Caveolin-1 Alleviates Acetaminophen-Induced Hepatotoxicity in Alcoholic Fatty Liver Disease by Regulating the Ang II/EGFR/ERK Axis.

Acetaminophen (APAP) is a widely used antipyretic analgesic which can lead to acute liver failure after overdoses. Chronic alcoholic fatty liver disease (AFLD) appears to enhance the risk and severity of APAP-induced liver injury, and the level of angiotensin II (Ang II) increased sharply at the same time. However, the underlying mechanisms remain unclear. Caveolin-1 (CAV1) has been proven to have a protective effect on AFLD. This study aimed to examine whether CAV1 can protect the APAP-induced hepatotoxicity of AFLD by affecting Ang II or its related targets. In vivo, the AFLD model was established according to the chronic-plus-binge ethanol model. Liver injury and hepatic lipid accumulation level were determined. The levels of Angiotensin converting enzyme 2 (ACE2), Ang II, CAV1, and other relevant proteins were evaluated by western blotting. In vitro, L02 cells were treated with alcohol and oleic acid mixture and APAP. CAV1 and ACE2 expression was downregulated in APAP-treated AFLD mice compared to APAP-treated mice. The overexpression of CAV1 in mice and L02 cells alleviated APAP-induced hepatotoxicity in AFLD and downregulated Ang II, p-EGFR/EGFR and P-ERK/ERK expression. Immunofluorescence experiments revealed interactions between CAV1, Ang II, and EGFR. The application of losartan (an Ang II receptor antagonist) and PD98059 (an ERK1/2 inhibitor) alleviated APAP-induced hepatotoxicity in AFLD. In conclusion, our findings verified that CAV1 alleviates APAP-aggravated hepatotoxicity in AFLD by downregulating the Ang II /EGFR/ERK axis, which could be a novel therapeutic target for its prevention or treatment.

Profiling and quantitation of alkaloids in different parts of Sophora alopecuroides L. extracts by high-performance liquid chromatography with electrospray ionisation ion mobility spectrometry detection.

INTRODUCTION: Ambient pressure electrospray ionisation ion mobility spectrometry coupled to high-performance liquid chromatography (HPLC) was used to detect alkaloids from different parts of Sophora alopecuroides L. extracts. Multiplexing ion mobility spectrometry (IMS) was used to improve the signal-to-noise ratio while maintaining high resolving power for the detecting of eluents from HPLC separation. MATERIAL AND METHODS: The alkaloids profile and distribution are demonstrated by retention time-drift time two-dimensional spaces, and the contents of five major alkaloids including sophoridine, sophocarpine, cytisine, aloperine, and matrine were determined in the leaf, skin, stem, seed kernel, and seed husk using the HPLC-IMS method. This method offers extra separation ability to isomers such as matrine and sophocarpine, which can be difficult to distinguish by mass spectrometry. RESULTS: The reduced mobilities for cytisine, sophoridine, sophocarpine, matrine, and aloperine are 0.828, 0.718, 0.731, 0.725, and 0.769 cm2 /V/s, respectively. The limits of detection are 0.553, 0.488, 0.479, 0.484, and 0.513 ug/mL. This method adds extra separation ability to HPLC to resolve co-eluted peaks and provides another qualitative parameter besides HPLC retention time.

Structural and physicochemical properties of lotus seed starch-chlorogenic acid complexes prepared by microwave irradiation.

Lotus seed (LS) has a high starch content and possesses many useful functional properties, which are mainly attributed to its phenolic compound content. The objective of this study was to investigate the effect of microwave irradiation (MW) treatment on the structural and physicochemical properties of a lotus seed starch-chlorogenic acid (CA) blend. MW treatment appeared to promote the formation of LS-CA complexes and the modified starch displayed more rougher structures than native starch. The particle size distribution of starch remained approximately constant when the microwave power was 200 W, but increased sharply with further increases in microwave power; a similar trend was observed in the swelling and solubility of starch. XRD and FT-IR spectra show that MW treatment degraded the ordered crystalline structure of starch, facilitating exposure of the starch chains originally buried in the crystalline and amorphous regions within the grains. During this treatment, CA interacted with starch molecules by hydrogen bonding and form a LS-CA complex, which inhibited the self-assembly process of starch chains. These findings demonstrated the potential use of MW treatment in controlling the storage and processing quality of lotus seed, or other starchy foods rich in polyphenols.

