Cyclophosphamide-induced Immunosuppressed Mice Research Articles

Molecular Mechanism of the Regulatory Effect of Schisandrol A on the Immune Function of Mice Based on a Transcription Factor Regulatory Network.

The molecular mechanism of the regulatory effed of schizandrol A (SA) on the immune function of cyclophosphamide-induced immunosuppressive mice was explored in this study. On the basis of 1619 differentially expressed genes related to the regulatory effect of SA on the immune function of mice screened in our previous study, transcription factors and their corresponding target genes were screened in the Transcriptional Regulatory Element Database (TRED), and a transcription factor target gene regulatory network was constructed. The key nodes of the network were statistically analyzed to clarify the role of transcription factors in the regulatory network. The correlation of network genes with diseases was analyzed with an online annotation tool through the Database for Annotation, Visualization and Integrated Discovery (DAVID). Finally, the key factors related to the regulatory effect of SA on the immune function of mice were screened and verified by animal experiments and the detection of related protein expression by western blot analysis. The results showed that SA could alleviate the immunosuppression induced by cyclophosphamide in mice and regulate the protein expression of Jun, Trp53, and Creb1 in the spleen tissue of mice, together with the transcription factors Atf4 and E2f2. SA may thus play a role in the alleviation of some immunity-related diseases (such as cancer) by regulating the immune function of mice through multiple genes and their transcription factors.

The Immunomodulatory Effects of Phellodendri Cortex Polysaccharides on Cyclophosphamide-Induced Immunosuppression in Mice.

Cyclophosphamide is a commonly used anticancer drug, and immunosuppression is one of the most common side effects. How to recover the immunological function is important for cyclophosphamide-treated patients. In the present study, Phellodendri Cortex polysaccharides (CPP) could enhance the proliferation of mouse spleen lymphocytes in vitro. The immunoregulatory function of CPP was then investigated in cyclophosphamide-induced immunosuppressed mice. In CPP-treated groups, mice were orally treated with CPP at doses of 1, 0.5, and 0.25 g/kg bodyweight from 1 to 11 d, respectively. The cyclophosphamide was administrated in CPP and cyclophosphamide groups from 12 to 14 d. In the cyclophosphamide and normal control groups, the mice received equal volume of saline from 1 to 14 d. The results showed that CPP (1 g/kg) could significantly increase the bodyweight of mice, even during cyclophosphamide treatment. The organ coefficients of the spleen and thymus were recovered by CPP treatment. CPP upregulated the contents of cytokines (IL-2, IL-6, IFN-γ, and TNF-α) in serum, which were downregulated by cyclophosphamide. The mRNA levels of these cytokines were also elevated by CPP treatment in the spleen. Cyclophosphamide upregulated the expressions of NF-κB p65, TLR4, and MyD88, suggesting that the NF-κB signaling pathway was activated by cyclophosphamide. After CPP treatment, it was recovered to normal level. These results indicated that CPP alleviated the cyclophosphamide-induced immunosuppression.

Immune-enhancing effects of β-lactoglobulin glycated with lactose following in vitro digestion on cyclophosphamide-induced immunosuppressed mice

β-Lactoglobulin (β-LG) is a major milk protein, making up more than 53% of the total whey proteins, and is seen as a valuable ingredient in food processing because of its high essential amino acid content and diverse functional applications. The Maillard reaction can occur during the storage and processing of food and generate various beneficial effects, including anti-allergenicity, antioxidant, and immunomodulatory effects. The addition of an β-LG-lactose conjugate (LGL) produced by the Maillard reaction was shown to have a strong immune-enhancing effect, increasing both nitric oxide generation and cytokine expression through activation of RAW 264.7 cells, even after in vitro digestion. Furthermore, daily LGL administrationresulted in the upregulation of several immune markers in a cyclophosphamide-induced immunosuppressive mouse model, indicating that this treatment stimulates multiple immune cells, including macrophages, natural killer cells, and lymphocytes, enhancing the proliferation and activation of both the innate and adaptive immune responses. Taken together, these findings indicate that consuming LGL on a regular basis can improve immunity by increasing the natural production of various immune cells.

