Green Synthesis of Gold Nanoparticles from Polygahatous Polysaccharides and Their Anticancer Effect on Hepatic Carcinoma through Immunoregulation.

Hepatic carcinoma is one of the leading causes of morbidity and mortality among all cancers, but no effective treatment measures have been developed. Herein, polystyrene polysaccharide (PSP) extracted from Polygonatum was used to synthesize gold nanoparticles (PSP-AuNPs) by heating and reduction methods, and the characteristics of the PSP-AuNPs were detected after successful synthesis. In vitro, the immunoregulatory effects of PSP-AuNPs were studied by testing the concentrations of NO, TNF-α, and IL-12p70 in the culture media of PSP-AuNPs-treated RAW264.7 macrophages, and the effect of biocompatibility on the viability of RAW264.7 macrophages and L02 cells was studied via a CCK-8 assay. In vivo, tumor-bearing mice were established and treated with PSP-AuNPs, and the anticancer effects were studied by detecting trends in tumor volume, tumor inhibition rate, and tumor cell proliferation index. Immunoregulation was assessed by evaluating the serum levels of TNF-α and IL-10, the CD4+/CD8+ lymphocyte ratio in peripheral blood and the spleen and thymus indices; toxicity was investigated by measuring body weight, liver and renal function indices. The results showed that PSP-AuNPs could regulate immune function both in vitro and in vivo with almost no toxicity. PSP-AuNPs exhibited excellent anticancer effects on hepatic carcinoma in vivo. The anticancer effect could be strengthened, and the toxicity could be reduced by the combined use of PSP-AuNPs and ADM. In conclusion, PSP-AuNPs could be effective as a therapy and adjuvant therapy for treating hepatic carcinoma, providing potential treatment strategies for this disease.

Synthesis of Pseudostellaria heterophylla polysaccharide-gold nanocomposites and their antitumor effect through immunomodulation.

Polysaccharides from natural sources have an excellent immune function and low toxicity; however, their limitations such as short half-life and instability limit their sustained pharmacological activity. In this context, the combination of polysaccharides and nanotechnology have been developed to promote the stability and prolong the immune activities of polysaccharides. To synthesize and explore the antitumor effect and immunomodulatory activity of PHP-AuNPs. Polysaccharides extracted from Pseudostellaria heterophylla were used to synthesize gold nanocomposites (PHP-AuNPs), and their physicochemical properties and immunoregulatory effect in vitro and in vivo were analyzed. The PHP-AuNPs were green synthesized with high biosafety. PHP-AuNPs can activate macrophages in vitro and decrease the tumor weight and volume, whereas they increase the immune organ index in vivo. Besides, PHP-AuNPs showed a beneficial effect for maintaining the immune balance of CD4+/CD8+ T cells and modulating the release of cytokines such as TNF-α increase and IL-10 decrease in mice. All these results suggested that PHP-AuNPs exhibit a remarkable antitumor effect and stronger immunomodulatory activity than that of free PHP-1. RESEARCH HIGHLIGHTS: The P. heterophylla polysaccharide-gold nanocomposites (PHP-AuNPs) were synthesized and physicochemical properties were characterized. The cytotoxicity in vitro and immunomodulatory effects of PHP-AuNPs on macrophages were analyzed. The immune-antitumor effects in vivo of PHP-AuNPs have also been confirmed.

A pairwise immune gene model for predicting overall survival and stratifying subtypes of colon adenocarcinoma.

