This study aimed to investigate the mechanism of total flavonoids from Smilacis Glabrae Rhizoma(SGF) against obesity-associated colorectal cancer(CRC) via regulation of fatty acid oxidation(FAO). In vivo, syngeneic subcutaneous and orthotopic xenograft models of CRC cells MC38 were established in obese mice induced by high fat diet(HFD), which was followed by the administration with SGF. The tumor volume and tumor weight were measured in the CRC subcutaneous model. The growth of CRC orthotopic tumors was monitored by a small animal in vivo imaging system. Hematoxylin-eosin(HE) staining was used to observe the pathological changes in tumors and peritumoral adipose tissues. Immunohistochemistry, TdT-mediated dUTP nick end labeling(TUNEL), and oil red O staining were separately conducted to detect Ki67 expression, apoptosis, and lipid accumulation in tumors. Western blot was used to detect the phosphorylation levels of adenosine monophosphate-activated protein kinase(AMPK) and acetyl-CoA carboxylase(ACC), as well as the expression of FAO-related proteins and cell death-related proteins in tumor tissues. In vitro, MC38 cells cultured under low-glucose and high-oleic acid conditions were taken as the research object and treated with varying concentrations of SGF for 48 h. Apoptosis rates were assessed by flow cytometry, and the expression of FAO-related and cell death-related proteins was analyzed by Western blot. Further, the prolyl hydroxylase domain-containing protein 3(PHD3) expression was subjected to RNA interference(RNAi), and the viability and apoptosis of PHD3-knockdown MC38 cells following SGF treatment were assessed by MTT assay and flow cytometry, respectively. The results showed that both high and low doses of SGF significantly inhibited tumor growth in two obese mouse xenograft models. The SGF-treated groups exhibited extensive, scattered necrotic areas within the tumor tissue, with marked destruction of tissue structure. Compared with the HFD group, the SGF-treated groups showed a significant reduction in the rate of Ki67-positive cells and a significant increase in the proportion of TUNEL-staining-positive apoptotic cells. In obese mice, subcutaneous tumors showed significantly enhanced oil red O staining, with large lipid droplets visible. The SGF-treated groups markedly reduced lipid deposition. In both models, peritumoral adipocytes were significantly enlarged after SGF treatment. The tumor tissues of obese mice exhibited significantly increased expression of FAO-related proteins cluster of differentiation 36(CD36), carnitine palmitoyltransferase 1(CPT1), and acyl-CoA dehydrogenase long chain(ACADL), as well as elevated adenosine triphosphate(ATP) content. The phosphorylation levels of upstream regulatory proteins AMPK and ACC were significantly increased, while the expression of the energy switch protein PHD3 was significantly decreased. SGF significantly reduced the protein expression of CD36, CPT1, and ACADL, decreased ATP content, inhibited AMPK and ACC phosphorylation, and increased PHD3 protein levels. This was accompanied by a significant increase in the apoptotic protein cysteine-dependent aspartate-specific protease-3(caspase-3) and significant decreases in the anti-apoptotic protein B-cell lymphoma-2(Bcl-2) and the cell cycle protein cyclin D1. In MC38 cells cultured under low-glucose and high-oleic acid conditions, SGF significantly induced apoptosis and inhibited lipid uptake. FAO-related protein expression was significantly suppressed. Caspase-3 expression was significantly increased, while Bcl-2 and cyclin D1 expression was significantly decreased. PHD3 expression was significantly elevated. In MC38 cells transfected with siPHD3, the inhibitory effect of SGF on cell viability and its pro-apoptotic effect were both significantly enhanced. In summary, SGF effectively reduces lipid transfer between adipose tissue and tumors under obese conditions, regulates the AMPK/ACC signaling pathway, and targetedly inhibits tumor FAO, thereby exerting potent antitumor effects against obesity-associated CRC.

