Qihuzha granule attenuated LPS-induced acute spleen injury in mice via Src/MAPK/Stat3 signal pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Qihuzha granule (QHZG), is one of traditional Chinese patent medicines composed of eleven edible medicinal plant, which has been used in the clinic for the treatment of indigestion and anorexia in children caused by deficiency of the spleen and stomach. Yet it is noteworthy that QHZG has therapeutic effect on recurrent respiratory tract infection (RRTI) in children. However, its potential molecular mechanisms remained unclear. AIM OF THE STUDY: The aim of this study was to investigate the therapeutic effect and potential mechanism of QHZG on lipopolysaccharide (LPS) induced acute spleen injury. MATERIALS AND METHODS: The acute spleen injury model was induced by intraperitoneal injection of LPS (10 mg/kg) and safe doses of QHZG was administered by gavage once a day for 23 days before LPS treatment. Serum inflammatory cytokines including interleukin-2 (IL-2), IL-1ß, IFN-γ, and tumor necrosis factor-α (TNF-α) were tested by ELISA. Related protein levels were detected by Western blotting. Hematoxylin-eosin (HE) staining was employed to observe the histological alterations. The distribution of macrophages and neutrophils in the mouse spleen was examined by immunofluorescence analysis. RESULTS: QHZG pretreatment significantly abolished the increased secretion of cytokines such as interleukin-2 (IL-2), IL-1ß, IFN-γ, and tumor necrosis factor-α (TNF-α), which were attributable to LPS treatment. Immunofluorescence staining and Histological analysis of spleen tissue revealed the protective effect of QHZG against LPS-induced acute spleen injury in mice. Further study indicated that pretreatment with QHZG significantly inhibited LPS-induced phosphorylation of Src. Accordingly, the increased phosphorylation of Src downstream components (JNK, ERK, P38 and STAT3) induced by LPS was remarkably diminished by QHZG, suggesting the involvement of Src/MAPK/STAT3 pathway in the inhibitory effects of QHZG on spleen injury in mice. CONCLUSION: Our study demonstrated that QHZG protected mice from LPS-induced acute spleen injury via inhibition of Src/MAPK/Stat3 signal pathway. These results suggested that QHZG might serve as a new drug for the treatment of LPS-stimulated spleen injury.

Oxaliplatin-induced increase in splenic volume: experiences from multicenter study in Japan.

BACKGROUND: Chemotherapy with oxaliplatin is known to induce sinusoidal obstruction syndrome (SOS). In a previous single-center study, we reported that oxaliplatin-induced increase in splenic volume (SV) is strongly indicative of SOS, and that this increase in SV persisted for > 1 year after completing chemotherapy. The aim of this study was to confirm the oxaliplatin-induced SV change in a multicenter study in patients with stage III colon cancer in Japan. METHODS: We enrolled 59 patients who underwent curative resection for stage III colon cancer in the FACOS study in a phase II multi-center clinical study. Participants received mFOLFOX6 or CAPOX as adjuvant chemotherapy. SV change was assessed three times by computed tomographic volumetry: before surgery, on completion of adjuvant chemotherapy, and 1 year after completing adjuvant chemotherapy. RESULTS: SV on completing and 1 year after chemotherapy was significantly higher than that before surgery (P < 0.001). Oxaliplatin-induced SOS persisted for > 1 year after the completion of adjuvant chemotherapy in half of the patients. There was no difference in 3-year disease-free survival with respect to the presence or absence of increased SV. An increase in SV was observed in 72% of patients treated with mFOLFOX6 and 94% of patients treated with CAPOX (P = 0.13). CONCLUSION: This study can be verified the findings observed in our previous single-center study, oxaliplatin-based adjuvant chemotherapy was associated with an increase in SV. Furthermore, this increase can persist for > 1 year. The continuous presence of SOS may have a negative impact on prognosis in patients that develop recurrent disease.

The effects of aqueous extract of Maca on energy metabolism and immunoregulation.

