Effects of Kampo medicine, Toki-shakuyaku-san (Tang-Kuei-Shao-Yao-San), on choline acetyltransferase activity and norepinephrine contents in brain regions, and mitogenic activity of splenic lymphocytes in ovariectomized mice

We investigated the effects of Toki-shakuyaku-san (TSS, Tang-Kuei-Shao-Yao-San in Chinese), Japanese traditional herbal medicine, on the nervous and immune systems in ovariectomized mice as a climacteric disorder model. Female C57BL/6 mice were ovariectomized (OVX) and TSS was given daily through the drinking water for either 10 or 20 days from the day after ovariectomy. After completion of experimental sessions, animals were sacrificed and specific brain regions were assayed for choline acetyltransferase (ChAT) activity and norepinephrine contents. The mitogenic activities, alkaline phosphatase activity and 3-(4, 5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H terazolium bromide (MTT) activity, in splenic lymphocytes has also measured. Furthermore, the effects of TSS on learning and memory ability were studied by the step-through type passive avoidance test. As the results, the administration of TSS significantly suppressed the decrease of ChAT activity in the cerebral cortex (CC) and the dorsal hippocampus (DH) of ovariectomized mice at 10 days after ovariectomy, however no significant effect was observed at 20 days after ovariectomy. Norepinephrine contents in OVX group were decreased at 10 and 20 days after ovariectomy in the CC and the ventral hippocampus (VH). The administration of TSS significantly suppressed the decrease of norepinephrine contents at 20 days after ovariectomy. The mitogenic activities of lymphocyte in spleen were increased at 10 days after ovariectomy, and decreased at 20 days after ovariectomy. However, the suppression of these changes was observed in the group given TSS. The mean latent period was also shortened in the passive avoidance test in the OVX group, but TSS treated group improved mean latency. From these observations, it is inferred that administration of TSS brings on the synthesis of acetylcholine and norepinephrine in the CC and hippocampus, and may improve the memory related behavior and the abnormalities in lymphocytes in the models of the climacteric disorder.     

Antiosteoporotic activity of Er-Xian Decoction, a traditional Chinese herbal formula, in ovariectomized rats

The antiosteoporotic effect of a herbal formula, Er-Xian Decoction (EXD), in ovariectomized (OVX) rats model of osteoporosis was investigated. The rats were divided into Sham and OVX groups. The OVX rats were further sub-divided into four groups administered orally with water, nylestriol (1 mg/kg, weekly) or EXD (300, 600 mg/kg, daily) for 12 weeks. In OVX rats, the increases of body weight, serum BGP and ALP were significantly decreased by EXD treatment. In OVX rats, atrophy of uterus and descent of BMD were suppressed by treatment with EXD and nylestriol. In addition, EXD completely corrected the decreased concentration of calcium, phosphorus, and estradiol in serum observed in OVX rats. EXD also significantly increased biomechanical strength comparable to the Sham group. This was also confirmed by histological results that showed its protective action. The findings assessed on the basis of biochemical, bone mineral density, biomechanical, and histopathological parameters strongly suggested that EXD had a definite antiosteoporotic effect, which is similar to estrogen.     

清胰化积方联合IL-2对胰腺癌小鼠脾淋巴细胞功能的影响

目的:研究清胰化积方联合白细胞介素-2(IL-2)对Panc02胰腺癌小鼠脾淋巴细胞功能的影响。方法:荷瘤小鼠随机分为对照组、清胰化积方组、清胰化积方联合不同剂量的IL-2组,治疗后称量瘤质量并计算抑瘤率,分别以淋巴细胞增殖活性和转化率评价小鼠脾淋巴细胞功能。结果:中药组、中药+IL-2小、中、大剂量组的瘤质量均小于对照组(P<0.01),抑瘤率分别为37.84%、44.14%、38.74%和39.64%;各治疗组脾淋巴细胞增殖活性和转化率均高于对照组(P<0.01),并呈现IL-2的剂量依赖性,其中中药+IL-2大剂量组最高,与中药组比较有显著性差异(P<0.05)。结论:增强脾淋巴细胞免疫功能是清胰化积方联合IL-2抗小鼠胰腺癌的机制之一;清胰化积方联合IL-2的免疫效应优于单用清胰化积方,具有协同效应。     

