Spleen Index Of Mice Research Articles

The selenoprotein from selenium-rich glutinous rice improves spleen immune protection by inhibiting the NF-κB pathway

Moderate amounts of selenium supplementation have a positive effect on the immune response. Selenium deficiency status can lead to reduced immunity, but the benefits of a natural selenium-rich diet for selenium deficiency are unknown. In this experiment, low selenium feed (0.02 mg Se/kg diet) was given to the mice for 5 weeks and selenium supplementation with selenoprotein (SeP) for 4 weeks, and the selenium-deficient mouse model was successfully established. SeP supplementation had significant protective effects, including increased antioxidant capacity and alleviation of inflammatory damage. SeP markedly elevated the activities of thioredoxin reductase (TrxR) and glutathione peroxidase (GSH-Px) in the serum, kidney, liver, and spleen of selenium-deficient mice. SeP can restore the condition of damaged spleen tissue, raise the spleen index in mice, and encourage an increase in immunoglobulin (IgA, IgG, and IgM) content. The levels of catalase (CAT), total antioxidant capacity (T-AOC), GSH-Px, and cytokines (IL-1β, IL-6, and TNF-α) were all significantly elevated by SeP. SeP showed superior immunomodulatory effects over inorganic selenium (Na2SeO3). Together, these results suggest that SeP may be a potential dietary selenium supplement that can reduce symptoms of spleen damage caused by selenium deficiency.

The differences of parasitemia in Plasmodium berghei infected mice treated with extract of mango parasite leaves with Artemisinin combination

Malaria is a disease transmitted through the bite of a female Anopheles mosquito caused by the Plasmodium parasite. Plasmodium has been reported to become resistant to artemisinin. Thus, the study on the ethanol extract of mango parasite leaves/Dendrophthoe pentandra (MP) on P. berghei-infected BALB/c mice with and without artemisinin combination (MP+A) was conducted. This study is experimental laboratory research with a post-test only design. The percent parasitemia in mice treated with ethanol extract of MP and MP+A leaves decreased. There was no difference in the spleen index in mice given ethanol extract of MP, and MP+A leaves with p = 0.203 and the spleen weight of mice with p = 0.134 (significance: p <0.005). Pearson Correlation test showed a correlation between spleen index with parasitemia and spleen weight with parasitemia; however, there was no correlation between body weight and parasitemia. The ethanol extracts of MP and MP+A leaves had significant antimalarial activity, and the difference in the percent parasitemia between groups was significant but not so far. The spleen index value was not affected by the ethanol extract of MP and MP+A leaves, but the percent parasitemia was affected.

Photothermal effects of CuS-BSA nanoparticles on H22 hepatoma-bearing mice.

The objective of this study was to evaluate the in vivo application and photothermal ablation effects and mechanism of copper sulfide nanoparticles (CuS NPs) in hepatocellular carcinoma (HCC). Sheet-like CuS-BSA NPs with a particle size of 30 nm were synthesized using bovine serum albumin (BSA) as a biological modifier, and were physically characterized. To provide a reference range for the biosafety dose of CuS-BSA NPs, 36 male Kunming mice were randomly assigned into six groups. Different one-time doses of CuS-BSA NPs were injected via tail vein injection, and the potential damages of liver, kidney and spleen were observed 14 days later. To evaluate the in vivo photothermal effect of CuS-BSA NPs, 48 male Kunming mice were used to establish the H22 hepatoma-bearing mouse model and were randomly assigned into six groups. CuS-BSA NPs (600 μg/kg) were injected via tail vein or intratumoral injection. Irradiations were performed 30 min after injection, with a 980 nm near-infrared laser (2.0 W/cm2) for 10 min once a week for 3 weeks. The results indicated that the CuS-BSA NPs had good dispersibility in three different solvents and had a strong absorption peak at 980 nm. The heating curves demonstrated that the photothermal effects of CuS-BSA NPs aqueous solution exhibited concentration dependence and power density dependence. In the in vivo experiment, when the doses of CuS-BSA NPs were in the range of 1800–7,200 μg/kg, the thymus index and spleen index of mice were not significantly different from those of the control group, and the structures of liver, kidney and spleen were intact without remarkable pathological changes. A lower dose of CuS-BSA NPs (600 μg/kg) could effectively inhibit tumor growth in H22 hepatoma-bearing mice at 980 nm NIR. Moreover, under the near-infrared laser irradiation, both in the tail vein injection group and the intratumoral injection group, a large area of necrosis in the tumor tissue, as well as the up-regulation of apoptotic proteins including cleaved caspase-3 and cleaved caspase-9 were observed. CuS-BSA NPs are promising photothermal agents in the photothermal therapy of cancer.