INTRODUCTION:Epidemiological studies have demonstrated higher incidence and mortality rate of nonalcoholic steatohepatitis (NASH) in the elderly population than in younger groups. However, the mechanisms underlying this age-related exacerbation remain poorly understood. OBJECTIVE:This study aimed to elucidate the specific pathways through which aging exacerbates NASH progression, using an integrated in vivo and in vitro model. METHODS:Aged (18-month-old) and young (6-week-old) mice were fed a high-fat diet (HFD) for 16 weeks to induce NASH. A senescence-associated cellular model of NASH was established by co-treating murine hepatocyte AML-12 with H2O2 and free fatty acid (FFA). Gene expression profiling of liver tissue was performed using RNA sequencing to identify molecular signatures. Interventions were as follows: (1) In vitro, BMAL1 overexpression plasmids were transfected into AML-12 cells, followed by treatment with 2-deoxy-D-glucose (2-DG, a glycolysis inhibitor) and 2-methoxyestradiol (2-ME2, a HIF-1α inhibitor); (2) in vivo, hepatocyte-specific BMAL1 overexpression was achieved in aged HFD-fed mice through adeno-associated virus serotype 8 (AAV8) delivery. Mechanism validation was performed using biochemical assays, Western blot, cell staining, molecular docking, and Co-IP. RESULTS:Aged HFD-fed mice exhibited more severe NASH phenotypes than young mice. Transcriptomic analysis identified NLRP3-related signaling and circadian rhythm pathways as central contributors to age-specific NASH pathogenesis. These mice also exhibited elevated NLRP3 inflammasome activity, enhanced glycolysis, and reduced BMAL1 expression. In senescent NASH cells, BMAL1 overexpression along with 2-DG or 2-ME2 treatment significantly downregulated NLRP3 expression and attenuated lipid accumulation, inflammation, oxidative stress, and fibrosis. Mechanistically, BMAL1 directly bound to HIF-1α, thereby suppressing glycolysis. Hepatocyte-specific BMAL1 overexpression in aged HFD-fed mice markedly inhibited glycolysis and NLRP3 activation, resulting in an improvement in NASH-related pathologies. CONCLUSION:This study revealed a novel mechanism in which BMAL1 downregulation under aging and HFD conditions promotes NASH progression by binding to HIF-1α and modulating the glycolysis-NLRP3 inflammasome axis.

The therapeutic effects of Quzhou Aurantii Fructus aqueous extract(QAFA) and Aurantii Fructus aqueous extract(AFA) on functional dyspepsia(FD) in rats were evaluated and compared, and their underlying mechanisms were investigated. The FD rat models were constructed by a combined method of ″pinching tail + irregular diet + drinking dilute acidic water″. The rats were orally administered QAFA at a dose of 2.05 g·kg~(-1)(equivalent to 4 g·kg~(-1) QAF) and AFA at a dose of 1.93 g·kg~(-1)(equivalent to 4 g·kg~(-1) AF), respectively. After 14-day administration, the gastric emptying and intestinal propulsion were assessed. The pathological changes in the gastric antrum and duodenum were analyzed via histology, and the levels of serum gastrin(GAS) and vasoactive intestinal polypeptide(VIP) were measured by enzyme-linked immunosorbent assay. The underlying mechanisms were investigated by using 16S rRNA sequencing and serum metabolomics. The results showed that QAFA and AFA could increase gastrointestinal motility. Hematoxylin-eosin(HE) staining and immunofluorescence analysis showed that both QAFA and AFA repaired the damaged intestinal barrier, and ELISA analysis showed that they regulated GAS and VIP levels in serum. These results suggest that both can improve symptoms in FD rats. The intestinal microbiota analysis revealed that QAFA and AFA reduced the abundance of the harmful microbe Ruminococcus. AFA increased the abundance of the beneficial bacteria Prevotella and Lactobacillus, and QAFA increased the abundance of Clostridia UCG-014. Metabolomic analysis showed that both QAFA and AFA restored serum metabolites primarily through regulating phospholipid metabolism. Correlation analysis between metabolites and gut microbiota showed that both QAFA and AFA decreased the abundance of Ruminococcus, and AFA increased the abundance of Prevotella, thereby restoring serum metabolite levels. These findings suggest that QAFA and AFA may improve FD symptoms in rats by modulating specific gut microbiota to restore gastrointestinal hormones and phospholipid metabolism. However, there are some differences in the specific microbiotas they regulate.

