NOP14-mediated ribosome biogenesis is required for mTORC2 activation and predicts rapamycin sensitivity.

The mechanistic target of rapamycin (mTOR) forms two distinct complexes: rapamycin-sensitive mTOR complex 1 (mTORC1) and rapamycin-insensitive mTORC2. mTORC2 primarily regulates cell survival by phosphorylating Akt, though the upstream regulation of mTORC2 remains less well-defined than that of mTORC1. In this study, we show that NOP14, a 40S ribosome biogenesis factor and a target of the mTORC1-S6K axis, plays an essential role in mTORC2 signaling. Knockdown of NOP14 led to mTORC2 inactivation and Akt destabilization. Conversely, overexpression of NOP14 stimulated mTORC2-Akt activation and enhanced cell proliferation. Fractionation and coimmunoprecipitation assays demonstrated that the mTORC2 complex was recruited to the rough endoplasmic reticulum through association with endoplasmic reticulum-bound ribosomes. In vivo, high levels of NOP14 correlated with poor prognosis in multiple cancer types. Notably, cancer cells with NOP14 high expression exhibit increased sensitivity to mTOR inhibitors, because the feedback activation of the PI3K-PDK1-Akt axis by mTORC1 inhibition was compensated by mTORC2 inhibition partly through NOP14 downregulation. In conclusion, our findings reveal a spatial regulation of mTORC2-Akt signaling and identify ribosome biogenesis as a potential biomarker for assessing rapalog response in cancer therapy.

1,8-cineole ameliorates colon injury by downregulating macrophage M1 polarization via inhibiting the HSP90-NLRP3-SGT1 complex.

Ulcerative colitis (UC) is characterized by chronic relapsing intestinal inflammation. Currently, there is no effective treatment for the disease. According to our preliminary data, 1,8-cineole, which is the main active compound of Amomum compactum Sol. ex Maton volatile oil and an effective drug for the treatment of pneumonia, showed remarkable anti-inflammatory effects on colitis pathogenesis. However, its mechanism of action and direct targets remain unclear. This study investigated the direct targets and mechanism through which 1,8-cineole exerts its anti-inflammatory effects using a dextran sulfate sodium salt-induced colitis mouse model. The effects of 1,8-cineole on macrophage polarization were investigated using activated bone marrow-derived macrophages and RAW264.7 cells. In addition, 1,8-cineole targets were revealed by drug affinity responsive target stability, thermal shift assay, cellular thermal shift assay, and heat shock protein 90 (HSP90) adenosine triphosphatases (ATPase) activity assays. The results showed that 1,8-cineole exhibited powerful anti-inflammatory properties in vitro and in vivo by inhibiting the macrophage M1 polarization and protecting intestinal barrier function. Mechanistically, 1,8-cineole directly interacted with HSP90 and decreased its ATPase activity, also inhibited nucleotide-binding and oligomerization domain-, leucine rich repeat-, and pyrin domain-containing 3 (NLRP3) binding to HSP90 and suppressor of G-two allele of SKP1 (SGT1) and suppressed NLRP3 inflammasome activation in macrophages. These results demonstrated that 1,8-cineole is a potential drug candidate for UC treatment.

Proanthocyanidins attenuates ferroptosis against influenza-induced acute lung injury in mice by reducing IFN-γ.

BACKGROUND: Acute lung injury (ALI) is associated with high morbidity and mortality and is partly driven promoted by ferroptosis. Proanthocyanidins (PAs) is a natural bioactive flavonoid with anti-inflammatory and antioxidant activities. PAs can also significantly protect against acute lung inflammation and ferroptosis in alveolar epithelial cells. However, it is unclear whether PAs can alleviate ALI by reducing ferroptosis. This study aimed to evaluate the protective effects of PAs and the potential mechanisms against Influenza A virus (IAV)-induced ALI. METHODS: Mice were inoculated nasally with IAV to induce ALI. IAV-induced pulmonary inflammation and ferroptosis was tested by measuring the levels of malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase 4 (GPX4), solute carrier family 7 member 11 (SLC7A11) and acyl-CoA synthetase long-chain family member (ACSL4) in lung tissue. The potential targets that PAs protect against IAV-induced ALI were determined via a systemic pharmacological analysis. The molecular mechanism of PAs in ALI treatment was investigated by assessing the level of inflammation and ferroptosis markers using Western Blot and quantitative real-time PCR. RESULTS: Systemic pharmacological analysis suggested that PAs protect against IAV-induced pneumonia thorough TGF-ß1 and its relative signaling pathway. PAs effectively alleviated histopathological lung injury, reduced inflammatory cytokines and chemokines secretion, which were increased in IAV-infected mice. Meanwhile, PAs further prevented mouse airway inflammation in ALI, concomitant with the decreased expression TGF-ß1, smad2/3, p-Smad2, p-Smad3 and ferroptosis mediator IFN-γ. Furthermore，IFN-γ promotes cell lipid peroxidation and ferroptosis，PAs significantly reduced MDA and ACSL4 levels and upregulated GSH, GPX4, and SLC7A11. CONCLUSION: Overall, PAs can attenuate ferroptosis against IAV-induced ALI via the TGF-ß1/Smad2/3 pathway and is a promising novel therapeutic candidate for ALI.

