胃肠安丸对肠易激综合征大鼠消化酶、水通道蛋白的影响

目的:探讨胃肠安丸对腹泻型肠易激综合征(IBS-D)大鼠止泻作用机制。方法:以喂饲高乳糖饲料加束缚应激相结合的方法复制IBS-D大鼠模型。将雌性Wistar大鼠,随机分为空白对照组、模型组及胃肠安(60 mg.kg-1.d-1)用药3组,每组各8只。通过比较体重增长率、腹泻潜伏期及腹泻指数,确定最佳用药周期。将45只大鼠随机等分为空白对照组、模型组、胃肠安80,60,40 mg.kg-1.d-1不同剂量等5组。计测胸腺指数、脾脏指数;小肠黏膜LDH,MDH及二糖(乳糖)酶活性;血清D-木糖含量;近端结肠组织AQP4含量。结果:用药后4 d胃肠安组与模型组体重增长率差值最大,腹泻指数差别有统计学意义(P0.05),确定4 d为最佳用药周期。模型组较对照组胸腺指数、脾脏指数缩小(P0.05),小肠黏膜LDH,MDH,二糖酶活性下降(P0.05),AQP4含量升高(P0.05)。与之相对,中剂量胃肠安组胸腺指数、脾脏指数增大(P0.05),小肠黏膜LDH,MDH,二糖酶活性升高(P0.05),但AQP4含量无显著差异。结论:胃肠安丸具有止泻作用,通过增强小肠黏膜消化酶活性,调节腹泻状态下机体糖分解代谢,是止泻机制之一。     

Effects on neuroendocrinoimmune network of Lizhong Pill in the reserpine induced rats with spleen deficiency in traditional Chinese medicine

Aim of the study

Lizhong Pill, composed of Radix Ginseng (Panax ginseng C.A. Meyer), Rhizoma Zingiberis (Zingiber officinale Roscoe), Rhizoma Atractylodis Macrocephalae (Atractylodes macrocephala Koidz.) and Radix Glycytthizae (Glycyrrhiza uralensis Fisch.), is a classical herbal product for curing spleen deficiency in traditional Chinese medicine (TCM), and reserpine treated rats show similar signs to TCM spleen deficiency pattern. This paper is aimed to explore the regulatory effect on neuroendocrinoimmune network by Lizhong Pill in reserpine induced TCM spleen deficiency rats.

Materials and methods

100 healthy adult male SD rats, with a mean weight of 200 g, were randomly divided into five groups in average: control group, reserpine treated group, atropine treated group, treatment groups with Lizhong Pill at high dose and low dose (equal to the dosage of crude drugs for 4 g/kg/d and 8 g/kg/d). Rats in reserpine treated group were induced by intraperitoneal injection of reserpine at 0.5 mg/kg d for 4 weeks. The levels of IL-1, IL-6 and gastrin were measured with radioimmunoassay, TNF-α and IFN-γ in serum were measured with ELISA, the level of vasoactive intestinal peptide (VIP) and substance P (SP) in small intestine were determined with radioimmunoassay, and the TNF-α and TGF-β positive cells in small intestine were detected by immunohistological staining. Data were analyzed with SAS 9.1 software package.

Results

The rats in reserpine treated group, body weight, concentrations of IFN-γ, IL-1 and TNF-α in serum, expression of TGF-β in small intestine, VIP in small intestine decreased (P < 0.05), and the level of IL-6 in serum, expression of TNF-α, SP in small intestine and gastrin were increased (P < 0.05). Administration of Lizhong Pill at high dose could increase the body weights at day 21, and the weights of rats in Lizhong Pill groups were much higher compared to reserpine treated group. At high dose of Lizhong Pill could increase the level of TNF-α in serum. Lizhong Pill at high dose and low dose could reverse the changes of IL-1, IL-6 and IFN-γ, gastrin, expression of TGF-β and TNF-α, VIP and SP in small intestine.

Conclusions

The rats treated with reserpine, with similar signs to TCM spleen deficiency, show neuroendocrinoimmune disorders, and the restoration of the neuroendocrinoimmune disorders might be the part of mechanism of Lizhong Pill for reinforcing TCM spleen deficiency.     

