Kaempferol and nicotiflorin ameliorated alcohol-induced liver injury in mice by miR-138-5p/SIRT1/FXR and gut microbiota.

Aims: Excessive alcohol consumption can lead to alcoholic liver diseases (ALDs). Tetrastigma hemsleyanum Diels et Gilg is a rare Chinese medicinal herb. Tetrastigma hemsleyanum Diels et Gilg has been validated to be highly effective for treating hepatitis. Kaempferol and nicotiflorin are two highly representative flavonoids, which have exhibit therapeutic effects on liver disease. Therefore, the protective mechanism of kaempferol and nicotiflorin on alcohol-induced liver injury were investigated. Main methods: Forty mice were used in this study. After treatment of Kaempferol and nicotiflorin, serum and liver were collected and used for determination of biochemical indicators, H&E staining, and molecular detection. The interaction of miRNAs from serum extracellular vehicles (EVs) with mRNAs and 16S rRNA sequencing of gut microbiota were also investigated. Key findings: The results showed that kaempferol and nicotiflorins significantly ameliorated alcohol-induced liver damage and observably regulated gut microbiota. Specifically, the levels of malondialdehyde (MDA) and CYP2E1 in the liver significantly reduced, and the activity of superoxide dismutase (SOD) and glutathione (GSH) in the liver evidently increased. They also significantly relieved liver oxidative stress and lipid accumulation by suppressing miR-138-5p expression, inversely enhancing deacetylase silencing information regulator 2 related enzyme-1 (SIRT1) levels and then decreasing farnesoid X receptor (FXR) acetylation, which then modulated Nrf2 and SREBP-1c signaling pathways to regulate oxidative stress and lipid metabolism induced by alcohol. Significance: Kaempferol and nicotiflorin reduced alcohol-induced liver damage by enhancing alcohol metabolism and reducing oxidative stress and lipid metabolism. The intestinal microorganism disorder was also ameliorated after oral kaempferol and nicotiflorin.

Modulation Mechanism of Wuniuzao Dark Tea Polysaccharide on Lipid Metabolism in Hyperlipidemic Mice Induced by High-Fat Diet.

The aim of this study was to investigate the functional mechanism of Wuniuzao dark tea polysaccharide (WDTP) that protect against hyperlipidemia in mice induced by high-fat diet. WDTP was extracted by hot water, isolated and purified by DEAE-52 chromatography and characterized by high-performance liquid chromatograph (HPLC), Fourier transform infrared spectroscopy (FT-IR) and scanning electron microscope (SEM). Different doses (200 or 800 mg/kg/day) of WDTP were orally administered to mice induced by high-fat diet to evaluate the mechanism of WDTP regulating lipid metabolism. And these results showed that average molecular weight of WDTP was nearly 63,869 Da. And WDTP intervention significantly reduced body weight, lipid accumulation, and modulated blood lipid levels. The mechanism of WDTP ameliorating lipid metabolism was associated with regulating the expression of lipid metabolism-related genes and serum exosomes miR-19b-3p, and modulating the community structure of gut microbiota in mice.

Cucurbitacin B Exerts Significant Antidepressant-Like Effects in a Chronic Unpredictable Mild Stress Model of Depression: Involvement of the Hippocampal BDNF-TrkB System.

BACKGROUND: Although depression has been a serious neuropsychiatric disorder worldwide, current antidepressants used in clinical practice have various weaknesses, including delayed onset and low rates of efficacy. Recently, the development of new antidepressants from natural herbal medicine has become one of the important research hotspots. Cucurbitacin B is a natural compound widely distributed in the Cucurbitaceae and Cruciferae families and has many pharmacological activities. The present study aimed to investigate whether cucurbitacin B possess antidepressant-like effects in mice. METHODS: The antidepressant-like effects of cucurbitacin B on mice behaviors were explored using the forced swim test, tail suspension test, open field test, sucrose preference test, and a chronic unpredictable mild stress model of depression together. Then, western blotting and immunofluorescence were used to examine the effects of cucurbitacin B on the brain-derived neurotrophic factor (BDNF)-tyrosine kinase B (TrkB) signaling cascade and neurogenesis in the hippocampus of mice. Furthermore, BDNF-short hairpin RNA, K252a, and p-chlorophenylalanine methyl ester were adopted together to determine the antidepressant mechanism of cucurbitacin B. RESULTS: It was found that administration of cucurbitacin B indeed produced notable antidepressant-like effects in mice, which were accompanied with significant promotion in both the hippocampal BDNF-TrkB pathway and neurogenesis. The antidepressant mechanism of cucurbitacin B involves the hippocampal BDNF-TrkB system but not the serotonin system. CONCLUSIONS: Cucurbitacin B has the potential to be a novel antidepressant candidate.