Effect of Traditional Chinese Medicine on Psoriasis Vulgaris: A Meta-Analysis and Systematic Review.

BACKGROUND: Psoriasis is a chronic, inflammatory autoimmune skin disease that affects 2-3% of the world's population. Lesions are mainly found on the limbs, trunk, and scalp, but may also affect other parts of the body, and the cause is not yet known. The chronic and relapsing nature of psoriasis makes it one of the most complex and important diseases in current dermatology research. METHODS: The search was conducted using PubMed, Embase, Cochrane Library, Web of Science, China National Knowledge Internet, Wanfang Data, VIP journals database, and Chinese biomedical literature database (CBM). The retrieval time limit was from the establishment of the database to January 2021. The quality of the selected literature was evaluated, and ReView Manager 5.3 was used for meta-analysis after randomized controlled trials were filtered. RESULTS: Finally, 16 randomized controlled trials involving 1,967 patients were included. The total effective rate (OR = 3.68, 95% CI [2.73, 4.95], p < 0.00001), cure rate (OR = 2.01, 95% CI [1.62, 2.49], p < 0.00001), and PASI score (OR = -1.83, 95% CI [-2.39, -1.26], p < 0.00001) of the traditional Chinese medicine (TCM) were superior to the Diyin tablet. CONCLUSION: In the treatment of psoriasis, TCM shows higher efficacy than the Diyin tablet. However, due to the limitations of the included literature, we still need more double-blind, placebo-controlled trials with large samples and multiple centers to provide high-quality clinical evidence.

Adlay (Coix lacryma-jobi L.) Polyphenol Improves Hepatic Glucose and Lipid Homeostasis through Regulating Intestinal Flora via AMPK Pathway.

SCOPE: Non-alcoholic fatty liver disease (NAFLD) is a type of metabolic syndrome characterized of abnormal lipid deposition in the liver. Adlay polyphenol (AP), an effective component extracted from Coix lacryma-jobi L., has been reported that it can be used as a dietary supplement to prevent NAFLD. In this study, the mechanism and action of AP on lipid metabolism and regulation of intestinal flora are investigated. METHODS AND RESULTS: AP significantly decreases the lipid accumulation in free fatty acid-treated HepG2 cells. Western blot results indicate that AP improves lipid metabolism via activating the p-AMPK/p-ACC pathway. In vivo experiments show AP treatment significantly decreases the body weight, liver weight, hepatic triglyceride, and total cholesterol contents, as well as the serum glucose levels in high fat diet-fed mice, which may affect lipid accumulation by activating AMPK pathway and changing intestinal bacterial communities and intestinal microbiome metabolism. CONCLUSION: AP can be used as a food supplement for improving lipid metabolic dysfunction and reducing the incidence of metabolic diseases.

Luteolin induces pyroptosis in HT-29 cells by activating the Caspase1/Gasdermin D signalling pathway.