Effect of polypeptide 2B1 on condition of dampness pattern in rats in terms of Traditional Chinese Medicine

Objective

This study investigated how polypeptide 2B1 is involved in regulating and governing dampness in rat models with dampness pattern defined in terms of Traditional Chinese Medicine.

Methods

We randomly divided 48 SPF 10-week-old male Sprague-Dawley (SD) rats into a normal group, normal + Aristolochic acid I (AA-I) for 5 min group, normal + AA-I for 60 min group, dampness pattern group (DS-Group), dampness pattern + AA-I for 5 min group, and dampness pattern + AA-I for 60 min group. Groups were then treated accordingly. We took out the lung, stomach, liver, spleen, kidney, large intestine, and small intestine tissues to detect gene and protein expression of organic anion transporter polypeptide 2B1 (OATP2B1).

Results

Gene expression of OATP2B1 in spleen, kidney, and small intestine of rats with dampness pattern was lower than that in normal rats (P<0.05). The gene expressions of OATP2B1 in liver, stomach, large intestine, and small intestine were lower than that in control rats at different time points after being stimulated by AA-I (P<0.05).

Conclusion

There is coordination among multiple viscera in handling the condition of dampness, and the mechanism underlying the action may rely on regulating the expression of OATP2B1.     

小儿腹泻外敷散对胃肠动力学的影响

该文主要探讨小儿腹泻外敷散对胃肠动力学的影响。首先采用炭末推进法观察小儿腹泻外敷散对正常和虚泻小鼠肠运动的影响,结果表明其对正常小鼠组的炭末推进并无显著性的抑制作用,对番泻叶致虚泻小鼠的炭末推进有显著性的抑制作用;然后以腹泻指数为指标,观察其对虚泻小鼠腹泻的影响,结果表明其对番泻叶所致的虚泻小鼠具有良好的抗腹泻作用;随后的大鼠离体肠平滑肌测试结果表明小儿腹泻外敷散可竞争性拮抗由乙酰胆碱、氯化钡引起的胃肠痉挛,且与阿托品具有一定协同作用;最后采用放射免疫法测定敷药前后大鼠小肠中P物质(SP)、血管活性肠肽(VIP)含量的变化,结果表明敷药后大鼠小肠中的SP降低,VIP含量降低,可抑制小肠运动。在体和离体实验均表明,小儿腹泻外敷散为一种良好的胃肠动力型外敷中药,对脾虚泄泻所致腹泻小鼠有抗腹泻作用。     

Comparisons on enhancing the immunity of fresh and dry Cordyceps militaris in vivo and in vitro

Ethnopharmacological relevance

The immunomodulatory capacities of fresh Cordyceps militaris (FCM) and dry Cordyceps militaris (DCM) were compared.

Materials and methods

In vivo immunomodulatory assay, different doses of FCM and DCM were orally administrated over a period of 15 days in a cyclophosphamide (CY) induced immunosuppression mice; in vitro testing, the spleen cells were extracted from healthy mice and treaded with CY, then cultured with different dose of FCM or DCM; the contents of Cordyceps militaris polysaccharide (CMP), cordycepin, adenosine, total polyphenol (TP) and total flavonoids (TF) in FCM and DCM were measured.

Results

Our studys indicated that, FCM was significantly stronger than DCM on increasing the spleen and thymus indexes, spleen lymphocyte activity, macrophage function, and promoting the levels of IL-2, IFN-γ in vivo and in vitro. The contents of immunomodulatory CMP and TF in FCM were markedly higher than in DCM.

Conclusion

All these results suggested that FCM was superior to DCM on enhancing immunity.     

Immunomodulatory effects of Abnormal Savda Munsiq,a traditional Uighur medicine,on the combined stress mice

Objective

To investigate the immunomodulatory effects of ASMq, a herbal preparation used in Traditional Uighur Medicine, on the combined stress mice.

Materials and methods

The combined stress was induced on mice by application of the electric-foot shock in a cold–dry environment and three different doses of ASMq were orally administered to the animals for 14 days. The effect of ASMq on the immune apparatus weight index, lymphocytes proliferation, serum levels of the cytokines, immunoglobulins, hemolysin and NK cells as well as the phagocytic activity of the macrophages were evaluated.

