Stem-leaf saponins from Panax notoginseng (SLSP) comprise numerous PPD-type saponins with diverse pharmacological properties; however, their role in Parkinson's disease (PD), characterized by microglia-mediated neuroinflammation, remains unclear. This study evaluated the effects of SLSP on suppressing microglia-driven neuroinflammation in experimental PD models, including the 1-methyl-4-phenylpyridinium (MPTP)-induced mouse model and lipopolysaccharide (LPS)-stimulated BV-2 microglia. Our findings revealed that SLSP mitigated behavioral impairments and excessive microglial activation in models of PD, including MPTP-treated mice. Additionally, SLSP inhibited the upregulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX2) and attenuated the phosphorylation of PI3K, protein kinase B (AKT), nuclear factor-κB (NFκB), and inhibitor of NFκB protein α (IκBα) both in vivo and in vitro. Moreover, SLSP suppressed the production of inflammatory markers such as interleukin (IL)-1β, IL-6, and tumor necrosis factor alpha (TNF-α) in LPS-stimulated BV-2 cells. Notably, the P2Y2R agonist partially reversed the inhibitory effects of SLSP in LPS-treated BV-2 cells. These results suggest that SLSP inhibit microglia-mediated neuroinflammation in experimental PD models, likely through the P2Y2R/PI3K/AKT/NFκB signaling pathway. These novel findings indicate that SLSP may offer therapeutic potential for PD by attenuating microglia-mediated neuroinflammation.
Animals ; Panax notoginseng/chemistry* ; Saponins/pharmacology* ; Microglia/immunology* ; Mice ; NF-kappa B/immunology* ; Signal Transduction/drug effects* ; Proto-Oncogene Proteins c-akt/immunology* ; Phosphatidylinositol 3-Kinases/genetics* ; Male ; Parkinson Disease/immunology* ; Mice, Inbred C57BL ; Disease Models, Animal ; Plant Leaves/chemistry* ; Neuroinflammatory Diseases/drug therapy* ; Humans

Sjögren's syndrome (SS) is an autoimmune disease characterized primarily by oral and periocular dryness. Astragalus-Salvia (AS) and Ophiopogon-Dendrobium (OD) represent two frequently utilized herb pairs in SS treatment. While the combination of AS-OD herb pairs demonstrates clinical efficacy in alleviating SS symptoms, its underlying mechanism remains unclear. This investigation sought to assess the therapeutic effects and elucidate the potential mechanisms of AS-OD in non-obese diabetic (NOD)/Ltj mice with SS. The study utilized NOD/Ltj mice as SS models, administering AS-OD treatment for 10 weeks at doses of 113.1, 226.2, and 339.3 mg·d^-1·20 g^-1. Results demonstrated that AS-OD improved SS symptoms, evidenced by enhanced salivary flow rate, decreased anti-SSA/Ro and anti-SSB/La antibody levels, increased swimming duration, and reduced lactate (LA) and blood urea nitrogen (BUN) levels in NOD/Ltj mice. AS-OD reduced lymphocyte infiltration, enhanced Aquaporin-5 (AQP5) expression in the submandibular gland, decreased inflammatory cytokine levels in the submandibular gland, and reduced the T helper type 17/regulatory T lymphocyte (Th17/Treg) cell ratio in the spleen. Transcriptomic and proteomic analyses indicated AS-OD's involvement in regulating phosphatidylinositol-3-kinase/protein kinase B (PI3K/AKT) and Janus kinase 3/signal transducer and activator of transcription 3 (JAK1/STAT3) pathways, with inhibitory effects validated in both NOD/Ltj mice submandibular gland and A-253 cells. Furthermore, AS-OD enhanced cell viability and reduced A-253 cell apoptosis through the PI3K/AKT pathway. In A-253 cells, AS-OD reduced inflammatory cytokine levels, CXC chemokine ligand 9/10 (CXCL9/10), and T-cell chemotaxis by inhibiting the JAK1/STAT3 pathway. AS-OD mitigates SS by suppressing inflammation and immune responses through the PI3K/AKT and JAK1/STAT3 pathways.
Animals ; STAT3 Transcription Factor/genetics* ; Sjogren's Syndrome/immunology* ; Mice, Inbred NOD ; Proto-Oncogene Proteins c-akt/genetics* ; Phosphatidylinositol 3-Kinases/genetics* ; Mice ; Drugs, Chinese Herbal/administration & dosage* ; Signal Transduction/drug effects* ; Janus Kinase 1/genetics* ; Humans ; Female ; Astragalus Plant/chemistry* ; Male