Chondroitin Sulfate and Fucosylated Chondroitin Sulfate as Stimulators of Hematopoiesis in Cyclophosphamide-Induced Mice

The immunosuppression and inhibition of hematopoiesis are considered to be reasons for the development of complications after intensive chemotherapy and allogeneic hematopoietic stem cell transplantation. Chondroitin sulfate (CS), isolated from the fish Salmo salar, and fucosylated chondroitin sulfate (FCS), isolated from the sea cucumber Apostichopus japonicus, were studied for their roles as stimulators of hematopoiesis in a model of cyclophosphamide-induced immunosuppression in mice. The recombinant protein r G-CSF was applied as a reference. The studied polysaccharides were shown to stimulate the release of white and red blood cells, as well as platelets from bone marrow in immunosuppressed mice, while r G-CSF was only responsible for the significant increase in the level of leucocytes. The analysis of different populations of leucocytes in blood indicated that r G-CSF mainly stimulated the production of neutrophils, whereas in the cases of the studied saccharides, increases in the levels of monocytes, lymphocytes and neutrophils were observed. The normalization of the level of the pro-inflammatory cytokine IL-6 in the serum and the recovery of cell populations in the spleen were observed in immunosuppressed mice following treatment with the polysaccharides. An increase in the proliferative activity of hematopoietic cells CD34(+)CD45(+) was observed following ex vivo polysaccharide exposure. Further study on related oligosaccharides regarding their potential as promising drugs in the complex prophylaxis and therapy of hematopoiesis inhibition after intensive chemotherapy and allogeneic hematopoietic stem cell transplantation seems to be warranted.

Prevention of Cyclophosphamide-Induced Immunosuppression in Mice With Traditional Chinese Medicine Xuanfei Baidu Decoction.

Background and aims: Xuanfei Baidu decoction (XFBD), a traditional Chinese medicine formulation, was designed and successfully applied for COVID-19 disease treatment in China, while the mechanism is still not clear. Methods: To evaluate the protective effect of XFBD on immunosuppression in cyclophosphamide (CY)-treated mice, XFBD was orally administrated, the body weight was measured, and the immune organ index was calculated. HE staining was performed to analyze the pathological structures of the liver, spleen, and thymus. The levels of cytokines and immunoglobulin in the serum and spleen were evaluated by ELISA and RT-PCR. Splenic lymphocytes were isolated, and LPS-stimulated cell proliferation and the number of CD4+ and CD8+ T lymphocytes were evaluated. Results: XFBD significantly suppressed body weight loss and increased the indices of spleen and thymus. The pathological alteration was much improved after XFBD administration. The reductions of TNF-α, IFN-γ, IgG, and IgM levels in serum and IL-2, IL-4, and IL-6 expressions in the spleen were all significantly alleviated by XFBD. Splenic lymphocyte proliferation in response to LPS was further enhanced after treatment with XFBD. The reduction of CD4+ and CD8+ T lymphocytes in CY-treated mice was also highly increased in XFBD groups. Conclusion: Our findings suggested that XFBD played a crucial role in protection against immunosuppression in CY-treated mice and could be a potential candidate for immune modification and therapy.

Sulfated modification enhances the immunomodulatory effect of Cyclocarya paliurus polysaccharide on cyclophosphamide-induced immunosuppressed mice through MyD88-dependent MAPK/NF-κB and PI3K-Akt signaling pathways

The present study investigated the effect of sulfation on the immunomodulatory effect of Cyclocarya paliurus polysaccharide (CP) through a Cyclophosphamide (CTX)-induced immunosuppression mice model. The results showed that sulfated Cyclocarya paliurus polysaccharide (SCP3) had stronger immunomodulatory ability than CP. Administration of SCP3 alleviated immune organ atrophy and restored hematopoiesis in immunosuppressed mice, enhanced splenocyte proliferation, and promoted cytokines and nitric oxide (NO) production in splenocyte supernatants, as well as the number of CD3+, CD4+ and CD8+ T lymphocytes. Meantime, SCP3 significantly improved oxidative stress via increasing the activities of antioxidant enzymes and decreasing the levels of malondialdehyde (MDA) in liver. In addition, SCP3 significantly upregulated the phosphorylation expression of JNK, Erk 1/2, p38 of MAPKs signaling pathway at a dose of 50 mg/kg and accordingly showed increased phosphorylation of Akt, NF-κB (p65), IκB-α, and promoted the degradation of IkB-α. Furthermore, SCP3 significantly increased the expression of the upstream signaling molecule MyD88. All results demonstrated that sulfation can be an effective way to enhance the immunomodulatory effect of polysaccharides. SCP3 has high potential to be a functional food supplement candidate for alleviating chemotherapy drug-induced immunosuppression.