Results

Oral administration of ASMq was found to increase the thymus and spleen indices, lymphocytes proliferation induced by Con A and LPS, the percentage of CD4⁺ in thymus, spleen and peripheral blood and restore the CD4⁺/CD8⁺ ratio. The serum concentrations of INF-β, IL-2, IL-6, IgG, NK cells and hemolysin were also increased. The macrophage phagocytic activity was also enhanced.     

Pharmacological basis for the medicinal use of muskmelon base (Pedicellus Melo.) for abdominal distention and constipation

Ethnopharmacological relevance

Muskmelon base (Pedicellus Melo.) has a long history (Ming Dynasty) as a Chinese traditional medicine. According to traditional use, it was prepared as rectal suppositories for treating abdominal distention and constipation. The present study was carried out on the pharmacological basis for the medicinal use of muskmelon base.

Aim of the study

The objective of this study was to determine the pharmacological basis for the medicinal use of the ethanol extract from muskmelon base (EMB) for abdominal distention and constipation.

Materials and methods

In this study, we report the gastrointestinal prokinetic action of EMB following single rectal or large intestinal administration. Laxative activity, gastric emptying and small intestinal transit tests were examined in ICR mice. SD rats were used to determine changes in large intestinal transit and contractile effects of the proximal colon in vivo. Guinea pigs were used to evaluate the contractile effects of the proximal colon and the possible mechanism or mechanisms on proximal colon activity ex vivo. Moreover, the acute toxicity of EMB was evaluated.

Results

In the in vivo experiments, the acute toxicity test showed that the maximum tolerated dose (MTD) of EMB was 400 mg/kg. A laxative effect was observed in mice at different dosages (6.5, 13 and 26 mg/kg). EMB showed a dose-dependent acceleration of gastric emptying (13 and 26 mg/kg). It also promoted both small intestinal (6.5, 13 and 26 mg/kg) and large intestinal (4, 8 and 16 mg/kg) transit activity. In the SD rat model, single rectal administration of EMB (8 and 16 mg/kg) showed a significant increase in both the frequency and amplitude of proximal colon smooth muscle contractility. These increases in amplitude and frequency peaked 30–60 min after EMB administration and corresponded with the results of the laxative activity test. The ex vivo experiments showed that varying doses of EMB (11.5, 23 and 46 mg/kg) had a direct prokinetic effect that was sensitive to atropine.

Conclusions

Our results show that EMB is a low dosage, fast acting drug with a large therapeutic window (4–400 mg/kg) and shows significant gastrointestinal prokinetic action after single rectal or large intestinal administration. This gastrointestinal prokinetic effect was stronger in the intestines than in the stomach. This effect was sensitive to atropine, suggesting that EMB acts mainly through cholinergic mechanisms.     

The possible mechanisms of Fructus Mume pill in the treatment of colitis induced by 2,4,6-trinitrobenzene sulfonic acid in rats

Ethnopharmacological relevance

Fructus Mume pill (FMP) has been used as a folk remedy for gastrointestinal diseases in China over thousands of years. FMP was approved for the treatment of gastrointestinal diseases in 2001 by the State Food and Drug Administration (SFDA) of China. Although FMP had significant efficacy for treatment of the patients with inflammatory bowel disease (IBD) in the clinic, the mechanism of action is still unclear.

Aim of the study

In the present study, the effects and possible mechanism of FMP on colitis induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS) were investigated.

Materials and methods

Fifty-four SD rats were divided into six groups. Nine rats for each group from three independent experiments were investigated for the effects of FMP.

Results

FMP protected against diarrhea, colon weight increase, colonic accretion, ulceration and myeloperoxidase (MPO) activity elevation. The effects of FMP on recovery of colonic damage and restoration of the normal structures of colorectums were superior to dexamethasone (DEX). FMP promoted the restoration of abnormal cytokine secretion after TNBS treatment. FMP was effective in restoring the balance of intestinal bacteria population from the imbalance of G⁺/G⁻ in rats with colitis.

