This work was supported by S&#38;T Program of Hebei (NO.E2020100001, and NO.22372502D), High-level S &#38; T Innovation and Entrepreneurship Talent Project of Shijiazhuang (No. 07202203).

The animal experiments were conducted with the approval of the Committee on Animal Research and Ethics of the Hebei Yiling Medical Research Institute (approval number: N2022148). Male C57BL/6J mice weighing 18-22 g (see Table of Materials) were housed in the new drug evaluation center of Hebei Yiling Medical Research Institute. They were provided with normal food and clean water and exposed to artificial light for 12 h daily. The rooms maintained a controlled temperature range of 20-26 &#176;C and relati...

Cognitive Improvement in Chronic Hypoxia Mice with Acupuncture

Acupuncture, a non-pharmacological medical practice originating in China over 2,000 years ago, involves the insertion of thin needles into specific points on the body known as acupuncture points. These points are believed to be connected by channels or meridians through which the body&#39;s vital energy, or &#34;qi,&#34; flows24. By stimulating these points, acupuncture aims to restore balance and harmony to the body. It has been shown to effectively treat various conditions, including chronic pai...

The global population is currently facing a critical aging problem, resulting in a rapid increase in the prevalence of cognitive disorders. The worldwide incidence of cognitive impairment is approximately 53.97 per 1000 person-years1. Chronic cerebral hypoxia caused by vascular dysfunction or circulatory/respiratory disorders remains one of the major risk factors for age-related dementia2. Previous studies have demonstrated that cerebral hypoxia can increase amyloid-&#946; deposition by modifying BACE1 expression3. Additionally, hypoxia has been associated with glial-cell dysregulation and neuroinflammation4,5. Despite the growing magnitude of this issue, effective Western medicines for preventing chronic hypoxia-induced cognitive decline are currently lacking. Non-pharmacological traditional Chinese medicine, particularly acupuncture, has been used for thousands of years to treat cognitive disorders and has shown promising results in alleviating neurodegenerative diseases6,7. The Baihui, Shenting, and Zusanli acupoints are effective points for treating cognitive dysfunction8,9. Clinical studies have demonstrated that electro-acupuncture therapy significantly improves the Montreal Cognitive Assessment (MoCA) and Mini-Mental State Examination (MMSE) scores in patients with vascular cognitive impairment and effectively ameliorates cognitive dysfunction8. Although studies have suggested that acupuncture can significantly enhance the memory ability of rats with arterial ligation-an acute cerebral hypoxia models10, an acute cerebral hypoxia model, there is no report on the effects of acupuncture in any rodent model with chronic hypoxia-induced cognitive disorders. The lack of research into the mechanism has considerably impeded its clinical application. 
Previous research has demonstrated that subjecting rats to a hypoxic environment for a period of 8 weeks can significantly elevate levels of oxidative stress and inflammation in the brain, resulting in a decline in memory function11. The present study aims to investigate the impact of acupuncture on rodent models in order to further our understanding. It is worth noting, however, that anesthesia is typically required during acupuncture treatment in rodents due to the potential for agitation during repeated stimulation. Prolonged anesthesia can significantly impact cognitive function in mice, as most anesthetic drugs can suppress neural activity and impede information processing, leading to behavioral deficits12. Several studies have shown that administering 2.5% sevoflurane for a duration of 6 h can notably impair spatial memory, learning ability, and attention in mice13. Furthermore, evidence suggests that high doses of anesthesia may result in neuronal death or nerve damage in mice14. Therefore, it is imperative to identify a suitable approach to minimize the overall amount of anesthesia used. In this study, we introduce an effective acupuncture method for treating mice with cognitive impairment, along with behavioral tests to assess their memory abilities. Importantly, we present a modified pre-treatment anesthesia technique that can effectively reduce the total dosage of anesthesia administered during the experiment.

