To study the direct effect of somatostatin (SS) on calcium channel current ($I_{Ba}$) in guinea-pig gastric myocytes, $I_{Ba}$ was recorded by using whole-cell patch clamp technique in single smooth muscle cells. Nicardipine ($1{\mu}M$), a L-type $Ca^{2+}$ channel blocker, inhibited $I_{Ba}$ by $98{\pm}1.9$% (n=5), however $I_{Ba}$ was decreased in a reversible manner by application of SS. The peak $I_{Ba}$ at 0 mV were decreased to $95{\pm}1.5$, $92{\pm}1.9$, $82{\pm}4.0$, $66{\pm}5.8$, $10{\pm}2.9$% at $10^{-10}$, $10^{-9}$, $10^{-8}$, $10^{-7}$, $10^{-5}$ M of SS, respectively (n=3∼6; $mean{\pm}SEM$). The steady-state activation and inactivation curves of $I_{Ba}$ as a function of membrane potentials were well fitted by a Boltzmann equation. Voltage of half-activation ($V_{0.5}$) was $-12{\pm}0.5$ mV in control and $-11{\pm}1.9$ mV in SS treated groups (respectively, n=5). The same values of half-inactivation were $-35{\pm}1.4$ mV and $-35{\pm}1.9$ mV (respectively, n=5). There was no significant difference in activation and inactivation kinetics of $I_{Ba}$ by SS. Inhibitory effect of SS on $I_{Ba}$ was significantly reduced by either dialysis of intracellular solution with $GDP_{\beta}S$, a non-hydrolysable G protein inhibitor, or pretreatment with pertussis toxin (PTX). SS also decreased contraction of guinea-pig gastric antral smooth muscle. In conclusion, SS decreases voltage-dependent L-type calcium channel current ($VDCC_L$) via PTXsensitive signaling pathways in guinea-pig antral circular myocytes.
Tetrahydroisoquinoline (THI) alkaloids can be considered as cyclized derivatives of simple phenylethy-lamines, and many of them, especially with 6,7-disubstitution, demonstrate relatively high affinity for catecholamines. Two -OH groups at 6 and 7 positions are supposed to be essential to exert ?${\beta}-receptor$ activities. However, it is not clear whether -OH at 6,7 substitution of THIs also shows ?${\alpha}-adrenoceptor$ activities. In the present study, we investigated whether -OH or $-OCH_3$ substitutions of 6,7 position of THIs differently affect the ?1-adrenoceptor affinity. We synthesized two 1-naphthylmethyl THI alkaloids, $1-{\beta}-naphthylmethyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline$ HBr (YS 51) and $1-{\beta}-naphthylmethyl-6, 7-dimethoxy-1,2,3,4-tetrahydroisoquinoline$ HCl (YS 55), and their pharmacological actions on ?${\alpha}_1-adrenoceptor$ were compared. YS 51 and YS 55, concentration-dependently relaxed endothelium-denuded rat thoracic aorta precontracted with phenylephrine (PE, 0.1 ${\mu}M$) in which $pEC_{50}$ were $5.89{\pm}0.21$ and $5.93{\pm}0.19$, respectively. Propranolol (30 nM) did not affect the relaxation-response curves to YS 51 and YS 55. Concentration-response curves to PE were shifted to right by the pretreatment with YS 51 or YS 55. The $pA_2$ values of YS 51 and YS 55 showed $6.05{\pm}0.24$ and $5.88{\pm}0.16$, respectively. Both probes relaxed KCl (65.4 mM)-contracted aorta and inhibited $CaCl_2-induced$ contraction of PE-stimulated endothelium- denuded rat thoracic aorta in $Ca^{2+}-free$ solutions. In isolated guinea pig papillary muscle, 1 and 10 ${\mu}M$ YS 51 increased contractile force about 4- and 8- fold over the control, respectively, along with the concentration-dependent increment of cytosolic $Ca^{2+}$ ions. While, 10 ${\mu}M$ YS 55 reduced the contractile force about 50 % over the control and lowered the cytosolic $Ca^{2+}$ level, in rat brain homogenates, YS 51 and YS 55 displaced $[^3H]prazosin$ binding competitively with Ki 0.15 and 0.12 ${\mu}M$, respectively. However, both probes were ineffective on $[^3H]nitrendipine$ binding. Therefore, it is concluded that two synthetic naphthylmethyl-THI alkaloids have considerable affinity to ?1-adrenenoceptors in rat aorta and brain.
