Effects of nicotine on regional blood flow in the olfactory bulb in response to olfactory nerve stimulation.

This study examined the effect of olfactory nerve stimulation on regional cerebral blood flow and assessed the effect of intravenous nicotine administration on this response in anesthetized rats. Regional cerebral blood flow was measured with laser Doppler flowmetry or laser speckle contrast imaging. Unilateral olfactory nerve stimulation for 5 s produced current (≥ 100 µA) and frequency-dependent (≥ 5 Hz) increases in blood flow in the olfactory bulb ipsilateral to the stimulus. The increased olfactory bulb blood flow peaked at 30 ± 7% using stimulus parameters of 300 µA and 20 Hz. Nerve stimulation did not change frontal cortical blood flow or mean arterial pressure. The intravenous injection of nicotine (30 µg/kg) augmented the olfactory bulb blood flow response to nerve stimulation (20 Hz, 300 µA) by approximately 1.5-fold (60-s area after the stimulation). These results indicate that olfactory nerve stimulation increases olfactory bulb blood flow, and the response is potentiated by the activation of nicotinic cholinergic transmission.

Somatosensory regulation of resting muscle blood flow and physical therapy.

Somatosensory stimulation can affect skeletal muscle blood flow (MBF) at rest in anesthetized animals via pressor reflex response or antidromic and local vasodilation. Increase in MBF due to reflex pressor response occurs generally in the skeletal muscles of the entire body, while antidromic and local vasodilation are limited to the peripheral stimulation site. Since increased MBF improves several disorders (muscle stiffness, pain, etc.), it is reasonable to further explore the effective use of somatic stimulation in physical therapies, such as massage, acupuncture, anma, and shiatsu or acupressure, in treating skeletal muscle disorders.

Blood pressure-independent increase in the cortical cerebral blood flow induced by manual acupuncture of the auricular region in rats.

We aimed to determine whether acupuncture to the auricular region increases cortical regional cerebral blood flow (rCBF). The rCBF was measured using laser speckle contrast imaging in urethane-anesthetized rats. Acupuncture stimulation was performed manually at the auricular concha or abdomen. The former's stimulation significantly increased the rCBF of the bilateral cerebral cortex in the frontal, parietal, and occipital lobes without altering the systemic arterial pressure. In contrast, abdominal stimulation affected neither rCBF nor systemic arterial pressure. The increase in the rCBF was completely abolished by the severance of the somatic nerves that innervated the auricular region, comprising the trigeminal nerve, facial nerve, auricular branch of the vagal nerve, glossopharyngeal nerve, and great auricular nerve. Thus, application of acupuncture to the auricular region increases the rCBF without increasing arterial pressure.

Somatoautonomic reflexes in acupuncture therapy: A review.

Oriental therapies such as acupuncture, moxibustion, or Anma, have been used to treat visceral disorders since ancient times. In each of these therapies, stimulation of the skin or underlying muscles leads to excitation of afferent nerves. The sensory information is carried to the central nervous system, where it is transferred to autonomic efferents, thus affecting visceral functions. This neuronal pathway, known as the "somatoautonomic reflex", has been systematically studied by Sato and his colleagues for over a half century. Nearly all their studies were conducted in anesthetized animals, whereas human patients are conscious. Responses in patients or the events following therapeutic somatic stimulation may differ from those observed in anesthetized animals. In fact, it is increasingly apparent that the responses in patients and animals are not always coincident, and the differences have been difficult for clinicians to reconcile. We review the mechanism of the "somatoautonomic reflex" as described in anesthetized animals and then discuss how it can be applied clinically.

Effect of acupuncture-like stimulation on cortical cerebral blood flow in aged rats.