Hange-koboku-to, a Kampo medicine, modulates cerebral levels of 5-HT (5-hydroxytryptamine), NA (noradrenaline) and DA (dopamine) in mice

Cerebral monoamine systems play important pathogenic roles in various psychiatric and neurologic diseases, such as depression, anxiety and swallowing disturbance. Hange-koboku-to, a Kampo (Japanese herbal) medicine, has been successfully used for the treatment of these disorders. To elucidate the mechanisms underlying its clinical efficacy for these disorders, the effects of Hange-koboku-to (500 mg/kg, p.o.) on the cerebral monoamine systems were examined. Regional levels of 5-HT (5-hydroxytryptamine), NA (noradrenaline), DA (dopamine) and their metabolites in mouse brain were measured using a high-performance liquid chromatography system. Hange-koboku-to increased the 5-HT and NA levels and decreased 5-HIAA (5-hydroxyindole-3-acetic acid), thus decreasing 5-HT and NA turnover (metabolites/monoamine ratio) in the hypothalamus. The levels of DA, DOPAC (3,4-dihydroxyphenylacetic acid) and HVA (4-hydroxy-3-methoxy-phenylacetic acid) were all increased, resulting in a decreased DA turnover in the striatum. Since decreased 5-HT turnover has been observed after administration of various antidepressants, Hange-koboku-to-mediated reduction of 5-HT turnover may be related to the clinical efficacy of this Kampo medicine on certain psychiatric disorders. Furthermore, the beneficial therapeutic effects of Hange-koboku-to on swallowing disturbance may be related to the increased cerebral DA level brought about by this Kampo medicine.     

七宝美髯口服液对小鼠脑组织SOD活力及羰基化蛋白含量的影响

目的观察七宝美髯口服液(制何首乌、当归、补骨脂等)对SAMP8小鼠学习记忆功能、海马组织病理形态及SOD活力、羰基化蛋白含有量的影响。方法 27只SAMP8小鼠随机均分为模型对照组、盐酸多奈哌齐组及七宝美髯口服液组,另取9只SAMR1小鼠作为正常对照组,连续灌胃60 d,在给药第56天进行避暗训练,5 d后测定学习记忆能力;HE染色观察病理形态;ELISA法检测脑组织中SOD活力和羰基化蛋白含有量。结果与模型对照组比较,七宝美髯口服液组小鼠的逃避潜伏期显著延长,错误次数明显降低(P0.01),海马组织的病理形态显著改善,SOD活力显著升高,羰基化蛋白含有量明显降低(P0.01)。结论七宝美髯口服液能改善SAMP8小鼠的学习记忆功能,减轻海马组织的退行性病变程度。     

Clinical efficacy of EH0202, a Kampo formula, on the health of middle-aged women

To investigate the clinical effects of EH0202, a Japanese herbal supplement, on the menopausal symptoms and physical status of peri- and post-menopausal women. Thirty-two post-menopausal women (53.0+/-5.1 years) presented with menopausal complaints were enrolled in the clinical study. Patients were administered an herbal supplement, EH0202 (6 g/day for 6 months), and were assessed for improvement of their overall symptoms using the Greene Climacteric Scale and the Visual Analog Scale (VAS). Blood pressure, skin surface blood flow and plasma lipid profiles were measured before and 1,3 and 6 months after EH0202 administration. There was a significant decrease in the climacteric scale score (P < 0.001) and VAS (P < 0.0001) after 6 months of EH0202 treatment. There was a significant decrease in systolic (P < 0.001) and diastolic (P < 0.05) blood pressure, and a significant decrease in facial skin surface blood flow (P < 0.05) after 3 months of EH0202 administration. We observed a significant decrease in plasma TG and LDL-cholesterol concentrations after 3 months of EH0202 administration (P < 0.05). EH0202 (MACH) reduced blood pressure, excess facial skin blood flow (flushing) and abnormal lipid levels, as well as clinically improved menopausal symptoms in post-menopausal women. In post-menopausal women, this product appears to help maintain normal biological function and improves quality of life.     