Immunomodulatory effects of selenium-enriched peptides from soybean in cyclophosphamide-induced immunosuppressed mice.

In this study, selenium‐enriched soybean peptides (<3 kDa, named Se‐SPep) was isolated and purified from the selenium‐enriched soybean protein (Se‐SPro) hydrolysate by ultrafiltration. The in‐vivo immunomodulatory effects of Se‐SPep were investigated in cyclophosphamide‐induced immunosuppressed mice. Se‐SPep treatment could alleviate the atrophy of immune organs and weight loss observed in immunosuppressive mice. Besides, Se‐SPep administration could dramatically improve total protein, albumin, white blood cell, immunoglobulin (Ig) M, IgG, and IgA levels in blood. Moreover, Se‐SPep strongly stimulated interleukin‐2 (IL‐2), interferon‐gamma (IFN‐γ), nitric oxide (NO), and cyclic guanosine monophosphate productions by up‐regulating mRNA expressions of IL‐2, IFN‐γ, and inducible NO synthase in spleen tissue. Furthermore, Se‐SPep exhibits more effective immunomodulatory activity compared to Se‐SPro and SPep. In conclusion, Se‐SPep could effectively enhance the immune capacity of immunosuppressive mice. These findings confirm Se‐SPep is an effective immunomodulator with potential application in functional foods or dietary supplements.

Effects of Lycium barbarum Polysaccharides on Immunity and the Gut Microbiota in Cyclophosphamide-Induced Immunosuppressed Mice.

The mechanism of immunoregulation by Lycium barbarum polysaccharides (LBPs) was assessed by studying the effect of LBP on the immunity and the gut microbiota. LBP isolated and purified in this study was composed of nine monosaccharides, with an Mw 1,207 kDa. LBP showed immunomodulatory activity in cyclophosphamide (Cy)-treated mice by restoring the damaged immune organs and adjusting the T lymphocyte subsets. We also found that LBP increased the diversity of the gut microbiota and the relative abundances of bacteria, such as Rickenellaceae, Prevotellaceae, Bifidobacteriaceae, and so on, which were positively associated with immune traits. In addition, Caco2 cells model was used to explore the intestinal absorption of LBP. Results showed that LBP was hardly absorbed in the intestine, which suggesting that most LBP may interact with gut microbiota. These findings suggest that the immune response induced by LBP is associated with the regulation of the gut microbiota.

Analysis of Acanthopanax giraldii Harms Polysaccharide II Composition and Its Immune-Protective Role in a Cyclophosphamide-Induced Immunosuppressive Mice Model

Acanthopanax giraldii Harms is commonly used in traditional Chinese medicine to treat rheumatism, improve joints, and strengthen muscles and bones. The polysaccharides present in A. giraldii Harms contain major bioactive substances, which have antioxidant, anticancer, and antiviral activities. In this study, the structural characterization of the homogeneous polysaccharide isolated from A. giraldii Harms, known as AHP-II, and its immunomodulatory effects in vivo will be studied. High-performance ion chromatography (HPIC) and high-performance gel permeation chromatography (HPGPC) based analyses revealed that AHP-II was composed of various monosaccharides, which included rhamnose, arabinose, galactose, glucose, mannose, galacturonic acid, and glucuronic acid in molar ratios of 29.5 : 24.6 : 23.8 : 4.4 : 5.7 : 8.8 : 3.1, respectively, and had a collective molecular weight of 80.21 × 103 Da. Fourier-transform infrared (FTIR) spectroscopy indicated the presence of a pyranose ring and β-type glycosidic linkages in AHP-II. In addition, immunomodulatory effect analyses of AHP-II that used a cyclophosphamide-induced immunosuppressive mouse model demonstrated that its treatment could significantly restore spleen and thymus indices, promote the proliferation of splenic lymphocytes, elevate CD4+ T lymphocyte percentage and CD4+ : CD8+ ratio in the spleen, promote macrophage phagocytosis, and restore cytokines (IL-6, TNF-α, IgM, and IgG) levels. These results suggested that AHP-II could potentially be used as natural immunomodulator and as an alternative treatment to reduce chemotherapy-induced immunosuppression.

