Metabolic-associated fatty liver disease (MAFLD) is a serious condition that can progress to cirrhosis and liver cancer. Natural herbal therapeutics, characterized by their multi-constituent and multi-target properties, as well as favorable safety profiles—particularly lower hepatorenal toxicity—are attracting significant research interest for MAFLD management. In this review, we examine their ethnopharmacological applications, with a focus on autophagy regulation. Information was gathered from traditional medical texts and online databases (e.g., PubMed and CNKI) using keywords such as “MAFLD,” “autophagy,” and “natural herbal plants.” Incorporating herbal plants into MAFLD treatment offers several advantages. First, autophagy regulation involves multiple signaling pathways (e.g., PI3K/AKT/mTOR, AMPK/TFEB, PINK1/Parkin, and Unc-51-like autophagy activating kinase 1 (ULK1)/Beclin-1/VPS34). Single-target drugs often fail to modulate this complex network effectively, whereas various medicinal plants and their bioactive compounds can simultaneously interact with key targets such as mTOR, AMPK, TFEB, SIRT1, LC3B, Beclin-1, ATG5, ULK1, and PPARγ. Second, these plants demonstrate excellent safety profiles. Traditional Chinese compound preparations, such as Zexie Decoction and Shenling Baizhu Powder, have shown clinical efficacy over centuries. To elucidate their mechanisms, researchers are now isolating bioactive compounds from these formulas for cellular and animal studies, revealing their specific roles in modulating autophagy. In summary, plant-derived bioactive compounds—especially those targeting autophagy—have shown promising clinical results against MAFLD and represent valuable candidates for future drug development.

Myocardial ischemia-reperfusion injury (MIRI) involves complex molecular mechanisms. However, the roles of alternative splicing (AS) and RNA-binding proteins (RBPs) in its pathogenesis remain largely elusive. In this study, we conducted an integrated analysis of the public RNA sequencing dataset GSE214122 to identify regulated alternative splicing events (RASEs) and differentially expressed genes (DEGs) in a murine MIRI model. We identified 1262 DEGs (883 upregulated and 379 downregulated), among which 232 were RBPs. Notably, 42 RASE-related genes overlapped with the DEGs. Functional enrichment analysis revealed that aberrantly spliced genes were primarily involved in critical signaling pathways, including mechanistic target of rapamycin (mTOR) and mitogen-activated protein kinase (MAPK). Key genes identified within the mTOR pathway included Eif4e2, Atp6v1h, and Insr, while Traf6, Map4k4, and Nr4a1 were prominent in the MAPK pathway. Gene Ontology (GO) analysis further highlighted biological processes closely associated with MIRI, such as angiogenesis and cellular response to hypoxia. Co-expression network analysis demonstrated that the differentially expressed RBP LMNA was highly correlated with an alternative 5’ splice site (alt5p) event in Atp6v1h (clualt5p2389), the splicing ratio of which was significantly elevated in the MIRI group. Independent experimental validation confirmed the significant upregulation of splice isoforms for Eif4e2, Traf6, Insr, and Nr4a1. Furthermore, mRNA levels of seven RBPs (Anxa2, Fn1, Hyou1, Hif1a, Lmna, Myh9, and Stmn1) were significantly upregulated, whereas Crebrf was significantly downregulated. The Western blot results showed that the protein levels of HIF1A, FN1, LMNA, and EIF4E2 were increased in the MIRI group, while the expression of CREBRF protein was decreased. In conclusion, this study provides a systematic landscape of AS and RBP dysregulation in MIRI. We report for the first time that Lmna-regulated AS of Atp6v1h may participate in the hypoxic response and mTOR pathway modulation. These candidate RBPs and their associated AS events offer novel insights into the molecular mechanisms of MIRI and represent potential therapeutic targets.

