Hydrogen-rich saline treated neuropathic pain in rats by increasing autophagy

doi:10.11958/20230698

Abstract

Abstract:

Objective To evaluate the role of autophagy in the treatment of neuropathic pain (NP) with hydrogen-rich saline. Methods Forty adult male Sprague-Dawley rats with successful intubation were randomly divided into 5 groups (n=8) using a random number table: the sham operation group (group S), the neuropathic pain group (group C), the hydrogen-rich saline group (group H), the autophagy inhibitor group (group M) and the hydrogen-rich saline + autophagy inhibitor group (group HM). There were 8 rats in each group. The NP model was established by chronic constriction of the sciatic nerve (CCI) in rats. The autophagy inhibitor 3-methyladenine (3-MA) was intraperitoneally injected with 30μg/kg in the group M and the group HM. The hydrogen-rich saline (0.6 mmol/L) was intraperitoneally injected with 10 mL/kg in the group H and the group HM. The other groups were intraperitoneally injected with the same amount of normal saline twice a day for 7 consecutive days. Paw withdrawal threshold to mechanical stimulation (MWT) and paw withdrawal latency to thermal stimulation (TWL) were measured at 1 day before and 1, 3, 5, 7 and 14 days after modeling (T₀-T₅). After the last measurement of pain threshold, the L4-L6 segment of spinal cord was removed for determination of the expression of autophagy-related proteins microtubule-associated protein light chain 3 (LC3) Ⅱ, Beclin-1 and p62 proteins by Western blot assay. The expression levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in spinal cord tissue were detected. Results Compared with the group S, MWT and TWL were decreased in the group C at T_2-5, the expression levels of LC3 Ⅱ, Beclin-1 and p62 were increased, SOD activity was decreased, and MDA content was increased at T₅(P<0.05). Compared with the group C, MWT and TWL were increased in the group H at T_2-5, LC3 Ⅱ and Beclin-1 protein expression levels were increased, p62 protein expression levels were decreased, SOD activity was increased, and MDA content was decreased at T₅ (P<0.05). MWT and TWL were decreased in the group M at T_2-5, LC3 Ⅱ and Beclin-1 protein expression levels were decreased, p62 protein expression levels were increased, SOD activity was decreased, and MDA content was increased at T₅(P<0.05). Compared with the group M, MWT and TWL were increased in the group HM at T_2-5, LC3 Ⅱ and Beclin-1 protein expression levels were increased, p62 protein expression levels were decreased, SOD activity was increased, and MDA content was decreased at T₅ (P<0.05). Conclusion Hydrogen-rich saline can alleviate neuropathic pain and inhibit oxidative stress in spinal cord in rats, and the mechanism may be related to the increase of autophagy.

Key words: hydrogen, neuralgia, autophagy, spinal cord, oxidative

CLC Number:

R614

Figures/Tables 5

References 17

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组别	T₀	T₁	T₂	T₃	T₄	T₅	F
S组	17.4±1.3	16.2±1.7	16.0±1.6	17.4±0.7	16.2±0.8	17.1±0.8	1.925
C组	16.6±1.0	15.5±1.2	12.4±1.2^ABa	9.6±1.8^ABCa	7.8±1.1^ABCDa	7.4±1.0^ABCDa	101.919^＊
H组	17.3±0.9	16.2±1.8	14.3±1.5^ABb	12.1±1.2^ABCb	11.0±1.2^ABCb	11.4±1.5^ABCb	42.582^＊
M组	16.7±1.1	14.8±1.4^A	10.5±1.6^ABb	7.9±1.1^ABCb	6.1±0.7^ABCDb	5.9±0.7^ABCDb	128.416^＊
HM组	17.1±1.2	15.5±0.8^A	12.1±1.4^ABc	9.9±1.5^ABCc	7.9±1.2^ABCDc	7.9±1.1^ABCDc	99.278^＊
F	0.936	1.344	16.697^＊	61.703^＊	117.144^＊	144.049^＊