BACKGROUND: In the present work, we investigated the effects of aqueous extract of Maca (AEM) on energy metabolism and immunoregulation in spleen-deficient mice. METHOD: We established a cyclophosphamide-induced spleen-deficiency model with ginseng, a herb that strengthens splenic function, as a control. Sixty male Kunming mice were randomly divided among 5 groups: normal, model, ginseng control (1.5 g/kg), AEM high dose (1.5 g/kg), and AEM low dose (0.75 g/kg). All animals, except those in the normal group, were injected with cyclophosphamide to induce spleen deficiency. Furthermore, we investigated differences in the thermotropic behaviors of mice using the Animal Thermotropism Behavior Surveillance System to detect energy metabolism-related assays and immune regulation assays. RESULTS: Mice given AEM exhibited tropism in response to hot plate exposure. AEM inhibited loss of body weight and immune organ atrophy caused by cyclophosphamide, increased the cAMP/cGMP ratio in blood, and enhanced the activities of Na+-K+-ATPase, Ca2+-Mg2+-ATPase, lactate dehydrogenase, and hepatic glycogen. AEM significantly reversed declining white blood cells and platelet counts, and increased the hemoglobin content within peripheral blood cells. AEM improved the protein levels of IFN-γ, TNF-ß, IL-2, and IL-4 in the spleen. CONCLUSIONS: Maca possesses the Traditional Chinese Medicine (TCM) property of warm and appears to strengthen spleen function.

Dibutyl phthalate-mediated oxidative stress induces splenic injury in mice and the attenuating effects of vitamin E and curcumin.

Dibutyl phthalate (DBP) is a ubiquitous environmental contaminant that at certain levels can be harmful to human health. Although DBP has been widely linked to immunotoxicity, any association between DBP exposure and splenic injury remains unknown. The purpose of this study was to investigate whether DBP exposure can induce splenic injury and the antagonistic effects of two antioxidants, vitamin E (VitE) and curcumin (Cur), on DBP-induced splenic injury. The levels of ROS, GSH, T-AOC, IL-1ß, TNF-α, cytochrome C, caspase-8, caspase-9 and caspase-3 in the spleen homogenate of mice were measured. Any histopathological changes in the spleen were observed using H&E and toluidine blue staining. And the morphology of mitochondria was observed using Janus Green B staining. The results indicate that exposure to 50 mg/kg DBP could cause histopathological changes of the spleen and result in inflammation and apoptosis associated with oxidative stress, which may lead to splenic injury in mice. Moreover, both VitE and Cur could antagonize the oxidative stress induced by DBP to reduce splenic injury. These findings help to expand our understanding of DBP-mediated immunotoxicity, and to show that VitE and Cur can alleviate DBP-induced splenic injury and the possible DBP-associated decline in immune function.

Doxorubicin triggers splenic contraction and irreversible dysregulation of COX and LOX that alters the inflammation-resolution program in the myocardium.

Doxorubicin (DOX) is a widely used drug for cancer treatment as a chemotherapeutic agent. However, the cellular and integrative mechanism of DOX-induced immunometabolism is unclear. Two-month-old male C57BL/6J mice were divided into high- and low-dose DOX-treated groups with a maintained saline control group. The first group was injected with a high dose of DOX (H-DOX; 15 mg·kg-1·wk-1), and the second group was injected with 7.5 mg·kg-1·wk-1 as a latent low dose of DOX (LL-DOX). H-DOX treatment led to complete mortality in 2 wk and 70% survival in the LL-DOX group compared with the saline control group. Therefore, an additional group of mice was injected with an acute high dose of DOX (AH-DOX) and euthanized at 24 h to compare with LL-DOX and saline control groups. The LL-DOX and AH-DOX groups showed obvious apoptosis and dysfunctional and structural changes in cardiac tissue. Splenic contraction was evident in AH-DOX- and LL-DOX-treated mice, indicating the systems-wide impact of DOX on integrative organs of the spleen, which is essential for cardiac homeostasis and repair. DOX dysregulated splenic-enriched immune-sensitive lipoxygenase and cyclooxygenase in the spleen and left ventricle compared with the saline control group. As a result, lipoxygenase-dependent D- and E-series resolvin precursors, such as 16HDoHE, 4HDoHE, and 12-HEPE, as well as cyclooxygenase-mediated PG species (PGD2, PGE2, and 6-keto-PG2α) were decreased in the left ventricle, suggestive of defective immunometabolism. Both AH-DOX and LL-DOX induced splenic contraction and expansion of red pulp with decreased CD169+ metallophilic macrophages. AH-DOX intoxicated macrophages in the spleen by depleting CD169+ cells in the acute setting and sustained the splenic macrophage loss in the chronic phase in the LL-DOX group. Thus, DOX triggers a vicious cycle of splenocardiac cachexia to facilitate defective immunometabolism and irreversible macrophage toxicity and thereby impaired the inflammation-resolution program. NEW & NOTEWORTHY Doxorubicin (DOX) triggered splenic mass loss and decreased CD169 with germinal center contraction in acute and chronic exposure. Cardiac toxicity of DOX is marked with dysregulation of immunometabolism and thereby impaired resolution of inflammation. DOX suppressed physiological levels of cytokines and chemokines with signs of splenocardiac cachexia.