A narrative review of the role of exosomes and caveolin-1 in liver diseases and cancer.

Exosomes are nanoscale (40-100 nm) vesicles secreted by different types of cells and have attracted extensive interest in recent years because of their unique role in disease development. It can carry related goods, such as lipids, proteins, and nucleic acids, to mediate intercellular communication. This review summarizes exosome biogenesis, release, uptake, and their role in mediating the development of liver diseases and cancer, such as viral hepatitis, drug-induced liver injury, alcohol-related liver disease, non-alcoholic fatty liver disease, hepatocellular carcinoma, and other tumors. Meanwhile, a fossa structural protein, caveolin-1(CAV-1), has also been proposed to be involved in the development of various diseases, especially liver diseases and tumors. In this review, we discuss the role of CAV-1 in liver diseases and different tumor stages (inhibition of early growth and promotion of late metastasis) and the underlying mechanisms by which CAV-1 regulates the process. In addition, CAV-1 has also been found to be a secreted protein that can be released directly through the exosome pathway or change the cargo composition of the exosomes, thus contributing to enhancing the metastasis and invasion of cancer cells during the late stage of tumor development. In conclusion, the role of CAV-1 and exosomes in disease development and the association between them remains to be one challenging uncharted area.

Microwave-Induced Behavior and Digestive Properties of the Lotus Seed Starch-Chlorogenic Acid Complex.

The effect of chlorogenic acid (CA) on the dielectric response of lotus seed starch (LS) after microwave treatment, the behavior and digestive characteristics of the resulting starch/chlorogenic acid complex (LS-CA) at different degrees of gelatinization and the inhibition of α-amylase by chlorogenic acid were investigated. The variation in dielectric loss factor, ε″, and dielectric loss tangent, tanδε, of the microwave thermal conversion indicated that LS-CA had a more efficient microwave-energy-to-thermal-energy conversion efficiency than LS. This gelatinized LS-CA to a greater extent at any given temperature between 65 and 85 °C than LS, and it accelerated the degradation of the starch crystalline structure. The greater disruption of the crystal structure decreased the bound water content and increased the thermal stability of LS-CA compared to LS. The simulated in vitro digestion found that the presence of the LS-CA complex improved the slow-digestion property of lotus seed starch by increasing its content of resistant and slowly digested starch. In addition, the release of chlorogenic acid during α-amylase hydrolysis further slowed starch digestion by inhibiting α-amylase activity. These findings provide a foundation for understanding the correlation between the complex behavior and digestive properties of naturally polyphenol-rich, starch-based foods, such as LS, under microwave treatment, which will facilitate the development of starch-based foods with tailored digestion rates, lower final degrees of hydrolysis and glycemic indices.

Caveolin-1 alleviates acetaminophen-induced vascular oxidative stress and inflammation in non-alcoholic fatty liver disease.