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	<tbody>
		<tr>
			<td>10% paraformaldehyde solution</td>
			<td>Bioroyee (Beijing) Biotechnology Co., Ltd</td>
			<td>RL3234</td>
			<td></td>
		</tr>
		<tr>
			<td>ANY-maze</td>
			<td>Science</td>
			<td>&#160;SA201</td>
			<td>Video tracking system</td>
		</tr>
		<tr>
			<td>C75BL/6J mice</td>
			<td>BEIJING HFK BIOSCIENCE CO.,LTD</td>
			<td>No.110322220103041767</td>
			<td>Gender: Male,&#160; Weight: 18&#8211;22 g</td>
		</tr>
		<tr>
			<td>Electroacupuncture device</td>
			<td>Great Wall</td>
			<td>KWD-808 I</td>
			<td></td>
		</tr>
		<tr>
			<td>Hwato acupuncture&#160; needle</td>
			<td>Suzhou Medical Appliance Factory</td>
			<td>2655519&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>RWD Life Science Co.,Ltd</td>
			<td>R510-22</td>
			<td></td>
		</tr>
		<tr>
			<td>NanoZoomer Digital Pathology</td>
			<td>Hamamatsu Photonics K. K</td>
			<td>C9600-01</td>
			<td></td>
		</tr>
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			<td>Small animal anesthesia machine</td>
			<td>RWD</td>
			<td>YL-LE-A106</td>
			<td></td>
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acupuncture treatment in a mouse model of chronic hypoxia-induced cognitive dysfunction

The treatment of central nervous disorders has consistently posed significant challenges to the medical field. Acupuncture, a non-pharmacological practice rooted in traditional Chinese medicine, entails the insertion of fine needles into precise points on the body and is commonly employed for the management of diverse conditions. Recently, acupuncture has emerged as a promising therapeutic intervention for a range of neurological diseases, including anxiety and respiratory disorders. However, the potential of acupuncture in treating cognitive dysfunction induced by chronic hypoxia has not yet been explored. This paper presents a comprehensive protocol for establishing a mouse model of chronic hypoxia-induced cognitive impairment, administering mild anesthesia, performing acupuncture treatment, and assessing behavioral changes and memory abilities using open field tests and water mazes. The step-by-step protocol provides detailed instructions on accurately locating and positioning acupoints and needles for cognitive improvement. By employing this protocol, researchers can conduct systematic studies to thoroughly evaluate the therapeutic potential of acupuncture for cognitive dysfunction.

Author Spotlight: Improving Anesthesia Protocols for Enhanced Mouse Acupuncture Research

Acupuncture Treatment in a Mouse Model of Chronic Hypoxia-Induced Cognitive Dysfunction

Selecting the right anesthesia protocol is crucial for conducting mouse acupuncture. Due to potential cognitive impairment associated with prolonged anesthesia, it represents an essential initial challenge, demanding careful consideration to minimize cognitive function risks in mice during the procedure. In this study, we propose an improved protocol that helps to minimize functional and behavioral abnormalities caused by excessive anesthesia dosing, and enhances the accuracy of the experiments.In the future, our laboratory aims to study the underlying mechanisms of acupuncture despite its observable therapeutic effects on behavior and pathology. Our research will focus on unraveling acupuncture to gain a deeper understanding of its mechanisms.

Characterizing mouse locomotion trajectories in the open field experiment 
The trajectory map reveals that mice in the normal group exhibit a profound inclination for exploration in unfamiliar environments. Their activity trajectories are primarily concentrated in the corners while covering the entire open field (left panel). In contrast, the long-term hypoxia model group of mice displays a significantly diminished desire to explore novel surroundings. They predominantly linger in the corners withou...

Watch this Scientific Journal Video about Acupuncture Treatment in a Mouse Model of Chronic Hypoxia-Induced Cognitive Dysfunction at JoVE.com

Here, we describe a protocol for implementing mild anesthesia and acupuncture treatment on a chronic hypoxia mouse model and conducting behavioral tests to assess the cognitive alterations post-treatment.

The treatment of central nervous disorders has consistently posed significant challenges to the medical field. Acupuncture, a non-pharmacological practice ...

Acupuncture Treatment in a Mouse Model of Chronic Hypoxia-Induced Cognitive Dysfunction

Abstract