OBJECTIVES: There is increasing evidence for a close correlation between risk stratification, prognosis and the immune environment in colon adenocarcinoma (COAD). However, the efficacy of immunotherapy is different among different patients with COAD. Therefore, the current work tends to use immune-related gene to develop a gene-pair model to evaluate the COAD prognosis, and to develop a new method for risk stratification of COAD, which is conducive to better predict the immunotherapy effect of patients. METHODS: Specifically, from the TCGA and GEO (GSE14333 and GSE39582) databases, we first collected gene expression profiles, associated survival follow-up information of COAD patients. Through systematic bioinformatics analysis, we established a prognosis-related model of colon cancer with three pairs of "immune gene pairs", with uni- and multivariate and lasso cox regression analyses verifying the model stability. Most immune cells showed markedly different levels of infiltration between the two risk subgroups calculated by the model. More, single-cell RNA-seq analyses were also performed to validate the selected genes in the immune gene-pair model. RESULTS: A prognosis-related model of colon cancer with three pairs of "immune gene pairs" were built and validated by several datasets. The analysis of immune landscape of COAD revealed that low-risk subgroup obtained by the prognosis-related model for COAD can be further divided into three subclusters with different prognosis. Then, we applied the Tumor online Prognostic analyses Platform (ToPP) to construct a prognostic model using these five genes. Results show that APOD, ISG20 and STC2 are risk factors, while CXCL9 and IL7R are protection factors. We also found that only the five-gene model could also predict the prognosis of COAD patients, indicating the robustness of the gene-pair model. Among the five genes, including CXCL9, APOD, STC2, ISG20, and IL7R, in the gene-pair model, single-cell RNA sequencing reveals the high expression of CXCL9 and IL7R in inflammatory macrophages. Using cell-cell interaction and trajectory analysis, data indicate that CXCL9+/IL7R+ pro-inflammatory macrophages were capable of secreting and activating more anti-tumor pathways than CXCL9-/IL7R- pro-inflammatory macrophages. CONCLUSIONS: In short, we have successfully developed an "immune gene pair" related model that can judge the prognostic status of patients with COAD and may contribute to risk stratification and evaluate potential beneficiaries of immunotherapy, providing new ideas for the anti-COAD management and therapy.

Antitumor effect of a polysaccharide from Pseudostellaria heterophylla through reversing tumor-associated macrophages phenotype.

Tumor-associated macrophages (TAMs), which are predominant tumor-infiltrating immune cells in the tumor microenvironment, participate in promoting the occurrence and metastasis of tumor cells. Reprogramming TAMs has become a promising immunotherapeutic approach for novel cancer treatments. In this study, a homogeneous polysaccharide (PHP-1) was obtained from Pseudostellaria heterophylla, and its antitumor and immunological activities, as well as the underlying molecular mechanisms were explored. These findings suggested that PHP-1 can switch M2 macrophages to the M1 type, thereby promoting tumor cell apoptosis in vitro. In addition, PHP-1 can modulate the TAMs phenotype, maintain the CD4+/CD8+ lymphocyte balance, and exert antitumor effects in H22 tumor-bearing mice. Mechanistically, PHP-1 is recognized by the TLR4 receptor, promotes Ca2+ release, and activates the NF-κB and MAPK signaling pathways to reset the M2-type macrophages. These findings indicate that PHP-1 from P. heterophylla can function as a tumor immunotherapeutic modulator.

Green synthesized gold nanoparticles from Pseudobulbus Cremastrae seu Pleiones show efficacy against hepatic carcinoma potentially through immunoregulation.

Nanobiotechnology, the interface between biology and nanotechnology, has recently emerged in full bloom in the medical field due to its minimal side-effects and high efficiency. To broaden the application of nanobiotechnology, we composed gold nanoparticles from the extract of Pseudobulbus Cremastrae seu Pleiones (PCSP) using an efficient and green procedure. The biosynthesized Au nanoparticles containing PCSP (PCSP-AuNPs) were characterized by UV-vis spectroscopic, transmission electron microscopy (TEM), atomic force microscopy (AFM), dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FT-IR), and Energy Dispersive X-ray (EDAX). After verifying the stability of PCSP-AuNPs, we detected its biosafety and immune-modulatory effects on RAW264.7 in vitro using NO assay, ELISA (TNF-α, IL-12p70, and IL-1ß), and CCK-8 test. Furthermore, we examined the direct in vitro effects of PCSP-AuNPs on hepatocellular carcinomas (HCCs). Finally, we evaluated the immune regulation of PCSP-AuNPs using a mouse model with H22-tumor by testing the index of immune organs, splenic lymphocyte proliferation, cytokines levels (TNF-α and IL-10), and the CD4+/CD8+ cell ratio in the peripheral blood. Immunohistochemical analyses including H&E and PCNA staining were performed to investigate the anti-cancer efficacy and biocompatibility of PCSP-AuNPs. We found that PCSP-AuNPs not just possessed low toxicity, but also improved the immune-mediated antitumor response as compared to PCSP alone, suggesting its potential as a novel and efficient drug for liver cancer therapy.

