• The Journal of Korean Physical Therapy
• /
• 제19권2호
• /
• pp.41-55
• /
• 2007

Purpose: This study aimed to compare the effect on nerve regeneration of ultrasound irradiation in rats with peripheral nerve injury. Methods: To investigate alterations of the NCAM immunoreactivity in non-crushed part and crushed part of the spinal cord, the unilateral sciatic nerve of the rats were crushed. The expression of NCAM was used as the marked of peripheral nerve regeneration, and also plays an important role in developing nerve system. Experimental animals were sacrificed by perfusion fixation at post-injury 1, 3, 7, 14 days after ultrasound irradiation. The pulsed US was applied at a frequency of 1MHz and a spatial average-temporal average Intensity of 0.5W/of (20% pulse ratio) for 1 mins. The Luxol fast blue-cresyl violet stain were also done to observe the morphological changes. Results: Alteration of NCAM immunoreactivity in the crushed part and the non-crushed part of lower lumbar spinal cord were observed. NCAM-immunoreactivity cells were some increased in the dorsal horn lamina I, III and cell ventral horn at 1 day after unilateral sciatic nerve injury. However, there was not significant difference in the relationship between crushed part and non-crushed part. NCAM-inmmunoreactivity was remarkably increased at 3 days after unilateral sciatic nerve injuryin the gray matter and white matter. NCAM-immunoreactivity was increased in the ventral horn and post horn of experimental crushed part. Also, NCAM-immunoreactivity in large motor neurons in ventral horns lamina VIII, IX were increased at 7 days after unilateral sciatic nerve injury. At 14 days after sciatic nerve crushed injury, there was no significant difference. All group were decreased for 14 days. In the time course of NCAM expression, all groups showed a significant difference at 3day groups(p<0.05). Whereas, CC group was noted a significant difference between 3day and 7 day group respectively. In NCAM expression, there were significantly increased in all group. In the relationship between CNC group and ENC group, significant difference was detected among 3, 7, 14 day group(p<0.05). The difference between CC group and ENC group were noted in all groups(p<0.05). Conclusion: It is consequently suggested that the effects of the ultrasound irradiation may increase the NCAM immunoreactive neurons and glial cell in the spinal cord after unilateral sciatic nerve crushed injury. Therefore, the increased NCAM immunoreactivity in the spinal cord may reflect the neuronal damage and healing process induced by a ultrasound irradiation after peripheral nerve injury in rat.

• Journal of Acupuncture Research
• /
• 제18권2호
• /
• pp.51-66
• /
• 2001

The purpose of this morphological studies was to investigate the central neural pathway projecting to the acupoints $B_{62}$ and $K_6$ using the neuroanatomical method following injection of transsynaptic neurotropic virus, pseudorabies virus(PRV-Ba and PRV-Ga) into the $B_{62}$ and $K_6$. After survival times of 96 hours following injection into the twenty rats with PRV-Ba(Bartha strain) and PRV-Ga(Bartha strain, ${\beta}$-galacidodase insertion). They were perfused, and their spinal cord and brain were frozen sectioned($30{\mu}m$). These sections were stained by X-gal histochemical and PRV immunohistochemical staining method, and observed with light microscope. The results were as follows : 1. In spinal cord, overlaped PRV-Ba and PRV-Ga labeled neurons projecting to the $B_{62}$ and $K_6$ were founded in thoracic, lumbar and sacral spinal segments. In thoracic spinal segments, Densely labeled areas were founded in lamina IV, V, VII(intermediolateral nucleus) and X areas. In lumbar segemnts, labeled areas were founded in lamina II, IV, V and X areas. In sacral spinal segments, labeled areas were founded in lamina IV, V and VI areas. 2. In brain, overlaped PRV-Ba and PRV-Ga labeled neurons projecting to the $B_{62}$ and $K_6$ were founded in the $A_1$ noradrenalin cells/$C_1$ adrenalin cells/caudoventrolateral reticular nucleus, rostroventrolateral reticular nuclens, nucleus tractus solitarius, area postrema, raphe obscurus nucleus, raphe paltidus nucleus, raphe magnus nucleus, lateral paragigantoceltular nucleus, lateral rcticular nucleus, gigantocellular nucleus, locus coeruleus, subcoeruleus nucleus, motor trigeminal nucleus, Kolliker-Fuse nucleus, $A_5$ cell group, central gray matter, oculomotor nerve, paraventricular hypothalamic nucleus, median eminence, amygdaloid nucleus, frontal cortex, forelimb area, hindlimb area, 1, 2 areas of parietal cortex and granular and agranular cortex. This results were suggest that overlaped PRV-Ba and PRV-Ga labeled areas projecting to the $B_{62}$ and $K_6$ may be related to the emotional relay pathway in the central autonomic center.

