Astragaloside inhibits Fas/FasL signaling pathway to reduce neural dysfunction and neuronal apoptosis in traumatic brain injury of rats

doi:10.11958/20231226

Abstract

Abstract:

Objective To explore the effects of astragaloside on neural dysfunction and neuronal apoptosis of traumatic brain injury (TBI) in rats through Fas/FasL signaling pathway. Methods Fifty rats were randomly divided into 5 groups: the sham operation group, the model group, the astragaloside group (20 mg/kg), the Fas silencing group [4 μg Fas small interfering RNA (siRNA) lentiviral vector] and the astragaloside+Fas silencing group (20 mg/kg astragaloside+4 μg Fas siRNA lentiviral vector). Each group consisted of 10 rats. Except the sham operation group, the other groups of rats were established TBI rat model. Each group was received medication intervention according to the corresponding dosage, once a day, for 7 days. Water maze test was used to detect the nerve function defect in rats. Fluorescence quantitative PCR was used to detect expression levels of Fas and FasL messenger RNA (mRNA) in brain tissue. Hematoxylin-eosin, β-tubulin III (Tuj1) immunofluorescence and TUNEL staining were used to observe pathological changes, neuronal activity and apoptosis of brain tissue, respectively. Western blot assay was used to detect expression levels of Fas/FasL pathway, Caspase-3, Bcl-2 associated X protein (Bax) and B lymphoblastoma-2 (Bcl-2) protein in brain tissue. Results Compared with the sham operation group, the cell gap was increased and degeneration was obvious in brain tissue in the model group. The escape latency of rats on days 1-5, neuronal apoptosis rate, Caspase-3, Bax protein, Fas, FasL mRNA and protein levels were increased (P<0.05). The number of times crossing platforms, the number of Tuj1 positive cells and Bcl-2 protein were decreased (P<0.05). Compared with the model group, in the astragaloside group and the Fas silencing group, the cell gap in brain tissue was narrowed and degeneration was reduced, the escape latency of rats on days 1-5, neuronal apoptosis rate, Caspase-3, Bax protein, Fas, FasL mRNA and protein levels were decreased, and the number of crossing platforms, the number of Tuj1 positive cells and Bcl-2 protein level were increased (P<0.05). There were no significant differences in pathological changes of brain and the above indexes between the astragaloside group and the Fas silencing group (P>0.05). Compared with the astragaloside group and the Fas silencing group, the pathological damage of brain tissue of rats was further reduced in the astragaloside+Fas silencing group, neuronal apoptosis rate, Caspase-3, Bax protein, Fas, FasL mRNA and protein levels were decreased, and the number of Tuj1 positive cells and Bcl-2 protein were increased (P<0.05). Conclusion Astragaloside may reduce nerve function deficit and neuronal apoptosis in TBI rats by inhibiting Fas/FasL signaling pathway mediated apoptosis pathway.

Key words: brain injuries, traumatic, astragaloside Ⅳ, Fas receptor, neurons

CLC Number:

R651.15，R286.1

Figures/Tables 10

References 20

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基因名称	引物序列（5'→3'）	产物大小/bp
Fas	上游：TGCACAGAATTGAAGGAGTA	216
	下游：ATGGTTTCACGACTGGAGGT	216
FasL	上游：GACAGCAGTGCCACTTCATC	159
	下游：TTAAGGCTTTGGTTGGTGAA	159
GAPDH	上游：GGGAATAGTAGCTGTCAAATT	91
	下游：AATGACAGCTACCAGTCCCTT	91

基因名称	引物序列（5'→3'）	产物大小/bp
Fas	上游：TGCACAGAATTGAAGGAGTA	216
	下游：ATGGTTTCACGACTGGAGGT	216
FasL	上游：GACAGCAGTGCCACTTCATC	159
	下游：TTAAGGCTTTGGTTGGTGAA	159
GAPDH	上游：GGGAATAGTAGCTGTCAAATT	91
	下游：AATGACAGCTACCAGTCCCTT	91

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	第5天	穿越平台次数/次
假手术组	49.28±9.51	43.76±8.62	38.52±7.73	31.75±6.59	28.64±5.67	7.25±1.34
模型组	83.71±16.08^a	77.68±15.26^a	71.54±14.36^a	65.29±13.54^a	51.37±10.63^a	3.58±0.73^a
黄芪甲苷组	53.66±10.64^b	48.72±9.58^b	41.65±8.31^b	35.94±7.24^b	27.86±5.29^b	5.34±1.06^b
Fas沉默组	52.63±10.59^b	46.28±9.27^b	43.16±8.51^b	35.22±6.98^b	27.06±4.32^b	5.55±1.29^b
黄芪甲苷+Fas沉默组	46.59±9.35	41.20±8.27	34.59±6.85	28.61±5.54	22.36±4.15	6.52±1.27
F	17.217^＊＊	20.025^＊＊	23.807^＊＊	30.490^＊＊	31.068^＊＊	14.318^＊＊

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	第5天	穿越平台次数/次
假手术组	49.28±9.51	43.76±8.62	38.52±7.73	31.75±6.59	28.64±5.67	7.25±1.34
模型组	83.71±16.08^a	77.68±15.26^a	71.54±14.36^a	65.29±13.54^a	51.37±10.63^a	3.58±0.73^a
黄芪甲苷组	53.66±10.64^b	48.72±9.58^b	41.65±8.31^b	35.94±7.24^b	27.86±5.29^b	5.34±1.06^b
Fas沉默组	52.63±10.59^b	46.28±9.27^b	43.16±8.51^b	35.22±6.98^b	27.06±4.32^b	5.55±1.29^b
黄芪甲苷+Fas沉默组	46.59±9.35	41.20±8.27	34.59±6.85	28.61±5.54	22.36±4.15	6.52±1.27
F	17.217^＊＊	20.025^＊＊	23.807^＊＊	30.490^＊＊	31.068^＊＊	14.318^＊＊

组别	凋亡率/%	凋亡相关蛋白表达
组别	凋亡率/%	Caspase-3	Bax	Bcl-2
假手术组	1.91±0.37	1.24±0.26	0.67±0.13	2.05±0.40
模型组	16.52±3.06^a	3.06±0.61^a	2.38±0.46^a	0.53±0.11^a
黄芪甲苷组	10.57±2.43^b	1.80±0.37^b	1.46±0.27^b	0.94±0.22^b
Fas沉默组	10.63±2.55^b	1.83±0.32^b	1.39±0.22^b	0.96±0.20^b
黄芪甲苷+ Fas沉默组	6.72±0.83^cd	1.05±0.19^cd	0.88±0.16^cd	1.45±0.34^cd
F	64.597^＊＊	43.080^＊＊	58.528^＊＊	45.138^＊＊