Green synthesis and characterization of gold nanoparticles from Pholiota adiposa and their anticancer effects on hepatic carcinoma.

Gold nanoparticles (AuNPs) were successfully fabricated by Pholiota adiposa polysaccharide (PAP-1a) without employing any other chemicals. The physical and chemical properties of PAP-AuNPs were determined using transmission electron microscopy (TEM), dynamic light scattering (DLS), energy-dispersive X-ray spectroscopy (EDXR), Fourier-transform infrared spectroscopy (FT-IR), and atomic force microscopy (AFM). In an attempt to analyze the immune regulation, antitumor effect, and biological safety, the production of NO and TNF-α, IL-12p70, and IL-1ß from RAW264.7 as well as the proliferation of RAW264.7 were detected in vitro. Flow cytometry was conducted to determine the ratio of the CD4+/CD8+ cell in peripheral blood and immunohistochemical analysis involving hematoxylin and eosin (H&E) and proliferating cell nuclear antigen (PCNA) staining were conducted in vivo. The results of this study showed that PAP-AuNPs had a significantly improved immune regulation and anti-tumor effect in comparison to PAP-1a alone. PAP-AuNPs showed no toxicity both in vivo and in vitro. This study demonstrates a useful application of PAP-AuNPs as a novel nanomedicine for hepatic carcinoma.

Study on the physicochemical properties and immunomodulatory anti-tumor effect of the Pholiota adiposa polysaccharide.

In this study, the extraction, purification, physical and chemical properties, and biological activity of the Pholiota adiposa (PAP) polysaccharide were investigated. One fraction (PAP-1a) of Pholiota adiposa polysaccharides was isolated using DEAE Sepharose™ Fast Flow and Sephacryl™ S-300 High-Resolution columns. The HPLGPC results revealed that the molecular weight of PAP-1a was 16.453 kDa. PAP-1a was composed of mannose, ribose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, xylose, arabinose, and fucose and their molar % was 33.41, 0.53, 1.33, 0.07, 0.27, 5.28, 38.31, 0.83, 18.04 and 2.23, respectively. PAP-1a could activate macrophages to secrete NO and cytokines such as TNF-a, IL-6, and IL-12p70. When hepatocellular carcinoma cells (HCCs) and macrophages were co-cultured, it was observed that PAP-1a inhibited the growth of Hep-G2, Hep-3B, and Huh7 via immunoregulation. It triggered cell apoptosis by blocking the cell cycle in the G0/G1 stage. Furthermore, PAP-1a had no direct cytotoxicity against the hepatocyte cell line L02 and macrophages RAW264.7.

Multifunctional liposome for photoacoustic/ultrasound imaging-guided chemo/photothermal retinoblastoma therapy.

Retinoblastoma (RB) is a malignant intraocular neoplasm that occurs in children. Diagnosis and therapy are frequently delayed, often leading to metastasis, which necessitates effective imaging and treatment. In recent years, the use of nanoplatforms allowing both imaging and targeted treatment has attracted much attention. Herein, we report a novel nanoplatform folate-receptor (FR) targeted laser-activatable liposome termed FA-DOX-ICG-PFP@Lip, which is loaded with doxorubicin (DOX)/indocyanine green (ICG) and liquid perfluoropentane (PFP) for photoacoustic/ultrasound (PA/US) dual-modal imaging-guided chemo/photothermal RB therapy. The dual-modal imaging capability, photothermal conversion under laser irradiation, biocompatibility, and antitumor ability of these liposomes were appraised. The multifunctional liposome showed a good tumor targeting ability and was efficacious as a dual-modality contrast agent both in vivo and in vitro. When laser-irradiated, the liposome converted light energy to heat. This action caused immediate destruction of tumor cells, while simultaneously initiating PFP phase transformation to release DOX, resulting in both photothermal and chemotherapeutic antitumor effects. Notably, the FA-DOX-ICG-PFP@Lip showed good biocompatibility and no systemic toxicity was observed after laser irradiation in RB tumor-bearing mice. Hence, the FA-DOX-ICG-PFP@Lip shows great promise for dual-modal imaging-guided chemo/photothermal therapy, and may have significant value for diagnosing and treating RB.

Structure characteristics and immunomodulatory activities of a polysaccharide RGRP-1b from radix ginseng Rubra.

