Expression Of PI3K Research Articles

Myocardial Protective Effect of Xuebijing Injection on Extracorporeal Membrane Oxygenation Perfused Isolated Langendorff Heart Based on the PI3K/AKT Pathway

Objective: This study aimed to investigate the myocardial protective effect of Xuebijing injection in a model of extracorporeal membrane oxygenation (ECMO) isolated heart perfusion and determine the role of the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) pathway. Methods: To establish an ECMO isolated heart perfusion model, Guangxi Bama miniature pigs were randomly divided into two groups: (1) a normal saline group (NS group), in which 50 mL of normal saline was added to the perfusion solution; and (2) a Xuebijing injection group (XBJI group), in which 10 mL of XBJI was added to the perfusion solution and then continuously pumped at 5 mL/h. Perfusion was maintained for 8 h. The hemodynamic changes, inflammatory response, and myocardial enzyme levels at T0, T2, T4, T6, and T8 were evaluated. The protein expression level of the PI3K/AKT pathway was analyzed by Western blot analysis and Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR). Hematoxylin and eosin staining and transmission electron microscopy were used to observe the pathological morphology and ultrastructure of T8 cardiomyopathy. Results: There was no significant difference in hemodynamics between the two groups. XBJI could reduce serum inflammatory factors and myocardial enzyme levels. XBJI also upregulated the expression of PI3K and AKT mRNA and the phosphorylation levels of PI3K and AKT proteins. The results of pathological and electron microscopy showed that XBJI effectively reduced myocardial cell and mitochondrial damage. Conclusion: XBJI can reduce myocardial inflammation and myocardial cell and mitochondrial damage in isolated heart perfusion. XBJI may play a role in the myocardial protection of ECMO isolated heart perfusion by activating the PI3K/AKT pathway.

Gynostemma pentaphyllum (Thunb.) Makino Affects Autophagy and Improves Diabetic Peripheral Neuropathy Through TXNIP-Mediated PI3K/AKT/mTOR Signaling Pathway.

TXNIP is closely associated with diabetic peripheral neuropathy (DPN). Gynostemma pentaphyllum (Thunb.) Makino (GP), a perennial herb with five leaves, is considered to have medicinal values. However, it is unknown whether GP alleviates DPN by modulating TXNIP-mediated autophagy. The aim of this study was to evaluate the effect of GP on Schwann cell injury during DPN and to investigate the mechanism of GP in DPN for the first time. High-fat diet-fed GK rats and high-glucose-cultured RSC96 cells were used to establish DPN models. The effects of GP on DPN were investigated by blood glucose assay, neurological function assay, pathology assay, and immunohistochemistry. To investigate the effect of GP on autophagy and upstream PI3K/AKT/mTOR signaling pathway in Schwann cells, Western blot and immunofluorescence assay were performed on RSC96 cells to detect the expression of beclin-1 and LC3. Western blot method was used to detect the expression of PI3K, p-Akt/Akt, p-mTOR/mTOR, and RT-qPCR method and was used to detect the expression of PI3K. Apoptosis was detected by flow cytometry. The effects of TXNIP on the above indicators were also detected in RSC96 cells. Finally, the mechanism of GP regulation of autophagy and apoptosis in RSC96 cells was verified. GP reduced blood glucose level, attenuated peripheral nerve myelin damage, and improved nerve function in DPN rats. In addition, GP enhanced autophagy activity and reduced apoptosis in RSC96 cells. GP promoted autophagy by regulating TXNIP-mediated PI3K/AKT/mTOR signaling pathway, and GP reduced apoptosis in RSC96 cells by promoting cellular autophagy. GP attenuates DPN myelin damage in RSC96 cells by enhancing autophagy, and its mechanism may be related to the inhibition of PI3K/AKT/mTOR signaling pathway by up-regulating the expression of TXNIP.

Efficacy of silver-doped Carbon dots in Chemical Castration: a rat model study

This study evaluates silver-doped carbon dots (AgCDs) as a novel agent for chemical castration using a rat model. Six groups of rats (five males and ten females each, except for the surgical group which had only males) were utilized to compare the effects of different concentrations of AgCDs. The groups included control, sham, and three experimental groups injected with 1.25, 50, and 200 µg/mL AgCDs, respectively, along with a surgical castration group. Testosterone levels, sperm parameters, fertility index, oxidative damage, histopathological parameters, and gene expression of P53, Bax, Bcl-2, caspase-3, AKT, and PI3K were analyzed. Results demonstrated that the high-dose AgCDs group significantly reduced testosterone levels, sperm concentration, and motility, resulting in a decreased fertility index. MDA and NO significantly increased, while CAT, SOD, GPx, and TAC significantly reduced in the chemically castrated groups. Histological and genes expression analysis also revealed apoptosis and testicular damage in the AgCDs groups, indicated by significant increases in P53, Bax, and Caspase-3 levels, and significant reductions in AKT, PI3K, and Bcl-2. Based on these findings, AgCDs could be considered a potent and efficient agent for chemical castration, offering a less invasive, cost-effective solution with potential applications for population control.