Effects of Low-dose Splenic Irradiation on T lymphocyte Immune Function.

Relevant studies have confirmed that the stimulation of spleen function caused by low-dose splenic irradiation can have positive effects on tumors and other diseases. This study aimed to determine radiation-induced changes in spleen index, lymphocyte subsets, spleen cell apoptosis, and pathological features of the spleen in mice. The mouse model was established by irradiating the spleen at different doses. The mice were divided into the following groups: blank control, low-dose, low-dose fractionated irradiation, and challenge dose irradiation. The mice were sacrificed under humanitarian conditions, and spleen tissue and peripheral blood were collected. The spleen index was calculated, and flow cytometry was used to analyze spleen T lymphocyte subsets and spleen apoptosis. The pathological changes in the spleen were determined by hematoxylin and eosin (H&E) staining. The spleen index of mice in the low-dose fractionated irradiation group was significantly increased compared with that in the blank control group. The spleen indexes of the low-dose irradiation and low-dose fractionated irradiation groups were much higher than that of the challenge dose irradiation group. Compared with the blank control group, the percentage of CD3+ and CD4+ T lymphocytes in the peripheral blood and spleen tissues in the low-dose irradiation and low-dose fractionated irradiation groups was significantly increased, whereas that from the challenge dose irradiation group was obviously decreased. CD8+ T lymphocytes in the peripheral blood and spleen tissues in the low-dose irradiation, low-dose fractionated irradiation, and challenge dose irradiation groups were significantly lower than those in the blank control group. The apoptosis rate of the spleen in the challenge dose irradiation group was significantly higher than that in the blank control, low-dose irradiation, and low-dose fractionated irradiation groups. H&E staining analysis of the spleen showed pathological changes in the different irradiation groups compared with the blank control group. Low-dose irradiation and low-dose fractionated irradiation can change the T lymphocyte subsets in the peripheral blood and spleen of mice, which can promote immune excitation and improve immune effects.

Inhibition of JAK1/STAT3 pathway by 2-methoxyestradiol ameliorates psoriatic features in vitro and in an imiquimod-induced psoriasis-like mouse model

Psoriasis is characterized by hyperproliferative keratinocytes, dilated capillaries and leukocyte infiltration. 2-Methoxyestradiol (2-ME) has shown significant inhibition on proliferation, angiogenesis and inflammation. To evaluate the anti-psoriatic potential of 2-ME, psoriasis-like dermatitis was induced by topical application of imiquimod (IMQ) on the dorsal skin of C57BL/6 mice for seven consecutive days, followed by treatment of vehicle or 2-ME ointment from Day 4 on. The psoriasis area and severity index (PASI) was assessed daily. On Day 8, skin histology and spleen index were assessed. The effects of 2-ME on the proliferation, apoptosis, cell cycle, vascular endothelial growth factor A (VEGFA), and Janus kinase (JAK)-signal transducer and activator of transcription (STAT) pathways of HaCaT cells stimulated by interleukin-17 (IL-17A) were detected, together with its effect on the proliferation, tube formation and VEGF receptor expression of human umbilical vein endothelial cells (HUVECs). We found that topical 2-ME treatment significantly improved IMQ-induced psoriasis-like dermatitis and decreased the PASI scores, the activation of STAT3 in the skin (P < 0.05), and the spleen index in mice (P < 0.01). In vitro, 2-ME inhibited the proliferation of HaCaT cells by inducing apoptosis and G2/M phase arrest (P < 0.01). Moreover, 2-ME suppressed IL-17A-induced VEGFA (2.5 μM: P < 0.05; 5 μM: P < 0.01) and phosphorylation of STAT3 by blocking p-JAK1 in HaCaT cells and prevented tube formation (P < 0.01) and proliferation by targeting VEGF receptors 1 (VEGFR1) and 2 (VEGFR2) in HUVECs. We conclude that 2-ME alleviated psoriasis in vivo and in vitro by inhibiting JAK1/STAT3 pathway and was a promising therapeutic agent for psoriasis.