Acetaminophen (APAP) is one of the most widely used drugs in the world. The literature shows that excessive or long-term use of APAP can lead to increased cardiovascular dysfunction. An acute increase in angiotensin Ⅱ (Ang Ⅱ) caused by APAP use in fatty liver disease may increase the risk and severity of vascular injury. However, the underlying mechanism remains unclear. Caveolin-1 (CAV1) is a broad-spectrum kinase inhibitor that significantly determines endothelial function. This study aimed to observe the effects of APAP on the vasculature in non-alcoholic fatty liver disease (NAFLD) and to determine whether CAV1 could alleviate vascular oxidative stress and inflammation by targeting Ang Ⅱ or its downstream pathways. In this study, 7-week-old C57BL/6 male mice (18-20 g) were administered APAP by gavage after eight weeks of a high-fat diet. Any resulting vascular oxidative stress and inflammation were assessed. Levels of Ang Ⅱ, CAV1, and other related proteins were measured using ELISA and western blotting. In APAP-treated NAFLD mice, CAV1 expression was downregulated and Ang Ⅱ expression was upregulated compared to normal APAP-treated mice. In vitro, HUVECs were incubated with Ang Ⅱ (300 nM) for 48 h. Overexpression of CAV1 in HUVECs attenuated Ang Ⅱ-induced oxidative stress and inflammation and downregulated the expression of Protein kinase C (PKC) and p-P38/P38. After intervention with CAV1-siRNA, immunofluorescence results showed that the fluorescence intensity of PKC on mitochondria was further increased, and flow cytometry results showed that the mitochondrial membrane potential increased. PKC inhibitors alleviated Ang Ⅱ-induced endothelial injury. In conclusion, our findings confirmed that CAV1 exerts a protective effect against vascular injury by inhibiting oxidative stress and inflammation through the PKC/MAPK pathway. Therefore, restoration of CAV1 may have clinical benefits in reducing APAP-induced vascular damage in NAFLD patients.

Caveolin-1 ameliorates acetaminophen-aggravated inflammatory damage and lipid deposition in non-alcoholic fatty liver disease via the ROS/TXNIP/NLRP3 pathway.

The overuse of acetaminophen (APAP) may cause more severe hepatotoxicity in patients with non-alcoholic fatty liver disease (NAFLD). Caveolin-1 (CAV1), is an essential regulator of metabolic function, which can alleviate liver damage by scavenging reactive oxygen species (ROS). Evidence suggests that the NOD-like receptor family pyrin domain-containing 3 (NLRP3) -mediated pyroptosis is involved in the development of NAFLD. Moreover, thioredoxin-interactive protein (TXNIP) activation is a key event linking ROS to NLRP3 inflammasome. However, whether CAV1 alleviates APAP-aggravated hepatotoxicity in NAFLD via the ROS/TXNIP/NLRP3 pathway remains unclear. An in vivo fatty liver model was established by feeding mice a high-fat diet for 56 days. Additionally, using in vitro approach, AML-12 cells were incubated with free fatty acids for 48 h and APAP was added during the last 24 h. We found that the overuse of APAP in NAFLD not only induced oxidative stress, but also increased TXNIP expression, NLRP3-mediated pyroptosis, and lipid deposition. In addition to inhibiting ROS generation and lipid deposition, overexpression of CAV1 reduced the elevated levels of TXNIP expression and NLRP3-mediated pyroptosis. However, the effect of CAV1 on TXNIP expression, NLRP3-mediated pyroptosis, and lipid deposition was reversed by CAV1 small interfering RNA (siRNA) intervention. Finally, N-acetyl cysteine (NAC) treatment reduced CAV1 siRNA-mediated changes in TXNIP expression and NLRP3-mediated pyroptosis levels. These results demonstrate that the inhibitory effect of CAV1 on NLRP3-mediated pyroptosis may be mediated through the ROS/TXNIP axis. Moreover, the current study provides novel mechanistic insights into the protective effects of CAV1 on APAP-aggravated hepatotoxicity in NAFLD.

Caveolin-1 protects against liver damage exacerbated by acetaminophen in non-alcoholic fatty liver disease by inhibiting the ERK/HIF-1α pathway.