Quinoa ameliorates polycystic ovary syndrome via regulating gut microbiota through PI3K/AKT/mTOR pathway and autophagy

BackgroundPolycystic ovary syndrome (PCOS) is a unity of endocrine and metabolic disorders, associated with PI3K/AKT/mTOR, autophagy, and gut microbiota. Quinoa is a valuable food source, which contains rich minerals, unsaturated fatty acids, and has a positive modulating effect on metabolic diseases. However, its effects and potential mechanisms on PCOS have not been reported yet. Therefore, the purpose of this study is to investigate the effect of quinoa on PCOS rats by regulating PI3K/AKT/mTOR, autophagy, and gut microbiota.MethodsTen–week-old female Sprague-Dawley (SD) rats have received letrozole for 24 days for induction of PCOS and subsequently were treated with a quinoa diet for 8 weeks. Vaginal smears were used to analyze the estrous cycle of rats. Hormone and biochemical indexes were analyzed by kit assays and glucometer. The pathological changes of ovary, pancreas, duodenum and colon were observed by HE staining. PI3K, AKT, mTOR and autophagy-related proteins in the ovary and colon were measured by western blot and immunohistochemistry staining. Tight junction proteins in colon were measured by immunohistochemistry staining. 16 s rDNA sequencing was used to detect the changes of intestinal microbiota in rats. Network pharmacology and molecular docking were used to study the possible targets and mechanisms of quinoa on PCOS. Spearman correlation analysis was used to study the relationship between intestinal microbial abundance and hormone levels of PCOS rats at the phylum and genus level.ResultsQuinoa significantly improved estrous cycle and biochemical parameters of PCOS-like rats, and the pathological state of ovary, pancreas, duodenum and colon tissues. Especially, quinoa significantly regulated the expression of PI3K, AKT, mTOR and autophagy-related proteins in the ovary. Quinoa may repair the intestinal barrier by upregulating the expression of tight junction proteins in the colon, and regulate autophagy-related factors in colon. Additionally, quinoa increased the abundance of Lactobacillu, Bacteroides and Oscillospira, and decreased the Firmicutes/Bacteroidetes ratio and the Blautia, and Prevotella, reversing the dysregulation of the gut microbiota. Correlation analysis showed that there is a strong correlation between gut microbiota with significant changes in abundance and hormone related to PCOS.ConclusionOur result indicated that effect of quinoa on PCOS maybe associated with activation of the PI3K/AKT/mTOR signaling pathway, inhibition of autophagy, and regulation of intestinal flora.

Erxian decoction ameliorates myocardial tissue damage through activating PI3K/AKT signaling pathway in ovariectomized rats

Background Erxian decoction (EXD) is an empirical formula for treating cardiovascular disease, our previous work has shown that EXD could improve the cardiovascular structure and function in ovariectomized (OVX) rats, but its pharmacological mechanism is still unclear. Materials and Methods Network pharmacology was utilized to assess the key active components and central targets of EXD in treating postmenopausal cardiovascular disease. Then, an OVX rat model was established, HE staining and transmission electron microscope were utilized to observe myocardial tissue morphology, TUNEL staining was utilized to detect cardiomyocyte apoptosis, western blot, and ELISA were used to confirm efficacy and pathway of EXD. Results The network pharmacology prediction results showed that 129 common targets were identified by intersecting EXD targets and postmenopausal cardiovascular disease targets, including AKT1, TNF, IL-6, IL-1β, PTGS2 and other core targets, apoptosis, PI3K/AKT, and other signaling pathways may be closely related to postmenopausal cardiovascular disease. After ovariectomy, the myocardial tissue of rats was damaged, the expression level of PI3K/AKT pathway-related molecules in the myocardial tissue were decreased, the apoptosis index of cardiomyocytes was increased, and the levels of inflammatory factors (TNF-α, IL-6, and IL-1β) were enhanced. EXD intervention could improve myocardial tissue injury, EXD could up-regulate the protein expression of PI3K and p-AKT in myocardial tissue, and thereby prevent myocardial cell apoptosis. At the same time, EXD downregulated the levels of inflammatory factors in serum of ovariectomized rats. Conclusion EXD may prevent myocardial tissue damage through induction of the PI3K/AKT signaling pathway, thereby reducing cardiomyocyte apoptosis and inflammation. EXD may be a potential drug for the treatment of postmenopausal cardiovascular disease.