The present study was undertaken to explore the structure characteristics, immune regulation, and anti-cancer abilities of polysaccharides in radix ginseng Rubra (RGR). For this purpose, RGR polysaccharides (RGRP) were purified through DEAE and S-300 chromatography. Monosaccharide composition, methylation, and GC-MS analyses, as well as field emission scanning electron microscope (FESEM), atomic force microscope (AFM), Fourier-transformed infrared resonance (FT-IR), and nuclear magnetic resonance (NMR) spectra, were used to establish the structure of RGRP-1b. Our results revealed that RGRP-1a and RGRP-1b possess different molecular weights (21.3 kDa and 10.2 kDa, respectively). RGRP-1a was found to be composed of glucose, while RGRP-1b was composed of glucose, galactose, and arabinose. The main chain structure of RGRP-1b was composed of 1,4-α-Glcp, with a 1,4,6-α-Glcp branch unit. Its side chains were branched at the O-4 position of 1,4,6-α-Glcp, namely 1)-ß-Galp-(4 â†’ 1)-α-Araf-(5 â†’ α-Araf and 1)-ß-Galp-(6 â†’ α-Glcp. The changes in the nitric oxide (NO) levels and cytotoxicity revealed that macrophages probably get activated by RGRP-1b. The expressions of IL-6, IL-12, and TNF-α were found to be upregulated after treatment with RGRP-1b. RGRP-1b thus possesses the potential to arrest the growth of Huh7 through immunoregulation. Our cumulative findings indicate that RGRP-1b obtained from radix ginseng Rubra can function as a strong immune modulator.

Investigation of Potential Molecular Biomarkers for Diagnosis and Prognosis of AFP-Negative HCC.

BACKGROUND: Alpha-fetoprotein (AFP) is the most important diagnostic and prognostic index of hepatocellular carcinoma (HCC). AFP-positive HCC can be easily diagnosed based on the serum AFP level and typical imaging features, but a number of HCC patients are negative (AFP < 20 ng/mL) for AFP. Therefore, it is necessary to develop novel diagnostic and prognostic biomarkers for AFP-negative HCC. METHODS: RNA data from TCGA and differential expression of lncRNAs, miRNAs, and mRNAs were downloaded to analyze the differential RNA expression patterns between AFP-negative HCC tissues and normal tissues. A lncRNA-miRNA-mRNA ceRNA regulatory network was constructed to elucidate the interaction mechanism of RNAs. Functional enrichment analysis of these DEmRNAs was performed to indirectly reveal the mechanism of action of lncRNAs. A PPI network was built using STRING, and the hub genes were identified with Cytoscape. The diagnostic value of hub genes was assessed with receiver operating characteristic (ROC) analysis. And the prognostic value of RNAs in the ceRNA was estimated with Kaplan-Meier curve analysis. RESULTS: A total of 131 lncRNAs, 185 miRNA, and 1309 mRNAs were found to be differentially expressed in AFP-negative HCC. A ceRNA network consisting of 12 lncRNA, 23 miRNA, and 74 mRNA was constructed. The top ten hub genes including EZH2, CCNB1, E2F1, PBK, CHAF1A, ESR1, RRM2, CCNE1, MCM4, and ATAD2 showed good diagnostic power under the ROC curve; and 2 lncRNAs (LINC00261, LINC00482), 3 miRNAs (hsa-miR-93, hsa-miR-221, hsa-miR-222), and 2 mRNAs (EGR2, LPCAT1) were found to be associated with the overall survival of AFP-negative patients. CONCLUSION: This study could provide a novel insight into the molecular pathogenesis of AFP-negative HCC and reveal some candidate diagnostic and prognostic biomarkers for AFP-negative HCC.

Green synthesis of gold nanoparticles from Dendrobium officinale and its anticancer effect on liver cancer.

A novel gold nanoparticle (Do-AuNP) was successfully synthesized from water extracts of traditional Chinese medicine Dendrobium officinale (DO) without using any extra chemicals regents. The physicochemical properties of Do-AuNPs were analyzed by transmission electron microscopy, dynamic light scattering, energy-dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy, and atomic force microscopy. The amount of DO extract on the AuNPs was about 13%. In order to evaluate the anti-tumor efficiency and biosafety, the inhibitory rate of HepG2 cells and survival rate of L02 cells were performed in vitro, and the immunohistochemical analysis of H&E, Ki-67, and TUNEL staining were conducted in vivo. Our results demonstrated that Do-AuNP had better anti-tumor efficiency compared with DO extraction alone without increasing toxicity in vivo and in vitro. The present study provides useful information for Do-AuNP as a new nanomedicine for liver cancer.