The protein kinase A/protein kinase G/protein kinase C-family (AGC kinase family) of eukaryotes is involved in regulating numerous biological processes. The 3-phosphoinositide- dependent protein kinase 1 (PDK1), is a conserved serine/threonine kinase in eukaryotes. To understand the roles of PDK1 homologous genes in cell death and immunity in tetraploid Nicotiana tabacum, the previuosly generated transgenic CRISPR/Cas9 lines, in which 5-7 alleles of the 4 homologous PDK1 genes (NtPDK1a/1b/1c/1d homologs) simultaneously knocked out, were used in this study. Our results showed that the hypersensitive response (HR) triggered by transient overexpression of active Pto (Pto^Y207D) or soybean GmMEKK1 was significantly delayed, whereas the resistance to Pseudomonas syrangae pv. tomato DC3000 (Pst DC3000) and tobacco mosaic virus (TMV) was significantly elevated in these partial knockout lines. The elevated resistance to Pst DC3000 and TMV was correlated with the elevated activation of NtMPK6, NtMPK3, and NtMPK4. Taken together, our results indicated that NtPDK1s play a positive role in cell death but a positive role in disease resistance, likely through negative regulation of the MAPK signaling cascade.
Nicotiana/virology* ; Disease Resistance/genetics* ; Plant Diseases/immunology* ; Plants, Genetically Modified/genetics* ; Gene Knockout Techniques ; Plant Proteins/genetics* ; CRISPR-Cas Systems ; Protein Serine-Threonine Kinases/genetics* ; 3-Phosphoinositide-Dependent Protein Kinases/genetics* ; Pyruvate Dehydrogenase Acetyl-Transferring Kinase ; Tobacco Mosaic Virus/pathogenicity*

The plant F-box protein more axillary growth 2 (MAX2) is a key factor in the signal transduction of strigolactones (SLs) and karrinkins (KARs). As the main component of the SKP1-CUL1-FBX (SCF) complex ubiquitin ligase E3, MAX2 is responsible for specifically recognizing the target proteins, suppressor of MAX2 1/SMAX1-like proteins (SMAX1/SMXLs), which would be degraded after ubiquitination. It can thereby regulate plant morphogenesis and stress responses. There exist homologous genes of MAX2 in the important grain and oil crop soybean (Glycine max). However, its role in plant defense responses has not been investigated yet. Here, GmMAX2b, a homologous gene of MAX2, was successfully cloned from stressed soybean. Bioinformatics analysis revealed that there were two MAX2 homologous genes, GmMAX2a and GmMAX2b, with a similarity of 96.2% in soybean. Their F-box regions were highly conserved. The sequence alignment and cluster analysis of plant MAX2 homologous proteins basically reflected the evolutionary relationship of plants and also suggested that soybean MAX2 might be a multifunctional protein. Expression analysis showed that plant pathogen infection and salicylic acid treatment induced the expression of GmMAX2b in soybean, which is consistent with that of MAX2 in Arabidopsis. Ectopic expression of GmMAX2b compensated for the susceptibility of Arabidopsis max2-2 mutant to pathogen, indicating that GmMAX2b positively regulated plant disease resistance. In addition, yeast two hybrid technology was used to explore the potential target proteins of GmMAX2b. The results showed that GmMAX2b interacted with SMXL6 and weakly interacted with SMXL2. In summary, GmMAX2b is a positive regulator in plant defense responses, and its expression is induced by pathogen infection and salicylic acid treatment. GmMAX2b might exert its effect through interaction with SMXL6 and SMXL2. This study expands the theoretical exploration of soybean disease resistant F-box and provides a scientific basis for future soybean disease resistant breeding.
Glycine max/metabolism* ; Disease Resistance/genetics* ; Plant Diseases/immunology* ; Plant Proteins/genetics* ; Cloning, Molecular ; Gene Expression Regulation, Plant ; F-Box Proteins/genetics* ; Arabidopsis/genetics* ; Phylogeny

Herbal extracts have been extensively used worldwide for their application on memory improvement, especially among aged and memory-deficit populations. In the present study, the memory loss induced by human Abeta protein over-expression in fruitfly Alzheimer's disease (AD) model was rescued by multiple extracts from Gardenia jasminoides. Three extracts that rich with gardenia yellow, geniposide, and gardenoside components showed distinct rescue effect on memory loss. Further investigation on adding gardenoside into a formula of Ganoderma lucidum, Panax notoginseng and Panax ginseng (GPP) also support its therapeutic effects on memory improvement. Interestingly, the application of GPP and gardenoside did not alter the accumulation of Abeta proteins but suppressed the expression of immune-related genes in the brain. These results revealed the importance and relevancy of anti-inflammation process and the underlying mechanisms on rescuing memory deficits, suggesting the potential therapeutic use of the improved GPP formulation in improving cognition in defined population in the future.
Alzheimer Disease ; drug therapy ; Animals ; Antimicrobial Cationic Peptides ; genetics ; Brain ; drug effects ; immunology ; Cognition ; drug effects ; Disease Models, Animal ; Drosophila ; Drosophila Proteins ; genetics ; Gardenia ; chemistry ; Gene Expression Regulation ; drug effects ; Immunity, Innate ; drug effects ; Iridoids ; chemistry ; isolation & purification ; pharmacology ; Plant Extracts ; chemistry ; isolation & purification ; pharmacology ; Polymerase Chain Reaction