Therapeutic Effects of Althaea officinalis L. and Metformin on Estradiol-Induced Polycystic Ovary Syndrome in Rats: Insights into The PI3K/AKT Pathway, Inflammation, and Oxidative Stress.

Polycystic ovary syndrome (PCOS) is one of the most important causes of infertility, irregular menstrual cycles, and anovulation in women. The current study aimed to investigate the therapeutic effects of Althaea officinalis L. (A. officinale) extract on PCOS in rats. In this experimental study, 70 rats in 7 groups (n=10/group) were studied for three weeks as follows; healthy control (HC), patient (PCOS), metformin (PCOS+MET), A. officinale treatment (PCOS+250 and 500 mg/kg A. officinale) and synergistic (PCOS+MET+250 and 500 mg/kg A. officinale) groups. Luteinizing hormone (LH), folliclestimulating hormone (FSH), progesterone (P) and testosterone (T) levels as well as inflammatory cytokines were measured. Total antioxidant capacity and lipid peroxidation levels were analyzed in ovarian tissue. The expression of GLUT-4, AKT, PI3K, PTEN genes and Ki-67 was assessed by real-time polymerase chain reaction (PCR) and immunohistochemistry, respectively. A. officinale alone and especially in combination with MET moderated inflammatory and antioxidant parameters compared to the PCOS and MET groups. A. officinale in synergistic groups increased the apoptosis of granulosa cells by activating the PI3K/AKT pathway, resulting in a rise in the number of Ki-67 positive cells (P<0.05). Furthermore, following A. officinale treatment the LH/FSH rate decreased and FSH and P increased (P<0.05). Also, A. officinale extract could effectively normalize estrus cycle duration close to the normal group. The extract of A. officinale, in combination with metformin, can enhance the hypothalamic-pituitaryovary (HPO) axis with synergistic anti-inflammatory and antioxidant effects. Additionally, the extract showed apoptotic effect on cystic granulosa cells.

A novel strategy to enhance inhibition of Hela cervical cancer by combining Lentinus β-glucan and autophagic flux blockage

Lentinus β-D-glucan (LNT), derived from artificially cultured mushrooms of Lentinus edodes, shows an important yet incompletely understood biological functions in cancer. In this work, the chemical structure of the refined LNT comprising a β-D-(1, 6)-branched β-D-(1,3)-glucan was further clarified via 1D- and 2D-NMR with high resolution, and its drug resistance resulted from autophagy in human cervical cancer (CC) Hela cells besides its anti-cancer function were revealed in vitro and in vivo. In detail, LNT destroyed cellular homeostasis by significantly increasing the intracellular Ca2+ levels and promoted autophagic flux in vitro Hela cells, which was found to at least partially depend on the PI3K/Akt/mTOR-mediated pathway by up-regulating LC3-II levels and down-regulating the expression of p62, PI3K, p-Akt, and mTOR in Hela cells-transplanted BALB/c nude mice. In particular, LNT-induced autophagy led to a drug resistance against LNT-induced proliferation inhibition and apoptosis in Hela cells, and the co-treatment of autophagy inhibitors and LNT significantly enhanced the inhibition of Hela cells and tumor growth in vitro and in vivo. Therefore, the combination of LNT and autophagy inhibitors will be a novel therapeutic strategy to reduce the resistance and improve the prognosis of CC patients in the clinical.

Synergic effects of DL-limonene, R-limonene, and cisplatin on AKT, PI3K, and mTOR gene expression in MDA-MB-231 and 5637 cell lines