ETHNOPHARMACOLOGICAL RELEVANCE:Diabetes is a chronic metabolic disorder characterized by a variety of etiological factors that progressively damage various tissues and organs due to prolonged metabolic dysregulation and elevated blood glucose levels. This condition poses significant risks to both physical and mental health. Natural botanical therapeutics are characterized by their "multi-component, multi-target, and multi-pathway" properties, and their safety profiles have been well established, particularly due to their relatively low hepatotoxic and nephrotoxic effects. Consequently, there is a growing preference among researchers for these agents. AIM OF THE REVIEW:The current review aims to conduct a comprehensive analysis of the ethnopharmacological applications of natural herbaceous plants to mitophagy within the context of diabetic complications. METHODS:An investigation into natural herbal remedies for diabetic complications was conducted, with a particular emphasis on the regulation of mitophagy. This study utilized traditional medical texts and ethnomedicinal literature as primary sources. Furthermore, relevant information related to ethnobotany, phytochemistry, and pharmacology is obtained from online databases, including PubMed and the China National Knowledge Infrastructure (CNKI), among others. "Diabetic complications", "mitophagy", "natural botanical drugs", "traditional Chinese medicine compounds", "single herbs extracts", and "active metabolites" were used as keywords when searching the databases. Consequently, pertinent articles published in recent years were collected and analyzed. RESULT:The integration of herbal plants into managing diabetic complications offers several notable advantages, which can be examined from two primary perspectives. Firstly, the molecular mechanisms that regulate mitophagy are influenced by various signaling pathways, including, but not limited to PINK1/Parkin, PI3K/Akt/mTOR, and AMPK/ULK1. Consequently, pharmacological agents targeting a single pathway may encounter challenges in effectively engaging multiple biological processes. Furthermore, an expanding body of research suggests that numerous herbal plants and their bioactive constituents can modulate various biological targets. These compounds appear to interact with several critical targets associated with mitophagy during diabetic complications, such as PINK1, Parkin, LC3B, Beclin1, p62, ATG5, and Drp1. Secondly, herbal plants and their bioactive compounds have exhibited a favorable safety profile, particularly in terms of diminished hepatotoxicity and nephrotoxicity when compared to conventional Western pharmacotherapy. For example, various compound formulations, including Huangqi Danshen decoction, Ginseng Dingzhi decoction, Shexiang Baoxin pill, Tangzhiqing decoction, and Jianpi Xingqi Huoxue decoction, have been utilized in China for centuries, demonstrating promising clinical efficacy. Recently, an increasing number of researchers have sought to isolate active constituents from these clinically effective compound formulations using diverse chemical methodologies. This endeavor is driven by the necessity to address challenges related to complex ingredient compositions and intricate processing methods. The isolated active compounds have been utilized in cellular and animal studies to elucidate the molecular mechanisms that underlie the efficacy of these formulations. CONCLUSIONS:Numerous traditional compound formulations from China have emerged as promising candidates for the development of pharmacological agents aimed at addressing diabetic complications. Notably, several of these formulations, which focus on the regulation of mitophagy, are currently the subject of extensive research by an increasing number of scholars.

