Effects of rutin on hippocampal neuron injury and RhoA/ROCK signaling pathway in rats with chronic cerebral hypoperfusion

doi:10.11958/20220710

Abstract

Abstract:

Objective To investigate the effects of rutin on hippocampal neuron injury and Ras homolog gene family member A/Rho-associated coiled-coil protein kinase (RhoA/ROCK) signaling pathway in chronic cerebral hypoperfusion rats. Methods SD rats were randomly divided into the sham operation group, the model group, the rutin low dose group (20 mg/kg), the rutin medium dose group (40 mg/kg), the rutin high dose group (80 mg/kg) and RhoA inhibitor group (Rhosin hydrochloride, 40 mg/kg), with 9 rats in each group. Except the sham operation group, rats in the other groups were subjected to chronic cerebral hypoperfusion by ligating carotid artery. Intervention treatment was carried out according to the dosage of each group. The drug was administered once a day for 4 weeks. Morris water maze test was used to determine the cognitive function and memory ability of rats. Hematoxylin eosin staining was used to observe the status of hippocampal neurons. TUNEL staining was used to detect the apoptosis of hippocampal neurons. Transmission electron microscope was used to observe the ultrastructure of hippocampal neurons. Western blot assay was used to detect the expression levels of apoptosis related proteins [Caspase-3, B cell lymphoma-2 associated X protein (Bax)] and RhoA/ROCK pathway related proteins in hippocampus of rats in each group. Results Compared with the sham operation group, the number of neurons in hippocampus decreased in the model group, the ultrastructure and organelle structure of neurons were seriously damaged, the escape latency, apoptosis rate of hippocampal neurons, expression levels of apoptosis related proteins (Caspase-3, Bax), RhoA, ROCK1 and ROCK2 were significantly increased (P<0.05), and the number of platform crossings was significantly decreased (P<0.05). Compared with the model group, the number of hippocampal neurons significantly increased in the rutin treatment groups and RhoA inhibitor group, the ultrastructure of neurons and the damage of organelles recovered to a certain extent, the escape latency, apoptosis rate of hippocampal neurons, expression levels of apoptosis related proteins (Caspase-3, Bax), RhoA, ROCK1 and ROCK2 were significantly decreased (P<0.05), and the number of platform crossings was significantly increased (P<0.05). There was a dose-dependent effect in the rutin treatment groups. There were no significant differences in above indexes between the rutin high dose group and the RhoA inhibitor group (P>0.05). Conclusion Rutin can alleviate the neuronal injury induced by chronic cerebral hypoperfusion in rats, which may be achieved by inhibiting RhoA/ROCK signaling pathway.

Key words: rutin, neurons, rho-associated kinases, apoptosis, cognitive dysfunction, memory, chronic cerebral hypoperfusion

CLC Number:

R743

Figures/Tables 8

References 22

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组别	逃避潜伏期（s）				穿越平台次数（次）
组别	第1天	第2天	第3天	第4天	穿越平台次数（次）
假手术组	37.10±7.38	36.12±6.59	35.28±7.03	34.15±7.11	17.62±3.67
模型组	85.06±10.75^a	83.94±10.52^a	82.31±10.58^a	81.85±11.94^a	4.96±1.03^a
芦丁低剂量组	72.55±9.02^ab	71.83±9.87^ab	70.50±9.68^ab	69.33±10.52^ab	6.53±1.35^ab
芦丁中剂量组	62.83±8.74^abc	61.06±9.33^abc	59.22±8.36^abc	57.60±10.58^abc	9.18±2.15^abc
芦丁高剂量组	45.44±9.17^abcd	44.26±8.94^abcd	43.81±8.07^abcd	42.40±9.00^abcd	13.12±2.65^abcd
RhoA抑制剂组	47.08±8.99^abcd	46.52±7.99^abcd	45.60±9.70^abcd	44.28±8.94^abcd	14.61±3.10^abcd
F	36.835^＊＊	37.389^＊＊	35.664^＊＊	30.731^＊＊	34.785^＊＊

组别	逃避潜伏期（s）				穿越平台次数（次）
组别	第1天	第2天	第3天	第4天	穿越平台次数（次）
假手术组	37.10±7.38	36.12±6.59	35.28±7.03	34.15±7.11	17.62±3.67
模型组	85.06±10.75^a	83.94±10.52^a	82.31±10.58^a	81.85±11.94^a	4.96±1.03^a
芦丁低剂量组	72.55±9.02^ab	71.83±9.87^ab	70.50±9.68^ab	69.33±10.52^ab	6.53±1.35^ab
芦丁中剂量组	62.83±8.74^abc	61.06±9.33^abc	59.22±8.36^abc	57.60±10.58^abc	9.18±2.15^abc
芦丁高剂量组	45.44±9.17^abcd	44.26±8.94^abcd	43.81±8.07^abcd	42.40±9.00^abcd	13.12±2.65^abcd
RhoA抑制剂组	47.08±8.99^abcd	46.52±7.99^abcd	45.60±9.70^abcd	44.28±8.94^abcd	14.61±3.10^abcd
F	36.835^＊＊	37.389^＊＊	35.664^＊＊	30.731^＊＊	34.785^＊＊

组别	凋亡率（%）	凋亡相关蛋白
组别	凋亡率（%）	Caspase-3	Bax
假手术组	2.84±0.59	0.21±0.04	0.23±0.05
模型组	30.25±5.31^a	1.26±0.18^a	1.30±0.15^a
芦丁低剂量组	19.13±3.96^ab	0.90±0.13^ab	0.97±0.14^ab
芦丁中剂量组	11.26±2.73^abc	0.61±0.10^abc	0.66±0.12^abc
芦丁高剂量组	6.29±1.33^abcd	0.41±0.06^abcd	0.40±0.10^abcd
RhoA抑制剂组	5.87±1.24^abcd	0.39±0.07^abcd	0.43±0.10^abcd
F	35.114^＊＊	39.040^＊＊	36.955^＊＊

组别	凋亡率（%）	凋亡相关蛋白
组别	凋亡率（%）	Caspase-3	Bax
假手术组	2.84±0.59	0.21±0.04	0.23±0.05
模型组	30.25±5.31^a	1.26±0.18^a	1.30±0.15^a
芦丁低剂量组	19.13±3.96^ab	0.90±0.13^ab	0.97±0.14^ab
芦丁中剂量组	11.26±2.73^abc	0.61±0.10^abc	0.66±0.12^abc
芦丁高剂量组	6.29±1.33^abcd	0.41±0.06^abcd	0.40±0.10^abcd
RhoA抑制剂组	5.87±1.24^abcd	0.39±0.07^abcd	0.43±0.10^abcd
F	35.114^＊＊	39.040^＊＊	36.955^＊＊

组别	RhoA	ROCK1	ROCK2
假手术组	0.11±0.03	0.21±0.05	0.17±0.03
模型组	1.08±0.12^a	1.28±0.14^a	1.34±0.20^a
芦丁低剂量组	0.79±0.11^ab	0.99±0.12^ab	0.95±0.15^ab
芦丁中剂量组	0.53±0.09^abc	0.71±0.09^abc	0.63±0.11^abc
芦丁高剂量组	0.32±0.06^abcd	0.41±0.07^abcd	0.33±0.06^abcd
RhoA抑制剂组	0.30±0.05^abcd	0.43±0.07^abcd	0.36±0.07^abcd
F	55.651^＊＊	53.794^＊＊	42.043^＊＊