Rho/ROCK信号通路在阻塞性睡眠呼吸暂停低通气综合征大鼠急性缺血性卒中发生中的作用

doi:10.13481/j.1671-587X.20210514

Abstract

Abstract: Objective

To study the role of small GTP binding protein （Rho）/Rho-related coiled-coil forming protein kinase （ROCK） signaling pathway in the occurrence of acute ischemic stroke in the rats with obstructive sleep apnea hypopnea syndrome （OSAHS），and to provide reference for the prevention and treatment of acute ischemic stroke after OSAHS.

Methods

Fifty SD rats were randomly divided into sham operation group， OSAHS group， stroke group， post-OSAHS stroke group， and Rho inhibitor group （C3 transferase 5 μg·kg^－1·d^－1 intraperitoneal injection）， with 10 rats in each group. The OSAHS model was established by chronic intermittent hypoxia， and the middle cerebral artery was blocked by the suture method to establish an acute ischemic stroke model. The general situation and pathomorphology of cerebral cortex tissue of the rats after modeling were observed and the volumes of cerebral infarction were calculated. The respiratory frequencies and the neurological impairment scores of the rats in various groups were compared. The expression levels of myosin light chain （MLC）， Rho and ROCK in brain tissue of the rats in various groups were detected by Real-time fluorescence quantitative PCR （RT-qPCR）and Western blotting methods.

Results

Compared with sham operation group， the rats in other groups had changes such as poor mental state and cerebral cortex structure damage， among them， the rats in post-OSAHS stroke group changed the most and the rats in Rho inhibitor group changed slightly. Compared with sham operation group，the respiratory frequency of rats in OSAHS group was increased （P<0.05）；compared with OSAHS group， the respiratory frequency of rats in stroke group was decreased （P<0.05）； compared with stroke group， the respiratory rate and Longa score of rats in post-OSAHS stroke group were increased（P<0.05）， and the Longa score of rats in Rho inhibitor group were decreased （P<0.05）； compared with post-OSAHS stroke group， the respiratory frequency and Longa score of rats in Rho inhibitor group were decreased （P<0.05）. Compared with sham operation group， the escape latency of rats in OSAHS group was prolonged（P<0.05）， and the number of crossing platform was decreased （P<0.05）； compared with OSAHS group， the escape latency of rats in stroke group was prolonged（P<0.05） and the number of crossing platform was decreased （P<0.05）； compared with stroke group， the escape latency of rats in post-OSAHS stroke group was prolonged（P<0.05）， and the number of crossing platform was decreased（P<0.05）；compared with post-OSAHS stroke group， the escape latency of rats in Rho inhibitor group was shortened（P<0.05），and the times of crossing platform was decreased（P<0.05）. Compared with stroke group， the cerebral infarction volume of rats in post-OSAHS stroke group was increased（P<0.05）； compared with post-OSAHS stroke group，the cerebral infarction volume of rats in Rho inhibitor group was decreased（P<0.05）. Compared with sham operation group， the expression levels of Rho， rock mRNA and proteins and the P-MLC / MLC ratio in brain tissue of the rats in OSAHS group were increased （P<0.05）；compared with OSAHS group， the expression levels of Rho and ROCK mRNA and proteins and the p-MLC/MLC ratio in brain tissue of the rats in stroke group were increased （P<0.05）； compared with stroke group， the expression levels of Rho and ROCK mRNA and proteins in brain tissue of the rats in post-OSAHS stroke group and the p-MLC/MLC ratio were increased （P<0.05）；compared with post-OSAHS stroke group， the expression levels of Rho and ROCK mRNA and proteins in brain tissue of the rats in Rho inhibitor group and the p-MLC/MLC ratio were decreased （P<0.05）.

Conclusion

The intermittent hypoxia of OSAHS can damage the nerve function， and Rho/ROCK signaling pathway may play a role in the occurrence of acute ischemic stroke in the OSAHS rats.

Key words: obstructive sleep apnea hypopnea syndrome, acute ischemic stroke, small GTP binding protein, Rho associated coiled-coil forming protein kinase, myosin light chain

CLC Number:

R543.5

Peng JI,Xiangdong GUO,Gen SUN,Qixing KOU,Xueping QU,Yajing SUN,Limin JIANG. Role of Rho/ROCK signaling pathway in occurrence of acute ischemic stroke in rats with obstructive sleep apnea hypopnea syndrome[J].Journal of Jilin University(Medicine Edition), 2021, 47(5): 1178-1186.

Figures/Tables 8

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References 20

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Gene	Primer sequences
Rho	F:5′-AGTAGCCACGCGTGATTACGTTGC-3′
Rho	R:5′-TTAGCCTGAAGCTGATGTACGTGA-3′
ROCK	F:5′-TAGTCCTGTAGTATGAGTGTC-3′
ROCK	R:5′-CTGTAGAGGAGATTCTCGTGA-3′
MLC	F:5′-ATGACGATGTATGGCTCTG-3′
MLC	R:5′-TGTGCTATGGCATGTGACT-3′
β-actin	F:5′-TGTACTGAGCGTGTTATGTTGTA-3′
β-actin	R:5′-GAGCTGATGTGACTCGTGATCGG-3′

Group	n	Respiratory frequency （times·min^－1）	Longa score（points）
Sham operation	9	89.25±9.22	—
OSAHS	10	133.89±11.08^*	—
Stroke	9	92.32±8.50^△	1.03±0.25
Post-OSAHS stroke	8	134.50±10.74^*#	1.33±0.30^#
Rho inhibitor	9	105.32±9.25^*△#○	0.85±0.23^#○

Group	n	Escape latency（t/s）	Number of crossing platforms（times）
Sham operation	9	25.33±3.02	6.83±1.50
OSAHS	10	31.36±3.35^*	5.21±1.03^*
Stroke	9	38.89±4.50^*△	3.05±0.70^*△
Post-OSAHS stroke	8	48.48±5.12^*△#	2.08±0.52^*△#
Rho inhibitor	9	32.41±3.50^*#○	4.89±1.05^*#○

Group	Rho	ROCK	MLC
Sham operation	0.44±0.05	0.65±0.06	0.91±0.08
OSAHS	0.55±0.08^*	0.80±0.08^*	0.88±0.10
Stroke	0.82±0.10^*△	0.95±0.10^*△	0.92±0.10
Post-OSAHS stroke	1.08±0.12^*△#	1.22±0.14^*△#	0.95±0.12
Rho inhibitor	0.25±0.05^*△#○	0.32±0.05^*△#○	0.90±0.08

Group	Rho	ROCK	p-MLC/MLC
Sham operation	0.15±0.03	0.05±0.01	0.12±0.03
OSAHS	0.22±0.04^*	0.12±0.03^*	0.30±0.05^*
Stroke	0.35±0.05^*△	0.23±0.05^*△	0.45±0.05^*△
Post-OSAHS stroke	0.45±0.05^*△#	0.50±0.06^*△#	0.70±0.07^*△#
Rho inhibitor	0.09±0.03^*△#○	0.03±0.01^*△#○	0.06±0.02^*△#○

Role of Rho/ROCK signaling pathway in occurrence of acute ischemic stroke in rats with obstructive sleep apnea hypopnea syndrome

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