Copper-contaminated soil compromises thermal performance in the springtail Folsomia candida (Collembola).

The widespread agricultural and industrial emissions of copper-based chemicals have increased copper levels in soils worldwide. Copper contamination can cause a range of toxic effects on soil animals and influence thermal tolerance. However, toxic effects are commonly investigated using simple endpoints (e.g., mortality) and acute tests. Thus, how organisms respond to ecological realistic sub-lethal and chronic exposures across the entire thermal scope of an organism is not known. In this study, we investigated the effects of copper exposure on the thermal performance of a springtail (Folsomia candida), regarding its survival, individual growth, population growth, and the composition of membrane phospholipid fatty acids. Folsomia candida (Collembola) is a typical representative of soil arthropods and a model organism that has been widely used for ecotoxicological studies. In a full-factorial soil microcosm experiment, springtails were exposed to three levels of copper (ca. 17 (control), 436, and 1629 mg/kg dry soil) and ten temperatures from 0 to 30 °C. Results showed that three-week copper exposure at temperatures below 15 °C and above 26 °C negatively influenced the springtail survival. The body growth was significantly lower for the springtails in high-dose copper soils at temperatures above 24 °C. A high copper level reduced the number of juveniles by 50 %, thereby impairing population growth. Both temperature and copper exposure significantly impacted membrane properties. Our results indicated that high-dose copper exposure compromised the tolerance to suboptimal temperatures and decreased maximal performance, whereas medium copper exposure partially reduced the performance at suboptimal temperatures. Overall, copper contamination reduced the thermal tolerance of springtails at suboptimal temperatures, probably by interfering with membrane homeoviscous adaptation. Our results show that soil organisms living in copper-contaminated areas might be more sensitive to thermally stressful periods.

Antioxidative effect of Periplaneta americana extract on dextran sulfate sodium-induced ulcerative colitis through activation of the Nrf2 signal.

CONTEXT: Periplaneta americana L. (Blattariae) is used as a treatment for ulcerative colitis (UC) in Chinese traditional medicine. OBJECTIVE: To evaluate the antioxidative activity of P. americana whole body ethanol extract (PAE) on UC mice and whether glycine and proline could be used for quality control and identification of active PAE components. MATERIALS AND METHODS: NCM460 cells were pre-incubated in PAE, AA-L, AA-M, and AA-H (low, high and medium doses of proline and glycine), then treated with recombinant human TNF-α. The glutathione (GSH), malondialdehyde (MDA), superoxide dismutase (SOD) and reactive oxygen (ROS) levels were determined. UC mice were fed with water containing 2.5% dextran sulfate sodium (w/v) after pre-treatment with different doses of PAE once a day for 7 days. ELISA was used to detect the concentrations of inflammation-related factors. Colon tissues of mice were used to detect the activity of myeloperoxidase (MPO), GSH, MDA, and SOD. Histological changes were observed using H&E staining. The expression of target proteins was determined by western blotting. RESULTS: In vivo, PAE treatment reduced the DAI score more than in the model group, restoring the weight and colonic length. It also reduced the severity of colitis, and inflammatory and oxidative stress intensity. Additionally, western blotting showed that the Nrf2 pathway was activated by PAE. In vitro PAE significantly alleviated TNF-α-induced cell damage and oxidative stress, which is relevant to the activation of the Nrf2 pathway. CONCLUSIONS: PAE may relieve oxidative stress through the Nrf2 signaling pathway, and proline and glycine may be used as active components of its antioxidative stress activity.

