摘要：
Bacterial biofilms are usually resistant to antibiotics, thus powerful methods are required for removal. Nanomaterial involving a combination of treatment modalities recently has been recognized as an effective alternative to combat biofilm. However, its targeted and controlled release in bacterial infection is still a major challenge. Here, we present an intelligent phototherapeutic nanoplatform consisting of an aptamer (Apt), indocyanine green (ICG), and carboxyl-functionalized graphene oxide (GO–COOH), namely, [email protected], for targeted treatment of the biofilm formed by Salmonella Typhimurium. Since Apt-conjugated nanosheets (NSs) can specifically accumulate near abscess caused by the pathogens, they enhance greatly the local drug molecule concentration and promote their precise delivery. They can simultaneously generate heat and reactive oxygen species under near-infrared irradiation for photothermal/photodynamic therapy, thereby significantly enhancing biofilm elimination. The phototherapeutic [email protected] also displays a good biocompatibility. More importantly, the multifunction phototherapeutic platform shows an efficient biofilm elimination with an efficiency of greater than 99.99% in an abscess formation model. Therefore, [email protected] NSs with bacteria-targeting capability provide a reliable tool for clinical bacterial infection that circumvents antibiotic resistance.
摘要：
ETHNOPHARMACOLOGICAL RELEVANCE: Astragaloside IV (AST IV) is the active component of Astragalus membranaceus (Fisch.) Bunge, which regulates lipid and carbohydrate metabolism and improves insulin resistance. In this study, we investigated the effects of AST IV on insulin resistant cells and a non-alcoholic fatty liver disease (NAFLD) model induced by high-concentration insulin or oleic acid (OA) in HepG2 cells, as well as the associated regulatory markers. METHODS: First, the target of AST IV was predicted via pharmacophore model matching and molecular docking. Then, enzyme kinetics experiments were conducted in vitro to determine the effect of AST IV on the target protein. Next, AST IV's toxicity was tested on HepG2 cells in vitro, through an insulin resistance model and an NAFLD model, by high-concentration insulin or OA, respectively. To explore the effects of AST IV on insulin resistance and lipid metabolism, we detected the related indexes of glucose and lipid metabolism through commercially available kits. Relevant proteins were also detected by Western blot to provide future direction for study. RESULTS: Our preliminary results of pharmacophore model matching and molecular docking suggested that AST IV and protein tyrosine phosphatase 1B (PTP1B) can be well-combined through hydrogen bonding. Further, the enzyme kinetics experiment showed that AST IV was an effective and specific inhibitor to PTP1B. We found that the protein level of PTP1B in HepG2 cells was significantly increased after treating with high-concentration insulin or OA. Additionally, the intervention of AST IV significantly increased glucose consumption in an insulin resistance model and reduced the content of triglyceride (TG), total cholesterol (TC), and free fatty acid (FFA) in the NAFLD model. Moreover, the 2-N-(7-nitrobenze-2-oxa-1, 3 diazol-4-yl) (2-NBDG) uptake rate in the NAFLD model was also greatly improved. These results validated the effects of AST IV on improving insulin resistance and lipid accumulation. Furthermore, Western blot results illustrated that AST IV suppressed PTP1B and increased levels of phosphorylated insulin receptor (p-IR) and phosphorylated insulin receptor substrate-1 (p-IRS-1) in insulin-resistant HepG2 cells, while also decreasing protein levels of PTP1B and sterol element regulatory binding protein-1c (SREBP-1c) in the NAFLD model. CONCLUSION: This study demonstrated that AST IV inhibited PTP1B and effectively improved insulin resistance in insulin-resistant HepG2 cells and triglyceride accumulation in OA-treated HepG2 cells.
摘要：
The relationship between chemical structure and in vitro cytotoxic activities of a series of azastibocine-framework organoantimony(III) halide complexes against cancerous (HepG2, MDA-MB-231, MCF-7 and HeLa) and nonmalignant (HEK-293) cell lines was studied for the first time. A positive correlation between cytotoxic activity and the length of N-->Sb coordinate bond on azastibocine framework of same nitrogen substituent was observed. By comparison, the organoantimony(III) complex 6-cyclohexyl-12-fluoro-5,6,7,12-tetrahydrodibenzo[c,f][1,5]azastibocine (C4) exhibited the highest selectivity index, giving a IC50(nonmalignant)/IC50(cancerous) ratio of up to 8.33. The results of cell cycle analysis indicated that the inhibitory effect of C4 on the cellular viability was caused by cell cycle arrest mainly at the S phase. The necrosis induced by C4 was confirmed by the Trypan blue dye exclusion test and the increase of lactic dehydrogenase (LDH) released in the culture medium. Furthermore, evaluation of the levels of intracellular reactive oxygen species (ROS) in MDA-MB-231cells, by quantifying the relative fluorescence units (RFU) using spectrofluorometer, indicated that cytotoxic activity of C4 is dependent on the production of ROS. This work established the correlation between cytotoxic activity and N-->Sb inter-coordination, a finding that provided theoretical and experimental basis for in-depth design of antimony-based organometallic complexes as potential anticancer agents.