ETHNOPHARMACOLOGICAL RELEVANCE:Chinese herbal medicine constitutes a substantial cultural and scientific resource for the Chinese nation, attracting considerable scholarly interest due to its intrinsic characteristics of "multi-component, multi-target, and multi-pathway" interactions. Simultaneously, it aligns accurately with the intricate and continuously evolving progression of non-small cell lung cancer (NSCLC). Furthermore, contemporary pharmacological studies indicate that natural herbaceous plants and their bioactive compounds exhibit a diverse array of biological activities, including antioxidant, anti-inflammatory, and anti-tumor effects, among others. Additionally, these substances have been demonstrated to possess a degree of safety, particularly in terms of exhibiting comparatively lower levels of toxicity to the liver and kidneys when contrasted with conventional Western medicine. Thus, the development of herbal plants, which includes both single herbs and composite formulations, as well as their bioactive constituents, through the targeted regulation of ferroptosis and mitophagy, presents substantial potential and instills considerable hope for individuals diagnosed with NSCLC. AIM OF THE REVIEW:This review aims to conduct a critical analysis of the ethnopharmacological applications of natural herbaceous plants in relation to ferroptosis and mitophagy in NSCLC. The objective is to evaluate the potential advantages of prioritizing specific phytochemical constituents found in these plants, which may serve as novel therapeutic candidates informed by ethnobotanical knowledge. Additionally, this study seeks to enhance the current pharmacological applications of natural herbaceous plants. METHODS:An investigation into natural herbal remedies for NSCLC was conducted, with a particular emphasis on the ferroptosis and mitophagy pathways. This study utilized traditional medical texts and ethnomedicinal literature as primary sources. Furthermore, relevant information related to ethnobotany, phytochemistry, and pharmacology is obtained from online databases, including PubMed and the China National Knowledge Infrastructure (CNKI), among others. "Traditional Chinese medicine compound preparations", "single herb extracts", "active compounds", "NSCLC", "ferroptosis", and "mitophagy" were used as keywords when searching the databases. Consequently, pertinent articles published in recent years were collected and analyzed. RESULTS:Given the complex etiology of NSCLC, treatment strategies that concentrate exclusively on ferroptosis or mitophagy often demonstrate limitations. In this regard, the utilization of herbal plants offers unique benefits in the management of NSCLC. The rationale can be summarized within the following two dimensions: Firstly, due to the molecular mechanisms of ferroptosis and mitophagy involving multiple signaling pathways (including PINK1/Parkin, HMGB1, system Xc-/GPX4/GSH, FSP1/CoQ10/NAD (P) H, and so on), sometimes drugs with a single target are difficult to involve multiple pathways. Fortunately, there is an expanding body of evidence suggesting that various herbaceous plants and their bioactive compounds can affect multiple biological targets. Moreover, these compounds seem to interact with several targets associated with ferroptosis and mitophagy in NSCLC (such as NIX, BNIP3, FUNDC1, GPX4, FSP1, P53, Nrf2, LncRNA, and so on). Secondly, Herbaceous plants and their bioactive compounds have been shown to possess a favorable safety profile, particularly with respect to reduced hepatotoxicity and nephrotoxicity in comparison to conventional Western medicine. For example, Numerous compound formulations, such as Fangji Huangqi decoction, Mufangji decoction, Qiyu Sanlong decoction, and Fuzheng Kangai decoction, have been employed in China for millennia, and their clinical efficacy appears to be quite promising. Notably, In recent years, numerous researchers have sought to isolate active constituents from clinically effective compound formulations through the application of chemical methodologies. This endeavor has been driven by the necessity to tackle challenges related to complex ingredient compositions and sophisticated processing. These active compounds have been employed in cellular and animal studies to elucidate the molecular mechanisms underlying these formulations. CONCLUSIONS:The Asian region has a long-standing historical tradition of employing natural herbaceous plants for traditional medicinal purposes. Phytochemical and pharmacological studies have shown that various compound preparations derived from traditional Chinese medicine, along with individual herb extracts and their active constituents, display a range of bioactive effects. These effects encompass anti-tumor, anti-inflammatory, antibacterial, and antioxidant properties, among others. Numerous traditional compound formulations originating from China have emerged as promising candidates for the development of pharmacological agents targeting NSCLC. It is noteworthy that a variety of compound formulations aimed at the ferroptosis and mitophagy pathways, which demonstrate unique therapeutic effects on NSCLC, are presently under extensive investigation by an increasing number of researchers. Therefore, it is imperative to consider in vitro mechanistic studies, in vivo pharmacological evaluations, and assessments of clinical efficacy. Furthermore, it is essential to conduct a comprehensive assessment of plant resources, implement quality control measures, and engage in toxicological research to ensure that the data is appropriate for further examination.

