Saussurea involucrata (Snow Lotus) ICE1 and ICE2 Orthologues Involved in Regulating Cold Stress Tolerance in Transgenic Arabidopsis

As with other environmental stresses, cold stress limits plant growth, geographical distribution, and agricultural productivity. CBF/DREB (CRT-binding factors/DRE-binding proteins) regulate tolerance to cold/freezing stress across plant species. ICE (inducer of CBF expression) is regarded as the upstream inducer of CBF expression and plays a crucial role as a main regulator of cold acclimation. Snow lotus (Saussurea involucrata) is a well-known traditional Chinese herb. This herb is known to have greater tolerance to cold/freezing stress compared to other plants. According to transcriptome datasets, two putative ICE homologous genes, SiICE1 and SiICE2, were identified in snow lotus. The predicted SiICE1 cDNA contains an ORF of 1506 bp, encoding a protein of 501 amino acids, whereas SiICE2 cDNA has an ORF of 1482 bp, coding for a protein of 493 amino acids. Sequence alignment and structure analysis show SiICE1 and SiICE2 possess a S-rich motif at the N-terminal region, while the conserved ZIP-bHLH domain and ACT domain are at the C-terminus. Both SiICE1 and SiICE2 transcripts were cold-inducible. Subcellular localization and yeast one-hybrid assays revealed that SiICE1 and SiICE2 are transcriptional regulators. Overexpression of SiICE1 (35S::SiICE1) and SiICE2 (35S::SiICE2) in transgenic Arabidopsis increased the cold tolerance. In addition, the expression patterns of downstream stress-related genes, CBF1, CBF2, CBF3, COR15A, COR47, and KIN1, were up-regulated when compared to the wild type. These results thus provide evidence that SiICE1 and SiICE2 function in cold acclimation and this cold/freezing tolerance may be regulated through a CBF-controlling pathway.

Based on network pharmacology, using methods such as molecular docking and gene difference analysis, small molecule compounds for the treatment of lung adenocarcinoma and lung squamous cell carcinoma were screened in saussurea involucrata, and new therapeutic targets were discovered

Purpose: Based on systemic pharmacology and network pharmacology, to evaluate the therapeutic effect of saussurea involucrata (SAIN) on lung adenocarcinoma and lung squamous cell carcinoma (LUAD&LUSC) through clinical sample genetic difference analysis and compound-target molecular docking, and discover new The target of prevention or treatment of LUAD&LUSC. Materials and methods: Using the TCM System Pharmacology Database (TCMSP) as the starting point for preliminary selection of ingredients and targets (OB≥30%, DL≥0.18, n=9), with the GDC database as the end point, using Cytoscape 3.8, TBtools 1.082, AutoDock 4.2.6, R 4.0.4, PyMol and other tools have conducted a preliminary screening of the ingredients and targets of SAIN. In order to further screen the effective ingredients and targets, we used clinical samples from LUAD and LUSC from TCGA and GEPIA to perform genetic difference analysis (n=6), and perform biological process (BP) analysis (FDR) on these targets. ≤0.05, n=6), KEGG pathway analysis (FDR≤0.05, n=6), protein interaction network (PPI) analysis (n=6) and compounds-targets-pathways network analysis (n=6), obtain biological processes, disease pathways and various compounds regulated by targets-the relationship between targets and pathways. Through the precise molecular docking of ingredients and targets, we further screened the targets of the effective ingredients of SAIN (affinity≤-7.0 kcal/mol, H-Bond dist≤3.0, n=6) and visualized the data, Then these targets were verified by using PSORTⅡ, CELLO and BUSCA databases for subcellular localization prediction (n=6). Finally, use the large amount of TCGA clinical data provided by Cbiportal for the prognostic survival analysis of LUAD and LUSC for the genes obtained through the screening. , And consult a large number of documents to verify the results.Results: After screening, comparing, analyzing and verifying a series of data, it is finally confirmed that there are three main active ingredients in SAIN. They are Quercetin, Luteolin and Kaempferol, which mainly act on 6 protein targets. The target mainly regulates 20 signal pathways including Pathways in cancer, Transcriptional misregulation in cancer, EGFR tyrosine kinase inhibitor resistance, Adherens junction, IL-17 signaling pathway, Melanoma, Non-small cell lung cancer and MicroRNAs in cancer. The preventive or therapeutic effects of LUSC. Conclusion: There are three active compounds of Q, L and K in SAIN, which play a role in the treatment and prevention of NSCLC by directly or indirectly regulating the expression of genes such as MMP1, MMP3 and EGFR.