Luteolin, which is a natural flavonoid, has anti-inflammatory, antioxidant, and anticancer properties. Numerous studies have proven that luteolin inhibits the growth of many types of cancer cells by promoting apoptosis, autophagy, and cell cycle arrest in tumour cells. However, in vivo research on this topic has been limited. In addition, other studies have shown that luteolin exerts a good inhibitory effect on apoptosis-resistant cancer cells. While existing studies have not completely elucidated the mechanism underlying this phenomenon, we assume that luteolin, which is a natural compound that exerts its effects through various mechanisms, may have the potential to inhibit tumour growth. In our study, we proved that luteolin exerted a good inhibitory effect on the proliferation of colon cancer cells according to CCK8 and EdU fluorescence assays, and the same conclusion was drawn in animal experiments. In addition, we found that luteolin, which is an antioxidant, unexpectedly promoted oxidative stress as shown by measuring the levels of oxidative balance-related indicators, such as reactive oxygen species (ROS), SOD, H2O2 and GSH. However, the decreased oxidation of luteolin-treated HT-29 cells after treatment with the active oxygen scavenger NAC did not reverse the inhibition of cell growth. However, the Caspase1 inhibitor VX765 did reverse the inhibition of cell growth. Western blotting analysis showed that luteolin treatment increased the expression of Caspase1, Gasdermin D and IL-1ß, which are members of the pyroptosis signalling pathway, in colon cancer cells. We further intuitively observed NLRP3/Gasdermin D colocalization in luteolin-treated HT-29 cells and mouse tumour tissues by immunofluorescence. These results suggest that luteolin inhibits the proliferation of colon cancer cells through a novel pathway called pyroptosis. This study provides a new direction for the development of natural products that inhibit tumour growth by inducing pyroptosis.

Comparison Between Wedge Resection and Lobectomy/Segmentectomy for Early-Stage Non-small Cell Lung Cancer: A Bayesian Meta-analysis and Systematic Review.

BACKGROUND: Surgery has become an accepted method for the treatment of early-stage non-small cell lung cancer (NSCLC). The purpose of this Bayesian meta-analysis was to compare the overall survival (OS), disease-free survival (DFS), and relapse-free survival (RFS) between wedge resection and lobectomy/segmentectomy for treatment of early-stage NSCLC. METHODS: Eligible studies were retrieved from Web of Science, PubMed, MEDLINE, Cochrane Library, EMBASE, CNKI, and WanFang up to July 2021 and screened based on established selection criteria. The Bayesian meta-analysis was performed with the combination of the reported survival outcomes of the individual studies using a random-effect model. The OS, DFS, and RFS of the wedge resection group was compared with the lobectomy/segmentectomy group. The hazard ratio (HR) and standard error were extracted or calculated for each study using the Kaplan-Meier method. RESULTS: This study was registered with PROSPERO (INPLASY202080090).The pooled OS hazard ratio between segmentectomy and lobectomy was 1.1 [95% confidence interval (CI) 0.92-1.4], the pooled HR between lobectomy and wedge resection was 0.71 [95% CI 0.52-0.96], and the pooled HR between segmentectomy and wedge was 0.80 [95% CI 0.56-1.10]. The pooled HR of DFS or RFS was not statistically significant among the three surgical approaches. CONCLUSIONS: Patients with early-stage NSCLC received lobectomy had the lowest hazard ratio of OS than patients received wedge resection, indicating that the overall survival of patients received lobectomy was higher than patients received wedge resection. However, regarding DFS and RFS, the three surgical approaches showed no significant difference.

Higenamine Attenuates Neuropathic Pain by Inhibition of NOX2/ROS/TRP/P38 Mitogen-Activated Protein Kinase/NF-ĸB Signaling Pathway.

Oxidative stress damage is known as one of the important factors that induce neuropathic pain (NP). Using antioxidant therapy usually achieves an obvious curative effect and alleviates NP. Previous pharmacological studies have shown that higenamine (Hig) performs to be antioxidant and anti-inflammatory. However, the protective effect and mechanism of Hig on NP are still unclear. This study mainly evaluated the changes in reactive oxygen species (ROS) level, lipid peroxidation, and antioxidant system composed of superoxide dismutase (SOD) and glutathione (GSH) through chronic constrict injury (CCI) model rats and t-BHP-induced Schwann cell (SC) oxidative stress model. The expressions of two inflammatory factors, tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6), were also assessed. The possible molecular mechanism of Hig in the treatment of NP was explored in conjunction with the expression of mitochondrial apoptosis pathway and NOX2/ROS/TRP/P38 mitogen-activated protein kinase (MAPK)/NF-ĸB pathway-related indicators. Hig showed substantial antioxidant and anti-inflammatory properties both in vivo and in vitro. Hig significantly reduced the upregulated levels of ROS, malondialdehyde (MDA), TNF-α, and IL-6 and increased the levels of SOD and GSH, which rebalanced the redox system and improved the survival rate of cells. In the animal behavioral test, it was also observed that Hig relieved the CCI-induced pain, indicating that Hig had a pain relief effect. Our research results suggested that Hig improved NP-induced oxidative stress injury, inflammation, and apoptosis, and this neuroprotective effect may be related to the NOX2/ROS/TRP/P38 MAPK/NF-ĸB signaling pathway.