• Applied Microscopy
• /
• 제27권2호
• /
• pp.177-187
• /
• 1997

Hydrocephalus is induced experimentally in prenatal and suckling animals following an injection of 6-aminonicotinamide (6-AN). The most remarkable characteristic of these animals is aqueduct stenosis caused by swellings of the ependymal cells and subependymal cells in the periaqueductal gray matter and the central canal of the spinal cord. The present study was undertaken to investigate the morphological changes of the ependymal cells in the central canal of the spinal cord of 3.5 months old hamster after treatment with 6-AN. Intraperitoneal administrations of 6-AN (10 mg/kg body weight) every two days gave rise to partial central canal stenosis of the spinal cord after 27-29 days (13-l4th injection), but cilia and microvilli were located in the strictural area of the con#rat canal. The vacuolations in the ependymal cells were not observed and degenerating changes of intracellular organelles of the ependymal cells did not occur, so that the ependymal cells lining the central canal of the hamster spinal cord were not affected by 6-AN. But the present study demonstrate that 6-AN causes to create numerous vacuoles in the subependymal area of the central canal. Although the vacuoles were well developed in the neuroglial cells and the neuropils of the subependymal area, the neurons were not affected by 6-AN. These results strongly suggests that partial central canal stenosis occurred by 6-AN was due to vacuolations and swellings of the neuroglial cells and nueropils in the subependymal area.

• Korean Journal of Acupuncture
• /
• 제36권4호
• /
• pp.264-273
• /
• 2019

Objectives : Microglia play a crucial role in electroacupuncture (EA) analgesia on neuropathic pain. The role of chemokines in producing analgesic effects of EA, however, is largely unknown. In the present study, we investigated the role of chemokines in producing analgesic effects of EA in the neuropathic pain model. Methods : Sprague-Dawley rats were randomly assigned into three groups (anesthetized group (ANE), non-acupoint EA group (NAP), and ST36 - GB34 EA group (ACU)). Neuropathic pain was induced by tight ligation of L5 spinal nerve. Mechanical and thermal hypersensitivity of hind paw was tested. Western blot tests and immunofluorescence assay for C-C motif chemokine ligand 2 (CCL2) levels and microglia activation were performed on spinal cord L5/6. EA was treated once daily from the 3rd day after surgery for 5 days. Results : EA treatments applied to ST36 and GB34 significantly reduced both mechanical and thermal hypersensitivity after two and three times of treatment, respectively. While CCL2 expression significantly increased in neuropathic rats, it was significantly reduced in the ACU. In addition, co-localization of CCL2 and activated microglia significantly decreased in the ACU compared to those of ANE and NAP in the spinal cord L5/L6 dorsal horn. Conclusions : The present results suggest that EA applied to ST36 and GB34 modulates the reduction of CCL2 release from the injured neurons and consequently decreases microglia activation in the spinal cord. Regulation of chemokine induced spinal activation of microglia plays a key role in analgesic effects of EA in the rat model of neuropathic pain.