绞股蓝总苷对老化小鼠脑中MAO和Na/K-ATP酶活性的影响

目的：研究绞股蓝总苷对老化小鼠脑中MAO和Na/K-ATP酶活性的影响。方法：采用连续眼球后注射D－半乳糖1个月,造成小鼠老模型,观察绞股蓝总苷对老化鼠脑中单胺氧化酶（MAO）和Na/K-ATP酶的活性影响。结果：老化小鼠可造成脑中MAO活性的升高和Na/K-ATP活力的降低,而绞股蓝总苷在造模型同时ig给药,每日剂量75和150mg/kg下,能逆转小鼠因老化而产生的上述二种酶活性的改变。结论：绞股蓝革能对抗老化小鼠因衰老引起的MAO和Na/K-ATP酶的活性改变,从生化酶学角度,进一步阐明绞股蓝总苷的抗衰老作用。     

冰片对阿魏酸钠在小鼠血浆和脑区中分布的影响

目的 探讨剂量因素及冰片对阿魏酸钠在小鼠血浆和脑区中药动学的影响.方法 经ig给予小鼠单用高、低剂量阿魏酸钠或复合不同剂量冰片的低剂量阿魏酸钠,采用HPLC法测定高剂量阿魏酸钠在达峰时间及低剂量阿魏酸钠在不同时间点的血浆和脑区中药物浓度,采用PCNONLIN程序计算药动学参数.结果 单给阿魏酸钠低剂量组,其tmax为5.0 min,Cmax为44.μg/mL,AUC0-30为853.9μg/(mL·min);脑区中海马AUC0-30为11.8μg/(g·min).tmax为5.0 min,Cmax为1.4 μg/g;阿魏酸钠高剂量组与低剂量组比较,血药浓度仅增加2.5倍;脑区中浓度增加最多的小脑也仅增加1.9倍,其中海马没有增加;联用冰片后,冰片能加快阿魏酸钠在海马的分布,并显著提高其在海马中的量.阿魏酸钠200 mg/kg联用冰片50 mg/kg,海马Cmax与AUC0-30分别增加1.2倍和1.7倍.结论 单纯加大阿魏酸钠的给药剂量不能相应提高其在脑中的浓度,联用冰片可提高其脑内(特别是海马)的生物利用度,阿魏酸钠联用冰片在治疗脑缺血等中枢神经系统疾病中可能有显著的临床意义.     

Antiosteoporotic activity of OST-6(Osteocare), a herbomineral preparation in calcium deficient ovariectomized rats

The preventive effect of the herbal formulation OST-6 (Osteocare) on the progress of bone loss induced by ovariectomy and concurrent calcium deficiency was studied in rats. Ovariectomy (OVX) and calcium deficiency (CD) resulted in bone loss as evident from decreased femoral weight and density. Treatment with OST-6 at a dose of 250 mg/kg body wt twice a day orally for a period of 16 weeks significantly restored the femoral weight and density. The biomechanical strength of the tibia was decreased by OVX and CD and this was significantly suppressed by the administration of OST-6. Histologically decalcified bone sections revealed narrowed, and the disappearance of, trabeculae and widened medullary spaces. The total numbers of tartrate-resistant acid phosphates (TRAP) positive cells were increased significantly in OVX animals. Treatment with OST-6 significantly inhibited these histopathological changes and strongly suggested that OST-6 was effective in preventing the progress of bone loss induced by ovariectomy and concurrent calcium deficiency.     