Background: Testosterone deficiency is linked to an increased prevalence of non-alcoholic fatty liver disease (NAFLD), although the mechanisms underlying this association are not fully understood. Ferroptosis, a regulated cell death pathway driven by iron-dependent lipid peroxidation, has been suggested to play a role in NAFLD pathogenesis. Since testosterone deficiency is associated with lipid disorders and iron deposition, we hepothesize that ferroptosis may be involved in the pathogenesis of diet-induced NAFLD exacerbated by testosterone deficiency. Methods: Apolipoprotein E (APOE- /- ) mice were subjected to sham surgery or bilateral castration and subsequently fed a high-fat diet for 16 weeks. Liver gene expression was analyzed using RNA sequencing. Additional assessments included blood analysis, histological staining, measurement of iron and antioxidant enzyme levels, quantitative real-time PCR, Western blotting, and electron microscopy. The effects of testosterone on ferroptosis induced by free fatty acids (FFAs) and Erastin were further investigated in HepG2 cells in vitro. Results: Testosterone deficiency resulted in increased hepatic lipid accumulation and macrovesicular steatosis in high-fat diet-fed APOE- /- mice, accompanied by hepatic inflammation, fibrosis, and elevated liver enzyme levels. Transcriptomic analysis revealed that testosterone deficiency affects ferroptosis and circadian rhythmrelated signaling pathways. Castrated APOE- /- mice exhibited significantly higher hepatic iron deposition, lipid peroxidation, and expression of key ferroptosis-related proteins, along with decreased Brain and muscle ARNT-like gene 1 (BMAL1) protein expression. In vitro, testosterone treatment reduced lipid and iron accumulation and lipid peroxidation in HepG2 cells subjected to FFAs and Erastin. Moreover, BMAL1 knockdown negated the protective effects of testosterone against ferroptosis in hepatocytes. Conclusion: Our study demonstrated that testosterone deficiency exacerbates NAFLD induced by a high-fat diet by promoting hepatocyte ferroptosis through modulation of the circadian protein BMAL1.

Actinidia eriantha polysaccharide (AEPS) is a potent adjuvant with dual Th1 and Th2 potentiating activity. linc-AAM has been previously proved to facilitate the expression of immune response genes (IRGs) in AEPS-activated RAW264.7 macrophages. However, its role in mediating adjuvant activity of AEPS remains to be elucidated. In this study, bone marrow-derived macrophages (BMDMs) from wide-type (WT) and linc-AAM knockout C57BL/6J mice treated with AEPS were subjected to transcriptome sequencing and bioinformatic analysis. linc-AAM deficiency inhibited M1 and M2 immune responses in BMDMs induced by AEPS. In mechanisms, AEPS facilitated the expression of IRGs and activated BMDMs through NF-κB−linc-AAM−JAK/STAT axis. Furthermore, linc-AAM knockout inhibited cytokine and chemokine production, immune cell recruitment as well as immune cell migration to draining lymph nodes at peritoneal cavity in mice induced by AEPS. More importantly, linc-AAM deletion reduced the adjuvant activity of APES on antigen-specific cellular and humoral immune responses to ovalbumin in mice. This study has for the first time demonstrated the role of lncRNAs in regulating the adjuvant activity of polysaccharides and its mechanisms. These findings expanded current knowledge on the mechanism of action of adjuvant and provide a new target for the design and development of vaccine adjuvants.

The miniature pig is a suitable animal model for investigating human cardiovascular diseases. Nevertheless, the alterations in lipid metabolism within atherosclerotic plaques of miniature pigs, along with the underlying mechanisms, remain to be comprehensively elucidated. In this study, we aim to examine the alterations in lipid composition and associated pathways in the abdominal aorta of atherosclerotic pigs induced by a high-fat, high-cholesterol, and high-fructose (HFCF) diet using lipidomics and RNA-Seq methods. The results showed that the content and composition of aortic lipid species, particularly ceramide, hexosyl ceramide, lysophosphatidylcholine, and triglyceride, were significantly altered in HFCF-fed pigs. Meanwhile, the genes governing sphingolipid metabolism, iron ion homeostasis, apoptosis, and the inflammatory response were significantly regulated by the HFCF diet. Furthermore, C16 ceramide could promote iron deposition in RAW264.7 cells, leading to increased intracellular reactive oxygen species (ROS) production, apoptosis, and activation of the toll-like receptor 4 (TLR4)/nuclear Factor-kappa B (NF-қB) inflammatory pathway, which could be mitigated by deferoxamine. Our study demonstrated that dysregulated ceramide metabolism could increase ROS production, apoptosis, and inflammatory pathway activation in macrophages by inducing iron overload, thus playing a vital role in the pathogenesis of atherosclerosis. This discovery could potentially provide a new target for pharmacological therapy of cardiovascular diseases such as atherosclerosis.