THE EFFECT OF KESAMBI BARK EXTRACT ON MICE LYMPHOCYTE COUNT AND SPLEEN INDEX

Kesambi is a plant widely used by people of Timor Island. Flavonoid content of Kesambi is potential to be used as immunomodulators. This study aimed to study the effect of Kesambi bark extract on lymphocyte count and spleen index of male mice as an indicator of their immune responses. Research included identification of phytochemical compounds macerated from Kesambi bark, lymphocytes count and spleen index calculation. Mice injected with sheep erythrocytes were treated orally with three doses of Kesambi extract (0.113 mg/g BW, 0.491 mg/g BW, and 0.712 mg/g BW) for 10 days. Lymphocyte count using blood smear method was carried out on day 0, 5 and 10, while lymphocyte count using hematology analyzer and spleen index measurement were performed after mice sacrificed. Kesambi bark contains alkaloids, flavonoids, tannins, and steroids. Treatment with Kesambi bark extract reduced mice lymphocyte count. Administration of 0.491 mg/gr BW dose reduced the mice lymphocytes count close to the number of lymphocytes in the negative control group. The spleen index of mice had no correlation with changes in the number of lymphocytes. This preliminary study shows a potential use of Kesambi bark extract as a natural immunosuppressant.

Protective effects of flavonoids isolated from Agrocybe aegirita on dextran sodium sulfate‐induced colitis

Mushroom derived phytochemical has become the promising agent to treat inflammatory bowel disease (IBD). Here, we investigated the effect of flavonoids from Agrocybe aegirita (AAF) on dextran sodium sulfate‐induced colitis. Our results showed that flavonoids from Agrocybe aegirita had a certain effect on physical signs in mice (improving the weight loss of mice, reducing reducing the DAI index and the spleen index of mice). AAF could also significantly reduce the shortening of the colon, and improve the level of tissue damage and colon inflammation. Besides, AAF could alleviate the colon inflammatory status including reducing the levels of TNF‐α and IL‐1β and increasing the levels of IL‐10. In addition, AAF significantly promoted the growth of goblet cells and enhance the intestinal barrier function (the secretion of mucin in the colon were increased). In conclusion, flavonoids from Agrocybe aegirita has the potential to relieve the DSS‐induced colitis in mice and could be a novel therapy for combating with IBD.

Evaluation of Allicin Against Alveolar Echinococcosis In Vitro and in a Mouse Model

PurposeAt present, the chemotherapy for alveolar echinococcosis (AE) is mainly based on albendazole (ABZ). However, more than 20% of patients fail chemotherapy. Therefore, new and more effective treatments are urgently needed. Allicin has been reported to have antibacterial and antiparasitic effects. The objectives of the present study were to investigate the in vivo and in vitro efficacy of allicin against Echinococcus multilocularis (E. multilocularis).MethodsThe effects of allicin on protoscolex survival and structural changes were evaluated in vitro. The 4-week-old BALB/c male mice used for in vivo modelling underwent inoculation of E. multilocularis protoscoleces by intraperitoneal injection, followed by intragastric administration of allicin for 6 weeks. Then, the effects of allicin on lymphocyte subsets, metacestode growth and host tissue matrix metalloproteinase 2 (MMP2)/MMP9 expression around metacestodes in mice were evaluated. The toxicity of allicin was further evaluated in vivo and in vitro.ResultsAtt 40 μg/mL, allicin showed a killing effect on protoscoleces in vitro and treatment resulted in the destruction of protoscolex structure. Molecular docking showed that allicin could form hydrogen bonds with E. multilocularis cysteine enzymes. After 6 weeks of in vivo allicin treatment, the spleen index of mice was increased and the weight of metacestodes was reduced. Allicin increased the proportion of CD4+ T cells and decreased the proportion of CD8+ T cells in the peripheral blood and spleen. Pathological analysis of the metacestodes showed structural disruption of the germinal and laminated layers after allicin treatment. In addition, allicin inhibited the expression of MMP2 and MMP9 in metacestode-surrounding host tissues. At 160 μg/mL, allicin had no significant toxicity to normal hepatocytes but could inhibit hepatoma cell proliferation. At 30 mg/kg, allicin had no significant hepatorenal toxicity in vivo.ConclusionThese results suggest that allicin exerts anti-E. multilocularis effects in vitro and in vivo and can enhance immune function in mice, with the potential to be developed as a lead compound against echinococcosis.

