Sae Uchida

Neural mechanisms of the reflex inhibition and excitation of gastric motility elicited by acupuncture-like stimulation in anesthetized rats

The effects of acupuncture-like stimulation of the various segmental areas on gastric motility were examined in anesthetized rats. An acupuncture needle (diameter 340 microns) was inserted into the skin and underlying muscles at a depth of 4-5 mm and was twisted right and left once every second for 60 s. Gastric motility in the pyloric region was measured with the balloon method. Gastric motility was inhibited by acupuncture-like stimulation applied to the abdomen and lower chest region, and was often excited when the limbs were stimulated, in all cases in which stimuli were delivered to the skin and muscles, the skin alone, and the underlying muscles alone. The inhibitory gastric response to abdominal stimulation was accompanied by an increase in the activity of the gastric sympathetic efferent nerve and was abolished by severance of either the sympathetic nerve branches to the stomach or the lower thoracic spinal nerves. The abdominal stimulation enhanced the activity of the lower thoracic spinal afferent nerves. The excitatory gastric response to hindpaw stimulation was accompanied by an increase in the activity of the gastric vagal efferent nerve and was abolished by severance of either the bilateral vagi or the femoral and sciatic nerves. The hindpaw stimulation enhanced the activity of the femoral and sciatic afferent nerves. In the spinalized animals, the inhibitory gastric response elicited by abdominal stimulation was present, and the hindpaw stimulation did not produce any gastric response. We conclude that the inhibitory gastric response elicited by acupuncture-like stimulation of the abdomen is a reflex response. Its afferent nerve pathway is composed of abdominal cutaneous and muscle afferent nerves, the efferent nerve pathway is the gastric sympathetic nerve, and its reflex center is within the spinal cord. The excitatory gastric response elicited by acupuncture-like stimulation of a hindpaw is also a reflex response. Its afferent nerve pathway is composed of hindpaw cutaneous and muscle afferent nerves, the efferent nerve pathway is the gastric vagal efferent nerve, and its reflex center requires the presence of the brain. Furthermore, the excitatory and the inhibitory gastric reflex responses were not influenced by i.v. administration of naloxone (0.4-4 mg/kg), suggesting that endogenous opioids are not involved in the present reflexes.

The effect of electro-acupuncture stimulation on the muscle blood flow of the hindlimb in anesthetized rats.

The effect of electro-acupuncture stimulation (EAS) on blood flow in the muscle biceps femoris (MBF) and on mean arterial pressure (MAP) was investigated in anesthetized, artificially ventilated rats. EAS was applied to a hindpaw for 30 s at intensities of 0.1-10.0 mA and at frequencies of 1-20 Hz, and MBF was measured by laser Doppler flowmetry. EAS at less than 1.0 mA, which excited group II fibers maximally and III fibers partially in a saphenous nerve, had no significant effect on MBF or MAP, although both revealed variable responses. EAS at 1.5 mA, which additionally excited group III fibers almost maximally and was subthreshold for group IV fibers, produced a small but significant increase in MBF and MAP. These responses were further increased at 2.0 mA or more, which was suprathreshold for group IV fibers. The increased response of MBF at 10.0 mA was followed by a small decrease in MBF. EAS at 1.5 mA or more also elicited a decrease in renal blood flow (RBF) and an arterial pressor response. Following severance of the bilateral splanchnic nerves, EAS at 10.0 mA induced only a slight increase in MAP and a decrease in MBF. The decrease in MBF was abolished following further severance of the bilateral lumbar sympathetic trunks (LSTs). In conclusion, EAS to a hindpaw at a stimulus strength sufficient to excite group III and IV afferent fibers, particularly group IV afferent fibers, can produce a reflex decrease in MBF via a reflex activation of muscle sympathetic activity, although this decrease in MBF is overridden by an increase in MBF caused passively by a reflex MAP pressor response elicited by a reflex increase, at least in splanchnic sympathetic activity.