目的 探讨土茯苓总黄酮(SGF)对兔胸主动脉环的舒张作用及作用机制.方法 制备内皮完整或去内皮的兔离体胸主动脉环并检验其内皮完整性,利用MedLab-U/8c生物信号采集处理系统检测药物刺激后血管张力.①内皮完整胸主动脉环分为氯化钾(KCl)60 mmol·L-1、去氧肾上腺素(PE)1 μmol·L-1、去甲肾上腺素(NE)1 μmol·L-1、一氧化氮合酶抑制剂左旋硝基精氨酸甲酯(L-NAME)100 μmol·L-1和β2肾上腺素能受体阻滞剂ICI1185510.1 μmol·L-1组.KCl组、PE组和NE组分别预收缩胸主动脉环10 min,L-NAME组和ICI118551组分别预处理胸主动脉环30 min,并加入NE 1 μmol·L-1预收缩10 min.所有组间隔6 min加1次SGF,检测SGF 15.625～500 mg·L-1浓度作用下血管张力;②去内皮胸主动脉环分为PE 1 μmol·L-1、NE 1 μmol·L-1和氯化钙(CaCl2)组.PE组和NE组预收缩胸主动脉环10 min,间隔6 min加1次SGF,检测SGF 15.625～250 mg·L-1浓度作用下血管张力.CaCl2组在无钙环境中加入KCl 60 mmol·L-1预收缩胸主动脉环,同时加SGF 500 mg·L-1预处理20 min,间隔6 min添加1次CaCl2,检测CaCl20.15～2.4 mmol·L-1浓度作用下血管张力.同时设生理盐水组.结果 在内皮存在条件下,SGF 15.625～250 mg·L-1对KCl预收缩胸主动脉环无明显作用,而SGF 31.25～500 mg·L-1对PE和NE预收缩胸主动脉环有显著舒张作用(P<0.01),最大舒张率分别达(56±12)%和(76±13)%.SGF 500 mg·L-1对胸主动脉环的舒张功能减弱,舒张率为(40±10)%和(46±15)%.L-NAME 100 μmol·L-1预处理基本阻断SGF诱导的胸主动脉环舒张作用(P<0.01),而ICI1185510.1 μmol·L-1预处理仅部分阻断SGF的舒张效应(P<0.01).去除胸主动脉环内皮后,SGF对PE和NE预收缩胸主动脉环无舒张作用,但SGF 500 mg·L-1预处理能显著降低CaCl20.6～2.4 mmol·L-1诱导的胸主动脉环收缩(P<0.05).结论 SGF可能具有一氧化氮依赖性诱导血管舒张、钙离子通道阻滞剂和β受体阻滞剂作用.

Two polysaccharides from Crocus sativus petals (PCSPs), PCSPA and PCSPB have been previously reported to possess the immunopotentiation activity and improve innate immunity in mice. In this study, PCSPB was evaluated for the anti-tumor activity and explored its immunological mechanisms based on tumor microenvironment (TME) using S180 sarcoma-bearing mice. Although PCSPB showed the lower toxicity to a series of tumor cells, it significantly and dose-dependently suppressed the growth of S180 sarcomas transplanted in mice. HE staining, immunohistochemical analysis, and TUNEL assay revealed that PCSPB significantly induced tumor cell necrosis, apoptosis, and vessel disruption in sarcoma tissues. Meanwhile, PCSPB markedly decreased the levels of inflammatory factors TGF-β, IFN-γ, IL-10 and TNF-α and down-regulated the mRNA expression levels of TGF-β and TNF-α in tumor tissues. Flow cytometric analysis showed that PCSPB significantly increased the proportion of CD8+ T cells and NK cells, but decreased that of regulatory T cells (Tregs), total myeloid-derived suppressor cells (MDSCs), and tumor-associated macrophages (TAMs) in sarcoma tissues. Furthermore, immunofluorescence assay demonstrated that PCSPB noteworthily reprogrammed TAMs from a tumorigenic M2 towards an antitumorigenic M1 phenotype in S180 tissues. These findings demonstrated that PCSPB might exert the anti-tumor activity by reconstructing TME and could act as an anti-tumor candidate with low toxicity.