Effect of bushen qingre yuyin decoction on salivary secretion, spleen index, submandibular gland index, submandibular gland histomorphology, and aqp5 expression in the nonobese diabetic mouse model

Objective: The objective of this study was to observe the effect of Bushen Qingre Yuyin Decoction on the spleen index, submandibular gland index, salivary secretion, submandibular gland morphology, and aquaporin 5 (AQP5) expression levels in the submandibular gland. Materials and Methods: Fifty 8-week-old nonobese diabetic (NOD) female mice were randomly divided into the following five groups: model; hydroxychloroquine; and high-, medium-, and low-dose Bushen Qingre Yuyin Decoction prescription. The blank group consisted of ten 8-week-old Balb/C rats. Bushen Yuyin Qingre Decoction was administered by gavage at 20, 40, and 80 g/kg in the low-, medium-, and high-dose groups, respectively. The hydroxychloroquine group was administered hydroxychloroquine at a dose of 80 mg/kg by gavage. The normal and model groups were administered the same amount of saline. After the different administrations, the amount of salivary secretion in the mice was regularly observed. After 12 weeks, the mice were sacrificed, and their submandibular gland tissues were excised, stained with hematoxylin-eosin, and pathologically scored. AQP5 expression levels in the tissues were detected using real-time polymerase chain reaction. Results: The spleen index of mice in the high-dose group was higher than that in the model group (P < 0.05), and the submandibular gland index of the hydroxychloroquine group and all the Bushen Qingre Yuyin Decoction prescription groups was higher than that in the model group (P < 0.05). After 6 weeks, the salivary volume of the hydroxychloroquine group and all the Bushen Qingre Yuyin Decoction prescription groups was significantly higher than that of the model group (P < 0.05); the amount of lymphocyte infiltration in mice among each administration group was significantly reduced (P < 0.05); and the AQP5 expression levels in the submandibular glands of mice had significantly increased in the hydroxychloroquine and high-dose groups (P < 0.05), with the high-dose group showing the best effect. Conclusion: Bushen Qingre Yuyin Decoction can effectively increase the spleen and submandibular gland indexes of NOD mice, increase the amount of salivary secretion, reduce lymphocyte infiltration in submandibular gland tissue, and significantly increase AQP5 expression levels, which may be positively correlated with the administered dose.

Repetitive restraint stress changes spleen immune cell subsets through glucocorticoid receptor or β-adrenergic receptor in a stage dependent manner

Immune system is sensitive to stress. Spleen is the largest peripheral immune organ innervated with sympathetic nerves and controlled by adrenomedullary system in the body. However, the alterations and mechanism of spleen immune cell subsets caused by repetitive restraint stress (RRS) is poorly understood. In this study, we found that RRS reduced spleen index in mice, and induced an expansion of white pulp and involution of the red pulp. Meanwhile, the percentage of CD3+CD8+ T lymphocytes, CD11b+F4/80+ macrophages, CD11b+Ly-6G−Ly-6Chi monocytic myeloid derived suppressor cells (mMDSCs) and CD11b+Ly-6G+Ly-6Cint granulocytic myeloid derived suppressor cells (gMDSCs) in spleen were significantly changed by RRS. Mechanistically, we found that the expression of norepinephrine (NE) and β-adrenergic receptor (β-AR) in spleen were up-regulated after 21 days of RRS, but not 7 days. The expression of corticosterone (CORT) and glucocorticoid receptor (GR) in spleen were up-regulated after 7 days of RRS but were lower after 21 days of RRS, even though they were still higher than that in mice without stress. By treating the stressed mice with RU486 (antagonist of GR) or propranolol (antagonist of β-AR), we demonstrated that GR was responsible for the changes of spleen induced by 7 days of RRS and β-AR was for 21 days of RRS. Our data suggest that RRS changes spleen immune cell subsets through GR or β-AR in a stage dependent manner.