NS2 and NS3 are two post-transcriptional gene silencing suppressors that are encoded by Rice stripe virus. Gene silencing suppressors are always related to the pathogenicity of viruses. In this study, the cDNA of NS2 and NS3 were recombined by overlapping PCR assays, ligated to the RNAi vector, and inserted into the PXQ expression vector using Pst I; the expressed vector was transferred into calluses induced from seeds of the japonica rice cultivar, 'Nipponbare', using an Agrobacterium-mediated method. Thirty-one T0 transgenic plants were selected by G418 screening. PCR and southern blot analyses confirmed that the target gene was transformed into transgenic rice successfully, and different transgenic plants contained various copies of the gene. The disease resistance assay revealed that T0 transgenic rice had a delayed onset of RSV for approximately 10-20 d, and the accumulation of virus in the transgenic plants was reduced by 30%-50%. This was related to the delayed onset of disease.
Disease Resistance ; Oryza ; genetics ; immunology ; virology ; Plant Diseases ; genetics ; immunology ; virology ; Plants, Genetically Modified ; genetics ; immunology ; virology ; RNA Interference ; Tenuivirus ; genetics ; immunology ; Viral Nonstructural Proteins ; genetics ; immunology

Liposoluble cape jasmine proteins were used to immunize BALB/C mice. Indirect ELISA was utilized to develop one monoclonal antibody by integrating SP2/0 cells and spleen cells from immunized BALB/C mice. The subclass of the monoclonal antibody was identified as IgG2b, with Kappa chain as its light chain. The ascite titer of 2H8 monoclonal antibody was 1:204 080. Western-blot analysis proved that 2H8 reacted with cape jasmine proteins to identify specific liposoluble protein with molecuar weight of around 58.5 kDa. Dot-ELISA was established with 2H8 ascites as the primary antibody, showing the minimum detectable amount of 19.5 ng. This study lays a foundation for the development of protein kits of Reduning injection.
Animals ; Antibodies, Monoclonal ; analysis ; immunology ; Antibody Specificity ; Drug Hypersensitivity ; immunology ; Drugs, Chinese Herbal ; adverse effects ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Humans ; Mice ; Mice, Inbred BALB C ; Plant Proteins ; analysis ; immunology ; Reagent Kits, Diagnostic

OBJECTIVETo explore the influence of rosa roxburghii tratt preparation on immune function of arseniasis patients caused by burning coal.

METHODSAccording to the diagnosis standard for endemic arseniasis(WS/T 211-2001), 62 cases of arseniasis patients who resided in endemic arseniasis area in Guizhou province were selected and divided stratified randomly into two groups: rosa roxburghii tratt juice treatment group and superoxide dismutase(SOD)-enriched rosa roxburghii tratt juice treatment group, with 31 patients in each group.Each patient took 120 ml/d rosa roxburghii tratt preparation or SOD-enriched rosa roxburghii tratt orally for one month. Another 30 healthy residents from a neighbour township 12 km away where arsenic was not prevalent were selected as controls. A 2 ml blood and 50 ml urine samples were collected from individuals and the urine arsenic contents, peripheral blood T-lymphocyte subsets (CD3(+), CD4(+), CD8(+) T cell), serum immunoglobulin (IgG, IgM, IgA) and complement (C3, C4) were detected. The differences between more than two groups on above indicators were compared. The correlations between urinary arsenic and immune parameters were analyzed.

