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1.
Kampo Medicine ; : 143-149, 2013.
Article in Japanese | WPRIM | ID: wpr-376165

ABSTRACT

It has been reported that the effect of shoseiryuto, a traditional Japanese “Kampo” medicine, on allergic rhinitis depends on several mechanisms. Previously, we reported that shoseiryuto administered in an allergic rhinitis rat model, inhibited increases of substance P (SP) and calcitonin gene-related peptide (CGRP) in the nasal mucus, which in turn are involved in the exacerbation of rhinitis symptoms and allergic symptoms. In the present research, we looked into whether the secretion of SP and CGRP are directly inhibited by the administration of shoseiryuto.<br>Histamine solution 5µl (10 mg/ml) was instilled in both nostrils of healthy, male SD rats, thereby inducing allergic symptoms, and for the group to which shoseiryuto had been pre-administered, increase in the concentration of SP and CGRP in the nasal mucus was significantly inhibited. Moreover, upon investigating SP and CGRP production in the trigeminal ganglia by means of immunostaining, it was found to be similarly significantly inhibited. The above results indicate that shoseiryuto directly inhibits the secretion of SP and CGRP in the C-fibers of nasal mucosa.

2.
Kampo Medicine ; : 78-85, 2013.
Article in Japanese | WPRIM | ID: wpr-374573

ABSTRACT

Yokukansan (YKS), one of the traditional Japanese “Kampo” medicines, is a mixture of extract powders from seven kinds of medicinal herbs (<i>Atractylodis Lanceae Rhizoma, Hoelen, Cnidii Rhizoma, Uncariae Uncis Cum Ramulus, Angelicae Radix, Bupleuri Radix, and Glycyrrhizae Radix</i>). YKS has been administered to fragile habitus patients who show symptoms such as emotional irritability, neurosis and insomnia, and to infants who suffer from night crying and convulsions. In recent years, YKS has been reported to be effective against pain disorders such as headache and chronic pain, but the mechanism underlying these beneficial effects is still unclear. In this study, the effect of YKS on chronic inflammatory pain and stress caused by pain were investigated using rats with adjuvant arthritis.<br>Male Wistar rats were injected with complete Freund's adjuvant into the plantar surface of the right hindpaw, and then pain thresholds and stress markers were measured. The thermal pain threshold measured with the plantar test significantly decreased, and the level of salivary chromogranin A (CgA), which is used as a mental stress marker, was significantly increased in this model. The administration of YKS controlled the activation of spinal microglia involved in the expression of chronic pain, and significantly reduced a decrease in the pain threshold. Moreover, an increase in the level of salivary CgA was significantly inhibited. The authors concluded that YKS has effects in reducing chronic inflammatory pain and the stress caused by pain.

3.
Article in Japanese | WPRIM | ID: wpr-376537

ABSTRACT

<b>Objective:</b> In general, antiallergic drugs, steroids, and autonomic drugs are administered for patients with allergic diseases, but the chronic administration of these medicines may occasionally induce side effects. As such, several complementary and alternative medicine (CAM) are used, even though their effectiveness has not been established by a significant amount of evidence. Health food, herbal medicines, herbal tea, and aromatherapy, etc., are used for the treatment of allergic rhinitis (AR) as CAM. Rosmarinic acid is a type of polyphenol, and is reported to have antiallergic, antioxydative and anti-inflammatory properties. Rosmarinic acid is included in Perilla Herbs, which are used in health foods and herbal medicines (Kampo medicines), Rosemary and Lemon balm, which are used as herbal teas and essential oils of aromatherapy. This study was performed to evaluate the effect of rosmarinic acid on AR using AR model rats.<br> <b>Methods:</b> Six-week-old male Sprague-Dawley rats were sensitized with toluene 2,4-diisocyanate (TDI) to induce AR. Rosmarinic acid (1 mg/kg i.p. or 3 mg/kg i.p.) is administrated for 21 days. On day 22, the symptom of nasal allergy was evaluated by counting the number of sneezes within 10 minutes. Substance P (SP), calcitonin gene-related peptide (CGRP) and nerve growth factor (NGF) in the nasal lavage fluids were also measured.<br> <b>Result:</b> SP and CGRP take part in the exacerbation of rhinitis symptoms, and are secreted from the nerve ending following nerve stimulation by chemical mediators such as histamine. The administration of Rosmarinic acid had no effect on NGF, but significantly suppressed increases of sneezes, SP and CGRP dose-relatedly. Conclusion: Rosmarinic acid could control the symptom of nasal allergy, and one of the mechanisms was suppressing of the secretion of SP and CGRP.<br>

