Protective effect of baicalin on nerve injury in rats with cerebral microbleeds through PI3K/AKT/eNOS pathway

doi:10.11958/20250606

Abstract

Abstract:

Objective To explore the protective effect of baicalin on nerve damage in rats with cerebral microbleeds through the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/endothelial nitric oxide synthase (eNOS) pathway. Methods The rat model of cerebral microbleeds was established by intraventricular injection of lipopolysaccharide (LPS), and rats were separated into the model group, the baicalin group (20 mg/kg), the LY294002 group (PI3K inhibitor, 10 mg/kg) and the baicalin+LY294002 group (20 mg/kg baicalin and 10 mg/kg LY294002). Rats without LPS injection were served as the control group. The nerve function was evaluated in five groups of rats. Triphenyltetrazolium chloride (TTC) staining was used to evaluate the cerebral infarction status. Hematoxylin-eosin (HE) staining was used to observe the pathological morphology of brain tissue of rats in each group. TUNEL method was used to detect neuronal apoptosis in brain tissue. Enzyme linked immunosorbent assay (ELISA) was performed to measure tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-17 (IL-17) in brain tissue of rats in each group. In addition, Western blot assay was used to measure B-cell lymphoma 2 (Bcl-2), Caspase-3, Bcl-2 associated X protein (Bax), PI3K, phosphorylated (p)-AKT, AKT, p-eNOS and eNOS proteins in brain tissue of rats. Results Compared with the control group, neurons in the model group was sparse, with fewer cells and disordered arrangement, the nerve function score, infarct area, TNF-α, IL-1β, IL-17, apoptosis rate, Bax and Caspase-3 proteins were increased, and Bcl-2, PI3K, p-AKT and p-eNOS proteins were decreased (P<0.05). Compared with the model group, the baicalin group showed clear improvement in brain pathological damage, the nerve function score, infarct area, apoptosis rate, Bax and Caspase-3 proteins, TNF-α, IL-1β and IL-17 were decreased, while protein expressions of Bcl-2, PI3K, p-AKT and p-eNOS were increased (P<0.05). However, the brain tissue damage of the LY294002 group was further aggravated, and the nerve function score, infarct area, TNF-α, IL-1β, IL-17, apoptosis rate and the expression of apoptosis proteins Bax and Caspase-3 were increased, the Bcl-2, PI3K, p-AKT and p-eNOS proteins were decreased (P<0.05). Intervention with LY294002 on basis of baicalin treatment could reverse the improvement effect of baicalin on the above indicators in rats with cerebral microbleeds (P<0.05). Conclusion Baicalin may exert a protective effect on nerve damage in rats with cerebral microbleeds by activating PI3K/AKT/eNOS pathway.

Key words: baicalin, cerebral hemorrhage, PI3K/AKT/eNOS, nerve damage

CLC Number:

R743

Figures/Tables 8

References 21

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组别	细胞凋亡率/%	Bax	Caspase-3	Bcl-2
对照组	2.94±0.37	0.31±0.03	0.27±0.03	0.95±0.09
模型组	32.41±3.65^a	0.90±0.08^a	0.92±0.09^a	0.37±0.03^a
黄芩苷组	8.83±0.94^b	0.42±0.04^b	0.39±0.04^b	0.86±0.07^b
LY294002组	49.56±5.72^b	1.26±0.10^b	1.35±0.12^b	0.10±0.02^b
黄芩苷+LY294002组	31.02±2.96^c	0.87±0.08^c	0.88±0.09^c	0.41±0.04^c
F	161.497^＊＊	148.093^＊＊	144.464^＊＊	200.425^＊＊

组别	细胞凋亡率/%	Bax	Caspase-3	Bcl-2
对照组	2.94±0.37	0.31±0.03	0.27±0.03	0.95±0.09
模型组	32.41±3.65^a	0.90±0.08^a	0.92±0.09^a	0.37±0.03^a
黄芩苷组	8.83±0.94^b	0.42±0.04^b	0.39±0.04^b	0.86±0.07^b
LY294002组	49.56±5.72^b	1.26±0.10^b	1.35±0.12^b	0.10±0.02^b
黄芩苷+LY294002组	31.02±2.96^c	0.87±0.08^c	0.88±0.09^c	0.41±0.04^c
F	161.497^＊＊	148.093^＊＊	144.464^＊＊	200.425^＊＊

组别	TNF-α/（ng/L）	IL-1β/（ng/L）	IL-17/（μg/L）
对照组	24.63±2.89	21.54±2.36	18.47±2.51
模型组	65.48±6.33^a	78.25±6.73^a	52.82±5.03^a
黄芩苷组	29.85±3.16^b	26.39±3.24^b	23.61±2.54^b
LY294002组	86.73±6.81^b	97.42±7.92^b	68.93±5.78^b
黄芩苷+LY294002组	62.57±6.06^c	71.26±6.87^c	49.85±4.72^c
F	120.233^＊＊	162.874^＊＊	119.551^＊＊

组别	TNF-α/（ng/L）	IL-1β/（ng/L）	IL-17/（μg/L）
对照组	24.63±2.89	21.54±2.36	18.47±2.51
模型组	65.48±6.33^a	78.25±6.73^a	52.82±5.03^a
黄芩苷组	29.85±3.16^b	26.39±3.24^b	23.61±2.54^b
LY294002组	86.73±6.81^b	97.42±7.92^b	68.93±5.78^b
黄芩苷+LY294002组	62.57±6.06^c	71.26±6.87^c	49.85±4.72^c
F	120.233^＊＊	162.874^＊＊	119.551^＊＊

组别	PI3K	p-AKT	p-eNOS
对照组	1.34±0.12	0.95±0.08	0.91±0.09
模型组	0.47±0.04^a	0.39±0.03^a	0.36±0.03^a
黄芩苷组	1.19±0.11^b	0.91±0.08^b	0.83±0.07^b
LY294002组	0.21±0.02^b	0.16±0.02^b	0.12±0.01^b
黄芩苷+LY294002组	0.52±0.05^c	0.43±0.04^c	0.39±0.03^c
F	194.460^＊＊	191.115^＊＊	189.044^＊＊