This study aimed to examine the effect of acupuncture-like stimulation on cortical cerebral blood flow (CBF) in aged rats and the contribution of the intracranial cholinergic vasodilatory system on its response. In urethane-anesthetized rats of 30-37 months of age, manual acupuncture-like stimulation of a forepaw produced an increase in the CBF, independent of systemic arterial pressure. The increase in the CBF induced by forepaw stimulation was abolished by intravenous administration of cholinergic receptor antagonists. Manual acupuncture-like stimulation of a forepaw increased extracellular acetylcholine release in the cerebral cortex. These results suggest that natural somatic afferent stimulation, such as acupuncture-like stimulation, activates the intracranial - most likely, basal forebrain - cholinergic vasodilatory system in the cerebral cortex, even in extremely aged rats.

Cholinergic vasodilative system in the cerebral cortex: effects of acupuncture and aging.

This article presents a review of our studies on the cholinergic vasodilative system in the cerebral cortex in relation to the effects of acupuncture and aging. In anesthetized rats, manual acupuncture-like stimulation of the cheek, forepaw, upper arm, and hindpaw increases the cortical cerebral blood flow (CBF). The mechanism for the increased response of CBF due to forepaw stimulation has been found to be a reflex response whose afferents are Groups III and IV somatic afferent fibers and whose efferents are cholinergic fibers that originate in the nucleus basalis of Meynert. Although the cholinergic cortical vasodilation to nucleus basalis of Meynert stimulation at high intensities declines with age, the increased response of CBF induced by natural somatic afferent stimulation, such as an acupuncture-like stimulation of a forepaw, is well maintained even in very old rats (approximately 3 years of age). These findings in anesthetized rats may support the application of acupuncture to elderly people and patients with disturbances in the CBF by activating the intracranial cholinergic vasodilative system.

Neural mechanisms of reflex inhibition of heart rate elicited by acupuncture-like stimulation in anesthetized rats.

We briefly review our recent studies on the neural mechanisms of the reflex effects of acupuncture-like stimulation on heart rate in rats. In pentobarbital anesthetized rats, acupuncture-like stimulation of one of various segmental areas of the body (forelimb, chest, abdomen, hindlimb) invariably induces a decrease in heart rate. In the case of the hindlimb, the effect can be produced by stimulation of the muscles alone but not of skin alone, and is abolished by severance of the hindlimb somatic nerves. Electrical stimulation of groups III and IV nerve fibers (in the tibial nerve) decreases heart rate. Decrease in heart rate by acupuncture-like stimulation of a hindlimb is accompanied by a decrease in cardiac sympathetic nerve activity, and is abolished by cardiac sympathectomy but not by vagotomy. High spinal cord transection or infusion of the GABA(A) receptors antagonist, bicuculline, into the cisterna magna is effective in disrupting the reflex bradycardia. Opioid receptor blockade does not disrupt the reflex arc. We conclude that the reflex pathway involved in the decrease of heart rate by acupuncture-like stimulation comprises groups III and IV muscle afferent nerves whose activation stimulates GABAergic neurons in the brainstem and inhibits sympathetic outflow to the heart. When the sympathetic tone is high due to hypercapnia, the induced reduction in both cardiac sympathetic nerve activity and heart rate is not augmented, suggesting that the magnitude of sympatho-inhibitory response to acupuncture-like stimulation does not depend on pre-existing sympathetic tone.

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 Aß (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.

Sympatho-inhibitory response of the heart as a result of short-term acupuncture-like stimulation of the rat hindlimb is not augmented when sympathetic tone is high as a result of hypercapnia.

This study examined whether the sympatho-inhibitory response of the heart to acupuncture-like stimulation of a hindlimb in anesthetized rats depends on the sympathetic tone. Observed reductions in both cardiac sympathetic nerve activity and heart rate following short-term acupuncture-like stimulation of a hindlimb were not augmented when the sympathetic tone was high as a result of hypercapnia.

Mechanism of the reflex inhibition of heart rate elicited by acupuncture-like stimulation in anesthetized rats.

