Effects of calycosin on neuronal autophagy and apoptosis in rats with spinal cord injury

doi:10.11958/20231623

Abstract

Abstract:

Objective To explore the effect of calycosin (CAL) on neuronal autophagy and apoptosis in rats with spinal cord injury (SCI) by regulating PI3K/Akt signaling pathway and its mechanism. Methods A total of 32 male or female adult SD rats were randomly divided into the sham group, the SCI group, the CAL low (20 mg/kg) dose group and the CAL high (40 mg/kg) dose group with 8 rats in each group. The rat model of moderate SCI was established by modified Allen’s method. After successful modeling, rats were injected intraperitoneally immediately with different dosage of CAL or equal amount of saline once a day for 7 consecutive days. Basso, Beattie and Bresnahan (BBB) scores were used to evaluate the recovery of motor function of rats at 1, 3 and 7 d after surgery. At 7 d after surgery, Nissl staining was used to detect the surviving number of motor neurons in anterior horn of spinal cord. Western blot assay was used to assess expression levels of p62, Beclin-1, microtubule-associated protein 1 light chain 3 (LC3B), phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway， Cleaved-Caspase-3, B-cell lymphoma-2 (Bcl-2) and Bcl-2-associated X protein (Bax) proteins. Immunofluorescence staining was used to measure the expression of LC3B in anterior neurons of spinal cord. Results Compared with the sham group, BBB scores, the surviving number of motor neurons and levels of Bcl-2, Bcl-2/Bax, p-PI3K, p-PI3K/PI3K, p-Akt and p-Akt/Akt were significantly decreased (P<0.05), and levels of p62, Beclin-1, LC3B Ⅱ, LC3B Ⅱ/Ⅰ, Cleaved-Caspase-3 and Bax were significantly increased in the SCI group (P<0.05). Compared with the SCI group, BBB scores, the survival of anterior horn motor neurons and levels of LC3B Ⅱ, LC3B Ⅱ/Ⅰ, Bcl-2, Bcl-2/Bax, p-PI3K, p-PI3K/PI3K, p-Akt and p-Akt/Akt were increased in the CAL low dose group and CAL high dose group, and levels of p62, Cleaved-Caspase-3 and Baxcould were significantly decreased (P<0.05). Conclusion CAL could promote autophagy and inhibit apoptosis of neurons through activating PI3K/Akt signaling pathway, thereby conferring a protective role following SCI in rats.

Key words: calycosin, spinal cord injury, autophagy, PI3K/Akt signaling pathway

CLC Number:

R651.2，R285.6

Figures/Tables 9

References 26

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组别	n	1 d	3 d	7 d
Sham组	6	21.00±0.00	21.00±0.00	21.00±0.00
SCI组	6	0.44±0.18^a	1.83±0.75^a	3.94±0.77^a
CAL低剂量组	6	0.56±0.27^a	2.89±0.66^b	6.17±1.05^b
CAL高剂量组	6	0.67±0.42^a	3.95±0.88^b	8.61±1.18^bc
F		0.778	11.372^＊＊	31.740^＊＊

组别	n	1 d	3 d	7 d
Sham组	6	21.00±0.00	21.00±0.00	21.00±0.00
SCI组	6	0.44±0.18^a	1.83±0.75^a	3.94±0.77^a
CAL低剂量组	6	0.56±0.27^a	2.89±0.66^b	6.17±1.05^b
CAL高剂量组	6	0.67±0.42^a	3.95±0.88^b	8.61±1.18^bc
F		0.778	11.372^＊＊	31.740^＊＊

组别	n	p62/ β-actin	Beclin-1/ β-actin	LC3BⅡ/Ⅰ	LC3BⅡ/ β-actin
Sham组	4	0.49±0.10	0.28±0.05	0.49±0.05	0.47±0.07
SCI组	4	1.03±0.11^a	0.54±0.09^a	0.81±0.08^a	0.87±0.10^a
CAL低剂量组	4	0.70±0.09^b	0.63±0.05	1.06±0.06^b	1.10±0.10^b
CAL高剂量组	4	0.39±0.11^bc	0.94±0.11^bc	1.26±0.03^bc	1.30±0.15^bc
F		30.722^＊＊	47.307^＊＊	148.901^＊＊	42.684^＊＊

组别	n	p62/ β-actin	Beclin-1/ β-actin	LC3BⅡ/Ⅰ	LC3BⅡ/ β-actin
Sham组	4	0.49±0.10	0.28±0.05	0.49±0.05	0.47±0.07
SCI组	4	1.03±0.11^a	0.54±0.09^a	0.81±0.08^a	0.87±0.10^a
CAL低剂量组	4	0.70±0.09^b	0.63±0.05	1.06±0.06^b	1.10±0.10^b
CAL高剂量组	4	0.39±0.11^bc	0.94±0.11^bc	1.26±0.03^bc	1.30±0.15^bc
F		30.722^＊＊	47.307^＊＊	148.901^＊＊	42.684^＊＊

组别	Bax/ β-actin	Cleaved-Caspase-3/ β-actin	Bcl-2/ β-actin	Bcl-2/ Bax
Sham组	0.43±0.07	0.53±0.08	0.91±0.15	2.14±0.38
SCI组	0.92±0.08^a	0.85±0.14^a	0.49±0.06^a	0.54±0.09^a
CAL低剂量组	0.67±0.15^b	0.61±0.05^b	0.74±0.10^b	1.24±0.34^b
CAL高剂量组	0.46±0.10^bc	0.47±0.03^b	1.06±0.10^bc	2.25±0.36^bc
F	19.059^＊＊	16.447^＊＊	19.748^＊＊	26.671^＊＊