Albizia julibrissin saponin active fraction (AJSAF), is a prospective adjuvant with dual Th1/Th2 and Tc1/Tc2 potentiating activity. Its adjuvant activity has previously been proven to be strictly dependent on its spatial co-localization with antigens, highlighting the role of local innate immunity in its mechanisms. However, its potential targets and pathways remain unclear. Here, its intracellular molecular mechanisms of innate immune response were explored using mouse C2C12 myoblast by integrative analysis of the in vivo and in vitro transcriptome in combination with experimental validations. AJSAF elicited a temporary cytotoxicity and inflammation towards C2C12 cells. Gene set enrichment analysis demonstrated that AJSAF regulated similar cell death- and inflammatory response-related genes in vitro and in vivo through activating second messenger–MAPK–CREB pathways. AJSAF markedly enhanced the Ca2+, cAMP, and reactive oxygen species levels and accelerated MAPK and CREB phosphorylation in C2C12 cells. Furthermore, Ca2+ chelator, CREB inhibitor, and MAPK inhibitors dramatically blocked the up-regulation of IL-6, CXCL1, and COX2 in AJSAF-treated C2C12 cells. Collectively, these results demonstrated that AJSAF induced innate immunity via Ca2+–MAPK–CREB pathways. This study is beneficial for insights into the molecular mechanisms of saponin adjuvants.

3β-hydroxy-12-oleanen-27-oic acid (ATA), a cytotoxic oleanane triterpenoid with C14-COOH isolated from the rhizome of Astilbe chinensis, has been previously proven to possess antitumor activity and may be a promising antitumor agent. However, its molecular mechanisms of antitumor action were still unclear. This study explored the underlying mechanisms of cytotoxicity and potential target of ATA against human colorectal cancer HCT116 cells via integrative analysis of transcriptomics and network pharmacology in combination with in vitro and in vivo experimental validations. ATA significantly inhibited the proliferation of HCT116 cells in a concentration- and time-dependent manner and induced the cell cycle arrest at the G0/G1 phase, apoptosis, autophagy, and ferroptosis. Transcriptomic analysis manifested that ATA regulated mRNA expression of the genes related to cell proliferation, cell cycle, and cell death in HCT116 cells. The integrated analysis of transcriptomics, network pharmacology, and molecular docking revealed that ATA exerted cytotoxic activity via interactions with FDFT1, PPARA, and PPARG. Furthermore, FDFT1 was verified to be an upstream key target mediating the antiproliferative effect of ATA against HCT116 cells. Of note, ATA remarkably suppressed the growth of HCT116 xenografts in nude mice and displayed an apparent attenuation of FDFT1 in tumor tissues accompanied by the alteration of the biomarkers of autophagy, cell cycle, apoptosis, and ferroptosis. These results demonstrate that ATA exerted in vitro and in vivo antiproliferative effects against HCT116 cells through inducing cell apoptosis, autophagy, and ferroptosis via targeting FDFT1.

Recent studies have showed that thrombosis is closely related to leucocytes involved in immunity. Interfering with the binding of leukocyte integrin Mac-1 and platelet GPIbα can inhibit thrombosis without affecting physiological coagulation. Mac-1-GPIbα is proposed as a potential safety target for antithrombotic agents. Guanxinning tablet (GXNT) is an oral Chinese patent medicine used for the treatment of angina pectoris, which contains phenolic acid active ingredients, such as salvianolic acids, ferulic acid, chlorogenic acid, caffeic acid, rosmarinic acid, tanshinol, and protocatechualdehyde. Our previous studies demonstrated that GXN exhibited significant antithrombotic effects, and clinical studies suggested that it did not increase bleeding risk. In addition, GXN exerted a significantly regulatory effect on immune inflammation. In the current study, we intended to evaluate the effects of GXN on bleeding events and explore the safety antithrombotic mechanism of GXN based on leukocyte-platelet interaction. First, we established a gastric ulcer model induced by acetic acid in rats and found that GXN not only did not increase the degree of gastrointestinal bleeding when gastric ulcer occurred, but also had a certain promoting effect on the healing of gastric ulcer. Second, in vitroexperiments showed that after pretreatment with GXN and activation by phorbol 12-myristate-13-acetate (PMA), the adhesion and aggregation of leukocytes with human platelets were reduced. It was also found that GXN reduced the expression and activation of Mac-1 in leucocytes, and inhibited platelet activation due to leukocyte engagement via Mac-1. Overall, the results suggest that GXN may be a safe antithrombotic agent, and its low bleeding risk mechanism is probably related to inhibited leukocyte-platelet aggregation and its interaction target Mac-1-GPIbα.