Conclusions

The results indicated that FMP is effective in treatment of colitis in an experimental rat model. The possible mechanisms may be through down-regulation of Th1-polarized immune response and opsonic effect of intestinal commensal bacteria in this model system.     

Effect of plants used in Mexico to treat gastrointestinal disorders on charcoal–gum acacia-induced hyperperistalsis in rats

Ethnopharmacological relevance

A total of 28 plant extracts, belonging to 26 different plant species are commonly used in Traditional Mexican Medicine for the treatment of gastrointestinal disorders such as diarrhea.

Aim of the study

To evaluate the effect of medicinal plant extracts on induced hyperperistalsis in rats.

Materials and methods

Charcoal meal test was used in this study. Extracts were tested at a dose of 300 mg/kg.

Results

From all the plant extracts tested, only Geranium mexicanum (roots) showed 100% of inhibition. The extracts of Artemisia absinthium, Matricaria recutita, Caesalpinia pulcherrima, Lygodium venustum, Chenopodium ambrosoides (green variety), Aloysia triphylla, Artemisia ludoviciana, Chiranthodendron pentadactylon, and Cocos nucifera showed moderate inhibitory activity with values ranging from 30 to 57%. Their activities were greater than that of or equal to loperamide (34% of inhibition at doses of 10 mg/kg) drug used as control. The remaining plants exhibited marginal or null inhibitory effect on hyperpropulsive movement of the small intestine.

Conclusions

The results obtained in this study give some scientific support to the popular use of 23 of the plants tested for the treatment of gastrointestinal disorders such as diarrhea in Mexican traditional medicine. However, roots of Geranium mexicanum should be used in herbal medicine with care to avoid toxicity.     

The protective and healing effects of Si Shen Wan in trinitrobenzene sulphonic acid-induced colitis

Ethnopharmacological relevance

Si Shen Wan is a traditional Chinese herbal medicine formula for the treatment of diseases with diarrhea, such as ulcerative colitis, allergic colitis and chronic colitis. To investigate the protective and healing effects of Si Shen Wan in the experimental colitis induced by trinitrobenzene sulphonic acid, and to furture explore its mechanism of action.

Materials and methods

Rats with colitis treated with Si Shen Wan for 10 days. Colon wet weight, colon organ coefficient, colonic damage score and pathological change after trinitrobenzene sulphonic acid challenge were determined. The levels of MPO, MDA, GSH-PX, SOD and the expression of IL-4 and IL-10 mRNA in the colon were also measured.

Results

After treatment, colon wet weight, colon organ coefficient and colonic damage score were lower than that in the control group (p<0.05). MDA and MPO concentrations in the inflamed colonic tissues were decreased remarkably in the treated groups compared with that in the control group (p<0.05). But SOD level, IL-4 and IL-10 mRNA expression in the inflamed colonic tissues were obviously increased.

Conclusions

It is a potential path that protective effect of Si Shen Wan on impaired colonic mucosa rats with experimental colitis was accomplished by down-regulating the level of MDA and MPO, and up-regulating the level of SOD and the IL-4 and IL-10 mRNA expression in the colon mucosa.     

结肠灵汤对肠易激综合征大鼠肠组织及脊髓后角脑肠肽表达的影响

目的探讨结肠灵汤对肠易激综合征大鼠肠黏膜及脊髓后角P物质(SP)、血管活性肠肽(VIP)阳性神经纤维表达的影响。方法将实验动物分为对照组、模型组、西药组及中药低、高剂量组。采用结肠慢性刺激法制作肠道高敏性大鼠模型,观察各组大鼠肠道敏感性指标,用免疫组织化学法对各组大鼠回盲部、结肠及脊髓后角SP、VIP的表达进行半定量分析。结果各治疗组大鼠肠道敏感性指标均有明显改善,结肠灵汤能降低大鼠肠组织SP、VIP的免疫表达及脊髓后角SP的免疫表达,呈一定量效关系。结论结肠灵汤可改善肠道敏感性症状,其机制可能在于调节肠道初级感觉神经纤维SP、VIP的分泌,抑制伤害性初级传入末梢上行释放SP,减弱背角神经兴奋性,从而提高内脏痛阈,改善腹痛腹泻症状。     

Ethnopharmacology guided screening of traditional Indian herbs for selective inhibition of Plasmodium specific lactate dehydrogenase

Ethnopharmacological relevance

Medicinal plants traditionally used to treat malaria can provide quality leads towards identifying novel anti-malarial drugs. Here we combined this approach with target based drug discovery and explored Plasmodium specific lactate dehydrogenase (LDH) inhibitory activity of 8 Indian plants which are ethnically used to treat malaria.