Protective effect of paeoniflorin on H2O2 induced Schwann cells injury based on network pharmacology and experimental validation.

This study was to investigate the protective effect of paeoniflorin (PF) on hydrogen peroxide-induced injury. Firstly, "SMILES" of PF was searched in Pubchem and further was used for reverse molecular docking in Swiss Target Prediction database to obtain potential targets. Injury-related molecules were obtained from GeenCards database, and the predicted targets of PF for injury treatment were selected by Wayne diagram. For mechanism analysis, the protein-protein interactions were constructed by String, and the KEGG analysis was conducted in Webgestalt. Then, cell viability and cytotoxicity assay were established by CCK8 assay. Also, the experimental cells were allocated to control, model (200 µmol·L-1 H2O2), SB203580 10 µmol·L-1 (200 µmol·L-1 H2O2+ SB203580 10 µmol·L-1), PF 50 µmol·L-1 (200 µmol·L-1 H2O2+ PF 50 µmol·L-1), and PF 100 µmol·L-1 (200 µmol·L-1 H2O2+ PF 100 µmol·L-1) groups. We measured the intracellular ROS, Hoechst 33258 staining, cell apoptosis, the levels of Bcl-xl, Bcl-2, Caspase-3, Cleaved-caspase3, Cleaved-caspase7, TRPA1, TRPV1, and the phosphorylation expression of p38MAPK. There are 96 potential targets that may be associated with PF for injury treatment. Then, we chose the "Inflammatory mediator regulation of TRP channels" pathway for the experimental verification from the first 10 KEGG pathway. In experimental verification, H2O2 decreased the cell viability moderately (P < 0.05), and 100 µmol·L -1 PF increased the cell viability significantly (P < 0.05). Depending on the difference of intracellular ROS fluorescence intensity, PF inhibited H 2O2-induced reactive oxygen species production in Schwann cells. In Hoechst 33258 staining, PF reversed the condensed chromatin and apoptotic nuclei following H2O2 treatment. Moreover, Flow cytometry results showed that PF could substantially inhibit H2O2 induced apoptosis (P < 0.05). Pretreatment with PF obviously reduced the levels of Caspase3, Cleaved-caspase3, Cleaved-caspase7, TRPA1, TRPV1, and the phosphorylation expression of p38MAPK after H 2O2 treatment (P < 0.05), increased the levels of Bcl-2, and Bcl-xl ( P < 0.05). PF inhibited Schwann cell injury and apoptosis induced by hydrogen peroxide, which mechanism was linked to the inhibition of phosphorylation of p38MAPK.

Identification of Potential Biomarkers of Depression and Network Pharmacology Approach to Investigate the Mechanism of Key Genes and Therapeutic Traditional Chinese Medicine in the Treatment of Depression.

BACKGROUND: To explore the potential target of depression and the mechanism of related traditional Chinese medicine in the treatment of depression. METHOD: Differential gene expression in depression patients and controls was analyzed in the GEO database. Key genes for depression were obtained by searching the disease databases. The COREMINE Medical database was used to search for Chinese medicines corresponding to the key genes in the treatment of depression, and the network pharmacological analysis was performed on these Chinese medicines. Then, protein-protein interaction analysis was conducted. Prediction of gene phenotypes was based on Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment scores. RESULTS: The total number of differentially expressed genes in the GEO database was 147. Combined with the GEO dataset and disease database, a total of 3533 depression-related genes were analyzed. After screening in COREMINE Medical, it was found that the top 4 traditional Chinese medicines with the highest frequency for depression were Paeonia lactiflora Pall., Crocus sativus L., Bupleurum chinense DC., and Cannabis sativa L. The compound target network consisted of 24 compounds and 138 corresponding targets, and the key targets involved PRKACA, NCOA2, PPARA, and so on. GO and KEGG analysis revealed that the most commonly used Chinese medicine could regulate multiple aspects of depression through these targets, related to metabolism, neuroendocrine function, and neuroimmunity. Prediction and analysis of protein-protein interactions resulted in the selection of nine hub genes (ESR1, HSP90AA1, JUN, MAPK1, MAPK14, MAPK8, RB1, RELA, and TP53). In addition, a total of four ingredients (petunidin, isorhamnetin, quercetin, and luteolin) from this Chinese medicine could act on these hub genes. CONCLUSIONS: Our research revealed the complicated antidepressant mechanism of the most commonly used Chinese medicines and also provided a rational strategy for revealing the complex composition and function of Chinese herbal formulas.