Calcitonin gene-related peptide produces skeletal muscle vasodilation following antidromic stimulation of unmyelinated afferents in the dorsal root in rats

In anesthetized rats, the contribution of calcitonin gene-related peptide (CGRP) to antidromic vasodilation of skeletal muscle blood flow (MBF) following electrical stimulation of muscle afferent was investigated by measuring biceps femoris MBF using laser Doppler flowmetry. Repetitive antidromic electrical stimulation of unmyelinated C fibers in ipsilateral dorsal roots at the 3rd-5th lumbar segments for 30 s caused an increase in MBF for 3-15 min (mean 4.5 min) without significant change in systemic arterial blood pressure. The increase in skeletal MBF started about 10 s after the onset of stimulation, and peaked at approximately 130% of the control value at about 30 s after the end of the 30 s period of stimulation. The MBF response was totally abolished by topical application of hCGRP (8-37), a CGRP receptor antagonist. It is concluded that antidromic vasodilation in skeletal muscles following stimulation of unmyelinated C afferents in dorsal roots is independent of systemic blood pressure and is mediated essentially by CGRP. It is suggested that this CGRP-related antidromic vasodilation may be important in the clinical improvement of skeletal MBF produced by physical therapy, e.g. acupuncture.

Effect of stimulation of nicotinic cholinergic receptors on cortical cerebral blood flow and changes in the effect during aging in anesthetized rats

The effect of intravenous injection of nicotine on cortical cerebral blood flow (CBF) was examined in urethane anesthetized rats. Nicotine (3-30 microg/kg) increased cortical CBF, independent of mean arterial pressure. This response was attenuated to about a half of the control one after lesioning the nucleus basalis of Meynert (NBM) bilaterally. The response was not significantly influenced after blocking the muscarinic receptors, but was abolished after blocking the nicotinic receptors in the parenchyma of the brain. It is concluded that the nicotine-induced cortical vasodilation was mediated by activation of the nicotinic receptors in the NBM and also in the cortex of the brain. The threshold dose of nicotine for increasing cortical CBF was shifted in aged rats of 23-26 months, and the nicotine-induced increase in cortical CBF was much reduced in aged rats of 32-36 months. Activation of nicotinic receptors in the brain may be of therapeutic value in aged subjects in facilitating the cholinergic neural vasodilative system.

Activation of the intracerebral cholinergic nerve fibers originating in the basal forebrain increases regional cerebral blood flow in the rat's cortex and hippocampus.

In the rat, activation of the intracerebral cholinergic system originating in the basal forebrain and projecting to the cortex and hippocampus releases acetylcholine in the cortex and hippocampus, which results in vasodilation and an increase in regional cerebral blood flow (rCBF) in the cortex and hippocampus. The augmentation of rCBF is independent of both systemic blood pressure and regional metabolism. The intracerebral cholinergic fibers are able to act as autonomic nerve fibers for the regulation of cortical and hippocampal blood flow.

Regulation of regional cerebral blood flow by cholinergic fibers originating in the basal forebrain.

The intracranial neural vasodilative system of cholinergic fibers projecting from the basal forebrain to the cortex was discovered by Biesold, Inanami, Sato and Sato (Biesold, D., Inanami, O., Sato, A., Sato, Y., 1989. Stimulation of the nucleus basalis of Meynert increases cerebral cortical blood flow in rats. Neurosci. Lett. 98, 39–44) using laser Doppler flowmetry in anesthetized rats. This cholinergic vasodilative system, which operates by increasing extracellular ACh release, relies upon activation of both muscarinic and nicotinic cholinergic receptors in the parenchyma of the cortex. Further, the involvement of nitric oxide in this cholinergic vasodilation, indicates the necessity to this system of neurons, which contain nitric oxide synthase. The increase in cortical blood flow elicited by this cholinergic vasodilative system is independent of systemic blood pressure and is not coupled to cortical metabolic rates. This cholinergic vasodilative system may be activated by somatic afferent stimulation. Most of the data presented here were obtained in anesthetized animals.