Inhibitory effects of sanguinarine on monoamine oxidase activity in mouse brain

The effects of benzophenanthridine alkaloids, such as sanguinarine and chelidonine, on monoamine -oxidase (MAO) activity in mouse brain were investigated. Sanguinarine showed an inhibitory effect on MAO activity in a concentration dependent manner (53.4% inhibition at 25 microM). However, chelidonine did not inhibit MAO activity. The IC(50) value of sanguinarine was 24.5 microM. Sanguinarine inhibited non-competitively MAO activity using kynuramine as a substrate. The K(i) value for sanguinarine was 22.1 microM. These results suggest that sanguinarine partially contributes to the regulation of catecholamine content.     

Effect of Shimotsu-to (a Kampo medicine, Si-Wu-Tang) and its constituents on triphasic skin reaction in passively sensitized mice

Previous studies have reported that mice passively sensitized with anti-DNP (dinitrophenol) IgE antibody exhibited IgE-mediated skin reaction with an immediate phase response (IPR) at 1 h and a late phase response (LPR) at 24 h after the challenge of DNFB (dinitrofluorobenzene). We recently found that a third phase inflammatory reaction with intense and persisting infiltration of eosinophils, named very late phase response (vLPR), was induced by DNFB challenge peaking at 8 days. In this study, we examined the effects of a Kampo medicine, Shimotsu-to (Si-Wu-Tang), and its constituent crude drugs on triphasic skin reaction in passively sensitized mice. Shimotsu-to inhibited ear swelling in LPR and vLPR after DNFB challenge in a dose-dependent manner, and slightly diminished the scratching behavior considered to be associated with pruritis in IPR. The inhibitory effect on LPR and vLPR was partly due to Cnidii Rhizoma (Senkyu) in Shimotsu-to formulation, especially its fraction 5 containing cnidilide. On the other hand, Angelicae Radix (Toki) rather than Cnidii Rhizoma (Senkyu) in Shimotsu-to, inhibited the scratching behavior, although it did not inhibit the ear swelling in IPR. These findings indicate that the Shimotsu-to formulation is useful for the inhibition of cutaneous inflammatory diseases.     

Effects of Fuhe decoction on behaviors and monoamine neurotransmitters in different brain regions of CUMS combined with social isolation depression model rats

ObjectiveTo investigate the effect of Fuhe decoction on the behavior and levels of monoamine neurotransmitters in different brain regions in a depression rat model induced by chronic unpredictable mild stimulation (CUMS) combined with social isolation.MethodsFifty male SD rats were randomly divided into a blank group, model group, fluoxetine group, Chaiqinwendan decoction group, and Fuhe decoction group. Chronic unpredictable mild stimulation combined with a social isolation method was used to replicate the depression rat model. After 42 days of administration, a tail suspension test and high-performance liquid electrochemical detection (HPLC-ECD) were used to detect the behavioral changes and changes in the content of monoamine neurotransmitters norepinephrine (NE), dopamine (DA), 5-hydroxytrytamine (5-HT), and metabolites in different brain regions of rats in each group before and after treatment.ResultsCompared with the model group, the epinephrine (E) content in the Fuhe decoction group was highly significantly increased (P < .01). Compared with the model group, the 5-HT content of the prefrontal cortex in rats in the Fuhe decoction group was highly significantly increased (P < .01). Furthermore, compared with the model group, the 5-HT content in the hippocampus of rats in the Fuhe decoction group was significantly increased (P < .05).ConclusionFuhe decoction can improve the depression-like behaviors of model rats, and its antidepressant effect may be related to the increase in 5-HT content in the prefrontal cortex and hippocampus of rats.     

Effects of SYJN,a Chinese herbal formula,on chronic unpredictable stress-induced changes in behavior and brain BDNF in rats

Aim of the study

Suyu-Jiaonang (SYJN) is a Chinese herbal formula that contains four herbs: Bupleurum chinense DC, Curcuma aromatica Salisb., Perilla frutescens (Linn.) Britt., and Acorus tatarinowii Schott. Previous studies conducted in our laboratory have revealed an antidepressant-like effect of the formula in various mouse models of behavioral despair. The present study aimed to investigate whether SYJN could produce antidepressant-like effects in chronic unpredictable stress (CUS)-induced depression model in rats and its possible mechanism(s).