作者机构：
[Zhang, Mengxia] Hunan Univ Chinese Med, Dept Histol & Embryol, Changsha 410208, Hunan, Peoples R China.;[Tang, Shengsong] Hunan Univ Med, Hunan Prov Key Lab Antibody Based Drug & Intellig, Huaihua 418000, Hunan, Peoples R China.;[Zhang, Mengxia; Liu, Qi; Mo, Zhongcheng] Univ South China, Clin Anat & Reprod Med Applicat Inst, Dept Histol & Embryol, Hengyang 421001, Hunan, Peoples R China.;[Tang, Shengsong; Lei, Xiaoyong; Tu, Jian; Li, Lijun] Univ South China, Insitute Pharm & Pharmacol, Hengyang 421001, Hunan, Peoples R China.;[Ning, Jing; Tang, Shengsong] Hunan Univ Med, Dept Pharmacol, Huaihua 418000, Hunan, Peoples R China.
通讯机构：
[Tang, Shengsong] H;Hunan Univ Med, Hunan Prov Key Lab Antibody Based Drug & Intellig, Huaihua 418000, Hunan, Peoples R China.
关键词：
macrophage colony-stimulating factor;breast cancer;apoptosis;HIF-1 and BINP3
摘要：
Macrophage colony-stimulating factor (M-CSF), a tumour marker, is related to tumour cell anti-apoptosis and drug resistance. However, the role of M-CSF in MCF-7 cells is unknown. In the present study, the effect and mechanism of M-CSF on hypoxia-inducible factor-1 (HIF-1)/BCL2/adenovirus E1B 19 kDa-interacting protein 3 (BNIP3)/Apoptosis Regulator BAX signalling in human breast cancer MCF-7 cells were investigated. Western blotting revealed that the expression of HIF-1, BNIP3, Bax, caspase-3 and caspase-9 was lower in MCF-7-M cells compared to MCF-7 and MCF-7-C cells treated with adriamycin (ADM). Immunoprecipitation combined with western blotting was used to detect the interaction between Bcl-2 and BNIP3 or Bax protein. MCF-7-M cells had a higher amount of Bax binding to Bcl-2 compared to MCF-7 cells or MCF-7-C cells, while the amount of BNIP3 binding to Bcl-2 was decreased in MCF-7-M cells. Hoechst 33342 staining and flow cytometry were utilized to evaluate the effect of M-CSF on apoptosis in MCF-7 cells treated with ADM. Compared to ADM-treated MCF-7 cells, the apoptotic rate of MCF-7-M cells was significantly decreased. These effects were dependent on the concentration of ADM. In conclusion, cytoplasmic M-CSF suppressed apoptosis by inhibiting the HIF-1/BNIP3/Bax signalling pathway, which potentiated the dissociation of Bcl-2 from Bcl-2-BNIP3 compounds and the formation of Bcl-2-Bax compounds.
作者机构：
[李迎红; 王世强; 廖小华] Physical Education College, Hunan University of Technology, Zhuzhou 412000, Hunan Province, China;[李敏] Department of Basic Medicine, Hunan College of Traditional Chinese Medicine, Zhuzhou 412000, Hunan Province;[李强] Department of Rehabi litation and Health Care, Hunan College of Traditional Chinese Medicine, Zhuzhou 412000, Hunan Province;[郭义] Center for Experimental Acupuncuture Research, Tianjin University of Traditional Chinese Medicine, Tianjin 300193;[Luo, Xiang-Jun] Department of Acupuncture and Moxibustion, The First Affiliated Hospital of Hunan College of Traditional Chinese Medicine, Zhuzhou 412000, Hunan Province
期刊：
International Journal of Molecular Medicine,2019年43(5):2055-2063 ISSN：1107-3756
通讯作者：
Liu, Lu-Shan;Wang, Mei-Mei
作者机构：
[Jiang, Zhi-Sheng; Ren, Zhong; Xiao, Jun; Liao, Ling; Tang, Zhi-Han; Zhou, Min; Xiang, Qiong; Peng, Juan; Bai, Xue-Qin; Liu, Lu-Shan] Univ South China, Inst Cardiovasc Dis, Key Lab Arteriosclerol Hunan Prov, 28 West Changsheng Rd, Hengyang 421001, Hunan, Peoples R China.;[Wen, Hong-Yan] Hunan Univ Chinese Med, Med Coll, Changsha 410208, Hunan, Peoples R China.;[Wang, Mei-Mei] Univ South China, Affiliated Nanhua Hosp, Dept Pediat, 28 West Changsheng Rd, Hengyang 421001, Hunan, Peoples R China.