4.
Article in Japanese | WPRIM | ID: wpr-362844

ABSTRACT

In oriental medicine, the state in which blood is stagnant is called "oketsu, (Yu xie)"meaning preceding state or symptomatic of sickness. However, the pathological detail and mechanism of "oketsu"is not clear. Therefore we attempt to clarify the meaning of "oketsu"with Western medical knowledge and methods. The relationship between "oketsu"and blood fluidity (BF) was studied in human studies and the influence of several kinds of stress, "anti-oketsu"herbal medicine, electro-acupuncture stimulation (ACU), adrenoceptor agonist and antagonist on BF, platelet aggregation (PA), plasma ATP level, reactive oxygen metabolites level (d-ROM) and biological anti-oxidant potential level (BAP) were studied in animal studies using male Wistar rats. BF and PA were examined using a Micro Channel Array Flow Analyzer (MC-FAN) and a laser scattering platelet aggregometer (PA-20), respectively. D-ROM and BAP levels were studied using a Free Radical Elective Evaluator. In ACU study, 1Hz, 3-5 V stimulation was applied to the acupoints for 15 or 60 minutes. Stimulated acupoints were as follows:ZuSanli (ST-36), Sanyinjiao (SP-6), Hegu (L-I4), Neiguan (P-6) and Shenshu (BL-23). A significant decrease of BF was observed in "oketsu"patients compared to the "non-oketsu"patients in human studies. In the animal study, restrained and electrical shock stress applied to rats results in a quick decrease of BF, increase of PA, increase of plasma ATP level, increase of d-ROM level and decrease of BAP level. ACU applied to ST-36, SP-6 and L-14 revealed significant increases in BF, while PA activity decreased. The effect was observed even with 15 min stimulation. However, no significant changes were observed when ACU was applied to P-6 and BL-23. Furthermore, ACU applied to ST-36 represented a significant reduction to the stress induced blood changes such as BF, PA, plasma ATP level, d-ROM, and BAP level. These data suggest the close relation of BF with the "Oketsu"and that sympathetic nervous system activity and plasma ATP level have important roles in the changes of blood fluidity. Acupoints may have some role in the result that ACU affects BF depending on the stimulation loci. ACU has an immediate effect on BF since 15 min stimulation shows a significant effect. PA decrease by ACU suggests that an increase of BF is mainly associated with PA ability. The result that ACU reduced the stress induced BF and other changes of blood properties may imply that ACU has anti-stress activity.

5.
Kampo Medicine ; : 337-346, 2011.
Article in Japanese | WPRIM | ID: wpr-362626

ABSTRACT

Swelling and pain appear when blood flow is delayed in states of “<i>oketsu</i>.” Until now “<i>oketsu</i>” as a circulatory disturbance has been studied from profiles of vascular resistance characteristics and blood fluidity. Blood fluidity is influenced by blood cell function, plasma ingredients and reactive oxygen. In this study, we administered the various Chinese herbal medicines, tokishakuyakusan, saikokaryukotsuboreito, tokakujokito, keishibukuryogan and juzentaihoto to determine the effect of these medicines on “<i>oketsu</i>” by observing reactive oxygen dynamics and blood fluidity in rats.SPF male Wistar rats weighing 200 g were used. Each experimental group was given feed containing 3% extract of one Chinese herbal medicine for1week. Blood samples were mixed with heparin, EDTA-2K or sodium citrate to block coagulation. Blood fluidity was measured with a Micro Channel Array Flow Analyzer (MC-FAN). A PA-20 examined platelet aggregation by the reaction to ADP, which activates platelet aggregation. Reactive oxygen dynamics were measured with a Free Radical Elective Evaluator (FREE).As a result, anti-oxidative ability and blood fluidity increased in all Chinese herbal medicine-administered groups. Furthermore, an inverse correlation between the fluidity of erythrocyte suspensions and anti-oxidative ability was shown. We speculate that an anti-oxidative influence of Chinese herbal medicines affected erythroid deformability or stickiness. In addition, tokishakuyakusan, tokakujokito and keishibukuryogan decreased platelet aggregation ability.This study shows that “<i>oketsu</i>” improves blood properties and suggests that Chinese herbal medicine improves decrease in blood fluidity, the causes of conditions such as thrombosis or embolism.

