Acupuncture anesthesia

Evidence Details for Sqstm1

PMID	Title	Journal	Year	Abstract
36422883	Acupuncture promotes nerve repair through the benign regulation of mTOR-mediated neuronal autophagy in traumatic brain injury rats.	CNS Neurosci Ther. 2023 Jan;29(1):458-470. doi: 10.1111/cns.14018. Epub 2022 Nov 24.	2023 Jan	AIMS: Recent investigations have already proved the neuroprotective efficacy of acupuncture in clinical practice in the treatment of neurological diseases, such as traumatic brain injury (TBI). Since growing evidence has suggested that neuronal autophagy was involved in multiple stages of TBI, this study aims to clarify the autophagy mediating mechanism underlying the neuroprotective effect of acupuncture in TBI rats. METHODS: Three experiments were carried out to detect changes in neuronal autophagy and identify the potential molecular mechanism underlying the neuroprotective effect of acupuncture for TBI treatment. Feeney's free-falling epidural impingement method was used to establish the moderate TBI rat model; modified neurological severity scoring (mNSS) was used for neurological recovery evaluation. Nissl and HE staining were used to examine the histopathological changes. Immunofluorescence was used to detect the LC3-positive cell rate. The transmission electron microscope (TEM) was used to investigate the morphology and quantity of autophagosomes. Western blotting was used to determine the protein expressions of LC3, p62, beclin1, mTOR, ULK1, p-mTOR, and p-ULK1. Quantitative real-time polymerase chain reaction (qRT-PCR) was used for gene expressions analysis of LC3 mRNA and p62 mRNA. Co-immunoprecipitation (CO-IP) method was used to identify the protein interaction of mTOR and ULK1. RESULTS: On Day 3 after TBI, acupuncture accelerated the removal of damaged cellular structures by promoting neuronal autophagy; on Day 7 and Day 14 after TBI, acupuncture inhibited neuronal autophagy, preventing excessive autophagy and thus alleviated nerve damage. In addition, the simultaneous treatment with 3-MA or rapamycin at different stages after TBI attenuated the effect of acupuncture. CONCLUSION: Acupuncture has a benign regulatory effect on neuronal autophagy in different stages of TBI, possibly through the mTOR/ULK1 pathway."

Evidence Sentence:	Western blotting was used to determine the protein expressions of LC3, p62, beclin1, mTOR, ULK1, p-mTOR, and p-ULK1.
Evidence Sentence:	Quantitative real-time polymerase chain reaction (qRT-PCR) was used for gene expressions analysis of LC3 mRNA and p62 mRNA.
Evidence Sentence:	To confirm this hypothesis, we used the immunofluorescence assay to detect the LC3 positive cell rate of neurons in the injured side of the cerebral cortex, and the Western blot assay to measure the protein expression of autophagy marker LC3 and autophagy substrate p62.
Evidence Sentence:	At the same time, acupuncture downregulated the high level of p62 relative protein expression (Figure 2D) caused by brain injury; and promoted the expression of LC3II/I protein (Figure 2C).
Evidence Sentence:	Furthermore, acupuncture promoted the relative protein expression of p62 and downregulated the expression level of LC3II/I.
Evidence Sentence:	In the meantime, acupuncture treatment increased LC3-positive cells in neurons of TBI rats (Figure 4D), the multiplication of beclin1, conversion of LC3-I to LC3-II, and decreased the expression of autophagy substrate p62, but these effects were counteracted by autophagy inhibitor 3-MA (Figure 4E,F).