Objectives : The purpose of this study was to investigate the effects of Yanghyuljanggeungunbo-tang(Yangxuezhuangjinjianbu-tang, YGKT) and electrical acupuncture treatment in spinal cord injury(SCI)-induced rats. Methods : The subjects were divided into 5 groups ; Normal, Control-no treatment after SCI, Experimental I(Exp. I)-taken with YGKT 500 mg/kg $0.5m{\ell}$ daily after SCI. Experimental II(Exp. II)-taken with electrical acupuncture after SCI and Experimental III(Exp. III)-taken with YGKT 500 mg/kg $0.5m{\ell}$ and electrical acupuncture after SCI. After each operation, the present author observed cytological changes, the motor behavior recovery and nerve regeneration by analysis of the motor behavior tests, EMG, hematological(AST, ALT, WBC), histological and immunological changes. Rats were tested by Motor behavior test at 1st, 3rd, 7th, 14th and 21st day. Results : 1. All the experimental groups were improved compared with control group in the motor behavior tests including Tarlov test, Basso-Beattle-Bresnahan locomotor rating scale, modified inclined plane test, open field test, grid walk test and narrow beam test. Especially Exp. III was significantly improved among other groups. 2. In EMG test, H and M wave were significantly increased in Exp. III. 3. All the experimental groups were significantly decreased compared with control group in AST, ALT and WBC. 4. NGF, BDNF and Trk B of spinal cord gray matter in all the experimental groups were increased compared with control group. Especially, Exp. III was more effective. 5. In histological observations, muscle contraction and denaturation of gastrocnemius muscle of all the experimental groups were inhibited. Especially, those of Exp. III was more effective. On the observations of liver and kidney, cell atrophy and apoptosis of all the experimental groups were decreased compared with control group. Especially, those of Exp. III was more effective. Conclusions : It can be suggested that YGKT and electrical acupuncture may improve motor behavior, EMG, hematological, histological and immunological findings in SCI-induced rats. Especially, combination of these two treatments will be somewhat better in spinal nerve recovery and motor function improvement.
Objectives : The purpose of this study was to investigate the effects of Yanghyuljanggeungunbo-tang(Yangxuezhuangjinjianbu-tang) and Cervi Cornu Parvum pharmaco-acupuncture in Spinal Cord Injury(SCI)-induced rats. Methods : The subjects were divided into 5 groups ; Normal, Control no treatment after SCI, Experimental I taken with Yanghyuljanggeungunbo-tang (Yangxuezhuangjinjianbu-tang) 500 mg/kg $0.5m{\ell}$ daily after inducing SCI. Experimental II taken with Cervi Cornu Parvum pharmaco-acupuncture at Taegye(KI3) and $Yangnungch{\acute{o}}n$(GB34) after inducing SCI and Experimental III taken with Yanghyuljanggeungunbo-tang(Yangxuezhuangjinjianbu-tang) 500 mg/kg $0.5m{\ell}$ and Cervi Cornu Parvum pharmaco-acupuncture at KI3 and GB34 to SCI-induced rats. After each operation, the present author observed the motor behavior recovery and nerve regeneration by analysis of the motor behavior tests, EMG, hematological(AST, ALT, WBC), histological and immunological changes. Rats were tested at modified Tarlov test at the 1st, 2nd, 3rd, 4th day, and Motor behavior test at 1st, 3rd, 7th, 14th, 21st day. Results : Results are as follows. 1. All the experimental groups were improved compared with control group in the motor behavior tests including Tarlov test, Basso-Beattle-Bresnahan locomotor rating scale, modified inclined plane test, open field test, grid walk test and narrow beam test. Especially Experimental III was improved significantly among other groups. 2. In EMG test, H wave appeared weak only in Experimental III. And M wave was increased significantly in Experimental III. 3. All the experimental groups were significantly decreased compared with control group in serum AST, serum ALT and serum WBC tests. 4. significantly decreased in Tumor Necrosis Factor-${\alpha}$ test compared with the first day of SCI. 5. Muscle contraction and denaturation of all the experimental groups were inhibited in histological observations of gastrocnemius muscle. Especially, those of experimental III was more effective. 6. NGF and BDNF of spinal cord gray matter in all the experimental groups were increased compared with control group. Especially, those of experimental III was more effective. Conclusions : As above, it can be suggested that Yanghyuljanggeungunbo-tang(Yangxuezhuangjinjianbu-tang) and Cervi Cornu Parvum pharmaco-acupuncture may improve motor behavior, EMG, hematological, histological and immunological findings in Spinal Cord Injury(SCI)-induced rats. Especially, effects will be somewhat better in combination of these two treatments.