Methods

LDH from Indian Plasmodium falciparum and Plasmodium vivax strains, were cloned and expressed in Escherichia coli, followed by purification of recombinant enzymes (rPfLDH and rPvLDH respectively). Extracts of 8 plants in different organic and aqueous solvents, were screened for their inhibitory activity on rPfLDH, rPvLDH and mammalian LDHs. Phyllanthus amarus aqueous extract was further tested for in vitro parasiticidal activity.

Results

Aqueous extract of Phyllanthus amarus Schum. and Thonn. and chloroform extract of Murraya koenigii (L.) Spreng. exhibited profound and exclusive inhibitory effect on Plasmodium falciparum LDH (IC₅₀=11.2 μg/ml±0.4) and Plasmodium vivax LDH (IC₅₀=6.0 μg/ml±0.6) respectively. Moreover, Phyllanthus amarus aqueous extract also demonstrated antiplasmodial activity in vitro, on Chloroquine sensitive and resistant strains of Plasmodium falciparum (IC₅₀=7.1 μg/ml±0.5 and 6.9 μg/ml±0.7 respectively).

Conclusion

Target specific screening of traditional herbs used in malaria treatment has proffered Phyllanthus amarus and Murraya koenigii extracts as hits which can optimistically provide novel antimalarial drugs.     

Muscarinic-dependent inhibition of gastric emptying and intestinal motility by fractions of Maytenus ilicifolia Mart ex. Reissek

Ethnopharmacological relevance

Maytenus ilicifolia Mart. ex. Reissek (Celastraceae) is widely used in Brazilian folk medicine to treat gastric disturbances.

Aim of the study

This work intended to characterize the effects of Maytenus ilicifolia on gastrointestinal motility.

Materials and methods

Gastric emptying and intestinal transit were measured in the same animal. Mice received a semisolid marked with phenol red, half an hour after treatment with extracts. The amount of marker in the stomach and the distance reached in the intestine after 15 min were measured as index of gastrointestinal emptying and intestinal transit, respectively.

Results

Intraperitoneal administration of a flavonoid-rich extract potently reduced the gastric emptying (ED₅₀ = 89 mg/kg) and the intestinal transit (ED₅₀ = 31 mg/kg) of mice. Bio-guided purification of the flavonoid-rich extract by chemical partition with solvents of decreasing polarity yielded fraction insF with about 12–14 times higher activity than the initial flavonoid extract in both the gastric emptying and the intestinal transit. The inhibitory effects of the insF (9.7 mg/kg, i.p.) on gastric emptying and intestinal transit were reversed by co-administration of bethanechol (10 mg/kg, s.c.) but not by co-administration of metoclopramide (30 mg/kg, p.o.) indicating muscarinic but not dopaminergic interaction of the compounds of Maytenus.Chemical investigation of the insF fraction by HPLC–MS allowed the identification of 4 free flavonoids (catechin, epicatechin, quercetin and kaempferol), 29 flavonol glycosides and 8 tannins. The flavonol glycosides ranged from 1 to 4 monosaccharide units, having mainly quercetin and kaempferol as aglycone moieties, and the tannins were composed by catechin/epicatechin and/or afzelechin/epiafzelechin.

Conclusions

Overall, the results indicate that the components of Maytenus ilicifolia have a potential use in the treatment of gastrointestinal motility disturbances such as diarrhea.     

Neuroprotective effects of the Chinese Yi-Qi-Bu-Shen recipe extract on injury of rat hippocampal neurons induced by hypoxia/reoxygenation

Objective

To explore the protective effects of the Chinese Yi-Qi-Bu-Shen recipe (YB) against neuronal injury induced by hypoxia–reoxygenation, which has shown beneficial effect in improving the brain function of type 2 diabetics likely through its antihyperglycemic, antioxidant activity, and investigate its mechanisms.