6.
Article in Japanese | WPRIM | ID: wpr-362624

ABSTRACT

[Objective] Perspiration is almost only heat radiation mechanism under high temperature environments. And sudoriferous water is supplied from blood. Blood flow is determined by blood fluidity, blood volume and the cardiovascular system. It was reported that strong stress decreased blood fluidity.In this experiment, we investigated the relation between blood fluidity and water supply in rats loaded with forced exercise in high temperature environment.[Methods] SPF male Wistar rats weighing 250g were used. All animals were put in high temperature environment (Wet Bulb Globe Temperature; WBGT: 28°C) through whole experimental period. The rats were divided into four groups randomly; Suitable temperature environment-Exercise-Non water intake (SEN), High temperature environment-Exercise-Non water intake (HEN), High temperature environment-Exercise-Water intake (HEW) and Baseline (BL). In a group of water supply, distilled water was served before and later exercise by sonde forcibly. The blood was collected before or later of exercise and blood and erythrocyte suspension fluidity were measured.[Results] In the HEN, hydroperoxides, blood sodium, lactic acid and adrenaline increased while blood and erythrocyte suspension fluidity were decreased significantly compared with the BL. In addition, the hematocrit did not increase even if water equivalent to 4% of body weight lost it.[Conclusion] We speculate that exercise in high temperature environment decreases blood fluidity. However, the water supply in exercise that might not be sufficiently improve blood fluidity.

7.
Article in Japanese | WPRIM | ID: wpr-362618

ABSTRACT

[Objective] Physical exercises raise more or less body temperature. A body temperature is regulated constantly generally by homeostasis mechanism. Perspiration is only heat radiation mechanism under high temperature environments. And sudoriferous water is supplied from blood. Blood flow is determined by blood fluidity, blood volume and the cardiovascular system. It was reported that strong stress decreased blood fluidity.In this experiment, we investigated the relation between blood fluidity and water supply in rats loaded with forced exercise in high temperature environment.[Methods] SPF male Wistar rats weighing 150 g were used. All animals were put in high temperature environment (Wet Bulb Globe Temperature; WBGT: 28°C) through whole experimental period. In a group of water supply, distilled water was served before and later exercise by sonde forcibly. The rats were divided into five groups randomly; Rest-Non water intake (RN), Rest-Water intake (RW), Exercise-Non water intake (EN), Exercise-Water intake (EW) and Baseline (B). The blood was collected before or later of exercise and blood fluidity or platelet aggregation was measured.[Results] In the EN, platelet aggregation, lactic acid and corticosterone increased while blood fluidity were decreased significantly compared with the RN, RW and EW. In addition, the hematocrit did not increase even if water equivalent to 8 % of body weight lost it.[Conclusion] We speculate that exercise in high temperature environment decreases blood fluidity. However, the water supply that does not completely make up for quantity of depletion in exercise may improve blood fluidity.

8.
Article in Chinese | WPRIM | ID: wpr-340149

ABSTRACT

<p><b>OBJECTIVE</b>To observe the effect of nitric oxide (NO) on the differentiation of neural stem cells (NSCs) derived from subventricular zone (SVZ) of neonatal rats in vitro.</p><p><b>METHODS</b>Conventional method was used to isolate and culture the NSCs from SVZ. Diethylenetriamine/NO(DETA/NO) was used as NO donor and Nitro-L-arginine methylester (L-NAME) was used as inhibitor of nitric oxide synthase (NOS). The immunofluorescence was used to identify the expression of nestin (a marker of NSCs), beta-III-tubulin (Tuj-1, a marker of neurons), glial fibrillary acidic protein (GFAP, a marker of astrocytes) and nNOS. The concentration of NO in medium was measured by Greiss assay.</p><p><b>RESULTS</b>Cultured neurospheres were nestin-, BrdU- and nNOS-positive. After treatment with 40 micromol/L, 50 micromol/L and 60 micromol/L of DETA/NO for 5 days, the concentration of NO released was increased significantly (P < 0.01) as compared with that of the control group. The percentage of both differentiated neurons and astrocytes was increased significantly (P < 0.01 and P < 0.05) as compared with that of the control group. After treatment with 100 micromol/L, 150 micromol/L and 200 micromol/L of L-NAME for 5 days, the concentration of NO released was decreased as compared with that of the control group (P < 0.05). The percentage of both differentiated neurons and astrocytes were decreased as compared with that of the control group (P < 0.05).</p><p><b>CONCLUSION</b>NO could directly promote the differentiation of NSCs derived from rat subventricular zone in vitro.</p>