Objectives : In the present study, the author intended to investigate Gagam-jeonggitang(GJT), Gami-hwajeongjeon(GHJ) and Gami-tonggyutang(GTT) significantly affect in vivo and in vitro mucin secretion from airway epithelial cells. Methods : In vivo experiment, the author induced hypersecretion of airway mucin, hyperplasia of tracheal goblet cells and the increase in intraepithelial mucosubstances by exposing rats to SO2 during 3 weeks. Effects of orally-administered GJT, GHJ and GTT during 1 week on in vivo mucin secretion and hyperplasia of tracheal goblet cells were assesed using ELISA and staining goblet cells with alcian blue. For in vitro experiment, confluent HTSE cells were metabolically radiolabeled with 3H-glucosamine for 24 hrs and chased for 30 min in the presence of each agent to assess the effects of each agent on 3H-mucin secretion. Possible cytotoxicities of each agent were assessed by measuring lactate dehydrogenase release. Also, the effects of each agent on contractility of isolated tracheal smooth muscle and effects of each agent on MUC5AC gene expression in cultured HTSE cells were investigated. Results : GJT, GHJ and GTI inhibited hypersecretion of in vivo mucin: GJT and GHJ inhibited the increase of number of goblet cells. However, GTT did not affect the increase of number of goblet cells; GJT and GTT significantly increased mucin secretion from cultured HTSE cells, without significant cytotoxicity. GHJ increased mucin secretion and showed mild cytotoxicity at the highest concentration: GJT, GHJ and GTT chiefly affected the 'mucin' secretion; GJT, GHJ and GTT did not affect Ach-induced contraction of isolated tracheal smooth muscle; GTT did not significantly affect the expression levels of MUC5AC gene. However, GJT significantly. inhibit the expression levels of MUC5AC gene and GHJ significantly increased the expression levels of MUC5AC gene. These results suggest that GJT, GHJ and GTI can increase mucin secretion during short-term treatment(in vitro), whereas it can inihibit hypersecretion of mucin during long-term treatment(in vivo) and GJT and GHJ can not only affect the secretion of mucin but also affect the expression of mucin gene. Conclusions : The author suggests that the effects GJT, GHJ and GTT with their components should be further investigated and it is valuable to find, from oriental medical prescriptions, novel agents which might regulate hypersecretion of mucin from airway epithelial cells.
Propofol is known to cause vasorelaxation of several systemic vascular beds. However, its effect on the pulmonary vasculature remains controversial. In the present study, we investigated the effects of propofol on human pulmonary arteries obtained from patients who had undergone surgery. Arterial rings were mounted in a Multi-Myograph system for measurement of isometric forces. U46619 was used to induce sustained contraction of the intrapulmonary arteries, and propofol was then applied (in increments from $10-300{\mu}m$). Arteries denuded of endothelium, preincubated or not with indomethacin, were used to investigate the effects of propofol on isolated arteries. Propofol exhibited a bifunctional effect on isolated human pulmonary arteries contracted by U46619, evoking constriction at low concentrations ($10-100{\mu}m$) followed by secondary relaxation (at $100-300{\mu}m$). The extent of constriction induced by propofol was higher in an endothelium-denuded group than in an endothelium-intact group. Preincubation with indomethacin abolished constriction and potentiated relaxation. The maximal relaxation was greater in the endothelium-intact than the endothelium-denuded group. Propofol also suppressed $CaCl_2$-induced constriction in the 60 mM $K^+$-containing $Ca^{2+}$-free solution in a dose-dependent manner. Fluorescent imaging of $Ca^{2+}$ using fluo-4 showed that a 10 min incubation with propofol ($10-300{\mu}m$) inhibited the $Ca^{2+}$ influx into human pulmonary arterial smooth muscle cells induced by a 60 mM $K^+$-containing $Ca^{2+}$-free solution. In conclusion, propofol-induced arterial constriction appears to involve prostaglandin production by cyclooxygenase in pulmonary artery smooth muscle cells and the relaxation depends in part on endothelial function, principally on the inhibition of calcium influx through L-type voltage-operated calcium channels.