Ethnopharmacological relevance: Smilax glabra Roxb., the dry rhizome of Sarsaparilla, which is also known as Tu fuling (TFL) in China, is a well-known traditional CHINESE medicine that is widely used for detoxication, relieving dampness and as a diuretic. We have previously shown that the extracted TFL flavonoids (designated TFLF) possess anti-cardiac hypertrophy effects in vitro. However, the anti-cardiac hypertrophy effects of TFLF in vivo and the underlying mechanisms remain to be elucidated.Aim of the study: To reveal the underlying therapeutic mechanism of TFLF on cardiac hypertrophy by using transverse aortic constriction (TAC) model and cellular assays in vitro.Material & methods: Cardiac hypertrophy was replicated by TAC surgery in rats or by isoprenaline treatment of rat H9C2 myocardial cells in vitro. Cardiac structure and function were evaluated by echocardiographic and hemodynamic examinations in vivo and histological analysis of tissues ex vivo. Biochemical kits and quantitative PCR were used to analyze markers of cardiac hypertrophy. Expression and phosphorylation of key proteins in the Raf/MEK/ERK pathway were quantified by Western blotting. We further confirmed our findings in H9C2 rat cardiomyocytes treated with isoprenaline and the ERK inhibitor in vitro.Results: TFLF attenuated cardiac hypertrophy and fibrosis and improved cardiac dysfunction in TAC rats. TFLF treatment induced a strong reduction in serum NT-proBNP levels. Cardiac hypertrophy marker gene (ANP, BNP and beta-MHC) expression and the phosphorylation levels of c-Raf and ERK1/2 were decreased by TFLF treatment. TFLF also protected H9C2 cells from isoprenaline-induced hypertrophy in vitro via a similar molecular mechanism as that observed in the rat heart. Moreover, pretreatment with TRLF and the ERK inhibitor further inhibited the mRNA overexpression of hypertrophic genes in vitro.Conclusions: TFLFs may protect against pathological cardiac hypertrophy via negative regulation of the Raf/ MEK/ERK pathway. Thus, TFLFs are implicated as a potential pharmacological agent for treating cardiac hypertrophy in clinical practice.

非人哺乳动物细胞系在医学生物学研究中正起着越来越重要的作用.与人源细胞一样,非人哺乳动物细胞在使用中也面临着错用、污染和变异的问题.但与人源细胞相比,非人哺乳动物细胞受到的关注比较少,细胞身份鉴定技术发展较慢.目前对非人哺乳动物细胞系身份鉴定的研究主要集中在小鼠、中国仓鼠和非洲绿猴3个物种,方法以短串联重复序列(short tandem repeats,STR)标记检测为主,单核苷酸多态性(single nucleotide polymorphism,SNP)分析为辅.本文就非人哺乳动物细胞系身份鉴定技术在不同物种中的发展现状进行综述.

Diacid metabolite as the stable form of norcantharidin (DM-NCTD) derived from Chinese blister beetle (Mylabris spp.). The previous studies reported that DM-NCTD could enhance ABT-737-triggered cell viability inhibition and apoptosis in hepatocellular carcinoma (HCC) cell lines. To translate this synergistic therapy into in vivo anticancer treatment, a folate receptor-targeted lipid bilayer-supported chlorodimethyloctadecylsilane-modified mesoporous silica nanoparticle (FA-LB-CHMSN) with DM-NCTD loaded in CHMSN and ABT-737 in lipid bilayer was prepared, which could promote the cancer cell uptake of the drugs through folate receptor-mediated endocytosis. The structure and the properties of the nanoparticle were evaluated. FA-LB-CHMSN with DM-NCTD/ABT-737 loaded induced apparent tumor cell apoptosis and showed remarkably tumor inhibition in H22 tumor-bearing mice model, with significant cellular apoptosis in the tumor and no obvious toxicity to the tissues. We expect that this nanoparticle could be of interest in both biomaterial investigations for HCC treatment and the combination of chemotherapeutic drugs for synergistic therapies.