Subject(s)
Animals , Animals, Newborn , Cell Differentiation , Cells, Cultured , Cerebral Ventricles , Cell Biology , Neural Stem Cells , Cell Biology , Nitric Oxide , Pharmacology , Rats , Rats, Sprague-Dawley
9.
Neuroscience Bulletin ; (6): 229-235, 2007.
Article in English | WPRIM | ID: wpr-300959

ABSTRACT

<p><b>OBJECTIVE</b>To search novel genes or pathways involved in the recovery process after restraint stress in rats.</p><p><b>METHODS</b>We compared the hypothalamus transcriptional profiles of two different recovery patterns (fast recovery vs slow recovery) from restraint stress in rats using oligonucleotide microarray, the recovery pattern was determined by the decrement of plasma adrenocorticotropic-hormone (ACTH) and corticosterone levels during one hour recovery period after stress. A real-time quantitative RT-PCR was applied to validate the differential expressed genes.</p><p><b>RESULTS</b>Analysis of the microarray data showed that most of genes were not differentially expressed between fast recovery group and slow recovery group. Among the differentially expressed genes we found that talin, together with serine/threonine protein phosphatase PP1-beta catalytic subunit (PP-1B) and integrin alpha-6 precursor (VLA-6) genes, were at least 1.5 fold up-regulated in the fast recovery group, while junctional adhesion molecule 1 (F11r) was 1.5 fold down-regulated in the fast recovery group.</p><p><b>CONCLUSION</b>The results implied that integrin signaling pathway may be involved in the recovery from restraint stress in rats. The present study provided a global overview of hypothalamus transcriptional profiles during the process of recovery from the restraint stress in rats. The integrin signaling pathway seems to be involved in the recovery process, which deserves further study to clarify the integrin-mediated recovery mechanism after restraint stress.</p>


Subject(s)
Adrenocorticotropic Hormone , Blood , Animals , Corticosterone , Blood , Disease Models, Animal , Gene Expression Regulation , Physiology , Integrins , Genetics , Metabolism , Male , Oligonucleotide Array Sequence Analysis , Methods , RNA, Messenger , Rats , Rats, Sprague-Dawley , Recovery of Function , Physiology , Restraint, Physical , Methods , Reverse Transcriptase Polymerase Chain Reaction , Methods , Signal Transduction , Physiology , Stress, Psychological , Metabolism , Time Factors
10.
Neuroscience Bulletin ; (6): 1-6, 2006.
Article in English | WPRIM | ID: wpr-300977

ABSTRACT

Objective To explore the effects of exercise on dentate gyrus (DG) neurogenesis and the ability of learning and memory in hippocampus-lesioned adult rats. Methods Hippocampus lesion was produced by intrahippocampal microinjection of kainic acid (KA). Bromodeoxyuridine (BrdU) was used to label dividing cells. Y maze test was used to evaluate the ability of learning and memory. Exercise was conducted in the form of forced running in a motor-driven running wheel. The speed of wheel revolution was regulated at 3 kinds of intensity: lightly running, moderately running, or heavily running. Results Hippocampus lesion could increase the number of BrdU-labeled DG cells, moderately running after lesion could further enhance the number of BrdU-labeled cells and decrease the error number (EN) in Y maze test, while neither lightly running, nor heavily running had such effects. There was a negative correlation between the number of DG BrdU-labeled cells and the EN in the Y maze test after running. Conclusion Moderate exercise could enhance the DG neurogenesis and ameliorate the ability of learning and memory in hippocampus-lesioned rats.