The anticancer and cytotoxic effects of DL-Limonene and R-Limonene are well-documented. However, the role of natural compounds in enhancing the efficacy of platinum-based drugs like Cisplatin (CisPt) remains debated. This study aims to boost Cisplatin's impact on breast (MDA-MB-231) and bladder (5637) cancer cells using DL-Limonene and R-Limonene. Different concentrations of DL-Limonene, R-Limonene, and Cisplatin, combined, were used to treat MDA-MB-231 and 5637 cells in this experimental study. The cell's viability was evaluated using an MTT assay. AnnexinV- PI staining was applied to evaluate the percentage of apoptotic cells. Cytotoxicity results showed that combining DL-Limonene, R-Limonene, and Cisplatin significantly improved outcomes in MDA-MB-231 cells (P < 0.05). Annexin/PI staining revealed apoptosis rates of 74 %, 28 %, 43 %, 81 %, and 91 % for Cisplatin40, R-Limonen1000, DL-Limonen1000, R-Limonen1000/DL-Limonen1000, and the combined treatment, respectively, versus 13 % in the control. The combination also resulted in the greatest reduction of AKT, PI3K, and mTOR gene expression. Our results show that R-Limonene and DL-Limonene enhance Cisplatin's cancer-inhibiting effects in breast and bladder cancer cell lines. These compounds may be promising for combination therapy, potentially allowing for lower doses of chemotherapy and reducing side effects like nephrotoxicity.

Anti-Cancer Activity and Mechanism of Resveratrol Against Triple-Negative Breast Cancer

This research analyzed the antitumor of resveratrol in triple negative breast cancer (TNBC). TNBC were divided into control group and resveratrol group. Tnbc-associated TNBC cells were stimulated by different concentrations of resveratrol, and the proliferation, apoptosis, migration of the cells were analyzed. The amount of migration was measured by Transwell method. The PI3K, Akt, and mTOR were detected. Results showed that the cell apoptosis was obvious and the invasion ability was decreased in resveratrol group, and the effect was more obvious with the increase of dosage, and there was a statistical difference between groups (P &lt; 0.05). Resveratrol significantly decreased the expression of Akt, PI3K and mTOR, and there were statistical differences between groups (P &lt; 0.05). In summary, resveratrol reduces cell migration, promotes cell apoptosis, inhibits cell proliferation, and its mechanism is closely related to the blocking of PI3K/Akt/mTOR.

Isoliquiritigenin: A natural compound with a promising role in inhibiting breast cancer lung metastasis

Background and aimIsoliquiritigenin (ISL) is a dietary bioactive compound that is derived from the roots of licorice and various other plants. Accumulating evidence suggests that ISL has noteworthy anticancer activity in a variety of cancer cells, but it is currently unknown how ISL affects the invasion and migration of breast cancer (BC) cells. Hence, the purpose of this study to determine the inhibitory effect of ISL on cell invasion, migration, and metastasis of BC cells in 4T1-treated lung metastatic animal models. Experimental ProcedureISL was found to inhibit BC lung metastasis, as confirmed by MTT assay, clonogenic assay, wound healing assay, transwell assay, histological analysis, western blot and bioluminescence assay in the xenograft model. Results and conclusionThe in vitro studies revealed that ISL treatment significantly prevented BC cell invasion and migration for 24 h. ISL decreased the protein expressions of PI3K, Akt, p-Akt, mTOR, p-mTOR, matrix metalloproteinase (MMP)-2, MMP-9, and N-cadherin , thereby inhibiting BC cell invasion. In addition, ISL decreased MMP-2/9 and N-cadherin expression in lung tissues and reduced mouse 4T1 BC cell lung metastasis. Based on these findings, it suggests that ISL inhibits cell migration and invasion in BC cells by inhibiting the PI3K/Akt signaling pathway and decreasing the activation of MMP-2 and MMP-9 expression. Therefore, ISL may be useful in managing BC invasion and metastasis.

Bile acid conjugated chitosan nanoparticles promote the proliferation and epithelial-mesenchymal transition of hepatocellular carcinoma by regulating the PI3K/Akt/mTOR pathway

Bile acids have been known to play significant roles at certain physiological levels in gastrointestinal metabolism. Yet, they are known to be carcinogenic and aid in tumor progression in most cases, although the roles remain uncertain. Hence, we tested the cytotoxic potential of cholic acid (CA) loaded chitosan nanoparticles (CNPs) on Hep3B cells. The physicochemical properties of the CNPs synthesized with CA load (CA-CNPs) were determined using standard techniques such as ultraviolet–visible spectrophotometry (UV–Vis), fourier-transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), dynamic light scattering (DLS) and transmission electron microscopy (TEM). The characteristic peak for chitosan nanoparticles were observed for plain CNPs (pCNPs) and CA-CNPs at around 300 nm as per UV–Vis analysis. FTIR analysis indicated the possible trapping of CA onto CNPs as certain peaks were retained and some peaks were shifted. XRD analysis determined that the peaks representing CA and pCNPs were collectively obtained in CA-CNPs. As per DLS analysis, the particle size, PDI and ζ-potential of the CA-CNPs were 259 nm, 0.284 and 30.4 mV. Further, the CA-CNPs were non-cytotoxic on Hep3B cells at the maximum tested concentration of 500 μg/mL. The viability at 500 μg/mL of CA-CNPs was two-fold higher than 500 μg/mL of pCNPs. Also, the pCNPs were not hemolytic and therefore could not have played a role in the increase of viability after treatment with CA-CNPs, which indicates that CA posed a major role in increased viability of Hep3B cells. As per quantitative PCR (qPCR), the upregulated gene expressions of PI3K, Akt, mTORC2, cMyc, Fibronectin, hVPS34, Slug and ZEB1 and the downregulated expression of the tumor suppressor PTEN indicates that PI3K/Akt/mTOR pathway mediated the induction of epithelial-to-mesenchymal transition (EMT) in response to CA-CNPs treatment on Hep3B cells.