Due to the unavoidable impact of ionizing radiation on the heart located near the mediastinum, varying degrees of myocardial damage may occur. As a result, the clinical application of radiotherapy in cancer treatment is significantly limited. However, the molecular mechanisms underlying radiation-induced heart disease (RIHD) are not yet fully understood, and there is a lack of disease-specific treatment strategies. Astragalus polysaccharide (APS), is an active compound abundant in the traditional Chinese herb Astragalus membranaceus (Fisch.) Bunge (AS), has been shown to have cardioprotective effects against various cardiovascular diseases. Thus, this study aims to investigate the potential cardioprotective effect of APS on RIHD and its underlying molecular mechanisms. The network pharmacology results indicated that 9 core genes were identified from the biological network of the effective components of AS acting on RIHD. The results of GO enrichment analysis showed that these hub genes were mainly involved in biological processes such as cell apoptosis, cell proliferation, inflammatory response, and response to external stimuli. The results of KEGG enrichment analysis showed that these hub genes mainly regulated the occurrence of RIHD through pathways such as the EGFR signaling pathway, PI3K/Akt signaling pathway, IL-17 signaling pathway, and so on. In molecular docking analysis, we found that AKT1 and mTOR had good and stable binding abilities with the three types of glucosides rich in AS. The results of in vitro and in vivo experiments all showed that APS could not only improve cardiac dysfunction, myocardial injury, inflammatory response, and myocardial fibrosis in RIHD rats, but also alleviated apoptosis and atrophy of H9C2 cells under ionizing radiation stimulation. In addition, we also found that APS improved the accumulation of autophagic flux induced by ionizing radiation, which could be confirmed by the reversal of Beclin1, p62, LC3B proteins and accelerated degradation of accumulated autophagic vesicles. Rapamycin (Rap) was a classic autophagy flux inducer that could attenuate the improvement effect of APS on H9C2 cell apoptosis under ionizing radiation stimulation. Finally, we found that APS could reverse the inhibition of PI3K/Akt/mTOR signaling pathway activity by ionizing radiation in vitro, thereby improving ionizing radiation-induced autophagy flux accumulation, cardiomyocyte apoptosis, and atrophy. All in all, this study provides important evidence for understanding the molecular mechanisms of the cross-talk between autophagy and apoptosis, and provides new directions and insights for APS combined with autophagy regulators as a therapeutic strategy for RIHD.

Osteosarcoma (OS) has a high recurrence rate, disability rate, mortality and metastasis, it brings great economic burden and psychological pressure to patients, and then seriously affects the quality of life of patients. At present, the treatment methods of OS mainly include radiotherapy, chemotherapy, surgical therapy and neoadjuvant chemotherapy combined with limb salvage surgery. These treatment methods can relieve the clinical symptoms of patients to a certain extent, and also effectively reduce the disability rate, mortality and recurrence rate of OS patients. However, because metastasis of tumor cells leads to new complications, and OS cells become resistant with prolonged drug intervention, which reduces the sensitivity of OS cells to drugs, these treatments still have some limitations. More and more studies have shown that traditional Chinese medicine (TCM) has the characteristics of "multiple targets and multiple pathways," and can play an important role in the development of OS through several key signaling pathways, including PI3K/AKT, Wnt/β-catenin, tyrosine kinase/transcription factor 3 (JAK/STAT3), Notch, transforming growth factor-β (TGF-β)/Smad, nuclear transcription factor-κB (NF-κB), mitogen-activated protein kinase (MAPK), nuclear factor E2-related factor 2 (Nrf2), Hippo/YAP, OPG/RANK/RANKL, Hedgehog and so on. In this paper, the signaling pathways of cross-interference between active ingredients of TCM and OS were reviewed, and the development status of novel OS treatment was analyzed. The active ingredients in TCM can provide therapeutic benefits to patients by targeting the activity of signaling pathways. In addition, potential strategies for targeted therapy of OS by using ferroptosis were discussed. We hope to provide a unique insight for the in-depth research and clinical application of TCM in the fields of OS growth, metastasis and chemotherapy resistance by understanding the signaling crosstalk between active ingredients in TCM and OS.