Acetaminophen (APAP) is a common antipyretic and analgesic drug that can cause long-term liver damage after an overdose. Non-alcoholic fatty liver disease (NAFLD) increases susceptibility to APAP. In NAFLD, excessive accumulation of lipids leads to an abnormal increase in hypoxia-inducible factor-1α (HIF-1α). Caveolin-1 (CAV1) may protect against NAFLD by inhibiting HIF-1α. This research aimed to determine whether CAV1 could attenuate APAP-exacerbated liver injury in NAFLD by inhibiting oxidative stress involving HIF-1α. In this study, 7-week-old C57BL/6 mice were fed a high-fat diet (HFD) for eight weeks, followed by the instillation of APAP. Levels of oxidative stress and liver lipid deposition were determined, and p-ERK1/2 and HIF-1α protein expression were measured by the Western blot (WB) method. In the APAP-treated group, the level of CAV1 was decreased, while the levels of HIF-1α and reactive oxygen species (ROS) were significantly increased. AML12 cells were treated with a mixture of palmitic acid (PA) and oleic acid (OA) (1:2 mix) for 48 h, and APAP was added for the last 24 h. Overexpression of CAV1 in AML12 cells significantly inhibited the expression of ROS and HIF-1α. And the results of immunofluorescence after treatment with CAV1-SiRNA showed that the HIF-1α levels were significantly increased in mitochondria. In conclusion, our experimental results suggest that CAV1 has a protective function in the fatty liver based on preventing oxidative stress, which involves HIF-1α. Thus, upregulation of CAV1 may attenuate APAP-exacerbated liver injury in NAFLD.

Insights into the formation and digestive properties of lotus seed starch-glycerin monostearate complexes formed by freeze-thaw pretreatment and microfluidization.

The objective of this paper was to investigate the formation and digestive properties of lotus seed starch-glycerin monostearate complexes (LSG) formed by freeze-thaw pretreatment and microfluidization. The results showed that the preparation of LSG with six freeze-thaw cycles at 60 MPa had the highest complex index (69.92%). The formation of LSG led to the conversion of the crystalline pattern of lotus seed starch from C-type to V-type and increased the proportion of the microcrystalline region. In addition, the digestive results indicated that LSG had a high resistance to digestive enzymes, which was conducive to increasing the content of resistant starch. Based on the above investigation, the formation and digestive properties showed that the appropriate number of freeze-thaw cycles of pretreatment could facilitate the complexation of starch and lipid under low-pressure microfluidization, which made for the directional regulation of helical conformation and anti-digestion.

Effect of chlorogenic acid on the structural properties and digestibility of lotus seed starch during microwave gelatinization.

The structural evolution of lotus starch (LS)-chlorogenic acid (CA) complexes was investigated after microwave-heating treatment, to reveal the relationship between the interactions of lotus starch and chlorogenic acid molecules, and the digestive properties of the starch, after microwave gelatinization. During the early stage of microwave gelatinization (65, 70 °C), CA was mainly participating in the rearrangement of starch molecules in a weakly-bound form, and at that stage, the LS-CA complex acted as an inhibitor of digestion, under small intestine conditions, mainly through the release of CA, which inhibited amylase. However, during the late stage of microwave gelatinization (85 °C), many chlorogenic acid molecules entered the hydrophobic helical cavity of the starch, promoting formation of the V-type starch helical structure in the LS-CA complex, which made a major contribution to inhibiting digestion under oral digestion conditions.

Insights into the multi-scale structural properties and digestibility of lotus seed starch-chlorogenic acid complexes prepared by microwave irradiation.

By inspecting starch hierarchical structural evolution, this work explored how microwave irradiation tailored the digestion characteristics of lotus seed starch-chlorogenic acid mixtures. The results showed that after microwave treatment, the granular structure, short-range ordered structure, helical conformation, and lamellar structure of starch exhibited different degrees of disorganization. In this procedure, chlorogenic acid interacted with starch molecules to form lotus seed starch-chlorogenic acid complexes and participated in the reorganization of the matrixes of the starch substrate in three forms: V-type inclusion complex, non-inclusion complex, and simply physically entrapped. These structural changes, coupled with the inhibition of chlorogenic acid on carbohydrate hydrolyzing enzymes, contributed to the slowly digestible features of lotus seed starch-chlorogenic acid complexes. This study provided a basis for understanding the multi-scale structure-digestibility relationship of starchy foods rich in phenolic acids under microwave treatment.

A new chlorine-containing iridoid glycoside from Plantago maxima.

A phytochemical investigation on the whole plant of Plantago maxima Juss. ex Jacq led to the isolation of a new and rare chlorinated iridoid glycoside named plantomoside (1), along with three known compounds, geniposidic acid (2), 10-deoxygeniposidic acid (3), and viteoid II (4). The structure of 1 was determined through 1 D and 2 D NMR spectroscopic data analysis, HR-ESI-MS, and acid hydrolysis.