Gut microbial diversity in rat model induced by rhubarb.

Rhubarb is often used to establish chronic diarrhea and spleen (Pi)-deficiency syndrome animal models in China. In this study, we utilized the enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) method to detect changes in bacterial diversity in feces and the bowel mucosa associated with this model. Total microbial genomic DNA from the small bowel (duodenum, jejunum, and ileum), large bowel (proximal colon, distal colon, and rectum), cecum, and feces of normal and rhubarb-exposed rats were used as templates for the ERIC-PCR analysis. We found that the fecal microbial composition did not correspond to the bowel bacteria mix. More bacterial diversity was observed in the ileum of rhubarb-exposed rats (P<0.05). Furthermore, a 380 bp product was found to be increased in rhubarb-exposed rats both in faces and the bowel mucosa. The product was cloned and sequenced and showed high similarity with regions of the Bacteroides genome. AS a result of discriminant analysis with the SPSS software, the Canonical Discriminant Function Formulae for model rats was established.

Haematological and biochemical toxicity induced by methanol in rats: ameliorative effects of Opuntia vulgaris fruit extract.

The ameliorative effects of Opuntia vulgaris fruit extract (OE) were evaluated against methanol-induced haematological and biochemical toxicity in rats. The methanol-induced haematological and biochemical perturbation significantly decreased the levels of red blood cell (RBC), haemoglobin (Hb), haematocrit (Ht), serum total protein and increased glucose, cholesterol, and triglyceride levels in serum. Treatment of rats with methanol significantly increased lipid peroxidation (LPO) level and decreased the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) in erythrocytes. OE treatment could increase significantly the levels of RBC, Hb, Ht and total protein, and decrease glucose, cholesterol and triglyceride levels in serum, and increase the activities of SOD, CAT and GPx in erythrocytes, when compared with methanol-treated group. Spleen histopathology showed that OE could significantly reduce the incidence of spleen lesion induced by methanol. These results suggested that OE could exhibit a potential source of natural antioxidants against methanol-induced haematological and biochemical disruption in rats. The protective effects of OE may be due to the modulation of antioxidant enzymes activities and inhibition of LPO.

[Effects of Sijunzi Decoction on urine's xylose excretion rate and ATP in mucosa of spleen deficiency rats].

OBJECTIVE: To observe the effects of Sijunzi Decoction on D-xylose excretion rate and ATP content in the mucosa membranes of small intestines of rats with spleen deficiency. METHODS: Spleen deficiency model rats were made by reserpine injection. D-xylose excretion rate was measured with p-bromoaniline method, and the ATP content of small intestines mucosa was detected with bioluminescence method. The correlation between D-xylose excretion rate and ATP content of mucosa was also analyzed. RESULTS: Rats' body weight and D-xylose excretion rate decreased after reserpine injection (P < 0.01, vs control group), but increased after treated with Sijunzi Decoction (P < 0.05, vs model group). The ATP content of mucosa showed no significant difference between model group and control group. There was obviously positive correlation between the change of urine's D-xylose excretion rate and mucosa ATP content. CONCLUSION: Sijunzi Decoction has the activity of improving xylose absorption in spleen deficiency rats, but no obvious effect on their mucosa ATP content. The reducing of urine's D-xylose excretion rate in spleen deficiency rats is accompanied with the decrease of mucosa ATP content.