In recent years, the incidence of intestinal ischemia-reperfusion injury (II/RI), inflammatory bowel disease (IBD), and colorectal cancer (CRC) has been gradually increasing, posing significant threats to human health. Autophagy and endoplasmic reticulum stress (ERS) play important roles in II/RI. Damage caused by ischemia and cellular stress can activate ERS, which in turn initiates autophagy to clear damaged organelles and abnormal proteins, thereby alleviating ERS and maintaining the intestinal environment. In IBD, chronic inflammation damages intestinal tissues and activates autophagy and ERS. Autophagy is initiated by upregulating ATG genes and downregulating factors that inhibit autophagy, thereby clearing abnormal proteins, damaged organelles, and bacteria. Simultaneously, persistent inflammatory stimulation can also trigger ERS, leading to protein imbalance and abnormal folding in the ER lumen. The activation of ERS can maintain cellular homeostasis by initiating the autophagy process, thereby reducing inflammatory responses and cell apoptosis in the intestine. In CRC, excessive cell proliferation and protein synthesis lead to increased ERS. The activation of ERS, regulated by signaling pathways such as IRE1α and PERK, can initiate autophagy to clear abnormal proteins and damaged organelles, thereby reducing the negative effects of ERS. It can be seen that autophagy and ERS play a crucial regulatory role in the development of intestinal diseases. Therefore, the progress in targeted therapy for intestinal diseases based on autophagy and ERS provides novel strategies for managing intestinal diseases. In this paper, we review the advances in regulation of autophagy and ERS in intestinal diseases, emphasizing the potential molecular mechanisms for therapeutic applications.

The death of cells can occur through various pathways, including apoptosis, necroptosis, mitophagy, pyroptosis, endoplasmic reticulum stress, oxidative stress, ferroptosis, cuproptosis, and disulfide-driven necrosis. Increasing evidence suggests that mitophagy and ferroptosis play crucial regulatory roles in the development of stroke. In recent years, the incidence of stroke has been gradually increasing, posing a significant threat to human health. Hemorrhagic stroke accounts for only 15% of all strokes, while ischemic stroke is the predominant type, representing 85% of all stroke cases. Ischemic stroke refers to a clinical syndrome characterized by local ischemic-hypoxic necrosis of brain tissue due to various cerebrovascular disorders, leading to rapid onset of corresponding neurological deficits. Currently, specific therapeutic approaches targeting the pathophysiological mechanisms of ischemic brain tissue injury mainly include intravenous thrombolysis and endovascular intervention. Despite some clinical efficacy, these approaches inevitably lead to ischemia-reperfusion injury. Therefore, exploration of treatment options for ischemic stroke remains a challenging task. In light of this background, advancements in targeted therapy for cerebrovascular diseases through mitophagy and ferroptosis offer a new direction for the treatment of such diseases. In this review, we summarize the progress of mitophagy and ferroptosis in regulating ischemia-reperfusion injury in stroke and emphasize their potential molecular mechanisms in the pathogenesis. Importantly, we systematically elucidate the role of medicinal plants and their active metabolites in targeting mitophagy and ferroptosis in ischemia-reperfusion injury in stroke, providing new insights and perspectives for the clinical development of therapeutic drugs for these diseases.

