Epimedium Polysaccharide Research Articles

Epimedium polysaccharides ameliorate ulcerative colitis by inhibiting oxidative stress and regulating autophagy.

Epimedium polysaccharide (EPS) is a bioactive compound derived from the traditional Chinese herb Epimedium brevicornum. The objective of this study was to investigate the protective effects of EPS on ulcerative colitis (UC) and to elucidate the underlying mechanisms involved. The findings showed that EPS treatment mitigated UC symptoms, including weight loss, anal bleeding, elevated disease activity index (DAI), and colon shortening. Hematoxylin and eosin (H&E) and Alcian blue-periodic acid-Schiff (AB-PAS) staining demonstrated that EPS alleviated histopathological damage and improved the integrity of the colonic mucosa. Mechanistically, EPS was found to substantially decrease inflammation by inhibiting the Toll-like receptor 4/nuclear factor-κB (TLR4/NF-κB) signaling pathway and to alleviate oxidative stress through modulation of the Kelch-like ECH-associated protein 1/nuclear factor erythroid-derived 2-like 2 (Keap1/Nrf2) pathway. Notably, EPS failed to exert protective effects against dextran sulfate sodium (DSS)-induced UC in Nrf2-knockout (Nrf2-/-) mice. Additionally, Western blotting and immunohistochemical analysis demonstrated that EPS facilitated autophagy via the adenosine monophosphate-dependent protein kinase/mammalian target of rapamycin (AMPK/mTOR) pathway. In vitro experiments revealed that EPS effectively suppressed lipopolysaccharide (LPS)-mediated cellular damage and oxidative stress by regulating Keap1/Nrf2 pathway. Transcriptomic analysis of LPS-treated Caco-2 cells following EPS treatment revealed a significant up-regulation of Nrf2 expression. In conclusion, the findings of this study suggest that EPS exerts protective effects against DSS-induced UC through the inhibition of the TLR4/NF-κB signaling pathway, regulation of the Keap1/Nrf2 pathway, and promotion of autophagy via the AMPK/mTOR pathway. Consequently, EPS may represent a potential therapeutic target for the treatment of UC. © 2024 Society of Chemical Industry.

Epimedium polysaccharides mitigates Porphyromonas gingivalis-exacerbated intestinal inflammation by suppressing the Th17 pathway and modulating the gut microbiota

This study aimed to investigate the potential alleviating effect of Epimedium polysaccharide (EP) on intestinal inflammation aggravated by Porphyromonas gingivalis (Pg). P. gingivalis, an oral pathogen, may play a role in intestinal inflammation, highlighting the necessity to explore substances capable of inhibiting its pathogenicity. Initially, in vitro screening experiments utilizing co-culturing and quantitative polymerase chain reaction revealed that EP significantly inhibited the growth of P. gingivalis and the levels of virulence genes, including Kgp and RgpA. Subsequent mouse experiments demonstrated that EP notably ameliorated Pg-aggravated weight loss, disease activity index, histopathological lesions, and disruption of intestinal barrier integrity, evidenced by a reduction in tight junction protein levels. Flow cytometry analysis further illustrated that EP attenuated Pg-induced Th17 differentiation and Th17-related cytokines, such as IL-17 and IL-6. Additionally, 16S rRNA amplicon sequencing analysis elucidated that EP significantly mitigated Pg-induced gut microbiota dysbiosis, enriching potentially beneficial microbes, including Akkermansia and Bifidobacterium. The metabolomic analysis provided further insight, indicating that EP intervention altered the accumulation of relevant intestinal metabolites and exhibited correlations with disease indicators. In conclusion, our research suggested that EP holds promise as a prospective therapeutic agent for alleviating P. gingivalis-aggravated intestinal inflammation.