Kim, Nari;Lee, Sang-Kyeong;Chung, Joon-Yong;Seog, Dae-Hyun;Kim, Euiyong;Jin Han
한국생물물리학회:학술대회논문집
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한국생물물리학회 2003년도 정기총회 및 학술발표회
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pp.23-23
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2003
It has been suggested that the impairment of smooth muscle cell (SMC) function by alterations in the $Ca^{2+}$-activated $K^{+}$ ( $K_{Ca}$ ) channels accounts for the reduction in coronary reserve during left ventricular hypertrophy (LVH). However, this hypothesis has not been fully investigated. The main goal of this study was to assess whether the properties of $K_{Ca}$ channels in coronary SMCs were altered during LVH. New Zealand white rabbits (0.8-1.0 kg) and Sprague-Dawley rats (300-400 g) were randomly selected to receive either an injection of isoproterenol (300 $\mu\textrm{g}$/kg body weight) or an equal volume of 0.9% saline (1 mL/kg body weight). The animals developed LVH 10 days after injection. In patch-clamp experiments, the unitary current amplitude and open probability for the $K_{Ca}$ channels were significantly reduced in LVH patches compared with control patches. The concentration-response curve of the $K_{Ca}$ channel to [C $a^{2+}$]$_{i}$ was shifted to the right. Inhibition of the $K_{Ca}$ channels with TEA was more pronounced in LVH cells than in the control cells. The whole-cell currents of $K_{Ca}$ channels were reduced during LVH. Western blot analysis indicated no differences in $K_{Ca}$ channel expression between the control and LVH coronary SM membranes. In contraction experiments, the effect of a high $K^{+}$concentration on the resting tension of the LVH coronary artery was greater than on that of the control. The effect of TEA on the resting tension of the LVH coronary artery was reduced as compared with the effect on the control. Our findings imply a novel mechanism for reduced coronary reserve during LVH.ing LVH.
Purpose: The purpose of this study was to help enhance the quality of life for women with urinary incontinence. Self-esteem, urinary symptoms, peak pressure and duration in seconds for vaginal contraction after pelvic muscles exercise were examined. Method: One-group pretest-posttest design was employed participants were 27 married women from G city. The instrument for this study were : the self-esteem scale developed by Rosenberg(1965). the Urinary Symptom Questionnaire, a subjective measurement of urinary incontinence, developed by Jackson et al.(1996) and the Perineometer used as an objective measurement of peak pressure and duration in seconds. Result: 1. Self-esteem showed significant improvement after the exercise (t=-3.832, p= .001). 2. Comparison of results before the pelvic muscles exercise and after showed that there was a statistically significant difference for several urinary symptoms including enuresis (t=2.833, p=.009), frequency of incontinence (t=2.964, p= .006), incontinence volume (t=2.280, p= .031), incontinence before getting to the restroom (t=3.035, p= .006), incontinence with no reason or feeling (t= 3.051, p= 005) burning sensation (t= 2.132, p=.043), and a sense of residual urine (t=2.267 p=.032). The mean scores showed improvement in the urinary symptoms of management of incontinence (M=$0.04{\pm}0.22$), daily frequency of incontinence management (M=$0.13{\pm}0.85$), urinary control (M=$0.15{\pm}0.86$), running to the restroom (M=$0.15{\pm}0.60$) incontinence caused by coughing or physical activities (M=$0.19{\pm}0.57$) and hesitancy (M=$0.07{\pm}0.55$). Overall urinary symptoms decreased significantly (t =3.073. p= .007). 3. Peak pressure showed an increase from a mean of $24.26{\pm}16.20mmHg$ before the exercise to a mean of $28.63{\pm} 17.79mmHg$ after (t=-2.399, p=.024). Duration in seconds also increased from a mean of $6.00{\pm}4.95sec$ to $9.15{\pm}5.83sec$ (t=-4.400, p= 000). Conclusion: These findings suggest that pelvic muscle exercise serves to decrease urinary problems, improve self-esteem and increase peak pressure and duration in seconds.