Characterization of a Novel Polysaccharide-Iron(III) Complex and Its Anti-Anemia and Nonspecific Immune Regulating Activities.

Dioscorea opposita Thunb is the famous food and traditional medicine in China and it was rich in polysaccharides. Polysaccharides of Dioscorea Opposita Thunb possess immunoregulatory activity, free radical scavenging activity and anti-diabetic activity. A novel polysaccharide- iron(III) complex (CYPIC) was synthesized by using crude polysaccharide extracted from Dioscorea opposita Thunb. The component, structure, morphology and molecular weights of CYPIC were analysed, and the anti-anemia, acute toxicity and nonspecific immune regulating activities of CYPIC were assayed. The results showed that CYPIC could increase red blood cell count (RBC), hemoglobin (Hb), hematocrit (HCT), thymus and spleen index of mice with iron deficiency anemia (IDA). Although the structure and deeper mechanisms of CYPIC should be further studied, CYPIC has the potential to be used as an iron supplement for the treatment of iron deficiency anemia. The large scale industrial production was suggested due to the simple preparation processing of CYPIC.

Protective effect of earthworm active ingredients against endoplasmic reticulum stress-induced acute liver injury in mice

To study the protective effect of earthworm active ingredients(EWAs) against endoplasmic reticulum stress(ERS)-induced acute liver injury in mice. The model of liver injury was induced through intraperitoneal injection of 10%CCl4. Serum glutamic-pyruvic transaminase(ALT), glutamic-oxaloacetic transaminase(AST), superoxide dismutase(SOD) and glutathione peroxidase(GSH-PX) activity and malondialdehyde(MDA) concentration were detected by colorimetric method. Histological examination was performed through hematoxylin-eosin staining and light microscopy, and apoptosis was detected using terminal transferase dUTP nick end labeling. The expressions of ERS related proteins, including glucose regulated protein 78(GRP78), protein kinase R-like ER kinase(PERK), eukaryotic transcription initiation factor 2α(eIF2α), active transcription factor-4(ATF4) and CCAAT/enhancer binding homologous protein(CHOP), were measured by immunohistochemistry and Western blot. According to the results, compared with the model group,serological indexes in the high, middle and low doses of EWAs were significantly improved (P<0.05 or P<0.01), the extent of liver lesion was decreased and the degree of injury was significantly reduced, and that the liver index and the spleen index of mice were significantly changed(P<0.05 or P<0.01). In liver tissue, the expressions of GRP78 and CHOP were significantly decreased(P<0.05 or P<0.01). The protein expressions of GRP78, CHOP and its upstream signaling pathway PERK-eIF2-ATF4 were significantly decreased in each dose group(P<0.05 or P<0.01). In summary, EWAs has a significant protective effect on ERS-induced acute liver injury, and its mechanism may be correlated with the inhibition of oxidative stress and ERS, and down-regulation of ERS marker protein CHOP expression, andinhibition of apoptosis.

Study on the therapeutic mechanisms of pseudolaric acid in mice with allergic contact dermatitis.