11.
Acta Physiologica Sinica ; (6): 415-420, 2006.
Article in English | WPRIM | ID: wpr-265435

ABSTRACT

To investigate the effect of forced running in motor-driven wheel on neurogenesis in the hippocampal dentate gyrus (DG) of adult rats, 5-bromo-2-deoxyuridine (BrdU), a thymidine analog was applied to mark cell proliferation. Neuroepthelial stem cell protein (nestin) expression was used to identify neural stem/precursor cells. The BrdU- and nestin-positive cells were examined by immunohistochemical technique. The ability of learning was evaluated by Y-maze test to explore the functional role of the newborn cells in the DG after forced running. It was found that the number of BrdU- and nestin-positive cells in the DG in running groups was significantly increased compared to that in the control group (P<0.05). The effect of forced running on neurogenesis was intensity-dependent. In addition, an improvement of learning ability in Y-maze test was observed after forced running. These findings suggest that forced running in motor-driven wheel could enhance neurogenesis in the hippocampal DG of adult rats and improve learning ability.


Subject(s)
Animals , Bromodeoxyuridine , Metabolism , Cell Survival , Dentate Gyrus , Cell Biology , Physiology , Intermediate Filament Proteins , Learning , Male , Maze Learning , Nerve Tissue Proteins , Nestin , Neurons , Physiology , Physical Conditioning, Animal , Rats , Rats, Sprague-Dawley , Running
12.
Kampo Medicine ; : 645-648, 2004.
Article in Japanese | WPRIM | ID: wpr-368471

ABSTRACT

Scientific elucidation of “oketsu (blood stasis)” states is very important to understanding the diagnostic methods of oriental medicine. We investigated the correlation of blood fluidity to oketsu states, by micro channel array flow analyzer (MC-FAN). Twenty-seven female patients were divided into three groups: a non-oketsu group, a mildly affected group, and a severely affected group according to the diagnostic criteria of their oketsu syndrome. Immediately after collecting their venous blood, whole blood passage time was measured by MC-FAN. And we compared and investigated the correlation with degree of oketsu state, diagnostic criteria and changes of whole blood passage time after the administration of ku-oketsu drugs. As a result, whole-blood passage time of the mildly affected group and the severely affected group were significantly longer than that of the non-oketsu group, while the [an] improvement of blood fluidity was observed after medication. This study confirms that reduced blood fluidity is seen using MC-FAN in oketsu states, and that ku-oketsu drugs are able to improve these states.

13.
Kampo Medicine ; : 763-771, 2003.
Article in Japanese | WPRIM | ID: wpr-368434

ABSTRACT

Collagen-induced arthritis (CIA) in DBA/1J mice is an autoimmune model driven by the combination of humoral and cellular immunity against cartilage type II collagen. It has been proven to be a useful model for human rheumatoid arthritis (RA). This study was designed to obtain experimental evidence of Kampo medicine Dokkatsu-kisei-to (DKT) in the treatment of RA by using mouse CIA. As positive control, prednisolone was given. The significant suppressive effect of DKT as measured by arthritis score persisted upon cessation of treatment on day 42 and remained until the termination of the experiment on day 63. The amelioration of inflammatory articular lesions was confirmed by radiological and histo-pathological findings. We also observed significantly decreased concentrations of serum TNF-α on day 28 and levels of serum IgG antibody against bovine type II collagen on day 49. These results suggest that DKT has important effects on immuno-inflammatory response and may be useful in the prevention of the onset and severity of arthritis.

14.
Article in Japanese | WPRIM | ID: wpr-376867

ABSTRACT

The influence of exercise on the endogenous levels of both β-endorphin (BE) and corticosteroid (CS) in the hypothalamus and plasma was examined in rats. F344 male rats, five weeks of age, were divided into three groups: involuntary exercised (IE), non-exercised (NE) and control (C) groups. Rats in the IE group were exercised every day for one hour at the speed of 20 m mm n for 7 to 28 days. After training, the hypothalamus and plasma were obtained from rats. BE levels in water soluble extracts of hypothalamus were examined by rat BE ELISA test kits; and CS contents in plasma was examined by RIA. The amount of BE in the hypothalamus obtained from rats in the IE group was much higher than that in the NE and C groups. Although there was no significant difference between rats exercised for 7, 14, and 21 days, BE levels in the hypothalamus from rats exercised for 28 days was significantly higher than for 7 days. The amount of BE in the hypothalamus of NE rats was nearly identical to that of C rats. On the other hand, there were no significant changes in plasma CS levels. These results may suggest that exercise, especially running, causes on increase in BE levels in the hypothalamus.