Systems Pharmacology–Based Dissection of Anti-Cancer Mechanism of Traditional Chinese Herb Saussurea involucrata

Cancer has the highest mortality in humans worldwide, and the development of effective drugs remains a key issue. Traditional Chinese medicine Saussurea involucrata (SI) exhibits a series of effects, such as anti-cancer, but the action mechanisms are still unclear. Here, systems pharmacology was applied to reveal its anti-cancer mechanism. First, we screened the active compounds of SI. Then, the compound–target network, target–disease network, and target–pathway network were constructed. DAVID was applied for GOBP analysis and KEGG pathway enrichment analysis on cancer-related targets. Seven potential compounds and 187 targets were identified. The target–disease classification network showed that compounds mainly regulated proteins related to cancer, nervous system diseases, and cardiovascular system diseases. Also, SI anti-tumor effect mainly associated with the regulation of NO production, angiogenesis, MAPK, and PKB from GOBP enrichment. Additionally, KEGG pathway enrichment indicated that targets involved in anti-inflammatory action, inhibiting angiogenesis and anti-proliferation or inducing apoptosis. Experimental validation showed that four active compounds could inhibit cell proliferation and promote apoptosis in A549 (except for kaempferol), PC-3, and C6 cells. This study not only provides experimental evidence for further research on SI in cancer treatment but also promotes the development of potential drugs of SI in modern medicine.

A New Flavonol From Saussurea involucrata With Anti-Inflammatory Activity

A new flavonol named 6-(2'',3''-epoxy-3''-methylbutyl)-resokaempferol (1), together with five known compounds (2-6) were isolated from the EtOAc-soluble extract of the aerial part of Saussurea involucrata. Their structures were elucidated on the basis of spectroscopic methods. All compounds were evaluated for their anti-inflammatory effects by measuring the production of nitric oxide (NO) and TNF-α in vitro. Among them, compound 1 showed potential inhibitory activity on the production of NO and TNF-α in LPS-induced RAW 264.7 cells with IC50 values of 48.0 ± 1.5 and 41.4 ± 1.7 µM, respectively.

A Study on the Anti-obesity Effect of Caffeoylquinic Acids From the Cell Culture of Saussurea Involucrata

Backgroud: Saussurea involucrata Kar. et Kir. (Compositae) (CCSauI) cells are rich in caffeoylquinic acids (CQAs), which have plasma lipid reducing properties and anti-obesity effects, although the mechanisms remain unclear. Clarify CQA’s anti-obesity mechanism and provide new treatments for obesity. Methods: Sprague Dawley (SD) rat were fed a high-fat diet (HFD), then CQAs was intragastric administrated (0.1, 0.5 or 1 mg/mL). Rats were randomly divided into five groups (n=30): NC, MC, TPL (0.1 mg/mL), TPM (0.5 mg/mL) and TPH (1 mg/mL). Digestive enzyme inhibition was obtained by measuring the inhibition rate of α-amylase, α-glucosidase, and pancreatic lipase in vitro. Anti-obesity function was detected by determining the content of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), malondialdehyde (MDA) superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). To analyze the related mechanisms qRT-PCR was employed. Results: From in vitro enzyme activity assays, the IC50 values of the CQAs extract for α-amylase, α-glucosidase, and lipase were 0.631, 0.31, and 0.438 mg/mL, respectively. CQAs administration for 8 weeks decreased retroperitoneal fat and serum and liver TC, TG, LDL-C, and MDA. Comparatively, levels of HDL-C, SOD, and GSH-Px were increased. Real-time fluorescent quantitative PCR showed inhibited expression of fatty acid synthase and 3-hydroxy-3-methyl glutaryl and coenzyme A reductase, peroxisome proliferator-activated receptor-α activation, and 7α-hydroxylase promotion. Conclusion: This study explored the role of CQAs in inhibiting digestive enzyme activities and up-regulating the expression of lipase, significant for prevention and treatment of diabetes and obesity.

SiFBA5, a cold-responsive factor from Saussurea involucrata promotes cold resilience and biomass increase in transgenic tomato plants under cold stress

Background Saussurea involucrata survives in extreme arctic conditions and is very cold-resistant. This species grows in rocky, mountainous areas with elevations of 2400–4100 m, which are snow-covered year-round and are subject to freezing temperatures. S. involucrata’s ability to survive in an extreme low-temperature environment suggests that it has particularly high photosynthetic efficiency, providing a magnificent model, and rich gene pool, for the analysis of plant cold stress response. Fructose-1, 6-bisphosphate aldolase (FBA) is a key enzyme in the photosynthesis process and also mediates the conversion of fructose 1, 6-bisphosphate (FBP) into dihydroxyacetone phosphate (DHAP) and glycerol triphosphate (GAP) during glycolysis and gluconeogenesis. The molecular mechanisms underlying S. involucrata’s cold tolerance are still unclear; therefore, our work aims to investigate the role of FBA in plant cold-stress response. Results In this study, we identified a cold-responsive gene, SiFBA5, based on a preliminary low-temperature, genome-wide transcriptional profiling of S. involucrata. Expression analysis indicated that cold temperatures rapidly induced transcriptional expression of SiFBA5, suggesting that SiFBA5 participates in the initial stress response. Subcellular localization analysis revealed that SiFBA5 is localized to the chloroplast. Transgenic tomato plants that overexpressed SiFBA5 were generated using a CaMV 35S promoter. Phenotypic observation suggested that the transgenic plants displayed increased cold tolerance and photosynthetic efficiency in comparison with wild-type plants. Conclusion Cold stress has a detrimental impact on crop yield. Our results demonstrated that SiFBA5 positively regulates plant response to cold stress, which is of great significance for increasing crop yield under cold stress conditions.