[The experimental research of Atractylodes macrocephala and Poria cocos on the meridian tropism theory in animals].

OBJECTIVE: The research was designed to study the meridian tropism theory of traditional Chinese drug through experiments in animals. METHOD: We used the mouse model of deficient spleen as the object. After administering Atractylodes macrocephala and Poria cocos respectively, we measured the indexes of MDA, SOD, NO to observe the effects of the drugs on various organs and compared the results with the traditional meridian tropism theory. RESULT: The two drugs had selective effects on the quantity or activity of MDA, SOD, NO in the organs for the normal group and the model group. CONCLUSION: The selective influence of the two drugs has close relativity with the traditional meridian tropism theory.

[Effect of sijunzi decoction on motilin and pharmacokinetic characteristics of tetramethylpyrazine in rat model of spleen deficiency syndrome].

OBJECTIVE: To explore relationship between the essence of Spleen Deficiency Syndrome (SDS) and pharmacokinetic characteristics (PK). METHODS: The level of motilin in serum and intestine, and PK of tetramethylpyrazine phosphate (TMPP) in rat model of SDS were observed simultaneously and treated with Sijunzi decoction (SJZD) for confirmation. RESULTS: The atrioventricular model of TMPP in SDS, normal and SJZD treated rats all belonged to the open double ventricular type; there were insignificant differences between normal and SJZD treated rats in motilin level and PK of TMPP, but the differences of these parameters between normal and SDS model rats were significantly (P < 0.01). The SDS state could markedly affect the absorption, distribution, metabolism and elimination of TMPP in rat; SJZD could normalize the abnormal PK of TMPP in SDS model rats. CONCLUSIONS: A supplementary scientific evidence for the hypothesis of "Pharmacokinetics of Syndrome Differentiation" is provided.

[Influence of stimulating zusanli with moxibustion of different quality and quantity on gastrointestinal motor function of reserpinized rats].

This article is focused on the observation of changes in body temperature, body weight, cholinesterase activity in blood, and gastrointestinal motility of reserpinized rats treated by stimulating Zusanli (ST 36) with moxibustion of different quality. (mugwort floss or pipe tobacco) and quantity (strong stimulation or weak stimulation). The experiment shows that better results are achieved with moxibustion not by burning tobacco; the result of strong stimulation with moxa-sticks is better than that of weak stimulation with the same material. Strong stimulation with moxa-sticks can obviously increase the activity of cholinesterase (P < 0.05), inhibit hyperactive gastrointestinal motility (P < 0.05), maintain normal body temperature (P < 0.05), and prevent body weight lossing. All these show that therapeutic results of moxibustion are closely related to the quality and quantity of moxibustion.

[Pharmacokinetic characteristics of tetramethylpyrazine and study on hemorheology in rat model of spleen deficiency syndrome].

UNLABELLED: In this experiment, the essence of Spleen Deficiency Syndrome (SDS) was explored with the rat model of SDS using tetramethylpyrazine (TMP). 120 Wistar rats were divided into two groups, 60 each for control and test group, they were treated with normal saline and reserpine respectively. The hemorheological parameters were also studied in 6 each of both groups. The pharmacokinetic properties were investigated in the remaining control group and test group. RESULTS: (1) Two-compartment open model of the control group was turned into that of one-compartment in test group; (2) The SDS model significantly affected the absorption and distribution of TMP in rats. AUC was much higher and serum concentration of TMP increased significantly than that of control. Hemorheological parameters (viscosity of whole blood, fibrinogen, etc.) increased significantly (P < 0.05-0.01), it demonstrated that the SDS model was in a state of Blood Stasis. It might be one of the pharmacokinetic mechanisms of TMP in the SDS model of rats.