Therapeutic Effect of Tetrapanax papyriferus and Hederagenin on Chronic Neuropathic Pain of Chronic Constriction Injury of Sciatic Nerve Rats Based on KEGG Pathway Prediction and Experimental Verification.

BACKGROUND: Hederagenin is one of the main components of Tetrapanax papyriferus, and Tetrapanax papyriferus is one of the ingredients of Danggui Sini decoction. To explore whether Tetrapanax papyriferus and hederagenin can alleviate mechanical pain, thermal hyperalgesia, and cold pain at the same time, we comprehensively investigated the effects of two drugs on the levels of p38 MAPK phosphorylation, TRP proteins, and IL1ß, IL6, and TNF-α in serum. METHODS: Firstly, we obtained pain-related targets and performed KEGG pathway enrichment on these targets. Then, 42 SD rats were separated randomly into six groups: sham operation group, CCI group, pregabalin group, mecobalamin group, Tetrapanax papyriferus group, and hederagenin group. All drugs were given orally. Rats in the sham operation group and CCI group were gavaged with saline. Rats in the pregabalin group were given pregabalin, while rats in the mecobalamin group were given mecobalamin. Rats in the Tetrapanax papyriferus group were given Tetrapanax papyriferus, while rats in the hederagenin group were given hederagenin. Besides, we conducted behavioral tests including acetone test, hot plate experiment, and von Frey filaments, and then dorsal root ganglion neurons were taken out on the 21st day after operation. Then, western blot, ELISA, and hematoxylin-eosin staining were conducted. RESULTS: Rats in the CCI group were more sensitive to hyperalgesia and allodynia to mechanical and thermal stimuli, as well as cold pain. All four drugs could relieve these pains. Pregabalin, mecobalamin, and Tetrapanax papyriferus can reduce the levels of IL1ß, IL6, and TNF-α in serum compared to those of the CCI group. The expression of TRPM8, TRPA1, TRPV1, TRPV4, and phosphorylated p38 MAPK in DRG increased evidently on the 21st day after the operation in the CCI group. All four drugs could reduce the expressions of TRPM8, TRPA1, TRPV1, TRPV4, and phosphorylated p38 MAPK in dorsal root ganglion compared to those of the CCI group. CONCLUSION: Tetrapanax papyriferus and hederagenin relieved sciatica by reducing inflammation levels, inhibiting p38 MAPK phosphorylation, and decreasing the levels of dorsal root ganglion proteins.

Higenamine exerts an antispasmodic effect on cold-induced vasoconstriction by regulating the PI3K/Akt, ROS/α2C-AR and PTK9 pathways independently of the AMPK/eNOS/NO axis.

The present study aimed to investigate the antispasmodic effect of higenamine on cold-induced cutaneous vasoconstriction and the underlying molecular mechanisms. A cold-induced cutaneous vasoconstriction rat model was established and different doses of higenamine were delivered by intravenous injection. The changes of cutaneous regional blood flow (RBF) between groups were analyzed. In vitro, the proliferation of human dermal microvascular endothelial cells was measured by MTT. The NO concentration was detected by a nitrate reductase assay. Flow cytometry was applied to measure reactive oxygen species (ROS) levels. The protein expression levels were detected by western blotting. The results demonstrated that in the model group, RBF declined compared with the normal control group, but was reversed by treatment with higenamine. The expression of endothelial nitric oxide synthase (eNOS), phosphorylated (p)-eNOS, protein kinase B (Akt1), p-Akt1, AMP-activated protein kinase (AMPK) α1 and p-AMPKα1 was upregulated by hypothermic treatment but was reversed by higenamine treatment. Treatment with higenamine significantly reduced the level of intracellular α2C-adrenoreceptor (AR) compared with the hypothermia group (P<0.05). Furthermore, the expression of twinfilin-1 (PTK9) was downregulated in the higenamine and positive control groups compared with the hypothermia group (P<0.05). Compared with the hypothermia group, the levels of ROS and α2C-AR (intracellular & membrane) were decreased in higenamine and the positive control group (P<0.05 and P<0.01, respectively). This study, to the best of our knowledge, is the first to assess the effects of higenamine on cold-induced vasoconstriction in vivo and its molecular mechanisms on the PI3K/Akt, AMPK/eNOS/nitric oxide, ROS/α2C-AR and PTK9 signaling pathways under hypothermia conditions. Higenamine may be a good therapeutic option for Raynaud's phenomenon (RP) and cold-induced vasoconstriction.