Real-time monitoring of the cell agglutination process with a quartz crystal microbalance.

The real-time monitoring of the agglutination process of human hepatic normal cells (L-02) at the quartz crystal microbalance (QCM) gold (Au) electrode was performed. Two lectins, concanavalin A (Con A) and wheat germ agglutinin (WGA), induced the cell agglutination, resulting in the different Deltaf(0) and DeltaR(1) responses from those caused by the normal cell attachment and growth. The cell-Con A-cell aggregates had higher affinity for the Au substrate due to the excellent adsorption ability of Con A, which was revealed by increased Deltaf(0) and DeltaR(1) shifts and the obvious mass effect of QCM. In contrast, the lower adsorption ability of cell-WGA-cell aggregates was related to the same characteristic of WGA, presenting the decreased Deltaf(0) and DeltaR(1) responses and the time-extended adhesion phase. Parallel microscopic observation experiments were also carried out and exhibited comparable results. The Deltaf(0) responses during the processes of cell growth and cell agglutination were analyzed using the equations Deltaf(0)=alpha(0)+alpha(1)e(-t/tau(1))+alpha(2)e(-t/tau(2))+alpha(3)e(-t/tau(3)) and Deltaf(0)=alpha(0)+alpha(1)e(-t/tau(1))+alpha(2)e(-t/tau(2)), respectively. Furthermore, the current work proved that the QCM measurement technique based on cell agglutination was useful for discriminating hepatic normal cells (L-02) and hepatic cancer cells (Bel7402).

Adsorption of Rhodamine B on two novel polar-modified post-cross-linked resins: Equilibrium and kinetics.

We employed two polar monomers, triallyl isocyanurate (TAIC) and butyl acrylate (BA), to copolymerize with divinylbenzene (DVB), and synthesized two starting copolymers labeled PDT and PDB. Then, the Friedel-Crafts alkylation reaction was performed for the two starting copolymers, and the residual pendent vinyl groups were consumed, and hence we obtained two novel polar-modified post-cross-linked resins PDTpc and PDBpc. The surface polarity greatly improved due to introduction of the polar monomers, and the Brunauer-Emmett-Teller (BET) surface area and pore volume significantly increased after the Friedel-Crafts alkylation reaction. Compared with the starting copolymers, the non-polar post-cross-linked resin PDVBpc and some other adsorbents in the references, PDTpc and PDBpc possessed a much enhanced adsorption to Rhodamine B, and the equilibrium capacity reached 578.2mg/g and 328.7mg/g, respectively, at an equilibrium concentration of 100mg/L, and the Freundlich model characterized the equilibrium data very well. The adsorption was a fast process and the kinetic data obeyed the micropore diffusion model. These results confirmed that PDTpc and PDBpc had the potential superiority in adsorptive removal of Rhodamine B from aqueous solution.

In vitro study on the individual and synergistic cytotoxicity of adriamycin and selenium nanoparticles against Bel7402 cells with a quartz crystal microbalance.

Selenium nanoparticles (Se NPs) were prepared based on the reduction of selenious acid (H(2)SeO(3)), by employing sodium alginate (SA) as a template. The real-time monitoring of the drug-inducing apoptosis process of human hepatic cancer cells Bel7402 was performed with the quartz crystal microbalance (QCM) measurement. The anti-tumor effect of adriamycin (ADM) used in combination with Se NPs was investigated. It is found that both drugs were able to inhibit cell proliferation in a dose-dependent way and the combined treatment with ADM and Se NPs was more effective in inhibiting cell growth than each of the two drugs alone. The cytotoxic effects of drug combination were evaluated with the modified Bürgi formula (Jin equation) based the Deltaf(0) responses. The grades gradually changed from apparent synergism to simple addition with the drug-treatment time increasing but the drug combination with lower concentrations still exhibited synergism after 24h, suggesting a potential application in cancer therapy.