Methods

The bilateral hippocampus was collected from newborn rats to establish single cell suspension. On the 10th day, the primarily cultured hippocampal neurons were randomly divided into five groups: the normal group (NG), the hypoxia/reoxygenation group (HG), and groups protected with small, medium and large dosages of YB (SG, MG and LG, respectively). The YB-protected groups were treated with different concentrations of YB containing serum before reoxygenation. The metabolic rate of MTT, the malondialdehyde (MDA) content, and the activity of superoxide dismutase (SOD) and lactate dehydrogenase (LDH) were measured with assay kits. The apoptosis rate of hippocampal neurons were tested using flow cytometry analysis. RT-PCR was used to evaluate the mRNA expressions of bcl-2 and bax genes.

Results

The SOD activity, the cell survival rate, the bcl-2/bax ratio, and the bcl-2 mRNA expression in the HG group were significantly lower (all P<0.01), but the levels of MDA and LDH, the apoptosis rate, and the bax mRNA expression were higher (all P<0.01) than those in the NG group. The SOD activity, the cell survival, the bcl-2 mRNA expression, and the bcl-2/bax ratio were significantly higher in all of the YB-protected groups (all P<0.01), but the level of MDA and LDH, the apoptosis rate, and the bax mRNA expression were lower (P<0.01, P<0.05) than those in the HG group in a dose dependent manner.

Conclusion

The YB extract has a protective effect on hippocampal neurons against injury induced by hypoxia/reoxygenation, through its antioxidant activity and the regulation of apoptosis.     

Antisecretory activity from the flowers of Chiranthodendron pentadactylon and its flavonoids on intestinal fluid accumulation induced by Vibrio cholerae toxin in rats

Ethnopharmacological relevance

The flowers of Chiranthodendron pentadactylon Larreat. (Sterculiaceae) has been traditionally used as folk medicine in Mexico for the treatment of gastrointestinal disorders such as diarrhea and dysentery.

Aim of the study

This study aimed to assess the antisecretory activity which supports the therapeutic use of Chiranthodendron pentadactylon and its flavonoids to treat diarrhea.

Materials and methods

The methanol extract of Chiranthodendron pentadactylon, subsequent fractions, and flavonoids were evaluated on cholera toxin-induced intestinal secretion in rat jejunal loops model.

Results

Three antisecretory flavonoids were isolated by bioassay-guided purification, namely, isoquercitrin 3, (+)-catechin 4 and (−)-epicatechin 5. Among them, epicatechin exhibited the most potent antisecretory activity with ID₅₀ of 8.3 μM/kg. Its potency was close that of to loperamide (ID₅₀ 6.1 μM/kg), drug used as control. Isoquercitrin (ID₅₀ 19.2 μM/kg) and catechin (ID₅₀ 51.7 μM/kg) showed moderate and weak activity, respectively.

Conclusion

The results of the present study lend some support to the anecdotal report for the traditional use of the flowers of Chiranthodendron pentadactylon in the control of dysentery.     

Effects of electroacupuncture of different frequencies on SP and VIP expression levels in colon of rats with slow transit constipation

Objective

To observe the effects of electroacupuncture (EA) of different frequencies on the expression levels of substance P (SP) and vasoactive intestinal peptide (VIP) in the colon of rats with slow transit constipation (STC).

Methods

One hundred healthy male Sprague-Dawley (SD) rats were randomly divided into a normal group, a model group, a low-frequency EA group, a high-frequency EA group and a variable-frequency EA group, with 20 rats in each group. The rats in the normal group were fed with normal diet. The rats in the other groups were fed with phenethylpiperidine in the diet at a dose of 8 mg/(kg·bw) per day, for 120 d, to establish the STC model. Rats in the normal group and the model group did not receive any treatment; rats in the low-frequency EA group were treated with 2 Hz continuous wave EA, rats in the high-frequency EA group were treated with 100 Hz continuous wave EA, and rats in the variable-frequency EA group were treated with 2 Hz/100 Hz sparse-dense EA. The current intensity of the EA was determined by the slight vibration of the rat limbs without painful screaming. The intervention was performed once a day, 15 min/time for continuous 15 d. After treatment, the intestinal transit function and the expression levels of SP and VIP in the colon of the rats in each group were determined.