Ginseng–Astragalus–oxymatrine injection ameliorates cyclophosphamide-induced immunosuppression in mice and enhances the immune activity of RAW264.7 cells

Ethnopharmacological significanceGinseng quinquefolium (L.), Astragalus membranaceus, and Sophora flavescens Aiton are popular folk medicines in many Asian countries and regions. These three traditional Chinese herbs and their extracts have been reported to considerably enhance the immune function. G. quinquefolium (L.) is considered the king of herbs in China. Traditionally, G. quinquefolium (L.) is believed to replenish vitality, which is considered as immune enhancement in modern Chinese pharmacy. One of the main uses of Astragalus is immunity enhancement; S. flavescens and oxymatrine obtained from its extract have been used to treat leukopenia. Considering the pharmacological properties of Ginseng, Astragalus, and oxymatrine, we evaluated the immunopotentiation effects of their combination, Ginseng–Astragalus–oxymatrine (GAO), in the present study. Aim of the studyThis study aimed to expand the clinical application of GAO and to preliminarily explore its mechanism of action by determining whether GAO injection can enhance immunity in vivo and in vitro. MethodsOverall, 17 major chemical components in GAO were analysed using HPLC and LC-MS. The immunity-enhancing effect of GAO was studied in the cyclophosphamide (CTX)-induced immunosuppressive mouse model and RAW 264.7 cells. ResultsQuantitative analysis showed that the potential active components of GAO include at least ginsenosides, astragaloside IV, and oxymatrine. GAO could significantly improve the nonspecific immunity including the indices of the thymus and spleen, number of peripheral blood leukocytes, levels of TNF-α and IL-6, phagocytic function of macrophages, and cytotoxic activity of natural killer (NK) cells. Additionally, GAO enhanced the humoural immunity, characterised by the antibody production ability of B cells, and cellular immunity, characterised by the activity of T cells, in immunosuppressed mouse. Moreover, GAO could enhance the phagocytic and adhesion functions of RAW 264.7 cells, which may be related to the activation of reactive oxygen species and NF-κB signalling pathway. ConclusionGAO could dramatically ameliorate CTX-induced immunosuppression in mouse and stimulate the immune activity in RAW 264.7 cells possibly by activating the NF-κB signalling pathway.

Modulating effects of the synergistic combination of extracts of herbal drugs on cyclophosphamide-induced immunosuppressed mice.

ObjectiveTaking leads from the available research, we aimed to develop a synergy-based herbal combination of Tinospora cordifolia (TC), Phyllanthus emblica (PE), and Piper nigrum (PN). Also, evaluating their synergistic effect on CP-induced immunosuppression in mice model and exploring the possible mechanisms involved in reversing the damage.MethodologyThe immunomodulatory activity of combination, of TC stem, PE fruits, and PN dried fruits, was determined by in vitro assays (splenocyte proliferation and pinocytic activity of peritoneal macrophages of mice) and in vivo study using CP-induced immunosuppression model in Swiss Albino mice. The ratio was optimized for combining three by in vitro MTT assay. The combination was further evaluated for anti-oxidant activity by DPPH scavenging method and quantified for its bioactive metabolites by HPTLC. Serum collected on day 0, 4, 7 and 14 was employed for estimation of haematogram (haematocrit, TLC, DLC, and haemoglobin, etc) and immune parameters (IL-10, IL-6 and TNF-α) by ELISA.ResultsThe study demonstrated, that combination of herbal extracts at an intermediate dose could inhibit the proliferation of spleen cells and peritoneal macrophages (P ≤ 0.0001) and induce suppression of pro-inflammatory mediators, and also certified that combination exerts synergized effects. The results showed that the combination possess potential antioxidant activity by DPPH scavenging method (IC50-113.5 µg/ml). It was identified that combination significantly (P ≤ 0.0001) improved the immune markers, haematogram parameters, and histological parameters, with maximum protection offered by an intermediate dose.ConclusionThe results suggested that present combination could be further explored clinically as potent synergy-based therapeutic approach for immune modulation.

Immunostimulatory Effect of Moringa oleifera Extracts on Cyclophosphamide-induced Immunosuppressed Mice

ABSTRACT In this study, the effect of Moringa oleifera extracts were investigated on an immunosuppressed model of cyclophosphamide-induced Swiss mice. M. oleifera seed and root extracts (2.0 g kg−1 body wt) were administered orally for 7 consecutive days; cyclophosphamide (CP) (200 mg kg−1 i.p.) was administered by IV on the 4th day. Results indicated that the extracts of the M. oleifera seeds and roots ameliorated the immunosuppressive effects of CP on delayed type hypersensitivity (DTH) response, leukocyte counts, relative organ weight, IgM concentration, serum TNF-α assay and micro-histological images. This study demonstrated in vivo the therapeutic effect of M. oleifera on the immune response in clinical application.

