRNA-30(miR-30)family in platelets,and the expression of the miR-30 family in platelets and PMVs was confirmed with qPCR.Ingenuity Pathway Analysis(IPA)software identified transcription factor 2(RUNX2)as a pivotal molecule linking miR-30 and cellular apoptosis.In vitro,PMVs delivered miR-30e,an important member of the miR-30 family,into VSMCs and increased cell apoptosis.A dual-luciferase assay was adopted to verify the interaction of miR-30e with RUNX2,and fluorescence in situ hybridization(FISH)proved the cytoplasmic localization of miR-30e in VSMCs.The apoptotic effect on VSMCs was further confirmed by using miR-30e mimics and RUNX2 siRNA.Furthermore,RUNX2 siRNA altered the expression of its downstream genes,Tnfrsf11b,Smad6 and Mmp13,which may participate in apoptosis,as suggested by IPA.Our results indicated that PMVs play important roles in the interactions between circulating elements and VSMCs by delivering miR-30e to VSMCs and then promoting apoptosis.PMVs and potential intercellular messages may be novel therapeutic targets for preventing pathological vascular remodeling after intimal injury.
