[SR 59230A对心力衰竭大鼠心肌中微小RNA表达的影响]

[SR 59230A on the expression of MicroRNAs in myocardium of heart failure rats].

作者信息

Jing Jia-Ni, Li Fan-Lu, Wang Xi, Wan Xin, Zhao Qian-Qian, Cui Xiang-Li

机构信息

Department of Physiology, Cell Physiology Key laboratory of Shanxi Province, Shanxi Medical University, Taiyuan, Shanxi 030001, China.

出版信息

Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2017 May 8;33(5):456-460. doi: 10.12047/j.cjap.5561.2017.109.

DOI:10.12047/j.cjap.5561.2017.109

PMID:29926593

Abstract

OBJECTIVE

To investigate the effects of β3-adrenoceptors(β3-AR) inhibitor SR 59230A on MicroRNAs expression in rat myocardium with chronic heart failure and the related mechanisms.

METHODS

One hundred male SD rats were randomly divided into sham operated group(40)and chronic heart failure(CHF)group(60). Coronary artery ligation was used to induce CHF. Then the rats in CHF group were further randomly divided into CHF control group and CHF+SR 59230A group (CHF+SR). Rats in the sham group were divided into sham control group and sham+SR 59230A group (Sham+SR). The rats in Sham+SR group and CHF+SR group were treated with 1 ml SR 59230A(85 mmoL/L in 0.9% saline)twice a day for seven weeks by intraperitoneal injection, while the rats in control groups were injected with the same amount of saline for seven weeks separately. miScript miRNA PCR Arrays were used to determine the expression profile of MicroRNAs. Immunohistochemistry was used to evaluate the distribution of the related proteins in the heart tissue sections. Western blot was used to detect the expressions of nuclear factor-kappaB(NF-κB),p53 and p53-Phospho-Serine 15 in the heart.

RESULTS

①After blockade of β3-AR by SR 59230A, there were 18MicroRNAs down-regulated in sham control group and CHF control group. Within them, 6 MicroRNAs were related to NF-κB signaling pathway, they were miR-125b-5p,miR-143-3p,miR-145-5p,miR-26a-5p,miR-30a-5p and miR-320-5p. ②Slides from the heart tissue showed that NF-κB was distributed both in nucleus and cytoplasm, while p53 in cytoplasm was more than that in nucleus in heart tissue sections. The expressions of NF-κB and p53 were higher in the CHF control group than those in the sham control group(<0.05), but were lower in CHF+SR group than those in CHF control group(<0.05),while they were elevated in Sham+SR group compared to the sham control group(<0.05). ③ Compared with the sham control group, the protein expression of NF-κB p65 was increased significantly in the CHF control group (<0.05). After treated with SR59230A ,the protein expressions of NF-κB and p53-Phospho-Serine 15 were decreased significantly in CHF rats(<0.05),while the protein expressions of NF-κB, p53 and p53-Phospho-Serine 15 proteins were increased in the sham rats (<0.05).

CONCLUSIONS

Blocking of β3-AR improved the damaged heart in CHF rats; β3-AR caused the change of MicroRNAs expression, and it related to NF-κB signal pathway.

摘要

目的

探讨β3-肾上腺素能受体（β3-AR）抑制剂SR 59230A对慢性心力衰竭大鼠心肌微小RNA表达的影响及其相关机制。

方法

将100只雄性SD大鼠随机分为假手术组（40只）和慢性心力衰竭（CHF）组（60只）。采用冠状动脉结扎法诱导CHF。然后将CHF组大鼠进一步随机分为CHF对照组和CHF+SR 59230A组（CHF+SR）。假手术组大鼠分为假手术对照组和假手术+SR 59230A组（Sham+SR）。Sham+SR组和CHF+SR组大鼠腹腔注射1 ml SR 59230A（85 mmol/L溶于0.9%生理盐水），每日2次，共7周，而对照组大鼠分别注射等量生理盐水7周。采用miScript miRNA PCR芯片检测微小RNA的表达谱。免疫组织化学法评估心脏组织切片中相关蛋白的分布。蛋白质印迹法检测心脏中核因子-κB（NF-κB）、p53和p53-磷酸化丝氨酸15的表达。

结果

①SR 59230A阻断β3-AR后，假手术对照组和CHF对照组中有18种微小RNA表达下调。其中，6种微小RNA与NF-κB信号通路相关，分别为miR-125b-5p、miR-143-3p、miR-145-5p、miR-26a-5p、miR-30a-5p和miR-320-5p。②心脏组织切片显示，NF-κB在细胞核和细胞质中均有分布，而心脏组织切片中p53在细胞质中的表达多于细胞核。CHF对照组中NF-κB和p53的表达高于假手术对照组（<0.05），但CHF+SR组低于CHF对照组（<0.05），而Sham+SR组与假手术对照组相比升高（<0.05）。③与假手术对照组相比，CHF对照组中NF-κB p65的蛋白表达显著增加（<0.05）。SR59230A处理后，CHF大鼠中NF-κB和p53-磷酸化丝氨酸15的蛋白表达显著降低（<0.05），而假手术大鼠中NF-κB、p53和p53-磷酸化丝氨酸15蛋白的表达增加（<0.05）。