• The Korean Journal of Physiology and Pharmacology
• /
• 제27권1호
• /
• pp.39-48
• /
• 2023

Spinal nerve injury causes mechanical allodynia and structural imbalance of neurotransmission, which were typically associated with calcium overload. Storeoperated calcium entry (SOCE) is considered crucial elements-mediating intracellular calcium homeostasis, ion channel activity, and synaptic plasticity. However, the underlying mechanism of SOCE in mediating neuronal transmitter release and synaptic transmission remains ambiguous in neuropathic pain. Neuropathic rats were operated by spinal nerve ligations. Neurotransmissions were assessed by whole-cell recording in substantia gelatinosa. Immunofluorescence staining of STIM1 with neuronal and glial biomarkers in the spinal dorsal horn. The endoplasmic reticulum stress level was estimated from qRT-PCR. Intrathecal injection of SOCE antagonist SKF96365 dose-dependently alleviated mechanical allodynia in ipsilateral hind paws of neuropathic rats with ED₅₀ of 18 ㎍. Immunofluorescence staining demonstrated that STIM1 was specifically and significantly expressed in neurons but not astrocytes and microglia in the spinal dorsal horn. Bath application of SKF96365 inhibited enhanced miniature excitatory postsynaptic currents in a dosage-dependent manner without affecting miniature inhibitory postsynaptic currents. Mal-adaption of SOCE was commonly related to endoplasmic reticulum (ER) stress in the central nervous system. SKF96365 markedly suppressed ER stress levels by alleviating mRNA expression of C/ EBP homologous protein and heat shock protein 70 in neuropathic rats. Our findings suggested that nerve injury might promote SOCE-mediated calcium levels, resulting in long-term imbalance of spinal synaptic transmission and behavioral sensitization, SKF96365 produces antinociception by alleviating glutamatergic transmission and ER stress. This work demonstrated the involvement of SOCE in neuropathic pain, implying that SOCE might be a potential target for pain management.

• International Journal of Oral Biology
• /
• 제45권4호
• /
• pp.225-231
• /
• 2020

Glial cells, including astrocytes and microglia, interact closely with neurons and modulate pain transmission, particularly under pathological conditions. In this study, we examined the excitability of substantia gelatinosa (SG) neurons of the spinal dorsal horn using a patch clamp recording to investigate the roles of microglial activation in the nociceptive processes of rats. We used xanthine/xanthine oxidase (X/XO), a generator of superoxide anion (O2·-), to induce a pathological pain condition. X/XO treatment induced an inward current and membrane depolarization. The inward current was significantly inhibited by minocycline, a microglial inhibitor, and fluorocitrate, an astrocyte inhibitor. To examine whether toll-like receptor 4 (TLR4) in microglia was involved in the inward current, we used lipopolysaccharide (LPS), a highly specific TLR4 agonist. The LPS induced inward current, which was decreased by pretreatment with Tak-242, a TLR4-specific inhibitor, and phenyl N-t-butylnitrone, a reactive oxygen species scavenger. The X/XO-induced inward current was also inhibited by pretreatment with Tak-242. These results indicate that the X/XO-induced inward current of SG neurons occurs through activation of TLR4 in microglial cells, suggesting that neuroglial cells modulate the nociceptive process through central sensitization.

• 대한중풍순환신경학회지
• /
• 제13권1호
• /
• pp.77-85
• /
• 2012

Amyotrophic lateral sclerosis(ALS) is a progressive disorder that causes degeneration of motor neurons of the brain and spinal cord. It is characterized by loss of motor neurons leading to muscle weakness, and sensation and mental function stay intact during the course of the disease. Diagnostic tests include magnetic response imaging(MRI), electromyogram(EMG), muscle biopsy, and blood tests. In order to a definitive diagnosis of ALS, damage must be evident in both upper and lower motor neurons. When three limbs are sufficiently affected, the diagnosis is ALS. There is no cure for ALS. We experienced one case of ALS, the patients was diagnosed as ALS by EMG and symptoms. We diagnosed her as ShinEumHur(SEH) and treated by Choakwiyeum(左歸飮). We report the change of her symptoms through oriental medical treatment.