Autophagy regulates many cell function related to cancer, including cell proliferation, invasion and apoptosis. Therefore, we investigated the potential value of crosstalk between autophagy and apoptosis. The present study demonstrated that seven autophagy related genes were screened from the biological network of salidroside (Sal) acting on liver cancer. The GO analysis showed that these genes were mainly involved in apoptosis and autophagy. The KEGG analysis showed that these genes regulated the process of liver cancer through Th17 cell differentiation, PI3K-Akt signaling pathway and other pathways. Moreover, seven genes were positively correlated with tumor purity, number of B cells, number of CD4(+) T cells, number of CD8(+) T cells, number of macrophages, number of dendritic cells and number of neutrophils. The overall survival time of liver cancer patients in the high expression group of BIRC5, HSP90AB1 and MTOR was lower than that in the low expression group (P < 0.05), while the overall survival time of the liver cancer patients in the high expression group of DLC1 and FOXO1 was higher than that in the low expression group (P < 0.05). In the pan-cancer analysis, we also found that BIRC5, HSP90AB1, MTOR, and ITGA6 were highly expressed in various cancers, while DLC1, FOXO1, and FOS were low expressed in various cancers. In the molecule docking analysis, we found that FOS, HSP90AB1, and MTOR had the best binding ability. Notably, in the vitro validation experiments, Sal was confirmed to induce autophagy and apoptosis, inhibite invasion and metastasis of liver cancer cells through the PI3K/Akt/mTOR signaling pathway. Meanwhile, inhibition of autophagy by chloroquine diphosphate (CQ) promoted Sal-induced mitochondrial apoptosis via corresponding cell and animal experiments. We speculated that Sal-induced autophagy might be a protective mechanism, inhibition of autophagy could further promote the progression of liver cancer. It may provide important insight into the molecular mechanism of crosstalk between autophagy and apoptosis, and provide a new theoretical basis of Sal combined with autophagy inhibitors as a adjuvant chemotherapeutic strategy for human liver cancer.

Abstract Background Salidroside (Sal) is a bioactive component extracted from the rhizome of Rhodiola rosea L. Pharmacological studies have shown that Sal has good anti-cancer properties in various cancers, but the exact mechanism is not clear. Method This study validated the efficacy and explored the potential mechanisms of Sal in treating hepatocellular carcinoma (HCC) by integrating network pharmacology analyses and experimental verification. The pharmacological effects and molecular mechanism of Sal on HCC were explored by network pharmacology approach. HepG2 cells were treated with Sal and/or chloroquine diphosphate (CQ). The cell counting kit-8 (CCK-8) assay, inverted microscope (IM) observation, transmission electron microscope (TEM) observation, various staining were used to detect the condition of autophagy and apoptosis, and the western blotting was used to detect related proteins. Moreover, Sal and/or CQ was also used to treat HCC mice, the hematoxylin and eosin (H & E) staining was used to observe the pathological change of tumor tissue, the immunohistochemistry and western blotting were used to detect the change of related proteins in tumor tissue. Results The network pharmacology approach successfully identified that Sal might adjust autophagy flux through PI3K/AKT/mTOR pathway, which might affect the occurrence and development of HCC. The in vitro experiments indicated that Sal induced HepG2 cells autophagy and apoptosis. The in vitro and vivo experiments indicated that inhibition of autophagy promoted mitochondrial damage and apoptosis induced by Sal. Moreover, Caspase cascade reactions might be involved in these processes, especially the increased expression of cleaved-caspase-3 and cleaved-caspase-9. Notably, Sal also inhibited the activation of PI3K/AKT/mTOR pathway, while CQ promoted the activation of this pathway. Conclusion These findings provide important view for the molecular mechanism of interaction between autophagy and apoptosis, and also provide new insights for monitoring, diagnosis and treatment of HCC.

Immunotherapy is currently the most promising clinical treatment for lung cancer, not only revolutionizing second-line therapy but now also approved for first-line treatment. However, its clinical efficiency is not high and not all patients benefit from it. Thus, finding the best combination strategy to expand anti-PD-1/PD-L1-based immunotherapy is now a hot research topic. The conventional use of chemotherapeutic drugs and targeted drugs inevitably leads to resistance, toxic side effects and other problems. Recent research, however, suggests that by adjusting the dosage of drugs and blocking the activation of mutational mechanisms that depend on acquired resistance, it is possible to reduce toxic side effects, activate immune cells, and reshape the immune microenvironment of lung cancer. Here, we discuss the effects of different chemotherapeutic drugs and targeted drugs on the immune microenvironment. We explore the effects of adjusting the dosing sequence and timing, and the mechanisms of such responses, and show how the effectiveness and reliability of combined immunotherapy provide improved treatment outcomes.