Acupuncture Affects Regional Blood Flow in Various Organs

In this review, our recent studies using anesthetized animals concerning the neural mechanisms of vasodilative effect of acupuncture-like stimulation in various organs are briefly summarized. Responses of cortical cerebral blood flow and uterine blood flow are characterized as non-segmental and segmental reflexes. Among acupuncture-like stimuli delivered to five different segmental areas of the body; afferent inputs to the brain stem (face) and to the spinal cord at the cervical (forepaw), thoracic (chest or abdomen), lumbar (hindpaw) and sacral (perineum) levels, cortical cerebral blood flow was increased by stimuli to face, forepaw and hindpaw. The afferent pathway of the responses is composed of somatic groups III and IV afferent nerves and whose efferent nerve pathway includes intrinsic cholinergic vasodilators originating in the basal forebrain. Uterine blood flow was increased by cutaneous stimulation of the hindpaw and perineal area, with perineal predominance. The afferent pathway of the response is composed of somatic group II, III and IV afferent nerves and the efferent nerve pathway includes the pelvic parasympathetic cholinergic vasodilator nerves. Furthermore, we briefly summarize vasodilative regulation of skeletal muscle blood flow via a calcitonin gene-related peptide (CGRP) induced by antidromic activation of group IV somatic afferent nerves. These findings in healthy but anesthetized animals may be applicable to understanding the neural mechanisms improving blood flow in various organs following clinical acupuncture.

Extracellular release of acetylcholine, noradrenaline and serotonin increases in the cerebral cortex during walking in conscious rats

The effect of walking on the extracellular release of acetylcholine (ACh), noradrenaline (NA) and serotonin (5-HT) in the parietal lobe of the cerebral cortex was examined in conscious rats using the microdialysis technique. Walking at the speed of 2.3 m/min for 5 min produced significant increases in ACh, NA and 5-HT release in the cerebral cortical extracellular space. The increase in ACh release was the most prominent among the transmitters examined. It is suggested that the increased release of ACh in the cerebral cortex may contribute at least partly to the increases in the cerebral cortical blood flow during walking.

Aging of the autonomic nervous system and possible improvements in autonomic activity using somatic afferent stimulation

There are significant age‐related changes in autonomic nervous system function that are responsible for an impaired ability to adapt to environmental or intrinsic visceral stimuli in the elderly. We review data on changes in autonomic nervous system regulation of cardiovascular and urinary function, as well as data on strategies to improve function. There are data showing alterations in peripheral and central autonomic nerve activity, and decreases in neurotransmitter receptor action that lead to diminished autonomic reactivity (e.g. blood pressure and cerebral blood flow regulation) and poorly coordinated autonomic discharge (e.g. bladder function). Simple strategies for autonomic function improvement and increasing cortical blood flow include walking and somatic afferent stimulation (e.g. stroking skin or acupuncture) to increase sympathetic, parasympathetic and central cholinergic activity. Geriatr Gerontol Int 2010; 10 (Suppl. 1): S127–S136.

Afferent nerve fibers and acupuncture

Acupuncture has been used for analgesia, for treating visceral function disorders and for improving motor functions. It is well established that stimulation of the skin and muscles, either electrically or with noxious or non-noxious stimuli, induces a variety of somato-motor and autonomic responses. This strongly suggests that acupuncture acts by exciting cutaneous and/or muscular afferent nerve fibers. A question of considerable scientific and practical interest is what kinds of somatic afferent fibers are stimulated by acupuncture and are involved in its effects. There are several types of afferent fiber: thick myelinated Aα and Aβ (group I and II), thin myelinated Aδ (group III) and thinner unmyelinated C (group IV) fibers. In recent studies we have tried to establish which ones of these types of somatic afferent fiber are stimulated by acupuncture. In this article we first review the experimental evidence showing that the effects of acupuncture are mediated by the activation of afferent nerve fibers innervating the skin and muscles. Secondly, we discuss what types of afferent nerve fiber are activated by electrical acupuncture, and what types are involved in its effects on somato-motor functions and on visceral functions. Finally, we present some new findings based on recordings from single afferent nerve fibers excited by manual acupuncture.