通讯机构：
[Liu, Lu-Shan; Wang, Mei-Mei] U;Univ South China, Inst Cardiovasc Dis, Key Lab Arteriosclerol Hunan Prov, 28 West Changsheng Rd, Hengyang 421001, Hunan, Peoples R China.;Univ South China, Affiliated Nanhua Hosp, Dept Pediat, 28 West Changsheng Rd, Hengyang 421001, Hunan, Peoples R China.
关键词：
HepG2 cells;Hydrogen sulfide;Lipid metabolism;Low-density lipoprotein receptor;Phosphoinositide 3-kinase/protein kinase B;Proprotein convertase subtilisin/kexin type 9;Sterol regulatory element-binding protein 2
摘要：
Hydrogen sulfide (H2S) is an endogenous gaseous signaling molecule that plays important roles in the cardiovascular system. In our previous studies, we demonstrated that H2S regulates lipid metabolism. In the present study, we aimed to explore the mechanisms through which H2S regulates lipid metabolism in HepG2 cells in vitro. Treatment of the HepG2 cells with H2S inhibited the expression of proprotein convertase subtilisin/kexin type 9 (PCSK9) and increased the level of lowdensity lipoprotein receptor (LDLR) in a time and dosedependent manner. The knockdown of PCSK9 by siRNA effectively increased the levels of LDLR and 1,1'dioctadecyl3,3,3',3'tetramethylindocarbocyanine perchloratelabeled LDL (DiILDL) uptake in the H2Streated HepG2 cells. Furthermore, the phosphoinositide 3kinase (PI3K)/protein kinase B (Akt)sterol regulatory element binding proteins 2 (SREBP2) signaling pathway was confirmed to be involved in H2Sregulated PCSK9 expression. Notably, the HepG2 cells were incubated with 30% serum and DiILDL for 24 h, and the results revealed that H2S increased lipid uptake, but caused no increase in lipid accumulation. On the whole, the findings of this study demonstrate that H2S is involved in the regulation of lipid metabolism in HepG2 cells through the regulation of the expression of PCSK9 via the PI3K/AktSREBP2 signaling pathway. To the very best of our knowledge, this study is the first to report that H2S can regulate the expression of PCSK9.
作者机构：
[唐标] Medical School, Hunan University of Chinese Medicine, Changsha 410028, China;[邓常清] Medical School, Hunan University of Chinese Medicine, Changsha 410028, China. dchangq@sohu.com
通讯机构：
[Lu, Fangguo] U;[Hu, J; Lu, FG] H;Hunan Univ Chinese Med, Dept Microbiol, Sch Med, Changsha 410208, Hunan, Peoples R China.;Univ Innovat Team Hunan Prov, Key Discipline Pathogen Biol, Changsha 410208, Hunan, Peoples R China.
关键词：
*16S rRNA;*Carboxyfluorescein;*DNA probe;*DNA/RNA hybrids;*Drug-resistant bacteria;*Enzymatic reaction;*Fluorescence resonance energy transfer;*Fluorometry;*Quenching
摘要：
The authors describe a method for the fluorometric determination of methicillin-resistant Staphylococcus aureus (MRSA) by exploiting target-triggered chain reactions and deoxyribonuclease I (DNase I)-aided target recycling. It is making use of a carboxy-fluorescein (FAM)-labeled single-stranded probe containing two sections. One is complementary to the 5' terminus of the target, while the 3' terminus of the other target is adsorbed on the surface of graphene oxide (GO) via pi-stacking interactions without the target (16S rRNA). This adsorption results in quenching of the fluorescence of the label and protects it from being cleaved by DNase I. However, upon addition of the target, DNA/RNA hybrids are repelled by GO. This leads to fluorescence recovery as measured at excitation/emission wavelengths of 480/514 nm due to a chain reaction that is triggered by the target. The signal is strongly amplified by using DNase I-mediated target recycling. The 16S rRNA of MRSA can be detected by this method in the 1 to 30 nM concentration range, and the detection limit is 0.02 nM. The method was applied to analyze bacterial samples, and the detection limit is as low as 30 CFU . mL(-1). The assay is highly sensitive and selective and in our percpetion has a large potential in diagnosis of drug-resistant bacteria. Graphical abstract Schematic of the graphene oxide-based fluorescent bioassay for Methicillin-resistant Staphylococcus aureus detection by using target-triggered chain reaction and deoxyribonuclease I-aided signal amplification.