Gut microbiota and serum metabolome reveal the mechanism by which TCM polysaccharides alleviate salpingitis in laying hens challenged by bacteria

This paper aimed to evaluate the effect of 3 kinds of TCM polysaccharides instead of antibiotics in preventing salpingitis in laying hens. After feeding the laying hens with Lotus leaf polysaccharide, Poria polysaccharide, and Epimedium polysaccharide, mixed bacteria (E. coli and Staphylococcus aureus) were used to infect the oviduct to establish an inflammation model. Changes in antioxidant, serum immunity, anti-inflammatory, gut microbiota, and serum metabolites were evaluated. The results showed that the 3 TCM polysaccharides could increase the expression of antioxidant markers SOD, GSH, and CAT, and reduce the accumulation of MDA in the liver; the contents of IgA and IgM in serum were increased. Decreased the mRNA expression of TLR4, NFκB, TNF-α, IFN-γ, IL1β, IL6, and IL8, and increased the mRNA expression of anti-inflammatory factor IL5 in oviduct tissue. 16sRNA high-throughput sequencing revealed that the 3 TCM polysaccharides improved the intestinal flora disturbance caused by bacterial infection, increased the abundance of beneficial bacteria such as Bacteroides and Actinobacillus, and decreased the abundance of harmful bacteria such as Romboutsia, Turicibacter, and Streptococcus. Metabolomics showed that the 3 TCM polysaccharides could increase the content of metabolites such as 3-hydroxybutyric acid and isobutyl-L-carnitine, and these results could alleviate the further development of salpingitis. In conclusion, the present study has found that using TCM polysaccharides instead of antibiotics was a feasible way to prevent bacterial salpingitis in laying hens, which might make preventing this disease no longer an issue for breeding laying hens.

Effects and mechanism of epimedium polysaccharide on solubility of icariin and baohuoside Ⅰ

This study evaluated the effects of epimedium polysaccharide(EPS) on the solubility of icariin and baohuoside Ⅰ so as to preliminary explore its solubilization function and the underlying mechanism. The solubility of these two insoluble flavonoids in water and polysaccharide solutions was compared by high performance liquid chromatography, and the mechanism was investigated by diffe-rential scanning calorimetry(DSC) and critical micelle concentration determination. The results indicated that their solubilization in crude EPS solutions was concentration-dependent. The solubility of icariin and baohuoside Ⅰ in 20 mg·mL~(-1) EPS-1-1 was 9.05 times and 5.76 times that in water, respectively; while their solubility in 20 mg·mL~(-1) EPS-2-1 was 10.55 and 8.39 times that in water, respectively. The change of the DSC thermograms suggested the formation of new complexes from icariin and baohuoside Ⅰ with polysaccharides. The critical micelle concentrations proved the micellar properties of both EPS-1-1 and EPS-2-1. In short, EPS can significantly increase the solubility of icariin and baohuoside Ⅰ, the mechanism of which may be related to the formation of micellar complexes between EPS and insoluble flavonoids.

Effect and Mechanism of Epimedium Polysaccharide on Bone Marrow Hematopoietic Function and Th17/Treg Balance in Aplastic Anemia

To observe the effects of Epimedium polysaccharides (EPS) on bone marrow hematopoietic function and Th17/Treg balance in aplastic anemia (AA) mice, and preliminarily explore its therapeutic mechanism. Forty BALB/C mice were randomly divided into control (control), model (model), stanozolol (stanozolol) and epimedium polysaccharide (EPS) group, with 10 mice in each group. Except for the control group, Acetophenazine, Gy irradiation and cyclophosphamide triple application were used to establish AA models for the other groups. After the model was established, the stanozolol group was intragastrically administered with 4 mg/kg stanozolol suspension, the EPS group was intragastrically administered with 100 mg/kg epimedium polysaccharide, while the control group and the model group were given an equal volume of 0.9% sodium chloride solution by gavage once a day, for 14 consecutive days. The automatic animal blood analyzer was used to detect the changes in peripheral blood hemoglobin (Hb), red blood cells (RBC), white blood cells (WBC) and platelets (PLT), flow cytometry was used to detect the proportion of Treg and Th17 cells, the levels of interleukin 2 (IL-2), interleukin 11 (IL-11) and tumor necrosis factor α (TNF-α) were detected by ELISA, the number of nucleated bone marrow cells was counted, HE staining and immunohistochemical staining were used to detect the number, the proliferation and apoptosis of bone marrow cells, Western blot was used to detect the expression of signal transducer and activator of transcription 3 (STAT3), retinoic acid receptor-related orphan receptor γ (RORγt), transducer and activator of transcription 5 (STAT5) and fork head transcription factor 3 (Foxp3). Compared with the model group, the levels of Hb, RBC, WBC and PLT in the peripheral blood of mice in stanozolol and EPS group significantly increased, the proportion of Th17 cells was significantly reduced, and the proportion of Treg cells significantly increased. The levels of IL-2 and TNF-α in serum were significantly reduced (P<0.05), the level of IL-11 significantly increased (P<0.05), the number of bone marrow nucleated cells significantly increased (P<0.05), the positive rate of Ki-67 significantly increased (P<0.05) and the positive rate of Caspase-3 was significantly reduced (P<0.05). At the same time, the protein expression of STAT3 and RORγt significantly decreased, and the protein expression of STAT5 and Foxp3 increased, the difference showed statistically significant (P<0.05). EPS can promote the recovery of bone marrow hematopoietic function in AA mice and improve Th17/Treg imbalance, the mechanism may be related to the inhibition of STAT3/RORγt expression and promotion of STAT5/Foxp3 expression.