Mobilization and distribution of beryllium over the course of chelation therapy with some polyaminocarboxylic acids in the rat.

The efficacy of three common polyaminocarboxylic acids in the treatment of experimental beryllium intoxication was investigated in male rats. N-(2-hydroxyethyl) ethylene diamine triacetic acid (HEDTA) was more effective than calcium disodium ethylenediamine tetraacetic acid (CaNa2EDTA) in reducing the beryllium concentration of the blood, kidneys and spleen and reducing beryllium-induced inhibition of hepatic alkaline phosphatase activity. HEDTA was also most effective in reducing histopathological lesions in the liver and spleen. Compared to these two chelators, the third amino chelator, calcium trisodium diethylene triaminepenta acetic acid (CaNa3DTPA) produced severe deleterious effects in the liver and systemic toxicity. The results suggest that HEDTA is a promising chelator for beryllium toxicity while DTPA enhances the toxic manifestation of beryllium.

[Immunohistochemical study on the enteroendocrine cells of the small intestine in experimental spleen deficiency syndrome in rats].

50 adult Wistar male rats were used and divided into 4 groups, i.e. normal control group, experimental Spleen Deficiency group induced by rhubarb, spontaneous recovery group and therapeutic group treated with Chinese recipe (Si Jun Zi decoction). All the animals of the 4 groups were killed simultaneously, and the jejunum and ileum were removed and processed for demonstration of gastrin cells and 5-HT cells according to immunohistochemical PAP technique. In addition, HE stained samples were prepared. The immunoreactivities of the two types of enteroendocrine cells were observed and semiquantitative estimation were performed under light microscopy. In addition, the immunoreactivities of 5-HT cells in normal control and experimental Spleen Deficiency group were measured by microspectrocytophotometer (MPV 2, Leitz). All the data were treated statistically. This study revealed that there were no obvious histological changes in the mucosa among the 4 groups. In the jejunum, the percentage of gastrin cells(+) in experimental Spleen Deficiency group was more than that of the normal control group, while the percentage of gastrin cells( ) was less than that of the normal control group. As compared with spontaneous recovery group, it showed contrary to the above result in the therapeutic group. No gastrin cells were found in the ileum in all the 4 groups. the percentage of 5-HT cells did not show significant changes in the jejunum and ileum among the 4 groups. But immuno-reactivity in the 5-HT cell was less than that of the normal control group in the jejunum of the Spleen Deficiency group.(ABSTRACT TRUNCATED AT 250 WORDS)

[Changes in cyclic nucleotides and its enzymes in the spleen and plasma of similar spleen deficiency rats induced by rhubarb and the readjusting function of yiqi jianpi decoction].

In order to investigate the nature of Spleen deficiency and the mechanism of immunodepression due to Spleen deficiency and explore the pharmacological action of Chinese drugs of Yiqi Jianpi decoction (YQJP), the authors had established the rats model by administration Rhubarb. The preliminary results demonstrated that the symptoms manifested in rats were similar to those of Spleen deficiency syndrome. The changes of cyclic nucleotides in the spleen and plasma were unanimous. The cAMP level and the ratio of cAMP/cGMP decreased significantly while cGMP level increased significantly. Adenylate cyclase (AC) activity in the spleen reduced remarkably while cAMP-PDE activity had little changes. After the administration of YQJP, the symptoms of Spleen deficiency improved to normal extent. YQJP elevated the cAMP level, the ratio of cAMP/cGMP and AC activity while it lowered the cGMP level. The results showed that the changes of cyclic nucleotides level and the ratio of cAMP/cGMP were important targets of Spleen deficiency and that the action of YQJP followed the change of the ratio of cAMP/cGMP. The results of this study indicated that immunodepression of Rhubarb was due to depressing AC activity and reducing the ratio of cAMP/cGMP. The readjusting action of YQJP was concerned with AC system. This study supplied Spleen deficiency and YQJP with certain data in biochemical mechanism and pharmacological function.