Effect of IMT-03, an Herbal Formulation in Cyclophosphamide-induced Immunosuppression in Mice

Abstract: Immumomodulatory actions of natural ingredients have been recognized from the time of early civilization. Though herbal medicines are equally used to treat deadly diseases, but till date there are lacks of evidences. In the present work, an herbal preparation IMT-03 was used to find out its immunostimulatory efficacy against chemotherapeutics. Although, most of the individual ingredients present in IMT-03 have been reported for their protective roles against chemotherapeutics, but there is no report in combination. Cyclophosphamide (300 mg/kg, intraperitoneally) induces immunosuppression through myelosuppression. Pretreatment with of IMT-03 at oral the dose of 100 mg/kg and 200 mg/kg in Cyclophosphamide induced immunosuppressive mice showed dose dependently and significantly (p&lt;0.001) improvement in total WBC and absolute neutrophil counts. It also protect the general health of animals from chemotherapeutic induced serious adverse events like signs of sickness, lethargy, immobility, reducing food habit, infections in nostrils and pinna etc. Moreover, IMT-03 has the abilities to counter the macrophages surge during the LPS challenge. The test drug also showed safe up to the oral dose of 2 g/kg. In vitro studies revealed IMT-03 has polyphenols and also radical scavenging actions. There are several evidences that polyphenols can able to modulate cytokines and chemokines signaling pathways in immune cells. Therefore, it is assumed that polyphenols present in IMT-03 either modulate the inflammatory signaling pathways or protect from oxidative stress related DNA damage in myeloid tissues.&#x0D;

Protective effect of exopolysaccharide fraction from Bacillus subtilis against dextran sulfate sodium-induced colitis through maintenance of intestinal barrier and suppression of inflammatory responses

We previously reported that an exopolysaccharide-enriched fraction from Bacillus subtilis J92 (B-EPS) could improve immune functions by regulating the immunological parameters of IFN-γ-primed macrophages, CD3/CD28-stimulated splenocytes, and in cyclophosphamide-induced immunosuppressed mice. In the present study, we investigated whether B-EPS contributes to the maintenance of intestinal barrier integrity in a dextran sodium sulfate (DSS)-induced colitis mouse model that mimics human inflammatory bowel disease (IBD). B-EPS treatment improved histological characteristics and common features including a high disease activity index (DAI), an increased spleen weight, and colon shortening in DSS-induced colitis. B-EPS also effectively restored intestinal barrier function by modulating tight junction-related proteins (claudin-1, claudin-2, and occludin) and epithelial-mesenchymal transition (EMT) marker proteins (E-cadherin, N-cadherin, and vimentin). Moreover, B-EPS downregulated immune cell infiltration and inflammatory responses including the production of inflammatory cytokines, such as IL-6 and IL-1β, and activation of nuclear factor-kappa B (NF-κB) and signal transducer and activator of transcription 3 (STAT3). Taken together, these results suggest that B-EPS could serve as a functional food ingredient for improving intestinal barrier function and alleviating colonic inflammation in IBD.

The increasing hematopoietic effect of the combined treatment of Korean Red Ginseng and Colla corii asini on cyclophosphamide-induced immunosuppression in mice

BackgroundHematopoiesis is the production of blood cells from hematopoietic stem cells (HSCs) that reside in the bone marrow. Cyclophosphamide (CTX) is a chemotherapy drug that suppresses the immune system. Korean Red Ginseng (KRG) and Colla corii asini (CCA) have been traditionally used for boosting the immune system. MethodsHSCs in the bone marrow, and immune cell subtype in splenocytes, PBMCs, and thymocytes were investigated. Serum levels of hematopoietic-related markers were analyzed using ELISA. Protein expression in spleen tissue was analyzed using western blot analysis. Hematoxylin & eosin staining in the femurs of mice were also conducted. ResultsThe combination of KRG and CCA with a ratio of 3:2 increased HSCs, CD3 and CD8+ T cells in the circulation, and CD3 T cells in the spleen. A ratio of 2:3 (KRG:CCA) increased the thymic regulatory T cells and recovered the CD3 T cells in the spleen and circulation while recovering proteins in the JAK-STAT pathway in the spleen. Overall, blood cell population and differentiating factors vital for cell differentiation were also significantly recovered by all combinations especially in ratios of 3:2 and 2:3. ConclusionA ratio of 3:2 (KRG:CCA) is the most ideal combination as it recovered the HSC population in the bone marrow of mice.

