Effects of LAMP1 on microglial cell injury and autophagy in rats with cerebral ischemia-reperfusion injury

doi:10.3877/cma.j.issn.2095-9141.2025.04.004

Abstract

Abstract:

Objective

To observe the role of lysosomal-associated membrane protein 1 (LAMP1) in microglial (MG) injury in rats subjected to cerebral ischemic-reperfusion injury (CIRI) and to investigate the impact and mechanism of LAMP1 on autophagy in this model.

Methods

Thirty adult healthy SD rats were selected and the rat model was established using transient middle cerebral artery occlusion (tMCAO) method. The rats were randomly divided into sham group (Sham group), surgical group (tMCAO group), and intervention group (tMCAO+CQ group) according to the random number table method, with 10 rats in each group. Three groups of rats were intraperitoneally injected with sterile physiological saline before surgery, while the tMCAO+CQ group was injected with chloroquine (CQ) lysosome inhibitor 2 h before surgery. CIRI rat model was established, and Sham group was the surgical control group. Five samples were taken from each group for TTC staining, and the remaining five brain tissue specimens were collected for experimental testing in sequence. After collecting brain tissue and extracting the ischemic penumbra tissue, the expression of LAMP1 protein in cells was detected. To investigate the effects of LAMP1 on MG injury and autophagy in a model, enzyme-linked immunosorbent assay (ELISA) was used to detect inflammatory factors [interleukin (IL)-6, tumor necrosis factor alpha (TNF-α), IL-10], and Western blot was used to detect the expression of p62, Cathepsin B, Cathepsin D, and LC3B. In addition, the Longa scoring system was used to evaluate the neurological function of rats 24 h after CIRI, TTC staining was used to measure the volume of cerebral infarction, HE staining was used to observe pathological changes in brain tissue.

Results

Compared with the Sham group, the Longa scores of the tMCAO group and the tMCAO+CQ group were significantly increased, and the cerebral infarction volume was significantly increased. The Longa score of the tMCAO+CQ group was higher than that of the tMCAO group, and the cerebral infarction volume was larger than that of the tMCAO group, with statistical significance (P<0.05). The results of ELISA and Western blot analysis showed that compared with the Sham group, the expression of LAMP1 protein was significantly decreased in the tMCAO group and the tMCAO+CQ group, while the levels of IL-6, TNF-α, LC3B, and Baclin1 protein expression were significantly increased. The levels of IL-10 and p62, LAMP1, Cathepsin B, and Cathepsin D protein expression were significantly decreased, and the differences were statistically significant (P<0.05); Compared with the tMCAO group, the expression of LAMP1 protein in the tMCAO+CQ group decreased significantly, while the levels of IL-6, TNF-α, LC3B, and Beclin1 protein increased. The expression levels of p62 and Cathepsin D protein decreased, and the differences were statistically significant (P<0.05). However, there was no statistically significant difference in the expression levels of Cathepsin B protein (P>0.05).

Conclusions

Inhibiting LAMP1 exacerbates glial cell damage in the rat model and this mechanism may be related to the inhibition of autophagy.

Key words: Lysosomal-associated membrane protein 1, Cerebral ischemic-reperfusion injury, Microglia, Autophagy, Injury

Tuerxun Aizimaitijiang, Mahemuti Yusufu, Shihao Jiang, Kadeer Kaheerman, Aisha Maimaitili, Riqing Su, Xiaojiang Cheng. Effects of LAMP1 on microglial cell injury and autophagy in rats with cerebral ischemia-reperfusion injury[J]. Chinese Journal of Neurotraumatic Surgery(Electronic Edition), 2025, 11(04): 230-237.

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References 30

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组别	只数	Longa评分[分，M（P₂₅，P₇₅）]	脑梗死体积占比（%，±s）
Sham组	5	0	0
tMCAO组	5	2.00（2.00，2.50）^a	28.87±4.52^a
tMCAO+CQ组	5	3.00（2.00，3.00）^ab	40.25±4.77^ab
H/F值		8.333	149.474
P值		0.004	<0.001

组别	只数	Longa评分[分，M（P₂₅，P₇₅）]	脑梗死体积占比（%，±s）
Sham组	5	0	0
tMCAO组	5	2.00（2.00，2.50）^a	28.87±4.52^a
tMCAO+CQ组	5	3.00（2.00，3.00）^ab	40.25±4.77^ab
H/F值		8.333	149.474
P值		0.004	<0.001

组别	只数	IL-6（±s）	IL-10（±s）	TNF-α[M（P₂₅，P₇₅）]
Sham组	5	22.18±4.53	433.10±86.42	4.30（3.05，8.20）
tMCAO组	5	90.10±6.86^a	159.10±23.44^a	21.50（16.56，23.59）^a
tMCAO+CQ组	5	128.90±14.52^ab	77.82±7.23^ab	29.56（26.85，31.18）^ab
F/H值		80.427	157.340	12.500
P值		<0.001	<0.001	0.002

组别	只数	IL-6（±s）	IL-10（±s）	TNF-α[M（P₂₅，P₇₅）]
Sham组	5	22.18±4.53	433.10±86.42	4.30（3.05，8.20）
tMCAO组	5	90.10±6.86^a	159.10±23.44^a	21.50（16.56，23.59）^a
tMCAO+CQ组	5	128.90±14.52^ab	77.82±7.23^ab	29.56（26.85，31.18）^ab
F/H值		80.427	157.340	12.500
P值		<0.001	<0.001	0.002

组别	只数	LAMP1	LC3B	p62	Beclin1	Cathepsin B	Cathepsin D
Sham组	5	2.765±0.074	0.649±0.024	1.574±0.091	1.072±0.084	2.060±0.141	1.972±0.068
tMCAO组	5	2.178±0.235^a	1.240±0.097^a	0.570±0.117^a	2.143±0.241^a	1.534±0.138^a	1.454±0.155^a
tMCAO+CQ组	5	0.862±0.034^ab	1.786±0.052^ab	0.261±0.036^ab	2.898±0.115^ab	1.513±0.094^a	1.277±0.117^ab
F值		230.679	382.255	123.738	160.126	29.907	46.149
P值		<0.001	<0.001	<0.001	<0.001	<0.001	<0.001

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