Physicochemical properties of polysaccharides from Ligusticum chuanxiong and analysis of their anti-tumor potential through immunoregulation.

We investigated the extraction, purification, physicochemical properties and biological activity of Ligusticum chuanxiong polysaccharides (LCXPs). Two polysaccharide fractions (Ligusticum chuanxiong [LCX]P-1a and LCXP-3a) were obtained by DEAE Sepharose™ Fast Flow and Sephacryl™S-300 high resolution column chromatography. The results showed that the molecular weight of LCXP-1a and LCXP-3a was 11.159 kDa and 203.486 kDa, respectively. LCXP-1a is composed of rhamnose, glucuronic acid, galacturonic acid, and glucose at a molar percentage of 0.52 : 1.88 : 1.06 : 95.36, But LCXP-3a has another molar percentage of mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, xylose, arabinose, and fucose of 0.64 : 6.69 : 1.03 : 43.74 : 2.20 : 26.90 : 0.82 : 15.94 : 1.80. Both LCXP-1a and LCXP-3a could stimulate macrophages to produce NO, TNF-α, IL-6, and IL-12p70. Co-culturing macrophages and hepatocellular carcinoma cells showed that LCXP-1a and LCXP-3a inhibited the growth of HepG2 and Hep3B through immunoregulation. They arrested the cell cycle at the G0/G1 phase and promoted apoptosis. Moreover, there was no cytotoxicity to the hepatocyte cell line, LO2. We also noted that the immunomodulatory activity and anti-tumor activity of LCXP-3a were significantly better than those of LCXP-1a. Our data demonstrate that LCXP-3a is potentially a well-tolerated and effective immunomodulatory adjuvant cancer treatment.

Immunomodulatory effects of a polysaccharide from Solanum nigrum Linne through TLR4-MyD88 signaling pathway.

Solanum nigrum Linne polysaccharide (SNLP), an active ingredient from Solanum nigrum Linne, has been proposed to inhibit tumor growth and display immunomodulatory activity. However, the molecular mechanism related to immune regulation remains unclear. In the present study, a homogeneous polysaccharide (SNLP-1) was extracted, the immune effects and the underlying molecular mechanisms were investigated. The immunomodulatory activity assay in vitro showed that SNLP-1 promoted the release of NO and TNF-α and IL-6 secretion in macrophages. In tumor-bearing mice, SNLP-1 could improve immune function including increasing the spleen index, thymus index and inducing Th1 responses mediated by IL-2, IFN-γ, and TNF, as well as decreasing the tumor weight. Furthermore, SNLP-1 elevated the expression of the critical nodes in the TLR4-Myd88 signaling pathways in vitro and in vivo. These results indicated that TLR4-MyD88 signal pathway may be one of the signal pathways of immune regulation of SNLP-1.

Immunomodulatory effects exerted by Poria Cocos polysaccharides via TLR4/TRAF6/NF-κB signaling in vitro and in vivo.

OBJECTIVE: Poria cocos polysaccharide (PCP) is the major active ingredients of P. cocos and possesses various pharmacological effects, including anti-oxidative and anti-apoptosis effects and activity against cancer. This study investigated the immunomodulatory mechanism by which PCP acts on RAW 264.7 macrophages and LLC tumors in mice. METHODS: The concentrations of nitric oxide, and Th1, Th2, and Th17 cytokines were examined by Griess reaction and using a bead-based cytokine assessment kit. qRT-PCR and western blotting were used to investigate relevant signaling molecule expression. RESULTS: Levels of nitric oxide, IL-2, IL-6, IL-17 A, TNF, and IFN-γ were increased by PCP while levels of IL-4 and IL-10 were unaffected. The addition of TAK-242 (TLR4 inhibitor) or assessment in C57BL/10ScNJ (TLR4-deficient) mice markedly reduced this effect. In C57BL/10 J (TLR4+/+wild-type) mice, the indices of organ immune activity were all elevated, and oral PCP delivery resulted in a significant reduction in tumor volume over a 25 day period. Relative to controls, TLR4, MyD88, TRAF-6, p-NF-κB and p-c-JUN expression significantly increased, while TRAM expression did not change. Nevertheless, there was no PCP-dependent activation of MyD88, TRAF-6, TRAM, p-NF-κB or p-c-JUN in TLR4-deficient mice. CONCLUSION: These results support the concept that PCP may exhibit immunomodulatory activity through TLR4/TRAF6/NF-κB signaling both in vitro and in vivo.