Immunomodulatory activity of puerarin in RAW264.7 macrophages and cyclophosphamide-induced immunosuppression mice

Context Puerarin, a natural isoflavone extracted from Radix puerariae, is famous for treating various cardiovascular and cerebrovascular diseases. However, little is known about its direct immunomodulatory activity. Objective This study was designed to investigate the in vitro and in vivo immunomodulatory effects of Radix puerariae by using the murine monocyte–macrophage cell line RAW264.7 and immunosuppressed cyclophosphamide-induced mice. Methods MTT and neutral red phagocytosis assays were conducted to evaluate the in vitro immunomodulatory activities of puerarin on cell viability and phagocytosis by measuring the proliferation, phagocytic, nitric oxide (NO) ability, and TNF-α production ability of stimulated and lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Immunosuppressed cyclophosphamide-induced mice were used to evaluate the in vivo immunomodulatory activities of puerarin by measuring IL-4 and IFN-γ, the serum half hemolysis value, spleen and thymus index, and proliferation assay for splenic lymphocytes. Results and discussion Results showed that puerarin improves immunomodulatory activity by increasing cell proliferation, cell phagocytosis, and NO secretion in RAW264.7 macrophages and reduces the abnormal immunologic activity by decreasing cell phagocytosis and NO secretion in LPS-stimulated RAW264.7 macrophages. In addition, puerarin enhanced the immunologic activity of cyclophosphamide-induced immunosuppression mice by increasing the secretion of NO, IFN-γ, and IL-4, the serum half hemolysis value (HC50), the spleen and thymus index, and proliferation for splenic lymphocytes. Conclusion Puerarin exhibited an upregulated immunomodulatory effect on RAW264.7 macrophages and immunosuppression mice. In addition, puerarin had a downregulated immunomodulatory effect on RAW264.7 macrophages. The results suggest that puerarin could be a promising immunomodulator to assist in the treatment of tumors.

The immune-enhancing effect of anthocyanin-fucoidan nanocomplex in RAW264.7 macrophages and cyclophosphamide-induced immunosuppressed mice.

Aronia, a healthy fruit well known as black chokeberry, has health-promoting effects on hypertension, oxidative stress, and diabetes. Despite many reports of bioactivities of aronia, there is little scientific research on the potential for immune-enhancement. So, anthocyanin-fucoidan nanocomplex (AFNC, a nanocomplex of aronia extract and fucoidan) has been developed to improve immune-enhancement. This study aimed to identify immunomodulatory effects and underlying mechanisms of AFNC using RAW264.7 macrophages and cyclophosphamide-induced immunosuppressed mice. As a result, AFNC-treated RAW264.7 macrophages elevated the production of IL-2, IL-6, tumor necrosis factor (TNF)-α, and nitric oxide (NO). AFNC-enhanced inducible NO synthase expression via nuclear factor-κB signaling pathways. AFNC dose-dependently increased levels of IL-2, IL-6, TNF-α, IL-10, IL-12, interferon-γ, or IL-4 in the serum and spleen of immunosuppressed mice. Taken together, AFNC encourages the immune-enhancing activity through immunostimulatory cytokine production by activation of macrophage. Therefore, these results suggest that AFNC is useful for immunodeficiency-related disorders. PRACTICAL APPLICATIONS: In these days of prevalence of infectious diseases, individual immunity is very important. AFNC has the immune-enhancing effects through immunostimulatory cytokine production by activation of macrophage. Therefore, AFNC could be widely applied to ameliorate a variety of diseases caused by immunosuppression such as infectious diseases.