To study the therapeutic mechanisms of pseudolaric acid on allergic contact dermatitis in mice. A total of 50 BALB/C mice were selected and randomly divided into control group, model group, and treatment A, B, C groups with 10 rats in each group. ACD model was established in model group, and treatment A, B, C groups but not in control group. Model group received no treatment, but treatment A, B, C groups were treated with external application of the concentration of 0.1%, 0.2% and 0.4% of the pseudolaric acid for the lesions of ear skin. And the weight gain and the swelling degree of the mice' ear were recorded, weight of thymus and spleen were measured. Spleen suspension was prepared to test T lymphocyte and B lymphocyte levels of mice in five groups. Changes in serum IFN-γ, IL-4 and IL-10 levels were tested through the enzyme linked immunosorbent assay (ELISA). The weight gain of mice in model group were significant lower than those of mice in the control group and the treatment A, B, C groups (P<0.05). Weight gain of mice in treatment A, B groups were significant lower than that of control group (P<0.05), but the difference in weight gain between treatment C group and control group showed no significant difference (P>0.05). The swelling degree and the weight of mice ears in model group were significant higher than those of mice in control group and treatment A, B, C groups (P<0.05). Swelling degree and the weight of mice ears of treatment A, B, C groups were obviously higher than that of control group (P<0.05). The swelling degree and weight of mice' ears in treatment A, B, C groups were decreased with the increase of the drug dosage, but comparison between A, B and C group showed statistically differences (P<0.05). The thymus and spleen index of mice in model group were significant higher than those of the other four groups (P<0.05), among the four groups, thymus and spleen index of treatment A and B group were higher than control group and treatment C group (P<0.05). The stimulation index of T and B cells of mice in model group was significantly higher than the rest four groups (P<0.05). The serum IFN-γ level of mice in control group and treatment A, B and C group was obviously lower than that of mice in model group (P<0.05). The serum IFN-γ level of mice in treatment A, B and C group were decreased with the increasing of the drug dosage, and the level of C group was obviously lower than that of A and B group (P<0.05). The pseudolaric acid has anti-inflammation and immune adjustment the effects showing a remarkable therapeutic effects for the ACD mice.

脑舒胶囊对衰老小鼠免疫功能的调节作用

Objective: To observe the effects of NaoShu capsule on immune function of aging mice . Methods: Aging mice was induced by subcutaneous injections model was induced by subcutaneous injections with 5% D-galactose at the neck for 6 weeks, IL-2 and IL-6 content in serum were detected by means of radio-immunity, and calculating the index of organ by a ratio of the weight of thymus and spleen and the weight of mice. Results: Compared with the control group, the thymus and spleen index of mice was decreased obvious; the IL-2 content was obvious decreased, and the IL-6 content was obvious increased in aging model group. Compared with the model group, the the IL-2 content was enhanced and the IL-6 content was obvious decreased in the therapeutic group with 1.33g/kg and 2.66g/kg Naoshu capsule ( P <0.05, 0.01). Conclusion: There has a certain correlation between aging and immune, and there are obvious accommodation of Naoshu capsule on immune function of aging mice.

Anti-radiation effect and mechanism studies of ethanol extracts from Spatholobus suberectus and its active component catechin

To study the anti-radiation effect and mechanism of ethanol extracts from Spatholobus suberectus and its active component catechin, ICR mice were exposed to 6Gy irradiation and randomly divided into normal group, model group, positive control group (amifostine, 43.6 mg•kg⁻¹, iv 30 min before irradiation), SSD group (10, 20, 40 g•kg⁻¹) and catechin group (50, 100, 200 mg•kg⁻¹). The mice were administered the appropriate drugs once a day after irradiation for 28 consecutive days. Blood samples were collected from the tail end and the number of peripheral blood cells was counted before irradiation and on day 1, 3, 7, 14, 21 and 28 using a microcell counter. Changes of thymus and spleen index of mice on day 7 were observed. The serum SOD, GSH-Px activity and MDA level were detected by the colorimetric method. The colony forming ability of bone marrow hematopoietic progenitor cells on day 7 was detected by semi solid culture method. The HE staining was adopted to observe the pathological changes. The apoptosis of bone marrow cells was detected by flow cytometry. The expression of cleaved caspase-3 and Bax of bone marrow cells were measured separately by western-blotting and immunohistochemistry method. SSD and catechin can both significantly revert the irradiated-induced decline in hematological parameters (RBC, WBC, PLT, Hb), improve thymus and spleen index, significantly enhance serum SOD and GSH-Px activity and decrease the MDA level. The proliferation and differentiation of hematopoietic progenitor cells in bone marrow were promoted, the apoptosis of bone marrow cells was significantly up-regulated and the expression of cleaved caspase-3 and Bax was significantly reduced in SSD and catechin group. SSD and catechin have significant anti-radiation effect and its mechanism may be related to hematopoietic promoting, antioxidant and anti-apoptotic effects.