Sea Buckthorn Oil Promotes the PI3K-Akt-ERK Signaling Pathway and Macrophage M2 Polarization to Reduce Radiation-induced Skin Injury.

In this work, we explored the role and mechanism of sea buckthorn oil in reducing radiation-induced skin damage. The radiation-induced rat skin injury model was established using strontium-90. Rats were treated with sea buckthorn oil twice a day postirradiation, and skin damage was observed at different times and evaluated using an injury score. Skin pathological changes were observed using hematoxylin and eosin (H&E) staining. Western blotting and immunohistochemistry were used to detect the expression of vascular growth and pathway proteins. ELISA was used to detect the secretion level of inflammatory factors. Immunohistochemistry was used to detect macrophage polarization marker proteins. We found that sea buckthorn oil can alleviate radiation-induced skin damage, accelerate skin vascular regeneration, and promote the up-regulation of vascular endothelial growth factor (VEGF) and its receptor (VEGFR). These results demonstrate the beneficial effects of sea buckthorn oil on radiation-induced skin damage. Furthermore, the levels of IL-1β and TNF-α in the sea buckthorn oil treatment group were significantly lower than those in the control group, while the levels of IL-4 and IL10 were significantly higher (P < 0.05). CD206 expression also increased in the sea buckthorn oil treatment group, while CD16 expression decreased compared to the control group (P < 0.05). Western blotting showed that PI3K, Akt and ERK expression increased in the sea buckthorn oil treatment group (P < 0.05). The beneficial effect of sea buckthorn oil in reducing the inflammatory response in irradiated rats was diminished when they were treated with PI3K inhibitor. We conclude that sea buckthorn oil may regulate macrophage M2 polarization by increasing the PI3K-Akt-ERK signaling pathway, thereby inhibiting the inflammatory response and promoting skin vascular regeneration to prevent and treat radiation-induced skin damage.

Integrating network pharmacology and experimental verification to study the mechanism of Polygonum hydropiper total flavonoids against stress-induced gastric mucosal damage

ContextPolygonum hydropiper L. (P. hydropiper) has outstanding clinical efficacy in treating both acute and chronic gastroenteritis. However, the definite mechanism remains unclear. ObjectiveThis study aimed to explore the potential mechanisms of the total flavonoid of P. hydropiper (FPH) in stress-induced gastric mucosal damage (SGMD) rats through a combination of network pharmacology, molecular docking, and animal experiments. MethodsNetwork pharmacology and molecular docking were utilized to predict the potential mechanisms of FPH against SGMD. In experimental studies, SGMD rat models were established using water-immersion restraint stress (WIRS). FPH was administered at doses of 140, 70, and 35 mg/kg, with ranitidine serving as a positive control, through gavage once daily for 6 consecutive days after model establishment. Stomach and serum specimens were analyzed using HE staining, Western blotting, qPCR, and ELISA to investigate the protective mechanism of FPH in SGMD. ResultsThe network pharmacology analysis identified 16 active ingredients and 183 common targets, with potential pathways including PI3K/Akt, MAPK and Keap1/Nrf2. In vivo experiments demonstrated that FPH intervention alleviated SGMD pathological changes, reduced elevated serum IL-6 and TNF-α levels, and enhanced SOD and GSH activity in rats. Additionally, FPH increased the protein expression of p62, Nrf2, HO-1, PI3K, and p-Akt, along with mRNA levels of Nrf2, p62, and HO-1. ConclusionsFPH exerts a gastric mucosal protective effect by upregulating antioxidant gene expression through the PI3K/Akt and Keap1/Nrf2 pathways. This study provides an experimental basis for the potential clinical treatment of SGMD with the traditional Chinese medicine P. hydropiper.