组别	T₀	T₁	T₂	T₃	T₄	T₅	F
S组	17.4±1.3	16.2±1.7	16.0±1.6	17.4±0.7	16.2±0.8	17.1±0.8	1.925
C组	16.6±1.0	15.5±1.2	12.4±1.2^ABa	9.6±1.8^ABCa	7.8±1.1^ABCDa	7.4±1.0^ABCDa	101.919^＊
H组	17.3±0.9	16.2±1.8	14.3±1.5^ABb	12.1±1.2^ABCb	11.0±1.2^ABCb	11.4±1.5^ABCb	42.582^＊
M组	16.7±1.1	14.8±1.4^A	10.5±1.6^ABb	7.9±1.1^ABCb	6.1±0.7^ABCDb	5.9±0.7^ABCDb	128.416^＊
HM组	17.1±1.2	15.5±0.8^A	12.1±1.4^ABc	9.9±1.5^ABCc	7.9±1.2^ABCDc	7.9±1.1^ABCDc	99.278^＊
F	0.936	1.344	16.697^＊	61.703^＊	117.144^＊	144.049^＊

组别	T₀	T₁	T₂	T₃	T₄	T₅	F
S组	14.6±1.0	13.7±0.8	14.8±1.3	14.4±1.7	15.2±1.6	14.6±1.4	1.709
C组	14.5±1.3	13.1±1.7^A	10.2±1.6^ABa	7.7±1.0^ABCa	6.2±1.1^ABCDa	6.6±1.0^ABCa	74.429^＊
H组	15.2±1.3	13.5±1.1^A	12.4±1.2^ABb	10.5±1.5^ABCb	9.6±1.1^ABCb	10.0±1.1^ABCb	28.789^＊
M组	14.3±1.3	12.5±1.0^A	8.6±1.3^ABb	6.0±0.9^ABCb	4.9±0.8^ABCDb	5.3±0.8^ABCb	96.627^＊
HM组	14.5±1.3	13.1±1.5^A	10.5±1.4^ABc	8.2±1.2^ABCc	6.5±1.0^ABCDc	6.6±0.9^ABCDc	71.564^＊
F	0.630	1.136	24.649^＊	51.482^＊	103.909^＊	105.028^＊

组别	T₀	T₁	T₂	T₃	T₄	T₅	F
S组	14.6±1.0	13.7±0.8	14.8±1.3	14.4±1.7	15.2±1.6	14.6±1.4	1.709
C组	14.5±1.3	13.1±1.7^A	10.2±1.6^ABa	7.7±1.0^ABCa	6.2±1.1^ABCDa	6.6±1.0^ABCa	74.429^＊
H组	15.2±1.3	13.5±1.1^A	12.4±1.2^ABb	10.5±1.5^ABCb	9.6±1.1^ABCb	10.0±1.1^ABCb	28.789^＊
M组	14.3±1.3	12.5±1.0^A	8.6±1.3^ABb	6.0±0.9^ABCb	4.9±0.8^ABCDb	5.3±0.8^ABCb	96.627^＊
HM组	14.5±1.3	13.1±1.5^A	10.5±1.4^ABc	8.2±1.2^ABCc	6.5±1.0^ABCDc	6.6±0.9^ABCDc	71.564^＊
F	0.630	1.136	24.649^＊	51.482^＊	103.909^＊	105.028^＊

组别	LC3Ⅱ	Beclin-1	p62
S组	0.115±0.013	0.150±0.005	0.051±0.005
C组	1.510±0.012^a	0.916±0.029^a	0.286±0.009^a
H组	1.919±0.062^b	1.127±0.014^b	0.140±0.006^b
M组	0.289±0.031^b	0.215±0.010^b	0.401±0.010^b
HM组	1.068±0.735^c	0.908±0.012^c	0.287±0.009^c
F	4 722.678^＊	6 088.196^＊	2 362.972^＊