RESULTSAmong the rosa roxburghii tratt juice group, SOD-enriched rosa roxburghii tratt juice before intervention group and the control group, the levels of urine arsenic were (76.55 ± 23.02) , (72.60 ± 25.91) and (26.33 ± 11.30) µg/g Cr respectively and IgG were (11.31 ± 1.68), (11.35 ± 1.94) and (9.23 ± 1.75) g/L respectively. The differences were statistically significant(F values were 82.01, 13.82, both P values < 0.05). After intervention with rosa roxburghii tratt preparation, the levels of urine arsenic were (53.21 ± 16.51) and (51.72 ± 17.70)µg/g Cr, both decreased than before intervention (t values were 5.80 and 3.78, both P values < 0.05). The levels of CD3(+) were (44.47 ± 7.14)%, (43.44 ± 6.61)% and (70.78 ± 5.26)%, CD4(+) were (29.87 ± 5.67)%, (29.42 ± 5.87)% and (46.08 ± 5.87)%, CD4(+)/CD8(+) were(1.25 ± 0.42), (1.22 ± 0.39) and (1.79 ± 0.26) and C4 were (0.13 ± 0.08), (0.13 ± 0.09) and (0.20 ± 0.11) g/L respectively among the two treatment group before intervention and the control group. The differences were significant (F values were 178.04, 76.71, 23.13 and 5.26, all P values < 0.05). After intervention, the levels of CD3(+) were (59.73 ± 7.38)% and (66.31 ± 7.57)%, CD4(+) were (34.00 ± 7.97)% and (39.11 ± 5.81)%, CD4(+)/CD8(+) were (1.41 ± 0.37) and(1.58 ± 0.26), all increased than before intervention(t values were 12.47, 25.18, 5.41, 10.47, 3.22 and 5.05, all P values < 0.05). The levels of urine arsenic and CD3(+), CD4(+), CD4(+)/CD8(+), C4 were inversely correlated correlation, while positive correlation existed between the level of urine arsenic and IgG(r values were -0.68, -0.56, -0.51, -0.43 and 0.36, all P values < 0.01).

CONCLUSIONSThe level of urinary arsenic level is closely related to immune function suppression in arseniasis patients caused by burning coal, rosa roxburghii tratt preparation can effectively improve immune function of arseniasis patients.

Adult ; Arsenic ; urine ; Arsenic Poisoning ; etiology ; immunology ; China ; Coal ; Complement System Proteins ; immunology ; Female ; Humans ; Immunoglobulins ; immunology ; Male ; Middle Aged ; Plant Extracts ; pharmacology ; Rosa ; chemistry ; Superoxide Dismutase ; pharmacology ; T-Lymphocyte Subsets ; immunology

Specific IgE to gliadin was proposed as a marker for wheat dependent exercise induced anaphylaxis, while Tri a 14 was found to induce IgE response in baker's asthma. We evaluated whether these components could be used for discriminating phenotypes of wheat allergy. Twenty-nine patients who were wheat-induced anaphylaxis and/or urticaria (n=21, group I) and baker's asthma (n=8, group II) were enrolled. The prevalence of serum specific IgE to Tri a 14 was higher in group II (25%) than in group I (4.8%), while the serum specific IgE to gliadin was significantly higher in group I (70%) than in group II (12.5%). The cutoff value for predicting the baker's asthma using the ratio of serum specific IgE to Tri a 14 to gliadin was 742.8 optical densityx1,000/(kU/L) with high sensitivity and specificity. These findings suggest that Tri a 14/gliadin may be a potential marker for predicting baker's asthma.
Adult ; Anaphylaxis/immunology ; Antigens, Plant/*immunology ; Asthma/blood/diagnosis/immunology ; Biological Markers/blood ; Carrier Proteins/*immunology ; Female ; Gliadin/*immunology ; Humans ; Immunoglobulin E/*blood/immunology ; Male ; Phenotype ; Triticum/immunology ; Urticaria/immunology ; Wheat Hypersensitivity/*diagnosis/*immunology

Genetic transformation was adopted to analyze the subcellular localization and the resistance to fungal pathogens of Arabidopsis lipid transfer protein AtDHyPRP1. The coding sequence of AtDHyPRP1 amplified by PCR from Ws ecotype was used to construct the plant binary expression vector pRI101-AN-AtDHyPRP1 and the fusion expression vector pCAMBIA1302-AtDHyPRP1-GFP. Transgenic tobacco and Arabidopsis plants were produced by leaf disc and floral dip protocols, respectively. AtDHyPRP1 could improve the resistance of tobacco to Botrytis cinerea remarkably and the infection sites on transgenic tobacco leaves accumulated large amounts of H2O2. Observation under laser scanning confocal microscope showed that AtDHyPRP1 was localized to cell surface. It suggested that AtDHyPRP1 might play special function after secretion to outside of the cell and was involved in plant defense system against pathogens.
Amino Acid Sequence ; Antigens, Plant ; genetics ; metabolism ; Arabidopsis ; genetics ; metabolism ; microbiology ; Arabidopsis Proteins ; genetics ; metabolism ; Botrytis ; Carrier Proteins ; genetics ; metabolism ; Disease Resistance ; Escherichia coli ; genetics ; metabolism ; Molecular Sequence Data ; Plant Diseases ; immunology ; microbiology ; Plant Proteins ; genetics ; metabolism ; Plants, Genetically Modified ; genetics ; metabolism ; microbiology ; Recombinant Proteins ; genetics ; metabolism ; Subcellular Fractions ; metabolism ; Tobacco ; genetics ; metabolism ; microbiology