• Advances in Traditional Medicine
• /
• 제2권1호
• /
• pp.36-40
• /
• 2002

Effects of hydrogen peroxide $(H_2O_2)-induced$ neurotoxicity were investigated in cultured newborn rat spinal dorsal root ganglion (DRG) neurons after DRG neurons were treated in the media containning various concentrations of $H_2O_2$. In addition, the protective effect of Herba Epimedii (HE) extract against $H_2O_2-induced$ neurotoxicity was examined. Cytotoxic values were determined by the cell viability of living cells using 3-(4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) assay. In the present study, exposure of neurons to $H_2O_2$ resulted in a significant cell death in a dose- and time-dependent manners in cultured DRG neurons. The decrement of cell viability by $H_2O_2$ was blocked by HE. These results suggest that the neuroprotective effect of HE against $H_2O_2-induced$ cytotoxicity may result from the prevention of injury induced by $H_2O_2$.

• International Journal of Oral Biology
• /
• 제36권2호
• /
• pp.83-89
• /
• 2011

Substantia gelatinosa (SG) neurons receive synaptic inputs from primary afferent $A{\delta}$- and C-fibers, where nociceptive information is integrated and modulated by numerous neurotransmitters or neuromodulators. A number of studies were dedicated to the molecular mechanism underlying the modulation of excitability or synaptic plasticity in SG neurons and revealed that second messengers, such as cAMP and cGMP, play an important role. Recently, cAMP and cGMP were shown to downregulate each other in heart muscle cells. However, involvement of the crosstalk between cAMP and cGMP in neurons is yet to be addressed. Therefore, we investigated whether interaction between cAMP and cGMP modulates synaptic plasticity in SG neurons using slice patchclamp recording from rats. Synaptic activity was measured by excitatory post-synaptic currents (EPSCs) elicited by stimulation onto dorsal root entry zone. Application of 1 mM of 8-bromoadenosine 3,5-cyclic monophosphate (8-Br-cAMP) or 8-bromoguanosine 3,5-cyclic monophosphate (8-Br-cGMP) for 15 minutes increased EPSCs, which were maintained for 30 minutes. However, simultaneous application of 8-BrcAMP and 8-Br-cGMP failed to increase EPSCs, which suggested antagonistic cross-talk between two second messengers. Application of 3-isobutyl-1-methylxanthine (IBMX) that prevents degradation of cAMP and cGMP by blocking phosphodiesterase (PDE) increased EPSCs. Co-application of cAMP/cGMP along with IBMX induced additional increase in EPSCs. These results suggest that second messengers, cAMP and cGMP, might contribute to development of chronic pain through the mutual regulation of the signal transduction.

• Applied Microscopy
• /
• 제36권4호
• /
• pp.299-305
• /
• 2006

Onuf 핵이란 척수 앞회색질뿔에 위치하는 운동핵으로 음부신경을 통해 방광과 항문괄약근을 조절하는 운동핵의 하나이다. Onuf핵은 앞회색질뿔내 다른 운동신경핵과는 달리 회색질척수염과 같은 병적인 상황에서도 상당기간 손상되지 않고 기능을 유지하며, 퇴행성변화의 정도가 미약한데 정확한 원인에 관해서는 논란의 여지가 많다. 본 연구는 흰쥐 척수회색질내 바깥요도조임근을 신경지배하는 Onuf핵의 위치를 HRP 추적법으로 확인하였으며, 이들 신경핵내 운동신경세포와 연접해 있는 zinc함유(ZEN)신경종말의 미세구조를 zinc selenium조직화학법(AMG)으로 염색하여 관찰하였다. HRP 추적법의 결과로는, Onuf핵은 랫드 척수회색질앞뿔의 내측에서 가지돌기의 무리와 거의 맞닿고 있었으며, 모양은 대개 구형 또는 난원형을 띠었다. 이들 신경핵내 운동신경세포의 세포체의 크기는 다른 운동핵의 신경세포보다 다소 작았다. 한편 AMG로 염색한 표본에서는 Onuf핵에 분포하는 ZEN신경종말은 다른 운동핵의 ZEN 신경종말과 비교하여 매우 높은 밀집도를 보였으나, 크기 면에서도 상대적으로 작았다. 미세구조 관찰로는 Onuf 핵내 ZEN신경종말은 운동핵의 세포체 및 가지돌기와 신경연접은 이루고 있었다. 이들 ZEN 신경종말은 주로 납작한 연접소포를 함유하였으며, 대칭적인 신경연접구조를 이루고 있었다.