Materials and methods

Rats were subjected to an experimental setting of CUS. The effect of SYJN treatment on CUS-induced depression was examined using behavioral tests including the sucrose consumption and open field tests. The mechanism underlying the antidepressant-like action of SYJN was examined by measuring brain-derived neurotrophic factor (BDNF) protein and mRNA expression in brain tissues of CUS-exposed rats.

Results

Exposure to CUS for 4 weeks caused depression-like behavior in rats, as indicated by significant decreases in sucrose consumption and locomotor activity (assessed in the open field test). In addition, it was found that BDNF protein and mRNA levels in the hippocampus and frontal cortex were lower in CUS-treated rats, as compared to controls. Daily intragastric administration of SYJN (1300 or 2600 mg/kg) during the 4-week period of CUS significantly suppressed behavioral changes and attenuated the CUS-induced decrease in BDNF protein and mRNA levels in the hippocampus and frontal cortex.

Conclusion

The results suggest that SYJN alleviates depression induced by CUS. The antidepressant-like activity of SYJN is likely mediated by the increase in BDNF expression in brain tissues.     

Inhibitive effect on apoptosis in splenic lymphocytes of mice pretreated with Lingzhi (Ganoderma Lucidum) spores

Objective

To investigate how the pretreatment of mice with Ganoderma spores affected the apoptosis of their splenic lymphocytes induced by dexamethasone after 19 days treatment.

Methods

Sixty Kunming mice were randomly divided into six groups: blank control groupdrenched with normal saline; a drug control group drenched with 150 mg/mL Ganoderma spores; a model group treated with saline; a low dose group with 50 mg/mL Ganoderma spores; a moderate dose group with 100 mg/mL Ganoderma spores; and a high dose group with 150 mg/mL Ganoderma spores. The effect of Ganoderma spores on apoptosis in spleen lymphocytes was analyzed. All groups were treated for 19 days. On day 20, the model group and the 3 treatment groups were intraperitoneally injected dexamethasone to induce apoptosis. Splenic index and apoptosis indes were employed to measure cell apoptosis.

Results

The results showed that Ganoderma spores reduced the splenic index to different degrees in each group and the best effect was seen in the high dose group (P<0.05). Terminal dexynucleotidyl transferase (TdT)-mediated 2′-Deoxyuridine 5′-Triphosphate nick end labeling staining revealed that the apoptotic index in all groups administered Ganoderma spores differed significantly from the model group, and a dose-response was observed. Flow cytometric analysis indicated that spleen lymphocyte apoptosis in the model group was extensive. Each dose of Ganoderma spores inhibited dexamethasone-induced apoptosis in spleen lymphocytes, and a dose-response was observed as well. The highest dose of Ganoderma spores decreased Malondialdehyde content in serum induced by dexamethasone (P<0.05).

Conclusion

The findings imply that the pretreatment of the mice with Ganoderma spores could reduce the apoptosis rate induced by dexamethasone in their splenic lymphocytes.     

Effect of monoamine neurotransmitters, enkephalin and morphine on substance P contents of several brain regions and pain threshold in rats]

The present work was carried out to observe the effect of intra-cerebroventricular (icv) injection of monoamine neurotransmitters, enkephalin and morphine on immunoreactive substance P(Ir-SP) contents in hypothalamus, striatum, hippocampus and pain threshold. The results were as follows: (1) After icv or intra-DR (dorsal raphe nucleus) injection of 5-HTP, the content of Ir-SP in hypothalamus significantly decreased and pain threshold markedly increased; After depletion of the 5-HT content in brain by pCPA or destruction of DR, the contents of Ir-SP were remarkably elevated in three brain regions by the former and in hypothalamus, striatum by the later. (2) The Ir-SP levels in the three brain regions and the pain threshold were not affected by the icv injection of NE, however, icv injection of DA caused a increase of Ir-SP concentration in striatum which was reversed by the DA receptor antagonist haloperidol, but without any change of the pain threshold. 7th day after icv injection of 6-OHDA, the content of Ir-SP in striatum significantly reduced. (3) Icv injection of met-enkephalin (MEK) or morphine could increase the Ir-SP levels in hypothalamus, striatum and the pain threshold, and above-mentioned effect of morphine could be prevented by the opioid receptor antagonist naloxone. Icv injection of leu-enkephalin (LEK) had no effects both on Ir-SP contents in three brain regions and the pain threshold.     