Effects of dietary primary bile acids on the growth, intestinal morphology, gene expression, serum metabolites and intestinal microbiota in pearl gentian grouper fed a high lipid diet

The grouper aquaculture industry flavors high lipid diets for economic reasons, yet this inevitably produces negative effects on the growth performance and intestinal function. Bile acids have been extensively reported to mitigate the negative effects on fish fed a high lipid diet, but it remains unclear whether this mechanism is bile acid type dependent. Therefore, an eight-week feeding trial was conducted to investigate the response of growth and intestinal health parameters in pearl gentian grouper Epinephelus fuscoguttatus♂ × E. lanceolatus♀ fed with high lipid diets to different types of bile acids. A high lipid diet (diet HL) containing 167 g/kg crude lipid served as the control diet, and three of primary bile acids glycocholic acid, taurocholic acid, and cholic acid were added at a dose of 900 mg/kg to generate the bile acid diets HBG, HBT, and HBC, respectively. Results revealed that dietary supplementation with glycocholic acid, taurocholic acid, or cholic acid does not affect the feed intake and growth of pearl gentian grouper fed a high lipid diet, while increasing the relative abundance of intestinal opportunistic pathogens such as Staphylococcus and Pseudomonas in this species. Dietary supplementation with glycocholic acid or cholic acid decreases muscle layer thickness and villus height, while increases intestinal injury score. Grouper fed with taurocholic acid showed higher villus width and serum antioxidant enzyme activity, along with lower intestinal injury score and serum lipid peroxidation level. For the protein and gene involved in the PI3K/AKT/mTOR pathways, S6K1 signaling, and bile acids metabolism, grouper fed with taurocholic acid exhibited an increase in the protein expression of PP70 and the mRNA expression of nrf2, pi3k, pdk1, tsc2, akt, mtorc1, fxr and tgr5. Moreover, the serum metabolites can be significantly influenced by the intestinal bacteria. Overall, dietary supplementation of 900 mg/kg taurocholic acid is recommended when feeding high lipid diets to grouper.

Regulating the lncRNA DSCR9/RPLP2/PI3K/AKT axis: an important mechanism of Xinfeng capsules in improving rheumatoid arthritis.

Rheumatoid arthritis (RA) is a systemic autoimmune disease characterized by chronic and symmetrical polyarthritis. RA patients often experience inflammatory reaction and hypercoagulable state, which together affect the self-perception of patient (SPP). Currently, inhibiting inflammation and hypercoagulable state are common treatment methods for alleviating RA symptoms. Xinfeng Capsules (XFC) has a long history of treating RA, and can effectively improve the inflammatory response and hypercoagulable state of RA. However, the potential mechanisms have not yet been determined. This study elucidated the action mechanism of XFC in RA inflammation and hypercoagulability through the lncDSCR9/RPLP2/PI3K/AKT axis. Clinical observations indicated that there was a strong link between XFC therapy and improvements in inflammatory and coagulation biomarkers, as well as SPP among RA patients. The subsequent network pharmacology analysis results identified the PI3K/AKT signaling pathway as a potential mediator for XFC treatment of RA. Furthermore, clinical validation and sequencing results revealed that lncRNA DSCR9 expression (a gene implicated in inflammation and coagulation) was negatively correlated with clinical markers of inflammation and coagulation, while positively correlated with SF-36 indicators. Notably, XFC treatment remarkably upregulated lncRNA DSCR9 expression and downregulated PI3K and AKT expressions, showing opposite expression trends to the untreated cases.The regulatory effect of XFC on the lncRNA DSCR9/RPLP2/PI3K/AKT axis in RA was investigated using techniques such as RNA pull-down assay, Western blot analysis, RT-PCR, and EdU assay. Moreover, the administration of the PI3K/AKT agonist RMH can counteract the effects of XFC on p-PI3K, p-AKT, inflammation, and hypercoagulability, reinforcing the role of pathway. Finally, animal studies utilizing HE staining and transmission electron microscopy (TEM) demonstrated that XFC notably decreased PI3K and AKT expressions in adjuvant-induced arthritis (AA) rats, mitigated inflammation and hypercoagulability, and enhanced the ultrastructure of synovial cells. These findings underscored the potential mechanisms of XFC in the treatment of RA. Regulating the lncRNA DSCR9/RPLP2/PI3K/AKT axis may be an important mechanism by which XFC improved RA inflammatory response and hypercoagulable state.

The Potential of Human Pulmonary Mesenchymal Stem Cells as Vectors for Radiosensitizing Metallic Nanoparticles: An In Vitro Study.