Immune-related effects of compound astragalus polysaccharide and sulfated epimedium polysaccharide on newborn piglets

This study aimed to evaluate the immune effects of compound astragalus polysaccharide and sulfated epimedium polysaccharide (APS-sEPS) on the peripheral blood lymphocyte and intestinal mucosa in newborn piglets. A total of 40 newborn piglets were randomly divided into four groups during a 25-day experiment, including APS-sEPS, APS, sEPS and control group. The results showed that supplementation with APS-sEPS to newborn piglets remarkably increased the physiological parameters, especially the WBC. In peripheral blood, piglets that received APS-sEPS showed the highest proliferation of T lymphocytes, the percentage of CD3 + CD4+ and CD3 + CD8+ cells were the highest on days 15 and 25 (p < 0.05). The serum concentrations of IFN-γ on days 7 and 15, and IL-4, IL-10, sIgA on days 7, 15 and 25 in APS-sEPS group were significantly higher than those in the control group (p < 0.05). Furthermore, the villus length and the ratio of villus length to crypt depth in APS-sEPS group were both significantly increased compared to that of control group (p < 0.05). In the duodenum, jejunum and illume, the concentrations of IFN-γ, IL-10, total IgG and sIgA in APS-sEPS group were all significantly higher than that in control group (p < 0.05). In intestinal mucosa, APS-sEPS significantly increased the expression of NF-κB and IRF-3 mRNA in each section of small intestine of piglets. Nevertheless, in the illume segment, the effect of APS-sEPS was more significant than that of APS and sEPS (p < 0.05). The expression of TLR4 was more significant than that of control group in duodenum only. The results from the present research provide evidence that the suckling piglets administered with APS-sEPS supplement exhibited enhanced immune function of peripheral blood lymphocyte and expression of specific antibodies, and ameliorated intestinal morphological development and increased activities of humoral immune response in the small intestine, which would be related to the activation of the TLR4-NF-κB signaling pathway and IRF3.

Epimedium Polysaccharide Ameliorates Benzene-Induced Aplastic Anemia in Mice.

Benzene (BZ) is an important occupational and environmental pollutant. Exposure to BZ may cause aplastic anemia which is characterized as bone marrow hematopoietic failure. In order to reduce the harmful effects of this pollutant, it is necessary to identify additional preventative measures. In this study, we investigated the protective effects of epimedium polysaccharide (EPS), a natural compound with antioxidant and immune-enhancing potency, on aplastic anemia induced by benzene exposure in mice. Male CD-1 mice were randomly divided into five groups including control, BZ (880 mg/kg), LE (EPS low-dose, 20 mg/kg + BZ), ME (EPS middle-dose, 100 mg/kg + BZ), and HE (EPS high-dose, 200 mg/kg + BZ) groups. Animals were exposed to BZ by subcutaneous injection in the presence or absence of EPS via oral administration. All mice were treated 3 times a week for 8 consecutive weeks to develop a mouse model of benzene-induced aplastic anemia (BIAA). Results showed that BZ induced a significant decrease in both white and red blood cells, platelet counts, and hemoglobin level compared with that in the control group (p < 0.01). Treatment of EPS led to a protective effect against these changes particularly in the highest-dose group (HE, p < 0.01). EPS also recovered the decreased number of nucleated cells in peripheral blood cell smears and femur biopsies by BZ exposure. The increased level of reactive oxygen species (ROS) in bone marrow mononuclear cells (BMMNCs) in mice from the BZ group was significantly lower (p < 0.01) in the mice from the highest concentration of EPS (HE) group when compared with that from the control group. In addition, BZ exposure led to a significant increase in the apoptosis rate in BMMNCs which was prevented by EPS in a dose-dependent manner (p < 0.01). The antiapoptosis effect of EPS was through reversing apoptotic proteins such as BAX, Caspase-9 and Caspase-3, and Bcl-2. Finally, EPS treatment partially restored the levels of T cells and the different subtypes except CD80+ and CD86+ compared with the BZ group (HE, p < 0.05). These results suggest that EPS has protective effects against BIAA via antioxidative stress, immune modulation, and antiapoptosis mechanisms.