15.
Article in Japanese | WPRIM | ID: wpr-372028

ABSTRACT

The influence of exercise on the endogenous levels of both β-endorphin (BE) and corticosteroid (CS) in the hypothalamus and plasma was examined in rats. F344 male rats, five weeks of age, were divided into three groups: involuntary exercised (IE), non-exercised (NE) and control (C) groups. Rats in the IE group were exercised every day for one hour at the speed of 20 m mm n for 7 to 28 days. After training, the hypothalamus and plasma were obtained from rats. BE levels in water soluble extracts of hypothalamus were examined by rat BE ELISA test kits; and CS contents in plasma was examined by RIA. The amount of BE in the hypothalamus obtained from rats in the IE group was much higher than that in the NE and C groups. Although there was no significant difference between rats exercised for 7, 14, and 21 days, BE levels in the hypothalamus from rats exercised for 28 days was significantly higher than for 7 days. The amount of BE in the hypothalamus of NE rats was nearly identical to that of C rats. On the other hand, there were no significant changes in plasma CS levels. These results may suggest that exercise, especially running, causes on increase in BE levels in the hypothalamus.

16.
Kampo Medicine ; : 357-366, 2002.
Article in Japanese | WPRIM | ID: wpr-368398

ABSTRACT

The Oriental medical therapies are increasingly used in medical treatment in conjunction with the Western medical health care system in our country. As Oriental medicine and Western medicine are based on different scientific systems, there is no curriculum of Oriental medicine in Western medical school education. In 1996, we investigated opinions about Oriental medicine among medical students and teachers at Showa University. A mini follow-up survey was done in 1999 and 2000. Awareness of Oriental medicine was still limited, but those surveyed were remarkably interested in Oriental medicine. Moreover, they recognized Oriental medicine as an important and useful tool for the care of illnesses that do not respond to Western medicine. About 40% of the medical doctors at Showa University hospitals used Oriental medicine with their patients in 1996, and traditional knowledge was learned in its own way. According to the pharmaceutical department of Showa University, both the number and variety of Kampo prescriptions increased between 1996 and 2000. These results suggest that it is necessary to properly learn Oriental medicine from the foundations during medical school. More research on Oriental medicine in terms of clinical and basic science is necessary, and the opportunity to exchange information about Oriental medical diagnosis and treatments should be included in postgraduate curricula.

17.
Article in Japanese | WPRIM | ID: wpr-365757

ABSTRACT

This study was designed to evaluate the myocardial protection with observation of the monophasic action potential (MAP) which was recorded by suction electrode. Using the isolated working rabbit hearts, amplitude, duration of MAP at 50% repolarization level (MAPD<sub>50</sub>), aortic flow and heart rate were measured after reperfusion. The comparative study obtained for all five groups under the following various conditions of the aortic cross clamping are stated as follows. Myocardial temperature were maintained at 20°C during aortic cross clamping. Group I was treated with St. Thomas' Hospital cardioplegic solution. The cardioplegic solution was infused every 20min during ischemia and kept at 20°C. The hearts of group I was divided into four sub-groups, all of which were infused with different concentration of diltiazem (D) in cardioplegia: group Ia D=0μg/ml (<i>n</i>=10), group Ib D=1μg/ml (<i>n</i>=5), group Ic D=5μg/ml (<i>n</i>=5). group Id D=10μg/ml (<i>n</i>=5), and in group II cardioplegic solution was not used. The amplitude of MAP following 30min working mode of reperfusion in group I showed a significantly higher recovery compared to those in group II. The MAPD<sub>50</sub> of MAP following 30min working mode of reperfusion in group I showed a significantly lower recovery compared to those in group II, and 10min Langendorff mode in group I a showed a significantly higher recovery compared to those in group Ib, group Ic and group Id. 20min working mode in group Ia and group Ib showed a significantly higher recovery compared to those in group Ic and group Id. The heart rate following 30min working mode of reperfusion in group Ia and group Ib showed a significantly higher recovery compared to those in group Ic and group Id. The aortic flow following 30min working mode of reperfusion in group Ia and group Ib showed a significantly higher recovery compared to those in group Ic, group Id and group II. We would like to conclude that the permeability of large amount of calcium across myocardial cell membrane seems to be depressed by diltiazem added to cardioplegia. But when the concentrations of diltiazem in cardioplegia was over 5μg/ml, it showed negative inotropic action and negative chronotropic action.

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