Identification of differentially expressed genes and enriched pathways in lung cancer using bioinformatics analysis.

Lung cancer is the leading cause of cancer­associated mortality worldwide. The aim of the present study was to identify the differentially expressed genes (DEGs) and enriched pathways in lung cancer by bioinformatics analysis, and to provide potential targets for diagnosis and treatment. Valid microarray data of 31 pairs of lung cancer tissues and matched normal samples (GSE19804) were obtained from the Gene Expression Omnibus database. Significance analysis of the gene expression profile was used to identify DEGs between cancer tissues and normal tissues, and a total of 1,970 DEGs, which were significantly enriched in biological processes, were screened. Through the Gene Ontology function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, 77 KEGG pathways associated with lung cancer were identified, among which the Toll­like receptor pathway was observed to be important. Protein­protein interaction network analysis extracted 1,770 nodes and 10,667 edges, and identified 10 genes with key roles in lung cancer with highest degrees, hub centrality and betweenness. Additionally, the module analysis of protein­protein interactions revealed that 'chemokine signaling pathway', 'cell cycle' and 'pathways in cancer' had a close association with lung cancer. In conclusion, the identified DEGs, particularly the hub genes, strengthen the understanding of the development and progression of lung cancer, and certain genes (including advanced glycosylation end­product specific receptor and epidermal growth factor receptor) may be used as candidate target molecules to diagnose, monitor and treat lung cancer.

Macrophage inhibitory cytokine-1 versus carbohydrate antigen 19-9 as a biomarker for diagnosis of pancreatic cancer: A PRISMA-compliant meta-analysis of diagnostic accuracy studies.

BACKGROUND: Because of the high malignant degree of pancreatic cancer (PC), the early diagnosis of PC is of great concern. Macrophage inhibitory cytokine-1 (MIC-1) was reported to be a potential diagnostic biomarker, but its diagnostic value is indeterminate. Therefore, we performed this meta-analysis to compare it to carbohydrate antigen 19-9 (CA19-9), the most frequently used serum biomarker in PC. MATERIAL AND METHODS: After a systematic review of the relevant studies, the pooled diagnostic indices, including sensitivity, specificity, positive/negative likelihood ratio (PLR/NLR), diagnostic odds ratio (DOR), summary receiver operating characteristic curve (sROC), and area under the SROC curve (AUC) were used to evaluate the diagnostic value of MIC-1 and CA19-9 for PC. These indices were pooled with random-effects models. We explored the heterogeneity by meta-regression. RESULTS: Fourteen studies comprising a total of 2826 subjects were included in our meta-analysis. The summary estimates for MIC-1 and CA19-9 are listed as follows: sensitivity, 80% [95% confidence interval (CI) 78-82] versus 71% (95% CI 68-73); specificity, 85% (95% CI 83-87) versus 88% (95% CI 86-90); DOR, 24.57 (95% CI 14.00-43.10) versus 17.65 (95% CI 11.65-26.76); area under sROC (AUC), 0.8945 versus 0.8322; PLR, 5.18 (95% CI 3.24-8.26) versus 5.34 (95% CI 3.78-7.54); and NLR, 0.23 (95% CI 0.19-0.29) versus 0.32 (95% CI 0.28-0.37). CONCLUSION: These data demonstrate that serum MIC-1 has a comparable diagnostic accuracy to CA19-9 for PC.

Polygonatum sibiricum polysaccharides play anti-cancer effect through TLR4-MAPK/NF-κB signaling pathways.

