Hideo Ohsawa

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.

Neural mechanism of depressor responses of arterial pressure elicited by acupuncture-like stimulation to a hindlimb in anesthetized rats.

The effects of acupuncture-like stimulation of a hindlimb on renal sympathetic nerve activity (RNA) as well as mean arterial blood pressure (MAP) were examined in anesthetized rats. An acupuncture needle (diameter of 160 microns) was inserted into the skin of a hindlimb and underlying muscles to a depth of 5 mm and was twisted at about 1 Hz. Under deep anesthetic condition, in about 70% of trials, acupuncture-like stimulation for 60 s induced a decrease in MAP which was accompanied by a decrease in RNA. Acupuncture-like stimulation applied to the muscles alone, but not to the skin alone, induced inhibition of RNA and MAP. Transection of sciatic and femoral nerves ipsilateral to the hindlimb stimulation completely abolished the responses of RNA and MAP. The hindlimb stimulation excited the femoral and common peroneal afferent nerves. In spinalized animals, the hindlimb stimulation did not produce any changes in RNA and MAP. The results indicate that the decrease in MAP induced by acupuncture-like stimulation of a hindlimb is a reflex response. The afferent pathway is composed of hindlimb muscle afferents while the efferent pathway is composed of sympathetic vasoconstrictors including the renal nerves. Endogenous opioids may not be involved in the present reflex, because an intravenous injection of naloxone, an antagonist of the opioid receptors, did not influence the reflex.

Somatocardiovascular reflexes in anesthetized rats with the central nervous system intact or acutely spinalized at the cervical level.

The effects of noxious mechanical stimulation of various segmental areas on heart rate and mean arterial blood pressure (MAP), as well as cardiac and renal sympathetic nerve activities were examined in anesthetized rats with the central nervous system (CNS) intact or acutely spinalized at the cervical level. In CNS-intact rats, pinching for 20 s applied to any segmental skin area, but particularly that of the paw, produced an increase in heart rate, blood pressure and the sympathetic nerve activities. In acutely spinalized rats, pinching the chest, abdomen and back of the body produced large increases, while hindlimb and perineum stimulation induced only a small increase or no increase in heart rate, blood pressure and the sympathetic nerve activities. Stimulation of the right side produced particularly large responses in heart rate and stimulation of the ipsilateral side produced large responses in cardiac and renal sympathetic nerve activities in spinalized rats. These results suggest the existence of the two types of reflex responses, supraspinal and propriospinal, in the somatocardiovascular reflex. The supraspinal one has characteristics of diffuse reflex organization, while the propriospinal one has strong segmental and lateral organization.

Neural mechanism of pupillary dilation elicited by electro-acupuncture stimulation in anesthetized rats

The neural mechanisms to reflex dilation elicited by electro-acupuncture stimulation were investigated in anesthetized rats. Two needles, with 160 microns diameter and about 5 mm apart, were inserted into the skin and underlying muscle of a hindpaw. Repetitive 20 Hz, 0.5 ms electrical pulses at various intensities were used for stimulation for 30s. The pupil size was magnified about 44 times via a microscope and was continuously recorded on a videotape. Electro-acupuncture stimulation at more than 0.5 up to 6 mA induced stimulus intensity-dependent pupil dilation. These responses were abolished by the severance of the sciatic and saphenous nerve of the stimulated hindlimb. Compound action potentials were recorded from the distal cut end of the tibial of a saphenous nerve following electro-acupuncture stimulation of the hindpaw. The mean threshold of the compound action potentials of the myelinated fibers in saphenous nerves was 0.18 mA, while that of unmyelinated fibers was 3.0 mA. The mean threshold of the compound action potentials of the myelinated fibers in the tibial nerve was 0.20 mA of unmyelinated fibers was 3.3 mA. Severance of bilateral trunks did not affect the response, while severance of the third cranial nerves abolished the responses. In conclusion, electro-acupuncture stimulation applied to the hindpaws of the anesthetized rats induced excitation of myelinated or of both myelinated and unmyelinated afferent fibers of the tibial and saphenous nerve, and involved a reflex response of pupil dilation through the third cranial parasympathetic efferent nerve.

Electro-acupuncture stimulation to a hindpaw and a hind leg produces different reflex responses in sympathoadrenal medullary function in anesthetized rats.

The effects of electro-acupuncture stimulation (EAS) of two different areas of a hindlimb with different stimulus intensities on sympathoadrenal medullary functions were examined in anesthetized artificially ventilated rats. Two needles of 160 microm diameter and about 5 mm apart were inserted about 5 mm deep into a hindpaw (Chungyang, S42) or a hind leg (Tsusanli, S36) and current of various intensities passed to excite various afferent nerve fiber groups at a repetition rate of 20 Hz and pulse duration of 0.5 ms for 30-60 s. Fiber groups of afferent nerves stimulated in a hindlimb were monitored by recording evoked action potentials from the afferents innervating the areas stimulated. The sympathoadrenal medullary functions were monitored by recording adrenal sympathetic efferent nerve activity and secretion rates of catecholamines from the adrenal medulla. EAS of a hindpaw at a stimulus strength sufficient to excite the group III and IV somatic afferent fibers produced reflex increases in both adrenal sympathetic efferent nerve activity and the secretion rate of catecholamines. EAS of a hind leg at a stimulus strength sufficient to excite the group III and IV afferent fibers produced reflex responses of either increases or decreases in sympathoadrenal medullary functions. All responses of adrenal sympathetic efferent nerve activity were lost after cutting the afferent nerves ipsilateral to the stimulated areas, indicating that the responses are the reflexes whose afferents nerve pathway is composed of hindlimb somatic nerves. It is concluded that electro-acupuncture stimulation of a hindpaw causes an excitatory reflex, while that of a hind leg causes either excitatory or inhibitory reflex of sympathoadrenal medullary functions, even if both group III and IV somatic afferent fibers are stimulated.

