脂联素通过抑制线粒体融合与分裂失衡减轻慢性间歇低氧所致大鼠胰岛细胞损伤

doi:10.24920/003834

摘要/Abstract

摘要：

目的探讨脂联素（adiponectin, APN）对慢性间歇低氧（chronic intermittent hypoxia, CIH）所致大鼠胰岛损伤的保护作用及相关机制。
方法 60只SD大鼠随机分为3组：对照（normal contral,NC）组、CIH组及CIH+APN组。5周的CIH暴露后,我们对大鼠进行口服葡萄糖耐量试验（oral glucose tolerance test, OGTT）及胰岛素释放试验（insulin released test, IRT）。提取胰岛,检测并比较三组间大鼠胰岛ATP水平,线粒体膜电位（mitochondrial membrane potential, MMP）水平,活性氧（reactive oxygen species, ROS）水平,线粒体三羧酸循环相关酶基因Ant1、Cs、Hmox1和Cox4i1表达水平,线粒体融合与分裂相关蛋白及基因DRP1、FIS1、MFN1和OPA1表达水平,及胰岛线粒体凋亡相关蛋白BAX、BCL-2、cleaved Caspase-3、cleaved PARP表达水平。
结果 CIH暴露5周后,OGTT及IRT试验显示各组大鼠在0分钟、20分钟、30分钟、60分钟及120分钟的血糖及胰岛素水平均无明显差异。然而我们发现CIH暴露后,大鼠胰岛ROS水平增加,ATP及MMP水平下降, Ant1、Cs、Hmox1和Cox4i1基因表达水平下降,MFN1和OPA1蛋白及基因表达水平下降,DRP1、FIS1蛋白及基因表达水平增加,cleaved Caspase-3和cleaved PARP蛋白表达水平增加,BCL-2/BAX蛋白表达水平比率下降,三组间差异均有统计学显著性。CIH+APN组上述胰岛损伤相关的指标测量值均明显改善。
结论 APN通过抑制线粒体融合与分裂失衡减轻CIH所致大鼠胰岛线粒体损伤,进而减轻大鼠胰岛损伤。

关键词: 阻塞性睡眠呼吸暂停低通气综合征, 慢性间歇低氧, 脂联素, 线粒体融合与分裂, 胰岛

Abstract:

Objective This study aimed to assess the protective value of adiponectin (APN) in pancreatic islet injury induced by chronic intermittent hypoxia (CIH).
Methods Sixty rats were randomly divided into three groups: normal control (NC) group, CIH group, and CIH with APN supplement (CIH+APN) group. After 5 weeks of CIH exposure, we conducted oral glucose tolerance tests (OGTT) and insulin released test (IRT), examined and compared the adenosine triphosphate (ATP) levels, mitochondrial membrane potential (MMP) levels, reactive oxygen species (ROS) levels, enzymes gene expression levels of Ant1, Cs, Hmox1, and Cox4i1 which represented mitochondrial tricarboxylic acid cycle function, the protein and gene expression levels of DRP1, FIS1, MFN1, and OPA1 which represented mitochondrial fusion and division, and the protein expression levels of BAX, BCL-2, cleaved Caspase-3, and cleaved PARP which represented mitochondrial associated apoptosis pathway of pancreatic islet.
Results OGTT and IRT showed blood glucose and insulin levels had no differences among the NC, CIH and CIH+APN groups (both P>0.05) at 0 min, 20 min, 30 min, 60 min, 120 min. However, we found that compared to NC group, CIH increased the ROS level, reduced ATP level and MMP level. The islets of CIH exposed rats showed reduced gene expression levels of Ant1, Cs, Hmox1, and Cox4i1, decreased protein and gene expression levels of MFN1 and OPA1, increased protein and gene expression levels of DRP1 and FIS1, increased protein expression levels of cleaved Caspase-3 and cleaved PARP, with lower ratio of BCL-2/BAX at protein expression level. All the differences among three groups were statistically significant. APN treated CIH rats showed mitigated changes in the above measurements associated with islet injuries.
Conclusion APN may ameliorate the pancreatic islet injury induced by CIH via inhibiting the imbalance in mitochondrial fusion and division.

Key words: obstructive sleep apnea hypopnea syndrome, chronic intermittent hypoxia, adiponectin, mitochondrial fusion and division, pancreatic islet

基金资助: 国家自然科学基金(81700090);南京市医学科技发展项目(YKK17237)

Can He, Xilong Zhang, Qiang Zhang, Luyao Ge, Wenxiao Ding. Adiponectin Ameliorated Pancreatic Islet Injury Induced by Chronic Intermittent Hypoxia through Inhibiting the Imbalance in Mitochondrial Fusion and Division[J].Chinese Medical Sciences Journal, 2021, 36(3): 225-233.

图/表 7

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Primer	Forward	Reverse
Drp1	5’- ACTGGCCCCCGTCCAGCTTT -3’	5’- AGCTCCACACAGCGGAGGCT -3’
Fis1	5’- GCACGCAGTTTGAATACGCC -3’	5’- CTGCTCCTCTTTGCTACCTTTGG -3’
Mfn1	5’- CCTTGTACATCGATTCCTGGGTTC-3’	5’- CCTGGGCTGCATTATCTGGTG -3’
Opa1	5’- CCTTGTACATCGATTCCTGGGTTC -3’	5’- CCTGGGCTGCATTATCTGGTG -3’
Hmox1	5’- GTCAAGCACAGGGTGACAGA -3’	5’- CTGCAGCTCCTCAAACAGC -3’
Cs	5’- GCACGCCAGTGCTTCTTC -3’	5’- CATGCTGCTGTCTGAAGGTC -3’
Ant1	5’- GTAGGATGATGATGCAGTCTGG -3’	5’- CGTCCTTCATCTTTTGCAATC -3’
Cox4i1	5’- CACTGCGCTTGTGCTGAT -3’	5’- CGATCAAAGGTATGAGGGATG -3’
Gapdh	5’- GCCCTCACCACGTGGCCTTG -3’	5’- TGCAGCCCAGGGCCTTTGAC -3’

Measurements	NC group	CIH group	CIH+APN group	F	P value
ATP (nmol/mg)	15.13±1.60	7.83±0.78*	11.37±1.41*^#	23.38	0.0015
MMP levels	25.96±1.33	13.22±1.03*	19.75±1.19*^#	86.04	<0.0001

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