Immunological modulation effects of an acid Epimedium polysaccharide on immune response in chickens

The purpose of the present study is to investigate the immunological activities of EPS-1 in the non-specific immune response and specific immune response of chickens. In vitro, the results showed that EPS-1 could increase the proliferation and cytokine secretion (IL-2, IL-4, IFN-γ and TNF-α) of spleen lymphocytes, expression of key surface molecules (MHC II, CD11c, CD40 and CD86) and cytokine secretion (TNF-α and IL-10) of matured chBM-DCs, phagocytic rate of matured chBM-DCs, and enhance the maturation and stimulating capacity of chBM-DCs. In vivo, EPS-1 could also prompt the HI antibody titer, boost the peripheral lymphocyte proliferation, enhance the release of cytokine products in blood (IFN-γ, IL-4 and IL-2) and duodenum (IL-17 and sIgA) of chickens. These results indicated that EPS-1 may have the potential as a powerful immune adjuvant in the treatment of chicken diseases.

The mucosal adjuvant effect of plant polysaccharides for induction of protective immunity against Helicobacter pylori infection

Some plant polysaccharides (PPSs) had been used as the adjuvants for systemic vaccination. In this study, we investigated whether PPSs could exhibit adjuvant effect at the mucosa. Groups of mice were intranasally immunized with Epimedium Polysaccharide (EPS), Trollius chinensis polysaccharide (TCPS), Siberian solomonseal rhizome polysaccharide (SSRPS) and Astragalus polysaccharides (APS) together with ovalbumin (OVA). Significantly higher levels of OVA-specific IgG in serum and secretory IgA in saliva, vaginal wash and intestinal lavage fluid were induced after immunization with OVA plus one of the four PPSs compared to OVA alone. Antigen absorption and TLR2 (Toll-like receptor 2) activation may be related to their mucosal adjuvant effect. Of note, when APS used as an adjuvant, intranasally vaccination with recombination UreB (rUreB, Urease subunit B) conferred more robust protection against Helicobacter pylori (H. pylori). Immunized with rUreB in combination APS resulted in mixed specific Th1 and Th17 immune response, which may contribute to the inhibition of H. pylori colonization. Though specific Th2-dominant responses were elicited when the other three PPS intranasally immunized with rUreB, no significant difference in the protective effect were found between those groups and rUreb alone group. Taken together, the four PPSs may be promising candidates for mucosal adjuvant, and APS could enhance rUreB-specific protective immunity against H. pylori infection.

EpimediumPolysaccharide Alleviates Polyglutamine-Induced Neurotoxicity inCaenorhabditis elegansby Reducing Oxidative Stress

Epimedium has been traditionally used to treat a variety of medical conditions, including neurological disorders. In this study, an acidic polysaccharide EbPS-A1 is isolated from Epimedium brevicornum and found to contain mainly galacturonic acid, galactose, and rhamnose but also arabinose and glucuronic acid. Using Caenorhabditis elegans models, we show that EbPS-A1 is capable of inhibiting behavioral dysfunction mediated by polyglutamine (polyQ), which is implicated in several neurodegenerative disorders such as Huntington's disease. Interestingly, EbPS-A1 does not inhibit polyQ aggregation or extend lifespan in the nematodes; it does, however, improve the survival under increased oxidative stress of both polyQ and wild-type nematodes intoxicated by paraquat. Further studies reveal that EbPS-A1 is capable of not only scavenging free radicals in vitro but also reducing reactive oxygen species levels, enhancing antioxidant enzyme activities, and decreasing lipid peroxidation product in C. elegans models. Together, these results suggest that the protective effect of Epimedium polysaccharide against polyQ-mediated neurotoxicity is likely due to its antioxidant function.