Immunomodulatory effect of the hydroalcoholic extract of Abrus precatorius L. leaves against cyclophosphamide-induced immunosuppression in mice

This study presents the immunomodulatory potential of Abrus precatorius Linn. (Indian wild licorice) leaves. A hydroalcoholic extract of A. precatorius leaves (EAPL) was prepared by maceration. Thirty male mice were divided into five groups as follows: control group, model control group (cyclophosphamide-treated), and three treatment groups (treated with EAPL at doses of 100, 200 and 300 mg/kg, per os, daily for 14 days). Parameters, including hematological, biochemical, organ indices, hemagglutination test (HA titer), delayed-type hypersensitivity (DTH), interleukin-2 (IL-2) level, splenocyte proliferation and liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) analysis were evaluated. Histopathological examination was carried out for the spleen, kidney and liver. Cyclophosphamide (CPMD)-induced changes in white blood cells, lymphocytes and platelets were improved in the treatment groups. Total protein and albumin levels in the treatment groups were significantly higher. EAPL treatment significantly stimulated splenic T-lymphocytemediated proliferation. Neutrophil adhesion was insignificantly decreased in the model control group compared to the normal control group, which was slightly improved by EAPL treatment. EAPL treatment significantly improved the HA titer and cell-mediated immunity, which is an indication of antibody production. The IL-2 level was significantly higher in the treatment groups that received 200 and 300 mg/kg EAPL. LC-QTOF-MS analysis of EAPL showed the presence of flavonoids, lignans, iridoids and phenolic glycosides. These results suggest that A. precatorius leaves are a good candidate for a new immunomodulatory herbal formulation.

The immunoenhancement effects of sea buckthorn pulp oil in cyclophosphamide-induced immunosuppressed mice.

In this study, the immunomodulatory effect of sea buckthorn (SBT) pulp oil was elucidated in immunosuppressed Balb/c mice induced by cyclophosphamide (CTX). The results showed that SBT pulp oil could reverse the decreasing trend of body weight, thymus/spleen index and hematological parameters induced by CTX. Compared with immunosuppressive mice induced by CTX, SBT pulp oil could enhance NK cytotoxicity, macrophage phagocytosis, and T lymphocyte proliferation, and regulate the proportion of T cell subsets in mesenteric lymph nodes (MLN), and promote the production of secretory immunoglobulin A (sIgA), IFN-γ, IL-2, IL-4, IL-12 and TNF-α in the intestines. In addition, SBT pulp oil can promote the production of short fatty acids (SCFAs), increase the diversity of gut microbiota, improve the composition of intestinal flora, increase the abundance of Alistipes, Bacteroides, Anaerotruncus, Lactobacillus, ASF356, and Roseburia, while decreasing the abundance of Mucispirillum, Anaeroplasma, Pelagibacterium, Brevundimonas, Ochrobactrum, Acinetobacter, Ruminiclostridium, Blautia, Ruminiclostridium, Oscillibacter, and Faecalibaculum. This study shows that SBT pulp oil can regulate the diversity and composition of intestinal microflora in CTX-induced immunosuppressive Balb/c mice, thus enhancing the intestinal mucosa and systemic immune response. The results can provide a basis for understanding the function of SBT pulp oil and its application as a new probiotic and immunomodulator.

Polysaccharide of Atractylodes macrocephala Koidz (PAMK) Alleviates Cyclophosphamide-induced Immunosuppression in Mice by Upregulating CD28/IP3R/PLCγ-1/AP-1/NFAT Signal Pathway.

The polysaccharide of Atractylodes macrocephala Koidz (PAMK) is recognized as an immune enhancer, with anti-cancer, anti-tumour, lymphocyte-activating and lymphocytes proliferation-inducing effects. For investigating the mechanism that PAMK alleviates the decline in T cell activation induced by CTX, 24 6-week-old BALB/c female mice were randomly divided into four groups (C, PAMK, CTX, PAMK + CTX). The spleen index, splenocytes morphology and death, cytokine concentration, T cell activating factors (CD25, CD69, CD71), mRNA expression levels related to the CD28 signal pathway were detected. Furthermore, the lymphocytes of mice was isolated and cultured, and then the Th1/Th2 ratio, activating factors, mRNA levels related to the CD28 signal pathway were detected. The results showed that PAMK significantly improved the spleen index, alleviated abnormal splenocytes morphology and death, maintained the balance of Th1/Th2 cells, increased the levels of IL-2, IL-6, TNF-α, and IFN-γ, and increased the mRNA levels of CD28, PLCγ-1, IP3R, NFAT, and AP-1. In conclusion, PAMK increased cytokines levels and alleviated the decline in activation level of lymphocytes induced by CTX through CD28/IP3R/PLCγ-1/AP-1/NFAT signal pathway.