Effect of Choto‐san,a Kampo medicine,on impairment of passive avoidance performance in mice

In mice with procephalic ischaemia loading, disrupted passive avoidance retention performance was dose‐dependently improved by Choto‐san, but this effect was antagonized by NAN‐190, a serotonin_1A­receptor antagonist. In mice with decreased intracerebral serotonin concentration, Choto‐san prevented disturbance in acquiring a passive avoidance response after scopolamine administration, but did not­influence the decreased serotonin concentration. These results suggested that Choto‐san showed the­anti‐amnestic effect based on the stimulation of serotonin_1A receptors. Copyright © 1999 John Wiley & Sons, Ltd.     

夏天无对小鼠学习记忆及脑内乙酰胆碱酯酶的影响

目的 阐明夏天无制剂对记忆障碍小鼠的学习记忆功能及对小鼠脑内乙酰胆碱酯酶(AchE)的影响。方法 以东莨菪碱造成小鼠记忆获得障碍和乙醇造成小鼠记忆再现障碍，同时测定了小鼠脑内AchE的活性。结果 夏天无口服液及胶囊剂0．5，1，2mg／kg对记忆障碍小鼠的学习记忆功能有不同程度的改善作用，并可抑制小鼠脑内AchE活性。结论 夏天无口服液及胶囊剂能增强小鼠的学习记忆功能，该作用可能与抑制AchE有关。     

Effects of five kampohozais on the mitogenic activity of lipopolysaccharide, concanavalin A, phorbol myristate acetate and phytohemagglutinin in vivo

Mitogenic activity of lipopolysaccharide (LPS), concanavalin A (Con A), phorbol myristate acetate (PMA) and phytohemagglutinin (PHA) were investigated using spleen cells prepared from C57BL/6 mice orally pretreated with one of five kampohozai extracts (Shosaikoto, Daisaikoto, Hochuekkito, Juzendaihoto and Tokishakuyakusan). Shosaikoto and Daisaikoto elevated the mitogenic activity of LPS and reduced those of Con A. The mitogenic activities of PMA and PHA were elevated by low doses of Shosaikoto and Daisaikoto and suppressed by high doses. Juzendaihoto and Tokishakuyakusan showed no effect on the mitogenic activity of LPS and Con A, but increased those of PMA and PHA. Hochuekkito increased the mitogenic activity of LPS, Con A, PMA and PHA. In the absence of mitogens, these five kampohozais showed no mitogenic activity. These results indicate that the kampohozais used in this experiment appear to possess the immunomodulating or immunostimulating activities that might be expected from clinical experiences in Japan and China.     

柴郁温胆汤对大鼠抑郁模型行为学及脑内单胺类神经递质的影响

目的:探讨中药复方柴郁温胆汤抗抑郁作用的机制.方法:以孤养加慢性轻度不可预见性应激方法建立大鼠抑郁症模型,用旷场行为测定及糖水消耗试验进行行为学评分,并用高效液相 - 电化学方法检测其脑内单胺类神经递质的含量,观察模型大鼠给药前后的变化.结果:抑郁症模型大鼠体质量增长缓慢,水平和垂直活动及糖水消耗量均明显减少(P<0.05,P<0.01);杏仁核NE、DA、5-HT、HVA含量减少(P<0.05);纹状体NE、DA、5-HT含量减少(P<0.05).柴郁温胆汤能改善模型大鼠行为学变化,升高杏仁核NE、5-HIAA、5-HT、HVA(P<0.05)及DA含量(P<0.01),升高纹状体DA和5-HT含量(P<0.05).结论:柴郁温胆汤具有抗抑郁作用,对中枢单胺类神经递质的调节作用是其疗效机制之一.