Abstract: BACKGROUND: Both acupuncture and moxibustion and rehabilitation training can effectively alleviate the symptoms of patients with cerebral ischemia. At present, it is unclear whether the combined treatment of the two has a regulatory effect on neurological dysfunction and intestinal flora imbalance in patients with cerebral ischemia and its role mechanism. 
OBJECTIVE: To explore the effect of acupuncture and moxibustion combined with rehabilitation therapy on neurological function and intestinal flora in rats with cerebral ischemia. 
METHODS: Sixty Sprague-Dawley rats were randomly divided into sham operation group, model group, acupuncture group, rehabilitation group, and acupuncture+rehabilitation group (n=12 per group). Animal models of cerebral ischemia were prepared in all groups except for the sham operation group. In the sham operation group, the left common carotid artery was only separated but not ligated. The acupuncture group was treated with scalp cluster acupuncture intervention, the rehabilitation group was given task-oriented treadmill training, and the acupuncture+rehabilitation group was given scalp cluster acupuncture and task-oriented treadmill training intervention. Treatments in each group lasted for 14 days. The sham operation and model groups were not intervened. The neurological function was assessed by Longa score at 4 hours, 1, 7, and 14 days after modeling. Changes in brain water content were detected after 14 days of intervention. Western blot was applied to detect the expressions of neuron growth-related proteins (growth-associated protein 43, neurofilament 200, repulsive guidance molecule a) in the ischemic penumbra. Kit detection was used for detecting the levels of lactate dehydrogenase, superoxide dismutase, and malondialdehyde in brain tissue. ELISA was applied to detect the levels of tumor necrosis factor-α and interleukin-1β in brain tissue. Real-time fluorescent quantitative PCR was used to detect the changes of Escherichia coli, Bifidobacterium, Lactobacillus, and Enterococcus in rat feces. 
RESULTS AND CONCLUSION: At 4 hours, 1, 7, and 14 days after modeling, there was no neurological dysfunction in the sham operation group but obvious neurological dysfunction in the model group (P < 0.05). Compared with the model group, the neurological deficits of rats in the acupuncture and rehabilitation groups were significantly improved with the prolonged intervention time (P < 0.05), and the neurological function of rats in the acupuncture+rehabilitation group improved more significantly (P < 0.01). Compared with the model group, the expressions of growth-associated protein 43 and neurofilament 200 in the ischemic penumbra were significantly up-regulated in the acupuncture and rehabilitation groups (P < 0.05), the expression of repulsive guidance molecule a was significantly down-regulated (P < 0.05), the brain water content and levels of lactate dehydrogenase, malondialdehyde, tumor necrosis factor-α and interleukin-1β in brain tissue were reduced (P < 0.01), the superoxide dismutase level was raised (P < 0.01), the counts of Escherichia coli and Enterococcus in feces were significantly increased (P < 0.01), and the counts of Bifidobacterium and Lactobacillus were significantly decreased (P < 0.01). The above-mentioned indicators were improved more significantly in the acupuncture+rehabilitation group (P < 0.01). To conclude, acupuncture and moxibustion combined with rehabilitation therapy can improve neurological function, relieve oxidative stress and inflammatory responses, and regulate intestinal flora disorders in rats with cerebral ischemia by stimulating the regeneration of neurons in the ischemic penumbra. The combine therapy has a protective effect against ischemic stroke.

Key words: acupuncture+rehabilitation method, acupuncture, cerebral ischemia, neurological function, brain water content, inflammation, intestinal flora, tissue engineering