Results

After treatment, the defecation duration of the first dark stool in the model group was significantly longer than that in the normal group (P<0.05); the defecation durations of the first dark stool in the low-frequency EA group, high-frequency EA group and variable-frequency EA group were significantly shorter than the duration in the model group (all P<0.05); compared with the low-frequency EA group, the first dark stool defecation duration of rats in the variable-frequency EA group was significantly shorter (P<0.05); compared with the normal group, the SP and VIP expression levels in the colon of the model group were significantly decreased (both P<0.01); the SP and VIP expression levels in the colon of the low-frequency EA group, the high-frequency EA group and the variable-frequency EA group were significantly higher than those in the model group (all P<0.05); compared with the high-frequency EA group, the SP expression levels in the colon in the low-frequency EA group and the variable-frequency EA group were significantly increased (both P<0.05); compared with the low-frequency EA group, the VIP expression levels in the colon in the high-frequency EA group and the variable-frequency EA group were significantly increased (both P<0.05).

Conclusion

EA improves the intestinal function of STC model rats by regulating the expression levels of SP and VIP in rat colon. The EA stimulation with 100 Hz continuous wave, 2 Hz/100 Hz sparse-dense wave shows a better improvement in the colonic transit function in STC rats, followed by 2 Hz continuous wave.

     

Rheinanthrone,a metabolite of sennoside A,triggers macrophage activation to decrease aquaporin-3 expression in the colon,causing the laxative effect of rhubarb extract

Ethnopharmacological relevance

Aquaporin-3 (AQP3) is expressed in mucosal epithelial cells in the colon and is important for regulating fecal water content. We examined the role of AQP3 in the laxative effect of rhubarb extract.

Methods

After orally administering rhubarb extract or its major component (sennoside A) to rats, the fecal water content, AQP3 expression and prostaglandin E₂ (PGE₂) concentrations in the colon were examined. The mechanism by which sennoside A decreases the expression of AQP3 was examined using the human colon cancer HT-29 cells and macrophage-derived Raw264.7 cells.

Results

During diarrhea by rhubarb extract administration, the PGE₂ levels in the colon increased while the AQP3 expression significantly decreased. Similar changes were also observed when sennoside A was administered. When sennoside A or its metabolites, rheinanthrone and rhein were added to Raw264.7 cells, a significant increase in the PGE₂ concentration was observed only in cells treated with rheinanthrone. Fifteen minutes after adding PGE₂ to the HT-29 cells, the AQP3 expression decreased to approximately 40% of the control. When pretreated with indomethacin, sennoside A neither decreased the AQP3 expression nor induced diarrhea.

Conclusions

Sennoside A may decrease AQP3 expression in the colon to inhibit water transport from the luminal to the vascular side, leading to a laxative effect. The decreases in the levels of AQP3 are caused by rheinanthrone, which is a metabolite of sennoside A, this metabolite activates the macrophages in the colon and increases the secretion of PGE₂; PGE₂ acts as a paracrine factor and decreases AQP3 expression in colon mucosal epithelial cells.     

Therapeutic effect observation on Nie-pinching the spine manipulation for infantile diarrhea due to spleen deficiency

Objective

To observe the influence of Nie-pinching the spine manipulation on the excretory rate of urine D-xylose in the infants with diarrhea due to spleen deficiency, and to assess the clinical effects.

Methods

Sixty infants in conformity with the diagnostic criteria of diarrhea due to spleen deficiency were randomly divided into a treatment group and a control group by the random digital table, 30 cases in each group. The infants in the treatment group were treated by Nie-pinching the spine manipulation and traditional infantile tuina, in addition to the routine basic treatment. The infants in the control group were treated by the same traditional infantile tuina in addition to the routine basic treatment. The infants in the two groups were treated once every day, 4 weeks as a course. Totally, the treatment was given for a course. The symptom integrals of spleen deficiency were used to assess the improvement in the symptoms. The colorimetry was used to determine the excretory rate of urine D-xylose.