Structural characterization of an acidic Epimedium polysaccharide and its immune-enhancement activity

One acidic polysaccharide named EPS-1 was isolated from the aqueous extract of the leaves of Epimedium acuminatum Franch. It may be composed of 1,4-linked α-d-GalpA, 1,3,4-linked α-d-GalpA, 1,6-linked β-d-Galp and terminal α-l-Rhap residues in a molar ratio of 11.0:1.0:1.0:1.0 by chemical and spectroscopic analysis. EPS-1 possessed immune modulation effects on peripheral T lymphocyte and immature chBM-DCs such as promoting the proliferation and cytokine secretion of these cells and increasing the phagocytosis ability of immature chBM-DCs.

Effects of Epimedium polysaccharide on female mouse (Mus musculus)ovarian and uterine development

This study was conducted to investigate the effects of Epimedium polysaccharide (EPS) on ovarian and uterine development in female mice and assess the in vitro maturation (IVM) rate. Healthy Kunming female mice were divided into 4 groups and administered 2, 4, 6, and 8 mg/0.1 mL of EPS, respectively; the control group was given 0.1 mL of 0.9% physiological saline solution. After 7 and 14 days, the mice were sacrificed and their reproductive tracts (uterus, oviduct, and ovaries) were removed. The reproductive tract was weighed; some ovaries were punctured to harvest oocytes for IVM culture and the others were used for histological analysis. EPS had virtually no effect on the body weight of the female mice. EPS at 4 mg/0.1 mL for 14 days of treatment had a significant effect (P < 0.05) on ovarian and uterine development when compared with 2, 6, and 8 mg/mL but no significant effect (P > 0.05) when compared with the control group. The number of follicles increased in the experimental groups and the arrangements of ovarian follicles were not negatively affected during histological observation. In conclusion, EPS at appreciable dosages has a positive effect on mouse ovarian development.

Effects of epimedium polysaccharide-propolis flavone oral liquid on mucosal immunity in chickens

A previous study found that epimedium polysaccharide (EP)-propolis flavonoid (PF) injection (EPI) produced reliable immunoenhancement. In this study, we investigate the effects of EP-PF oral liquid (EFO) on mucosal immunity in the chicken small intestine while using EPI, EP and PF as controls. Groups of fourteen-day-old chickens were given EFO orally at one of the three doses when they were vaccinated with ND vaccine. On days 7, 21 and 35 after the first vaccination, six chickens were selected randomly from each group for measurements of the sIgA and IL-17 contents of the washing liquors of the duodenum and jejunum, counts of the lymphocytes in the duodenal endothelium and counts of the IgA+ cells in the jejunal endothelium and cecum tonsil. The results indicated that EFO significantly promoted the secretion of sIgA and IL-17 and increased the numbers of lymphocyte and IgA+ cells. Furthermore, EFO was more efficient than EPI at the high and medium doses. These findings indicate that EPO may enhance intestinal mucosal immunity and may be exploited as an oral immunopotentiator.

Adjuvanticity of compound astragalus polysaccharide and sulfated epimedium polysaccharide per os

On the basis of previous researches, compound astragalus polysaccharide (APS) and sulfated epimedium polysaccharide (sEPS) oral liquid (AEO) was prepared. Three hundred and twenty 14-day-old chickens were randomly assigned into eight groups and vaccinated with ND vaccine except for blank control (BC) group, repeated vaccination at 28 days old. At the same time of each vaccination, the chickens in three experimental groups were taken orally with AEO, respectively, at three doses, in two component control groups with APS and sEPS, once a day for three successive days; in injection control group were injected with AEI once, and in vaccination control (VC) and BC groups were not administrated. On days 7, 14, 21, 28 and 35 after the first vaccination, peripheral lymphocyte proliferation, the serum antibody titer, IFN-γ and IL-2 concentrations and on day 35 immune organ index were measured. The results showed that AEO at high and medium doses could significantly promote lymphocyte proliferation and development of immune organ, enhance antibody titer and IFN-γ and IL-2 concentration, which was stronger than actions of AEI and two components. The results confirmed that AEO possessed reliable immunoenhancement and could be exploited into an oral immunopotentiator.