Autophagy is a conserved lysosomal pathway for the degradation of cytoplasmic proteins and organelles, which realizes the metabolic needs of cells and the renewal of organelles. Autophagy-related genes (ATGs) are the main molecular mechanisms controlling autophagy, and their functions can coordinate the whole autophagic process. Autophagy can also play a role in cardiovascular disease through several key signaling pathways, including PI3K/Akt/mTOR, IGF/EGF, AMPK/mTOR, MAPKs, p53, Nrf2/p62, Wnt/β-catenin and NF-κB pathways. In this paper, we reviewed the signaling pathway of cross-interference between autophagy and cardiovascular diseases, and analyzed the development status of novel cardiovascular disease treatment by targeting the core molecular mechanism of autophagy as well as the critical signaling pathway. Induction or inhibition of autophagy through molecular mechanisms and signaling pathways can provide therapeutic benefits for patients. Meanwhile, we hope to provide a unique insight into cardiovascular treatment strategies by understanding the molecular mechanism and signaling pathway of crosstalk between autophagy and cardiovascular diseases.

A microbial ecosystem is a complex community of multiple bacterial interactions. The potential role of gut microbiota in human health has already attracted the attention of many researchers. Dysregulation of the gut microbial community has been suggested to be closely associated with the progression of various chronic diseases. Malignant neoplasms represent a major global health burden and are now the leading cause of death. The formation of tumors is often thought to be influenced by genetic and environmental factors. Recent research advances have indicated that multiple malignancies may also be attributed to the gut microbiota. In this review, we highlight the complex interactions between gut microbes and their metabolites, as well as the potential impact of gut microecology on the occurrence and development of tumors. In addition, potential strategies for targeted therapy of tumors using gut microecology are discussed. In the near future, intestinal microecology is likely to be used for early screening of tumors and subsequent clinical treatment.

Abstract This study aims to investigate the effect of five-flavor sophora flavescens enteric-coated capsules (FSEC) on TNF-α-induced inflammatory bowel disease and its molecular mechanism. Wistar Rats were divided divided into 6 groups: Normal control group (group A): normal diet, drinking water; Model group (group B): 100 μg/L TNF-α; FSEC high-dose group (group C): 100μg/L TNF-α + FSEC (432 mg/kg); FSEC medium-dose group (group D): 100 μg/L TNF-α + FSEC (216 mg/kg); FSEC low-dose group (group E): 100 μg/L TNF-α + FSEC (108 mg/kg); Positive control group (group F): 100 μg/L TNF-α + 500 mg/kg sulfasalazine (SAZ). Animals in each group were intragastrically administered twice daily for 7 days. Animals were sacrificed 24 hours after the last treatment and colon tissues were collected for subsequent experiments. The results of HE staining showed that the colonic tissue of TNF-α-fed animals appeared damage, while the colonic tissue of animals treated with FSEC was improved to various degrees, and the histological characteristics of colon were basically recovered in the high-dose group, suggesting that FSEC could be used to treat TNF-α-induced colonic tissue damage. According to the results of ELISA and immunohistochemistry, the recovery of colonic tissue structure in rats treated with different doses of FSEC might be related to the decrease of TNF-α, IL-6, IL-17, TLR-4 and NF-κB proteins expression. According to the results of Western blotting, TNF-α-pretreated IEC-6 cells cultured with medicated serum decreased the expression of TRIF and IFN-γ proteins. These results suggest that FSEC has a protective effect on ulcerative colitis (UC), and the mechanism may be through inhibiting the activation of TLR-4/NF-κB signaling pathway and preventing the release of related inflammatory factors.