Acupuncture or acupuncture-like stimulation applied to different body areas can modify autonomic nerve activity to various organs, including gut, bladder, adrenal medulla, and the heart. We studied the reflex bradycardia in response to insertion into the skin and underlying muscles and twisting of an acupuncture needle in pentobarbital-anesthetized rats. We found that acupuncture-like stimulation of forelimb, hindlimb, chest, and abdomen all produced significant heart rate decreases. Rate minima were reached at the end of the 60-second stimulation episode and significant bradycardia persisted for about 40 s after stimulation ended. Heart rate decreases were paralleled by decreases in cardiac sympathetic nerve activity, and could be produced by electrical stimulation of group IV muscle afferent fibers (tibial nerve). Electrical stimulation of the tibial nerve at rates as low as 0.1-2 Hz was effective for eliciting heart rate decreases. Nerve fiber groups were defined by stimulation of and recording from tibial nerve. Activation of groups I, II, or III fiber was ineffective for eliciting the reflex bradycardia. Sympathectomy, high spinal transection, or infusion of the GABA(A) receptor antagonist, bicuculline, into the cisterna magna were all effective for disrupting the reflex bradycardia. Vagotomy and opioid receptor blockade were ineffective for disrupting the reflex pathway. We conclude that the reflex pathway to decrease heart rate by acupuncture-like stimulation consists of mainly group IV muscle afferent fibers whose activity (even very low rate of activity) leads to the activation of GABA-ergic neurons in the brainstem and an inhibition of sympathetic outflow to the heart.

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.

Neural mechanism of bradycardiac responses elicited by acupuncture-like stimulation to a hind limb in anesthetized rats.

The effects of acupuncture-like stimulation of a hind limb on heart rate were examined in anesthetized rats. An acupuncture needle, having a diameter of either 160 or 340 microm, was inserted into the skin and underlying muscles at a depth of about 5 mm and twisted right and left twice every second for 1 min. Stimulation by a needle with a diameter of either 160 or 340 microm produced a decrease in heart rate. Severance of the femoral and sciatic nerves ipsilateral to the hind-limb stimulation completely abolished the bradycardiac response. Also, heart rate was significantly decreased by acupuncture-like stimulation of the hind-limb muscles alone, but was not significantly influenced by the stimulation of the hind-limb skin alone. The bradycardiac response induced by acupuncture-like stimulation was not influenced by bilateral severance of the vagal nerves at the cervical level, but was abolished by bilateral stellectomy. Acupuncture-like stimulation of the hind limb induced a decrease in the activity of the cardiac sympathetic efferent nerve as well as a decrease in heart rate. These results indicate that the decrease in heart rate induced by acupuncture-like stimulation of a hind limb is a reflex response. The afferent pathway is composed of hind-limb muscle afferents, and the efferent pathway is composed of cardiac sympathetic nerves.

Effects of thermal stimulation, applied to the hindpaw via a hot water bath, upon ovarian blood flow in anesthetized nonpregnant rats.

The effects of thermal stimulation, applied to the hindpaw via a hot bath set to either 40 degrees C (non-noxious) or 49 degrees C (noxious), upon ovarian blood flow were examined in nonpregnant anesthetized rats. Ovarian blood flow was measured using a laser Doppler flowmeter. Blood pressure was markedly increased following 49 degrees C stimulation. Ovarian blood flow, however, showed no obvious change during stimulation, although a small increase was observed after stimulation. Ovarian blood flow and blood pressure responses to 49 degrees C stimulation were abolished after hindlimb somatic nerves proximal to the stimuli were cut. Heat stimulation (49 degrees C) resulted in remarkable increases in both ovarian blood flow and blood pressure in rats in which the sympathetic nerves supplying the ovary were cut but the hindlimb somatic nerves remained intact. The efferent activity of the ovarian plexus nerve was increased during stimulation at 49 degrees C. Stimulation at 40 degrees C had no effect upon ovarian blood flow, blood pressure or ovarian plexus nerve activity. Electrical stimulation of the distal part of the severed ovarian plexus nerve resulted in a decrease in both the diameter of ovarian arterioles, observed using a digital video microscope, and ovarian blood flow.The present results demonstrate that noxious heat, but not non-noxious warm, stimulation of the hindpaw skin in anesthetized rats influences ovarian blood flow in a manner that is attributed to reflex responses in ovarian sympathetic nerve activity and blood pressure.