The preparation optimization and immune effect of epimedium polysaccharide-propolis flavone liposome

The preparation conditions of epimedium polysaccharide-propolis flavone liposome (EPL) were optimized by response surface methodology taking entrapment rates of epimedium polysaccharide and propolis flavone as indexes. The immunoenhancement of EPL prepared with optimized condition was determined taking epimedium polysaccharide-propolis flavone suspension (EPS) and epimedium polysaccharide-propolis flavone watery solution (EPW) as control. The results showed that the optimized preparation condition was as follows: the ratio of drug to lipid was 14:1, the ratio of soybean phospholipid to cholesterol was 6:1, and the ultrasonic time was 19min. EPL could significantly promote the proliferation of T and B lymphocytes singly or synergistically with PHA or LPS, mRNA expression of IL-2 and IL-6 and secretion of IgG and IgM as compared with EPS and EPW. These results indicated that liposome could significantly improve the immunoenhancement of epimedium polysaccharide-propolis flavone immunopotentiator (EPI) and would be as the suitable dosage form of EPI.

Astragalus polysaccharide and sulfated epimedium polysaccharide synergistically resist the immunosuppression

The immunoenhancement of compound polysaccharides, APS–sEPS composed with astragalus polysaccharide (APS) and sulfated epimedium polysaccharide (sEPS), was observed in immunosuppressed model chicken induced by cyclophosphamide (Cy). 11-day-old chickens were injected with Cy once a day for three successive days except vaccine control group. At day-14-old, all chickens were vaccinated with ND vaccine, and in experimental groups simultaneously administrated with APS–sEPS at three dosages, APS and sEPS once a day for three successive days. On days 7, 14, 21 and 28 after the administration, the peripheral T-lymphocyte proliferation, serum antibody titers, IFN-γ, IL-2, IgG and IgM were determined. The results displayed that APS–sEPS could overcome Cy-induced immunosuppression, significantly promote T-lymphocyte proliferation and raised serum antibody titers, IFN-γ, IL-2, IgG and IgM levels, its high and medium doses were superior to single APS or sEPS. This demonstrated that APS and sEPS could synergistically resist the immunosuppression and APS–sEPS was an effective immunopotentiator.

Optimization on preparation condition of epimedium polysaccharide liposome and evaluation of its adjuvant activity

The aim of this strategy was to investigate whether the adjuvant activity of epimedium polysaccharide (EPS) could be further enhanced after encapsulated with liposome. In preparation of EPS liposome (EPSL) test, an orthogonal L9 (34) test design was used to optimize the preparation condition of EPSL. In adjuvant activity test, 350 14-day-old chickens were randomly assigned to 7 groups and vaccinated with Newcastle disease (ND) vaccine. Simultaneously, the chickens in experimental groups were injected with EPSL at three doses, EPS and blank liposome, respectively. The activity of lymphocytes proliferation, titer of serum antibody and concentrations of cytokines were determined. Results showed that the optimal preparation condition of EPSL was that ratio of drug to lipid, ratio of soybean phospholipid to cholesterol, ultrasonic time, and water bath temperature were 1:30, 4:1, 10min and 40°C, respectively. EPSL could significantly enhance the immune response of ND vaccine and promote cytokines secretion, and its high dose possessed the best efficacy. These findings indicated that liposome encapsulation could significantly improve the adjuvant activity of EPS.

Epimedium polysaccharide and propolis flavone can synergistically inhibit the cellular infectivity of NDV and improve the curative effect of ND in chicken