구강하악 근긴장이상증(oromandibular dystonia)은 국소적인 근긴장이상증의 한 형태로, 저작근, 안면근 또는 혀 근육의 지속적이고 반복적인 근경련이 발생하여 불수의적인 개구 및 폐구, 악골의 측방 및 후퇴운동이 나타나는 것으로 정의내릴 수 있다. 구강하악 근긴장이상증에 이환된 환자의 경우, 저작, 연하 및 발음을 하는데 불편감을 가지게 되고, 그 결과 하악 운동에 지장을 초래하게 된다. 현재까지는 근긴장이상증에 대한 병태생리가 뚜렷히 입증된 바가 없어, 원인에 관련한 치료는 이루어지지 않고 있는 상태이다. 약물요법, 행동요법, 외과적 처치 등 다양한 방법이 구강하악 근긴장이상증의 치료법으로 제시되고 있으나, 성공률이 그리 높은 편은 아니며, 많은 부작용을 초래할 수 있다. 최근 들어 근긴장이상증에 이환된 근육에 보툴리눔 톡신을 이용한 치료법이 주목을 받고 있는 추세이다. 본 증례를 통해 구강하악 근긴장이상증을 중심으로 한 구강안면 운동장애에 대하여 전반적으로 고찰을 시행하고 해당 질환에서의 보툴리눔 톡신 치료법에 대하여 알아보고자 한다.
Objectives In the present study, the author intended to investigate whether bojung-ikgitang-gamibang(BJGB) and seonbang-paedoktang(SBPT) significantly affect in vivo and in vitro mucin secretion from airway epithelial cells. Methods In vivo experiment, mice's mucin which is on a hypersecretion of airway mucin, mice's tracheal goblet cells in hyperplasia and mice's intraepithelial mucosubstances were exposed with SO2for3weeks. Effects of orally-administered BJGB and SBPT during 1 week on vivo mucin secretion and hyperplasia of tracheal goblet cells were assessed by using both enzyme-linked immunosorbent assay(ELISA) and staining goblet cells with alcian blue. In vitro experiment, confluent hamster tracheal surface epithelial(HTSE) cells were metabolically radiolabeled with 3H-glucosamine for 24hrs and chased for 30 min in the presence of each agent to figure out the effectiveness of 3H-mucin secretion. Total elution profiles of control spent media and treatment sample through Sepharose CL-4B column were analyzed. The effects of each agent on contractility of isolated tracheal smooth muscle and effects of each agent on MUC5AC gene expression in cultured HTSE cells were investigated. Also, possible cytotoxicities of each agent were assessed by measuring lactate dehydrogenase(LDH) release. Additionally, effects of BJGB and SBPT on both MUC5AC gene expression in cultured HTSE cells and TNF- or EGF-induced MUC5AC gene expression in human airway epithelial cells (NCI-H292) were investigated. Results (1) BJGB and SBPT inhibited hypersecretion of in vivo mucin. SBPT also inhibited the increase the number of goblet cells. However, BJGB did not affect the increase of number of goblet cells; (2) BJGB significantly increased mucin secretion from cultured HTSE cells, without significant cytotoxicity, and chiefly affected the 'mucin' secretion; (3) SBPT did not affect mucin secretion from cultured HTSE cells without significant cytotoxicity, and also did not affect the secretion of the other releseable glycoproteins; (4) BJGB and SBPT did not affect Ach-induced contraction of isolated tracheal smooth muscle; (5) SBPT significantly inhibit the expression levels of MUC5AC gene and BJGB significantly increased the expression levels of MUC5AC gene in both HTSE cells and NCI-H292 cells. Conclusions BJGB and SBPT can not only affect the secretion of mucin but also affect the expression of mucin gene. The author suggests that the effects BJGB and SBPT with their components should be further investigated and it is highly desirable to find from oriental medical prescriptions, novel agents which might regulate hypersecretion of mucin from airway epithelial cells.
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