Manual acupuncture needle stimulation of the rat hindlimb activates groups I, II, III and IV single afferent nerve fibers in the dorsal spinal roots.

Using single unit nerve recording techniques in rats, the present experiment aimed to determine which specific population of afferent nerve fibers (groups I, II, III and IV) in the dorsal roots at the 4th or 5th lumbar segments (L4 or L5) are activated during manual acupuncture needle stimulation. An acupuncture needle 300-340 microm in diameter was inserted into the skin and underlying muscles around the Zusanli acupoint (ST36) area in the hindlimbs, and was manually rotated right and left at a frequency of about 1 Hz for 1 min. The dorsal root of the L4 and L5 spinal nerve was cut close to the entrance into the spinal cord after laminectomy and dissected free to record unitary afferent nerve activity. A single afferent fiber activated by acupuncture stimulation was identified by the identical shape of the discharge spikes during stimulation and during electrically evoked action potentials induced by single pulse electrical stimulation of the sciatic nerve. The conduction velocity of the afferent fiber was calculated by the latency of the electrically evoked action potential. A total of 35 units were intentionally recorded from all animals in order to include all 4 afferent fiber groups. Units were spontaneously silent in the absence of stimulation, while all units responded to ipsilateral manual rotation of the acupuncture needle. The conduction velocity of all 35 units ranged between 0.8 and 86.0 m/s, thus belonging to groups I-V fibers. Mean conduction velocity of groups I, II, III and IV were 57.9 m/s (n = 13), 42.9 m/s (n = 11), 10.3 m/s (n = 6) and 1.2 m/s (n = 5), respectively. Mean discharge rates during acupuncture stimulation of groups I, II, III and IV afferents were 7.4 Hz, 6.2 Hz, 4.7 Hz and 0.4 Hz, respectively. Discharge rates of group IV afferent fibers were significantly lower than those of groups I, II and III afferents. It was concluded that manual acupuncture needle stimulation to the hindlimbs activated afferent nerve fibers belonging to all four groups of afferents in rats. It is suggested that all four groups of somatic afferents activated by manual acupuncture stimulation will elicit various effects when action potentials are delivered to the central nervous system.

Effect of moxibustion stimulation of various skin areas on cortical cerebral blood flow in anesthetized rats.

The effect of moxibustion stimulation of various skin areas (cheek, forepaw, upper arm, chest, back, lower leg, hindpaw and perineum) on cerebral blood flow (CBF) of the parietal cortex was examined in anesthetized rats after eliminating emotional influences. Moxibustion stimulation was performed by burning a moxa cone of about 4 mg weight placed on the shaved skin. CBF of the parietal cortex was measured using a laser Doppler flowmeter. Stimulation of the cheek, forepaw, upper arm and hindpaw produced significant increases in CBF, but stimulation of the other areas did not produce significant responses. Moxibustion stimulation of the forepaw and hindpaw produced an increase in the mean arterial pressure (MAP), while stimulation of the other areas did not. After spinal transection at the 2nd thoracic level, the MAP response to stimulation of the forepaw was abolished, whereas the CBF response to stimulation of the forepaw remained. The CBF response in spinalized rats was not affected by cutting cervical sympathetic and facial parasympathetic nerves, while it was almost abolished by intravenous administration of muscarinic and nicotinic cholinergic blocking agents. The CBF response was abolished by crushing the brachial plexus ipsilateral to the stimulated side. It is suggested that the increase in CBF, independent of MAP and emotional responses, elicited by moxibustion stimulation is a reflex response whose afferent pathway is composed of somatic afferent nerves, and whose efferent pathway involves intracerebral cholinergic nerves. A contribution of endogenous opioids in the present CBF responses was neglected, because naloxone did not influence the CBF responses.