Role of microRNA9 in spinal dorsal horn neurons in over-expression of calcium homeostasis modulator 1 in rats with diabetic neuropathic pain
10.3760/cma.j.issn.0254-1416.2017.08.015
- VernacularTitle:脊髓背角神经元miR-9在糖尿病神经痛大鼠钙稳态调节蛋白1过表达中的作用
- Author:
Wenjie LIU
1
;
Jun JIANG
;
Anchao LI
;
Wanyou HE
;
Xiaoling HU
;
Qulian GUO
Author Information
1. 南华大学附属第一医院麻醉科
- Keywords:
microRNAs;
Calcium channels;
Spinal cord;
Neurons;
Diabetes mellitus;
Neuralgia
- From:
Chinese Journal of Anesthesiology
2017;37(8):954-957
- CountryChina
- Language:Chinese
-
Abstract:
Objective To evaluate the role of microRNA9 (miR-9) in spinal dorsal horn neurons in over-expression of calcium homeostasis modulator 1 (CALHM1) in rats with diabetic neuropathic pain (DNP).Methods Ninety-three healthy male Sprague-Dawley rats,aged 2 months,weighing 180-200 g,were used in the study.Diabetes mellitus was induced by intraperitoneal 1％ streptozocin (STZ) 60 mg/kg.The experiment was performed in two parts.Experiment Ⅰ The rats were divided into control group (group C,n=10) and DNP group (n=83) using a random number table.The mechanical paw withdrawal threshold (MWT) was measured before STZ injection and at 1,2,3,4,5 and 6 weeks after STZ injection.The expression of miR-9 in the spinal dorsal horn was determined using the in situ hybridization at 6 weeks after STZ injection.Experiment Ⅱ The rats with DNP were selected at 6 weeks after STZ injection,and the spinal dorsal horn neurons were isolated and cultured.The neurons were seeded in culture plates at the density of 5×106 cells/ml (2 ml/well) and divided into 2 groups (n=18 each) using a random number table:control group (group C) and miR-9 antisense oligonucleotide group (group ASO).The neurons were cultured in normal culture atmosphere in group C.In group ASO,the single nucleotide sequence of miR-9 antisense oligonucleotide sequence 5'-UUCUCCGAACGUGUCACGUTT-3'was added with the final concentration of 100 pmol/L.The expression of miR-9 and CALHM1 mRNA was detected using quantitative real-time polymerase chain reaction at 48 h of incubation.The expression of CALHM1 was detected by Western blot at 72 h of incubation.Results Experiment Ⅰ Compared with group C,the MWT was significantly decreased at 2-6 weeks after STZ injection,and the expression of miR-9 in the spinal dorsal horn was up-regulated in group DNP (P＜0.05).Experiment Ⅱ Compared with group C,the expression of miR-9 and CALHM1 protein and mRNA in spinal dorsal horn neurons was significantly down-regulated in group ASO (P＜0.05).Conclusion miR-9 in spinal dorsal horn neurons probably mediates the over-expression of CALHM1 in rats with DNP.