Smilax glabra Roxb. (SG) is a well-known traditional Chinese medicine that has been extensively used as both food and folk medicine in many countries. Although many beneficial health effects of SG and its primary components have been reported, their action on adipocyte function remains unknown. In the present study, we investigated the effects of the total flavonoids from Smilax glabra Roxb. (SGF) on lipid accumulation in mouse 3T3-L1 adipocytes and further elucidated its potential mechanism using RNA-Seq transcriptome technique. Our results showed that SGF exposure significantly decreased the lipid droplet size and the levels of cellular free fatty acids, while triglyceride accumulation was not affected by SGF. Transcriptome analysis revealed that SGF induced the expression of genes involved in triglyceride storage, fatty acid β-oxidation and mitochondrial biogenesis. Furthermore, we also observed an increased cellular ATP level and mitochondrial mass after SGF exposure, indicating that SGF enhanced mitochondrial function. The other relevant transcriptional changes appeared to be involved in AMPK/PGC-1α signaling, inflammatory response, as well as PI3K/AKT and calcium signaling pathways, which might contribute to the beneficial metabolic effects of SGF on adipocyte function. The results of Western blotting confirmed that SGF could increase the phosphorylation of AMPK while decrease the phosphorylation of AKT in adipocytes. Altogether, our results provided novel information about the molecular mechanism responsible for the effects of SGF on fat storage in adipocytes and highlights the potential metabolic benefits of SGF on human obesity and its related chronic diseases.

OBJECTIVE:To investigate the mRNA, protein expression levels and the phosphorylation levels of key factors in rapidly accelerated fibrosarcoma/mitogen-activated protein kinase kinase/extracellular regulated protein kinases (Raf/MEK/ERK) pathway, and to clarify the regulatory function of Raf/MEK/ERK pathway in myocardial hypertrophy.METHODS:Twenty SD rats were divided into sham-operated group and model group. The myocardial hypertrophy model was established by transverse aortic constriction (TAC). At 12 weeks after TAC, blood samples were collected from the submandibular vein, and the serum was separated to detect the content of N terminal pro B type natriuretic peptide (NT-proBNP). After that, the rats were subjected to echocardiography and hemodynamic measurement. Then the pathological changes of myocardial tissue were observed. And the levels of mRNA, protein expression and the phosphorylation of key factors in Raf/MEK/ERK pathway were detected in myocardial tissue.RESULTS:Compared with sham-operated group, left ventricular end-diastolic interventricular septal thickness (IVSd), left ventricular end-systolic interventricular septal thickness (IVSs), left ventricular end-diastolic posterior wall thickness (LVPWd) and left vebtricular end-systolic posterior wall thickness (LVPWs) in TAC model group were increased significantly (P＜0.05,P＜0.01), left ventricular end-systolic diameter (LVIDs) was decreased significantly (P＜0.01), LV Mass and LW(LV Mass/Weight)were increased significantly (P＜0.05, P＜0.01). The levels of heart rate (HR), left ventricular pressure maximal rate of rise (+dp/dtmax), left ventricular pressure maximal rate of fall (-dp/dtmax) were decreased significantly (P＜0.01). The serum level of NT-proBNP in TAC rat was increased significantly (P＜0.01). The myocardial cells in TAC model group were arranged disorderly, myocardial cell hypertrophy, cytoplasm were increased significantly, and inflammatory cells infiltrated. A large amount of collagen fibers were deposited and large area of myocardial cells were stained blue in TAC rat. The expression levels of phospho-c-Raf (Ser259) and phospho-c-Raf (Ser338) in myocardial tissue were significantly increased (P＜0.01), meanwhile the expression levels of phospho- MEK1/2(Ser217/Ser221) and phospho-ERK1/2 (Thr202/Tyr204) were also significantly increased (P＜0.01).CONCLUSION:The regulatory role of Raf / MEK / ERK pathway in cardiac hypertrophy may be through the activation of phosphorylation of c-raf, MEK1, Mek2, ERK1 and ERK2 at specific sites.

In this study, we aimed to determine the effects of dietary supplementation with chitosan nanoparticles (CNP) on growth performance, immune status, gut microbiota and immune responses after lipopolysaccharide challenge in weaned pigs. A total of 144 piglets were assigned to four groups receiving different dietary treatments, including basal diets supplemented with 0, 100, 200 and 400 mg/kg CNP fed for 28 days. Each treatment group included six pens (six piglets per pen). The increase in supplemental CNP concentration improved the average daily gain (ADG) and decreased the feed and gain (F/G) and diarrhoea rate (p < .05). However, significant differences in the average daily feed intake (ADFI) among different CNP concentrations were not observed. CNP also increased plasma immunoglobulin (Ig)A and IgG, and C3 and C4 concentrations in piglets in a dose-dependent manner on day 28, whereas IgM concentration was not affected by CNP. A total of 24 piglets in the control diet and control diet with 400 mg/kg CNP supplementation groups were randomly selected for the experiment of immunological stress. Half of the pigs in each group (n = 6) were injected i.p. with Escherichia coli lipopolysaccharide (LPS) at a concentration of 100 μg/kg. The other pigs in each group were injected with sterile saline solution at the same volume. Plasma concentrations of cortisol, prostaglandin E2 (PEG2), interleukin (IL)-6, tumour necrosis factor (TNF)-α and IL-1β dramatically increased after LPS challenge. However, CNP inhibited the increase in cortisol, PEG2, IL-6 and IL-1β levels in plasma, whereas TNF-α level slightly increased. Moreover, the effects of CNP on the gut microbiota were also evaluated. Our results showed that dietary supplementation with CNP modified the composition of colonic microbiota, where it increased the amounts of some presumably beneficial intestinal bacteria and suppressed the growth of potential bacterial pathogens. These findings suggested CNP supplementation improved the growth performance and immune status, alleviated immunological stress and regulated intestinal ecology in weaned piglets. Based on these beneficial effects, CNP could be applied as a functional feed additives supplemented in piglets diet.