Metallic nanoparticles (NPs) exhibit interesting radiosensitizing effects, and finding a way to accurately deliver them appears to be crucial. Due to their tumor tropism, mesenchymal stem cells (MSCs) represent a strategic approach. Therefore, we aimed to evaluate the impact of core-shell Fe3O4@Au NPs on the functionality of human pulmonary MSCs (HPMSCs). The results showed that 100 µg/mL Fe3O4@Au NPs, accumulated in HPMSCs (revealed by Prussian blue staining), did not alter cell viability as assessed by cell counting, MTT, and LDH assays. However, caspase 9 and Bcl2 gene expression, evaluated by RT-qPCR, was regulated 72 h after exposure to the NPs. Moreover, the NPs also decreased proinflammatory cytokine/chemokine secretions, except for CXCL8 (ELISA). These modulations were associated with the downregulation of AMPK gene expression at 24 h. In contrast, the NPs did not modulate VEGF, PI3K, or PDGF gene expression. Nevertheless, a decrease in VEGF secretion was observed after 24 h of exposure to the NPs. Interestingly, the Fe3O4@Au NPs did not modulate Nrf2 gene expression, but they did regulate the expression of the genes encoding Nox4 and HMOX-1. Additionally, the NPs increased ROS production, suggesting a redox imbalance. Finally, the Fe3O4@Au NPs did not affect the HPMSCs' viability or proangiogenic/tumorigenic markers. These findings are encouraging for investigating the effects of Fe3O4@Au NPs delivered by HPMSCs to tumor sites in combination with radiation.

Ameliorative effects of HGF-overexpressed exosomes derived from ADMSCs on oxidative stress in hepatic fibrosis.

Hepatic fibrosis, ultimately causing hepatic sclerosis, remains significant health concerns. Adipose-derived mesenchymal stem cell (ADMSC)-derived exosomes (Exo) exhibit amelioration of liver injury. Hepatocyte growth factor (HGF) regulates hepatocyte growthn. However, its involvement during hepatic fibrosis remains unclear. Isolation of ADMSCs and Exo, transfection of HGF overexpression, and activation of hepatic stellate cells (HSCs) by Angiotensin II (AngII) were conducted. Cells were randomized into HSC, AngII-HSC, ADMSCs-Exo, ADMSCs<supblank</sup>-Exo, and ADMSCs<sup>HGF</sup-Exo, DPI, LY294002, and SB203580 groups. MTT for cell viability, cell migration, and flow cytometry for ROS were performed. BALB/c mice were treated with CCL4 for hepatic fibrosis models. The mice were randomized into Control, PBS, ADMSCs-Exo, ADMSCs^blank-Exo, and ADMSCs^HGF-Exo groups (n=6). HE, Sirius red, and Oil Red O staining, liver function indicators, and ELISA for oxidative stress were performed. ROS generation-related and PI3K/Akt/P38MAPK-related factors were detected by immunofluorescence, immunohistochemistry, and western blot. After identification of ADMSC-Exo and transfection, AngII increased cell viability, migration, Collagen I (CoLI), α-smooth muscle actin (α-SMA), ROS, NADPH oxidase 4 (NOX4), PI3K, p-Akt, p-P38MAPK, ras-related C3 botulinum toxin substrate 1 (RAC1), p47phox, and p22phox expression. However, ADMSCsHGF-Exo, DPI, LY294002, and SB203580 reversed the above effects. Moreover, ADMSCsHGF-Exo inhibited pathological damage, fibrosis, lipid accumulation, ALT, AST, TBIL, CoLI, α-SMA, NOX4, MDA, PI3K, p-Akt, and p-P38MAPK expression, and increased ALB, SOD, GPx, CAT, GSH, Mn-SOD, Na+-K+-ATPase, and Ca2+-Mg2+-ATPase levels in hepatic fibrosis mice. ADMSCsHGF-Exo attenuated hepatic fibrosis by inhibiting oxidative stress through activating the PI3K/Akt/P38MAPK pathway, providing valuable insights for potential treatment of liver fibrosis.