Effects of aging on numbers, sizes and conduction velocities of myelinated and unmyelinated fibers of the pelvic nerve in rats

The effects of aging on the conduction velocities, numbers and sizes of the myelinated and unmyelinated fibers of the pelvic nerve in Wistar rats, aged 3-9 and 30-37 months were examined using electrophysiological and ultrastructural techniques. The myelinated fibers did not show significant age-related changes in the maximum conduction velocities (9.7 +/- 1.1 m/s in adult vs. 11.0 +/- 1.5 m/s in aged rats), the number of fibers (662 +/- 27 vs. 625 +/- 56) and distribution of fiber diameters. In aged rats, the unmyelinated fibers did not show a significant change in maximum conduction velocity (1.5 +/- 0.1 m/s vs. 1.5 +/- 0.1 m/s), but showed a significant decrease in the number of fibers (4133 +/- 114 vs. 3113 +/- 456), specifically of fibers smaller than 0.7 microm in diameter. It is concluded that myelinated fibers of the pelvic nerve in rats preserve their conduction ability and fiber population during aging, but unmyelinated fibers, particularly fibers with smaller diameters decrease in number. Unmyelinated fibers with larger diameters maintain their conduction ability and numbers with aging.

Is there a difference between the effects of single and triple indirect moxibustion stimulations on skin temperature changes of the posterior trunk surface

Objectives To determine whether any difference exists in responses to indirect moxibustion (IM) relative to thermal stimulation duration. Methods In experiment 1, 9 subjects attended two experimental sessions consisting of single stimulation with IM or triple stimulation with IM, using a crossover design. A K-type thermocouple temperature probe was fixed on the skin surface at the GV14 acupuncture point. IM stimulation was administered to the top of the probe in order to measure the temperature curve. In addition, each subject evaluated his or her subjective feeling of heat on a visual analogue scale after each stimulation. Experiment 2 was conducted on 42 participants, divided into three groups according to the envelope allocation method: single stimulation with IM (n=20), triple stimulation with IM (n=11) and a control group (n=11). A thermograph was used to obtain the skin temperature on the posterior trunk of the participant. To analyse skin temperature, four arbitrary frames (the scapular, interscapular, lumbar and vertebral regions) were made on the posterior trunk. Result In experiment 1, no significant difference in maximum temperature was found in IM and subjective feeling of heat intensity between single and triple stimulation with IM. In experiment 2, increases in skin temperature occurred on the posterior trunk, but no differences in skin temperature occurred between the groups receiving single and triple stimulation with IM. Conclusion No difference exists in the skin temperature response to moxibustion between the single and triple stimulation with IM.

Neural mechanism of localized changes in skeletal muscle blood flow caused by moxibustion-like thermal stimulation of anesthetized rats

Moxibustion-like thermal stimulation (MTS) was applied to the gastrocnemius muscle to measure local muscle blood flow (MBF) in the stimulated region and the change in the MBF in the region, and its mechanism was examined. In the experiment, we used urethane-anesthetized rats under artificial respiration and observed the change caused by gastrocnemius MTS using a laser Doppler blood-flow meter. MTS applied to the gastrocnemius muscle caused a two-phase response in blood flow that showed a transient decrease followed by an increase without blood pressure change. It is suggested that the increase in response occurs because of an axon reflex that has a reflex arc below the spinal cord, and the decrease in response is caused by direct stimulation of postganglionic muscle sympathetic fibers.

Cutaneous afferents producing a reflex pupil dilation in anesthetized rats

Cutaneous afferents producing a reflex pupil dilation were examined using natural mechanical stimulation of the hindlimb skin and electrical stimulation of a sural nerve in anesthetized, artificially ventilated rats. Pupil diameter was continuously recorded after magnification using a microscope connected to a charge coupled device camera. Innocuous brushing, or weak pressing of the skin, did not have any effect on pupil diameter, while pressing the skin more than 720 g/cm2 produced a pressure-dependent pupil dilation. Pinching rather than pressing the skin induced the larger pupil dilation. Electrical stimulation of a sural afferent nerve with weak intensity, which was supra-threshold for Abeta-afferents and sub-threshold for Asigma-afferents, induced a reflex pupil dilation. This dilation continued to increase with further increases in stimulus intensity which involved excitation of Asigma afferents and C afferents. It is concluded that Abeta, Asigma and C afferents in the skin can work as afferents in eliciting reflex pupil dilation in anesthetized rats.