目的 观察磷脂酶A2(PLA2)在不同的人癌裸小鼠移植瘤模型中的蛋白表达及特征.方法 分别采用肺癌、鼻咽癌、肝癌、胰腺癌、胃癌、结肠癌、食管癌和卵巢癌肿瘤细胞株构建裸小鼠皮下移植瘤模型,待肿瘤生长至瘤体积达1 cm3后,采用酶联免疫吸附(ELISA)法检测肿瘤组织、正常组织及血清中4种主要类型PLA2(cPLA2、sPLA2、iPLA2和lp-PLA2)的蛋白表达,并以正常裸小鼠血清作对照.结果 在呼吸系统、消化系统和生殖系统的8种人肿瘤细胞裸小鼠移植瘤模型中,肿瘤组织cPLA2、sPLA2、lp-PLA2的蛋白表达均显著或极显著高于正常组织(P＜0.05,P＜0.01),肿瘤组织iPLA2的蛋白表达均高于正常组织,除人结直肠癌细胞HT-29移植瘤模型外,均呈现极显著性差异(P＜0.01).8种移植瘤模型中,荷瘤小鼠血清中4种类型PLA2的蛋白含量均极显著高于正常小鼠(P＜0.01).结论 4种类型PLA2在呼吸系统、消化系统和生殖系统不同的裸小鼠移植瘤模型的肿瘤组织和血清中均呈现高表达,无肿瘤类型特异性,表明磷脂酶A2可能参与肿瘤的发生、发展.

目的 制备鳖甲肽HGRFG脂质体以及聚乙二醇(polyethylene glycol,PEG)修饰后的长循环脂质体,考察经修饰的脂质体较未修饰脂质体在动物体内分布及滞留情况.方法 采用BLB/c裸鼠荧光活体成像实验,进行脂质体体内分布及代谢研究;采用药动学实验,初步探究2种载药脂质体在血浆内的滞留时间.结果 2种载药脂质体均能广泛分布于实验动物体内.与未经修饰的脂质体载药组相比,经PEG修饰的长循环脂质体载药组中裸鼠荧光全部消失的时间明显延长,药物在血浆滞留时间也明显延长.结论 经PEG修饰后的长循环脂质体可以延长药物在实验动物体内的滞留时间,延长药物半衰期.

Royal jelly (RJ), a hive product with versatile pharmacological activities, has been used as a traditional functional food to prevent or treat inflammatory diseases. However, little is known about the anti-inflammatory effect of RJ in microglial cells. The aim of this study is to assess the anti-inflammatory effects of RJ in lipopolysaccharide- (LPS-) induced murine immortalized BV-2 cells and to explore the underlying molecular mechanisms. Our results showed that in LPS-stimulated BV-2 cells, RJ significantly inhibited iNOS and COX-2 expression at mRNA and protein levels. The mRNA expression of IL-6, IL-1β, and TNF-α was also downregulated by RJ in a concentration-dependent manner. Additionally, RJ protected BV-2 cells against oxidative stress by upregulating heme oxygenase-1 (HO-1) expression and by reducing reactive oxygen species (ROS) and nitric oxide (NO) production. Mechanistically, we found that RJ could alleviate inflammatory response in microglia by suppressing the phosphorylation of IκBα, p38, and JNK and by inhibiting the nucleus translocation of NF-κB p65. These findings suggest that RJ might be a promising functional food to delay inflammatory progress by influencing the microglia function.