Exogenous VEGF promotes hematopoietic stem cell mobilization

This study was aimed to investigate the effect of exogenous VEGF on hematopoietic stem cell mobilization and immune system. The C57BL/6J mice were randomly divided into the normal control group, VEGF short-term group (5 d) and VEGF long-term group (27 d). Mice in the experimental group were injected ip with VEGF (100 ng/d); mice in control group were injected ip with PBS. The white blood cell (WBC) count and the ratio of lymphocyte in the peripheral blood at different time point were assayed by hemacytometer. The percentage of hematopoietic stem cell (HSC), lymphocyte subgroup, regulatory T cell (Treg), myeloid-derived suppressor cells (MDSC) in the peripheral blood and spleen of different groups were detected by flow cytometry. The morphological changes of spleen and spleen index of mice in the control and long-term group were observed by microscopy. The results showed that the absolute number of WBC in the peripheral blood of mice significantly increased after injection of VEGF, and the peak value was at day 3. The percentage of Lin(-)Sca-1(+)CD117(+) cells in the peripheral blood and spleen of the long-term group were significantly higher than that in the normal control group (P < 0.05). The spleen of the mice in VEGF long-term group was larger than that of the control group, the spleen index also increased (P < 0.05), and remarkable extramedullary hematopoietic signs were found in the HE stained sections. There was no significant change in the total ratio of lymphocytes in the peripheral blood after injection, but the percentage of CD3(+) cells and the CD3(+)/B220(+) ratio in the long-term group deceased; the percentages of Treg and Gr-1(+)CD11b(+) MDSC in the experimental groups increased (P < 0.05), which more significantly increased in the long-term group than that in the short-term group (P < 0.05). It is concluded that the exogenous VEGF promotes hematopoietic stem cell mobilization, and at same time up-regulates the many kinds of suppressive immune cell levels which leads to changes of immuno-function.

Antitumor effect and mechanism of Gecko on human esophageal carcinoma cell lines in vitro and xenografted sarcoma 180 in Kunming mice

To investigate the anti-tumor effect of Chinese medicine Gecko on human esophageal carcinoma cell lines and xenografted sarcoma 180 in Kunming mice and its mechanism. The serum pharmacological method was used in vitro. The growth rates of the human esophageal carcinoma cells (EC9706 or EC1) were measured by a modified 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The transplanted tumor model of the mouse S180 sarcoma was established. Fifty mice were randomly divided into five groups (n=10). Three Gecko groups were treated respectively with oral administration of Gecko powder at a daily dose of 13.5 g/kg, 9 g/kg, and 4.5 g/kg. The negative group (NS group) was treated with oral administration of an equal volume of saline and the positive group (CTX group) was treated with 100 mg/kg Cytoxan by intraperitoneal injection at the first day. After 2 wk of treatment, the anti-tumor activity was evaluated by tumor tissue weighing. The impact on immune organ was detected based on the thymus index, spleen index, phagocytic rate and phagocytic index. The protein expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) were detected by immunohistochemistry. The cell apoptotic rate was detected by terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay. The OD value in each group treated with Gecko after 72 h was reduced significantly in EC9706 and in EC1. The tumor weight in each group of Gecko was decreased significantly (1.087+/-0.249 vs 2.167+/-0.592; 1.021+/-0.288 vs 2.167+/-0.592; 1.234+/-0.331 vs 2.167+/-0.592; P<0.01, respectively). However, the thymus index and Spleen index of mice in Gecko groups had no significant difference compared with the NS group. The immunoreactive score of VEGF and bFGF protein expression of each Gecko group by immunohistochemical staining were lowered significantly. The apoptosis index (AI) of each group was increased progressively with increase of dose of Gecko by TUNEL. Gecko has anti-tumor effects in vitro and in vivo; induction of tumor cell apoptosis and the down-regulation of protein expression of VEGF and bFGF may be contributed to anti-tumor effects of Gecko.