Results

In the comparison of the same group before and after the treatment, the differences in the global score of spleen deficiency symptoms and the excretory rate of urine D-xylose in the two groups were statistically significant (all P<0.01). After the treatment, the differences in the global score of spleen deficiency symptoms and the excretory rate of urine D-xylose between the two groups were all statistically significant (both P<0.01).

Conclusion

Chiropractics can reduce the integrals of spleen deficiency symptoms and elevate the excretory rate of urine D-xylose in the infants with diarrhea due to spleen deficiency, so as to enhance the therapeutic effects by alleviating the symptoms of spleen deficiency and the absorptive function of the small intestine.

     

半夏泻心汤对氟尿嘧啶致腹泻小鼠模型肠道免疫功能的影响

目的: 研究半夏泻心汤对氟尿嘧啶致腹泻小鼠模型肠道免疫功能的影响,揭示其治疗化疗相关性腹泻的作用机制. 方法: 采用腹腔注射氟尿嘧啶(40 mg·kg^-1)连续6 d,建立小鼠腹泻模型.随机分为6组:正常组,模型组,阳性药组(蒙脱石散1.17 g·kg^-1),半夏泻心汤高、中、低剂量组(2.5,1.25,0.625 g·kg^-1),造模前给药2 d,连续给药11 d.观察小鼠每日活动状况、进食量、体重、腹泻变化情况,检测胸腺、脾脏质量,取回肠组织做病理切片,采用ELISA法测定小肠中免疫球蛋白A(IgA)、血管活性肠肽(VIP)、白细胞介素15(IL-15)的含量. 结果: 与正常组比较,模型组小鼠化疗后腹泻加重,体重、胸腺及脾脏指数减小,肠道IgA,VIP的含量降低,而IL-15含量升高;与模型组比较,半夏泻心汤各剂量组能显著抑制氟尿嘧啶造成的小鼠体重降低、胸腺、脾脏指数减小,降低腹泻发生率,同时增加肠道IgA,VIP的含量,减少IL-15的含量(P<0.05或P<0.01). 结论: 半夏泻心汤能治疗化疗相关性腹泻,可能与其调节肠黏膜免疫功能有关.     

Antioxidant properties of magnesium lithospermate B contribute to the cardioprotection against myocardial ischemia/reperfusion injury in vivo and in vitro

Objective

To determine the cardioprotective effect of magnesium lithospermate B (MLB) on myocardial ischemia/reperfusion (MI/R) injury and to investigate the antioxidant potential in vivo and in vitro.

Methods

MI/R injury was induced by the occlusion of left anterior descending coronary artery for 30 min followed by reperfusion for 3 h in rats. After reperfusion, hearts were harvested to assess infarct size, histopathological damages, the levels of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), reduced glutathione (GSH) and malondialdehyde (MDA). Blood samples were collected to determine serum levels of creatine kinase-MB (CK-MB), cardiac troponin (cTnI) and lactate dehydrogenase (LDH). Furthermore, simulated ischemia/reperfusion (SI/R) injury in vitro was established by oxygen and glucose deprivation (OGD) for 2 h followed by 24-hour recovery period in cardiomyocytes. The activity of LDH in the cultured supernatant and the levels of intracellular reactive oxygen species (ROS), SOD and MDA in cardiomyocytes were also measured. Finally, cardiomyocytes apoptosis was determined with flow cytometry.

Results

MLB significantly limited infarct size, ameliorated histopathological damages and prevented leakage of CK-MB, cTnI and LDH. Additionally, SOD, CAT, GPx and GSH activities were notably increased by MLB, along with the MDA content decreased as compared with the model group in rats. In vitro study, MLB also decreased LDH activity in the cultured supernatant, increased SOD activity in cardiomyocytes, reduced intracellular ROS and MDA levels, and significantly suppressed cardiomyocytes apoptosis.

Conclusion

MLB possessed remarkably cardioprotective effects on MI/R injury in vivo and in vitro. The protection of MLB may contribute to its antioxidant properties.