OBJECTIVE: To investigate the anti-cancer effect of Polygonatum sibiricum polysaccharides (PSP) and the underlying mechanism. METHODS: Tumor-bearing mice were randomly divided into normal saline (NS) group, adriamycin (ADM) group, PSP group and lipopolysaccharide (LPS) group. RAW264.7 cells were pre-treated with or without TLR4 inhibitor or MyD88 inhibitor. Quantitative RT-PCR and Western blot were performed to detect the mRNA and protein expressions, respectively. ELISA and Griess reaction was used to measure cytokines and NO levels. Flow cytometry was employed to examine T-lymphocyte subset and CCK-8 assay was used for cell viability. RESULTS: The in vivo experiment found that PSP inhibited tumor growth and improved the spleen index, thymus index, the cytokines secretion and CD4+/CD8+ lymphocytes ratio. Compared with the NS group, the mRNA and protein expressions of the critical nodes inTLR4-MAPK/NF-κB signaling pathways (except TRAM) significantly increased in PSP group, as well as the NO and cytokines levels. Nevertheless, PSP had no obvious effects on TRAM. Further analysis showed that PSP effects on the critical nodes in TLR4-MAPK/NF-κB signaling pathways were suppressed by inhibitor in vitro. CONCLUSION: The immunoenhancement effect of PSP against lung cancer is mediated by TLR4-MAPK/NF-κB signaling pathways.

Immunomodulatory effects of herbal formula of astragalus polysaccharide (APS) and polysaccharopeptide (PSP) in mice with lung cancer.

OBJECTIVE: This study is to investigate the immunomodulatory effects of the herbal formula of astragalus polysaccharide (APS) and polysaccharopeptide (PSP) in mouse models of immunosuppression and lung cancer. METHODS: Immune parameters were recorded for these model mice. Peripheral white blood cells (WBC) were detected with the automatic blood cell analyzer. Spleen and thymus indices, and tumor inhibition ratio were obtained. Percentage of peripheral blood CD4+ and CD8+ T lymphocytes were detected by flow cytometry. Serum levels of Th1 (IL-2, TNF, and IFN-γ), Th2 (IL-4, IL-6, and IL-10), and Th17 (IL-17A) were detected with the BD cytometric bead array (CBA) mouseTh1/Th2/Th17 cytokine kit. RESULTS: Compared with the NS group, the PSP and APS herbal formula significantly improved the WBC, thymus index, spleen index, CD4+/CD8+ ratio, TNF, IFN-γ, IL-2, andIL-17Ainimmunosuppressivemice and lung cancer mice (P<0. 05). On the contrary, IL-10 was relatively low in the PSP+APS herbal formula group (P<0. 05). Besides, the PSP+APS herbal formula group induced comparable tumor inhibiting effect with the AMD group (23.3% and 24.1%, respectively). CONCLUSION: The PSP+APS herbal formula have immunomodulatory effects and anti-tumor activity in mice with of lung cancer.

Astragalus polysaccharides exerts immunomodulatory effects via TLR4-mediated MyD88-dependent signaling pathway in vitro and in vivo.

Astragalus polysaccharides (APS), which is widely used as a remedy to promote immunity of breast cancer patients, can enhance immune responses and exert anti-tumor effects. In this study, we investigated the effects and mechanisms of APS on macrophage RAW 264.7 and EAC tumor-bearing mice. Griess reaction and ELISA assays revealed that the concentrations of nitric oxide, TNF-α, IL-1ß and IL-6 were increased by APS. However, this effect was diminished in the presence of TAK-242 (TLR4 inhibitor) or ST-2825(MyD88 inhibitor). In C57BL/10J (TLR4+/+wild-type) and C57BL/6J (MyD88+/+wild-type) tumor-bearing mice, the tumor apoptosis rate, immune organ indexes and the levels of TNF-α, IL-1ß and IL-6 in blood increased and the tumor weight decreased by oral administration of APS for 25 days. APS had no obvious effects on IL-12p70. However, these effects were not significant in C57BL/10ScNJ (TLR4-deficient) and C57BL/B6.129P2(SJL)-Myd88m1.1Defr/J (MyD88-deficient) tumor-bearing mice. qRT-PCR and Western blot indicated that APS stimulated the key nodes in the TLR4-MyD88 dependent signaling pathway, including TLR4, MyD88, TRAF-6, NF-κB and AP-1, both in vitro and in vivo. However, TRAM was an exception. Moreover, TRAF-6 and NF-κB were not triggered by APS in gene-deficient tumor-bearing mice. Therefore, APS may modulate immunity of host organism through activation of TLR4-mediated MyD88-dependent signaling pathway.