Four prescriptions, epimedium flavone plus propolis flavone (EF-PF), epimedium flavone plus propolis extracts (EF-PE), epimedium polysaccharide plus propolis flavone (EP-PF) and epimedium polysaccharide plus propolis extracts (EP-PE), were prepared and their antiviral effects were compared. In test in vitro, the four prescriptions within safety concentration scope and Newcastle disease virus (NDV) were added into cultured chick embryo fibroblast (CEF) in three modes, pre-, post-adding drug and simultaneous-adding drug and virus after being mixed, the cellular A570 values were determined by MTT method and the highest virus inhibitory rates were calculated to compare the antiviral activity of four prescriptions. In test in vivo, three hundred 21-day-old chickens were randomly divided into 6 groups and challenged with NDV except for blank control group. After 24h the chickens in four prescription groups were injected with corresponding drugs respectively, in virus control and blank control groups, with physiological saline, once a day for three successive days. On days 3, 7 and 14 after challenge, the serum antibody titer was determined. On day 15 after challenge, the mortality, morbidity and cure rate in every group were counted. The results showed that the most of A570 values in EP-PF group were numberly or significantly larger than those of the corresponding virus control group and the highest virus inhibitory rates of EP-PF at optimal concentration group were the highest among four prescription groups in three drug-adding modes, which confirmed that EP-PF could significantly inhibit the infectivity of NDV to CEF, its action was stronger than those of other three prescriptions; in EP-PF group, the antibody titers and cure rate were the highest and the mortality and morbidity were lowest presenting numberly or significantly differences in comparison with other three prescription groups. These results indicated that epimedium polysaccharide and propolis flavone possessed synergistical action, EP-PF prescription could significantly inhibit the cellular infectivity of NDV, improve the curative effect of ND in chicken and would be expected to exploit into a new-type antiviral drug.

Epimedium polysaccharide and propolis flavone can synergistically stimulate lymphocyte proliferation in vitro and enhance the immune responses to ND vaccine in chickens

Four prescriptions, epimedium flavone plus propolis flavone (EF–PF), epimedium flavone plus propolis extracts (EF–PE), epimedium polysaccharide plus propolis flavone (EP–PF) and epimedium polysaccharide plus propolis extracts (EP–PE), were prepared and their immune-enhancing effects were compared. In test in vitro, the effects of them on chicken peripheral lymphocyte proliferation were determined by MTT method. The results showed that EP–PF group presented the highest stimulating index at most concentrations. In immune test, 300 14-day-old chickens were randomly divided into six groups and vaccinated with ND vaccine except for blank control (BC) group, re-challenged at 28 days of age. At the same time of the first vaccination, the chickens in four experimental groups were injected, respectively, with four prescriptions. The changes of the lymphocyte proliferation and antibody titer were determined. On day 28 after the first vaccination, the chickens except for BC group were challenged with NDV, the immune protective effect was observed. The results displayed that in EP–PF group, the antibody titers, lymphocyte proliferation and protective rate were the highest, the morbidity and mortality were the lowest. In dose test, 14-day-old chickens were randomly divided into five groups. The treatment and determinations were the same as the immune test except that the chickens in experimental groups were injected, respectively, with high, medium and low doses of EP–PF. The results revealed that in medium dose group, the antibody titers, lymphocyte proliferation and protective rate were the highest, the morbidity and mortality were the lowest. These results indicated that EP and PF possessed synergistically immune enhancement, EP–PF had the best efficacy, especially at medium dose, and would be expected to exploit into a new-type immunopotentiator.

Compound Chinese herbal medicinal ingredients can enhance immune response and efficacy of RHD vaccine in rabbit

In order to validate the immune-enhancement efficacy of compound Chinese herbal medicinal ingredients (cCHMIs), made with astragalus polysaccharide (APS), epimedium polysaccharide (EPS), propolis flavone (PF) and ginsenosides (GS), as immune potentiator or vaccine adjuvants for rabbits, the effects of two cCHMIs on rabbit lymphocyte proliferation and IFN-γ and IL-10 mRNA expression of T lymphocyte in vitro were determined. At the same time, two cCHMIs were injected into 35-day-old rabbits after mixed with rabbit hemorrhagic disease (RHD) vaccine taking aluminum adjuvant and phosphate-buffered saline (PBS) as controls. On days 7, 14, 21, 35 and 49 after the vaccination, the dynamic changes of peripheral lymphocyte proliferation and serum antibody titers of the rabbits were analyzed. On day 63, all rabbits were challenged with RHD virus. The results showed that the two cCHMIs could significantly promote rabbit lymphocyte proliferation and IFN-γ and IL-10 mRNA expression of T lymphocyte in vitro. In vivo, two cCHMIs could significantly enhance serum antibody titers and lymphocyte proliferation. Their adjuvanticity was slightly superior to aluminum adjuvant. All the rabbits vaccinated with the cCHMIs adjuvant vaccine were protected. These findings confirmed that two cCHMIs possessed better immune-enhancement efficacy and would be used as effective immune adjuvant of RHD vaccine.