Structural simplification of Osimertinib to elaborate new indolyle-pyrimidine-5-carbonitrile derivatives with Anti-proliferative and Anti-SARS-CoV-2 activities assisted by molecular dynamic simulation

Based on the simplicity and modification of the medication osimertinib, two new series of indolyle-pyrimidine-5-carbonitrile scaffolds were created and synthesized with dual action as anti-SARS-Cov-2 and anticancer. The newly created heterocyclic compounds' chemical structures were effectively characterized. With IC50 values of 18.52, 20.89, and 19.85, respectively, compounds 9b, 10, and 15 had inhibitory actions against SARS-CoV-2 when compared to remdesivir and chloroquine, which served as pharmacological controls and had IC50 values of 3.38 μM and 24.9 μM, respectively.Furthermore, compounds 9a and 13 showed anti-proliferative activity against HepG2 cell lines with IC50s of 5.63 μM and 3.06 μM, respectively, in comparison to doxorubicin's IC50 of 7.4 μM. qRT-PCR revealed that HepG2 cells treated with compounds 9a and 13 showed increased p53 expression levels and decreased CDK1 and PI3K expression levels in comparison to doxorubicin. Molecular dynamics simulations of 20 ns duration were performed with PI3Kα, PI3Kγ, and CDK and active compound complexes. The results confirm that compound 13 has the potential to be a therapeutic candidate for additional preclinical and clinical research, as indicated by the molecular docking data.

L-carnitine cause to increase cell proliferation of C-Kit+ hematopoietic progenitor cells via decreasing the PI3K and FOXO-1 protein expression

BackgroundStem cell-based therapy has emerged as an attractive approach for regenerative medicine. Poor survival and maintenance of the cells used in regenerative medicine are considered as serious barriers to enhance the efficacy of the cell therapy. Using some antioxidants has been reported to prevent the aging of stem cells, and finding effective factors to reduce the senescence of these cells has impressive potential in cell therapy. The PI3K pathway adversely regulates the transcription factors known as FOXO, which are thought to have an inhibitory influence on cell proliferation. By downregulating FOXO and other targets, PI3K signaling controls the growth of cells. For this reason, the aim of the present study is to investigate the effect of L-carnitine (LC) as antioxidant on the cell proliferation and the protein expression of PI3K and FOXO. MethodsFor understanding the in vitro effect of LC on the PI3K and FOXO-1 expression of C-kit+ hematopoietic progenitor cells, the bone marrow mononuclear cells were isolated, and C-kit+ cells was enriched by the magnetic-activated cell sorting (MACS). Next, the identification of enriched C-kit+ cells were done by flowcytometry and immunocytochemistry. Then, C-kit+ cells were treated with 0.2 mM LC, the cells were collected at the end of the treatment period (48 h), and the proteins were extracted. In the following, the protein expression of PI3K and FOXO-1 was measured by western blotting. In addition, flowcytometry was done to assess the Ki-67 expression as a key marker for cell proliferation investigation. Results0.2 mM LC cause to significantly decrease in the protein expression of PI3K and FOXO-1 (*P<0.05 and **P<0.01, respectively). Also, the expression of Ki-67 was significantly increased in the presence of 0.2 mM LC (***P<0.001). ConclusionBriefly, LC can be considered an effective factor in increasing the proliferation of C-kit+ cells via some signaling pathways.

Maternal Gestational Diabetes Mellitus and High-Fat Diet Influenced Hepatic Polyunsaturated Fatty Acids Profile in the Offspring of C57BL/6J Mice.

This research examines the effects of maternal high-fat (HF) diet and gestational diabetes mellitus (GDM) on offspring lipid metabolism and polyunsaturated fatty acids (PUFA) profile. GDM is induced using the insulin receptor antagonist S961. Weaning offspring are categorized into HF-GDM, HF-CON, NC-GDM, and NC-CON groups based on maternal diet or GDM. Adult offspring are then grouped into NC-CON-NC, NC-CON-HF, NC-GDM-NC, NC-GDM-HF, HF-CON-NC, HF-CON-HF, HF-GDM-NC, and HF-GDM-HF according to dietary patterns. Gas chromatography determines PUFA composition. Western blot assesses PI3K/Akt signaling pathway-related protein expression. Feeding a normal chow diet until adulthood improves the distribution of hepatic PUFA during weaning across the four groups. PI3K expression is upregulated during weaning in HF-CON and HF-GDM, particularly in HF-CON-NC and HF-GDM-NC, compared to NC-CON-NC during adulthood. Akt expression increases in NC-GDM-NC after weaning with a normal diet. The hepatic PUFA profile in HF-CON-HF significantly distinguishes among the maternal generation health groups. Maternal HF diet exacerbates the combined impact of maternal GDM and offspring HF diet on hepatic PUFA and PI3K/Akt signaling pathway-related proteins during adulthood. Early exposure to HF diets and GDM affects hepatic PUFA profiles and PI3K/Akt signaling pathway protein expression in male offspring during weaning and adulthood.