关键词：
Apolipoprotein E;Apolipoprotein E receptor 2;Atherosclerosis;Inflammation;Proprotein convertase subtilisin kexin 9
摘要：
Atherosclerosis is characterized by chronic inflammation and lipid accumulation in arterial walls, resulting in several vascular events. Proprotein convertase subtilisin kexin 9 (PCSK9), a serine protease, has a pivotal role in the degradation of hepatic low-density lipoprotein receptor (LDLR). It can increase plasma concentrations of low-density lipoprotein cholesterol and affect lipid metabolism. Recently, PCSK9 has been found to accelerate atherosclerosis via mechanisms apart from that involving the degradation of LDLR, with an emerging role in regulating the inflammatory response in atherosclerosis. Apolipoprotein E receptor 2 (apoER2), one of the LDLR family members expressed in macrophages, can bind to its ligand apolipoprotein E (apoE), exhibiting an anti-inflammatory role in atherosclerosis. Evidence suggests that apoER2 is a target of PCSK9. This review aims to discuss PCSK9 as a potential regulator of apoE/apoER2 against inflammation in atherosclerosis.
摘要：
Adipose stem cell (ASC) transplantation is a promising therapeutic strategy for diabetic renal fibrosis. Hypoxia-inducible factor 1 alpha (HIF1 alpha) is a negative regulatory factor of mitochondrial function. In the current study, we aimed to explore if HIF1 alpha deletion protects against hyperglycemia-induced ASC damage and enhances the therapeutic efficiency of ASCs in diabetic renal fibrosis. Our data indicated that HIF1 alpha was upregulated in ASCs in response to high glucose stimulation. Higher HIF1 alpha expression was associated with ASC apoptosis and proliferation arrest. Loss of HIF1 alpha activated mitophagy protecting ASCs against high glucose-induced apoptosis via preserving mitochondrial function. Transplanting HIF1 alpha-deleted ASCs in db/db mice improved the abnormalities in glucose metabolic parameters, including the levels of glucose, insulin, C-peptide, HbA1c, and inflammatory markers. In addition, the engraftment of HIF1 alpha-modified ASCs also reversed renal function, decreased renal hypertrophy, and ameliorated renal histological changes in db/db mice. Functional studies confirmed that HIF1 alpha-modified ASCs reduced renal fibrosis. Collectively, our results demonstrate that ASCs may be a promising therapeutic treatment for ameliorating diabetes and the development of renal fibrosis and that the loss of HIF1 alpha in ASCs may further increase the efficiency of stem cell-based therapy. These findings provide a new understanding about the protective effects of HIF1 alpha silencing on ASCs and offer a new strategy for promoting the therapeutic efficacy of ASCs in diabetic renal fibrosis.
通讯机构：
[Zhang, Xi] H;Hunan Univ Chinese Med, Dept Pathol, 300 Xueshi Rd, Changsha 410208, Hunan, Peoples R China.
关键词：
Qianlongtong;BPH;apoptosis
摘要：
Qianlongtong is a compound made from traditional Chinese herbs and it has proven to be very effective to treat patients with benign prostate hypertrophy. However, its mechanism is still unknown. This study is designed to investigate the effect of Qianlongtong on proliferation and apoptosis of hyperplastic prostate cells. Flow cytometry (FCM) and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) were used to assess proliferation and apoptosis of hyperplastic prostate cells in the following groups: control group, tamoxifen group, and groups with low, moderate, and high dosage of Qianlongtong. Reverse transcription-polymerase chain reaction analysis was used to investigate the underlying mechanisms for increased apoptosis. Cells treated with Qianlongtong were mainly blocked in the G0/G1 phase. The apoptotic index of each group was significantly higher than that in the control group. The apoptotic index in the high- and moderate-dosage groups was similar to that in the tamoxifen group. The high- and moderate-dosage groups had lower Bcl-2 and higher Bax messenger RNA (mRNA) levels compared with the control group. Qianlongtong inhibits proliferation and promotes the apoptosis of hyperplastic prostate cells.