Hepatocellular carcinoma (HCC) is the leading cause of cancer-associated death in the world. Chemotherapy remains the primary treatment method for HCC. Despite advances in chemotherapy and modalities, recurrence and resistance limit therapeutic success. Salidroside (Sal), a bioactive component extracted from the rhizome of Rhodiola rosea L, exhibits a spectrum of biological activities including antitumor effects. In the present study, it was demonstrated that Sal could induce apoptosis and autophagy of 97H cells by using CCK-8 assay, transmission electron microscopy (TEM), Hoechst33342 staining, MDC staining, western blotting. Pretreatment with Sal enhanced apoptosis and autophagy via upregulation of expression levels of Bax, Caspase-3, Caspase-9, light chain (LC)3-II and Beclin-1 proteins and downregulation of expression levels of Bcl-2, LC3-I and p62 protein in 97H cells. Furthermore, Sal was demonstrated to inhibit activation of the PI3K/Akt/mTOR signaling pathway and, when combined with autophagy inhibitor chloroquine diphosphate (CQ), increased phosphorylation of PI3K, Akt and mTOR proteins. The combined treatment with Sal and CQ not only decreased Sal-induced autophagy, but also accelerated Sal-induced apoptosis. Therefore, Sal-induced autophagy might serve a role as a defense mechanism in human liver cancer cells and its inhibition may be a promising strategy for the adjuvant chemotherapy of liver cancer.

AbstractBackground Salidroside (Sal) has many pharmacological effects, including anti-fatigue, anti-aging, immunoregulation and so on. However, its effects and related regulatory mechanisms on coronary artery disease (CAD) are elusive. We hypothesize that Sal may play a role in CAD through autophagic pathway. Herein, we used autophagy agonist (sirolimus, Sir) and inhibitor (chloroquine diphosphate, CQ) as the autophagic control group. And network pharmacology prediction was employed to explore the pharmacological effects and molecular mechanisms of Sal on CAD using network pharmacology prediction, and validated the results through in vitro experiments. Methods Through a search in the Swisstarget Prediction database, chemical composition and targets were retrieved. The related-autophagic targets for CAD were then obtained from the GEO and human autophagy databases. The network was constructed that depicted the interactions between putative drugs (Sal, Sir and CQ) and known related-autophagic targets for CAD using Cytoscape 3.8.0. Analysis of protein-protein interaction (PPI) was achieved via STRING software, followed by gene ontology (GO) functional enrichment and kyoto enrichment of gene and genome (KEGG) pathway analyses. The 3D structure of target protein was downloaded from RSCB PDB database, the 2D structure of drug was obtained from PubChem database, then the molecular docking was constructed by using AutoDockTools 1.5.6 and AutoDock Vina software, and the results were visualized by using PyMoL sofeware. To validate the computer-predicted results, in vitro experiments based on an anoxic injury model were designed by using CoCl2-exposed H9C2 cells, and treated with various concentrations of Sal/Sir/CQ. The effects of drugs on cells were detected by using the CCK-8, inverted microscope (IM), transmission electron microscope (TEM), MDC staining and AO staining. Moreover, the expression profiles of related-proteins were analyzed via western blot. Results Three compounds were identified a total of 399 targets, GEO revealed a total of 576 CAD-related genes, and 36 overlapping genes of drug-disease were obtained. The PPI network was employed for the analysis of 36 target proteins, including CASP3, mTOR, VEGFA and EGFR. The enrichment analysis demonstrated candidate targets of three compounds were more frequently involved in inflammation, lipid response, oxidative stress, autophagy, apoptosis, diabetic cardiomyopathy and estrogen signaling pathways. In vitro experiments revealed that Sal could induce autophagy in CoCl2-exposed H9C2 cells, significantly reverse cell death in the early period, and decrease the levels of Caspase-3 protein in CoCl2-exposed H9C2 cells. However, Sal-induced autophagy also could promoted the process of CoCl2-exposed H9C2 cells death in the late period. Conclusions This study revealed that Sal existed the effect of treating CAD by regulating multi-targets and multi-channels through the method of network pharmacology. Furthermore, in vitro results confirmed that Sal-induced autophagy played a protective role in the early period of CAD, while aggravated the condition in the late period of CAD.