A biomineralized Prussian blue nanotherapeutic for enhanced cancer photothermal therapy.

Photothermal therapy is a promising tumor ablation technique that converts light into heat energy to kill cancer cells. Prussian blue (PB), a biocompatible photothermal reagent, has been widely explored for cancer treatment. However, the translational potential of PB is severely hampered by its low photothermal conversion efficiency (PCE) and poor stability. To tackle these issues, we adopted the biomineralization modality where PB was integrated with calcium phosphate (CaP) through the binding between calcium ions and PB. The mineralized PB (CaP&PB) demonstrated significantly improved PCE (40.2%), resulting from a calcium-induced bandgap-narrowing effect, and exhibited superior suspension stability. Using a 4T1 orthotopic breast cancer BALB/c mouse model, we observed that mineralized PB showed a significant temperature increase within the tumor, which led to better tumoricidal activity compared with CaP and PB when identical NIR treatment was applied. These achievements demonstrated the success of introducing calcium phosphate into Prussian blue by biomineralization to improve the PCE and stability of photothermal reagents, suggesting an alternative translational strategy for enhanced cancer photothermal therapy.

Growth, serum biochemical parameters, salinity tolerance and antioxidant enzyme activity of rainbow trout (Oncorhynchus mykiss) in response to dietary taurine levels.

This study evaluated the effect of dietary taurine levels on growth, serum biochemical parameters, salinity adaptability, and antioxidant activity of rainbow trout (Oncorhynchus mykiss). Four diets were formulated with taurine supplements at 0, 0.5, 1, and 2% w/v (abbreviated as T0, T0.5, T1, and T2, respectively). Rainbow trouts (initial weight of 80.09 ± 4.72 g) were stocked in tanks (180 L capacity), and were fed these diets for six weeks and subsequently underwent salinity acclimation. Physiological indicators were determined before salinity acclimation at 1, 4, 7, and 14 days afterwards. Results showed that there were no significant differences in growth performance (final mean weight ranged from 182.35 g to 198.48 g; percent weight gain was between 127.68% and 147.92%) of rainbow trout in freshwater stage, but dietary taurine supplement significantly increased serum-free taurine content. After entering seawater, the Na+-K+-ATPase activity of T2 group returned to its freshwater levels, and the serum cortisol content was significantly higher than T0 and T0.5 groups. At the end of this experiment, the liver superoxide dismutase activity in the T0 and T0.5 groups was significantly lower than in the T1 and T2 groups, and the liver catalase in the T0 group was the lowest whereas that in the T2 group was the highest. Muscle malondialdehyde content was the highest in the T0 group, and the lowest in the T2 group. Based on the results of this study, supplement of dietary taurine (0.5-2%) enhanced the salinity tolerance in rainbow trout, which increased with the higher taurine concentration.

Tetramethylpyrazine attenuates endotoxin-induced retinal inflammation by inhibiting microglial activation via the TLR4/NF-κB signalling pathway.

Ocular inflammation is a common pathological condition of a series of retinal degenerative diseases. Tetramethylpyrazine (TMP), a Chinese herbal extraction, is widely used in the treatment of several ocular diseases in Eastern countries. However, the exact mechanisms correlating the vision protective effects of TMP have not been elucidated. Thus, this study aimed to investigate TMP's molecular targets in anti-inflammatory activity in endotoxin lipopolysaccharide (LPS)-induced retinal inflammation both in vitro and in vivo. The primary cultured retinal microglial cells were pretreated with TMP and then activated by LPS. We found pretreatment with TMP significantly inhibited LPS-induced upregulation of CD68, a marker of mononuclear microglia activation. The morphological changes induced by LPS were also inhibited by the TMP pretreatment. Moreover, Toll like receptor 4 (TLR4), phosphorylation of inhibitor of NF-κB alpha (p-IκB-α) and the translocation of nuclear factor kappa B p65 (NF-κB p65) were significantly downregulated in retinal microglial cells with TMP pretreatment, which indicated that TMP might suppress LPS-induced retinal microglial activation through TLR4/NF-κB signalling pathway. And these results were confirmed in vivo. Pretreatment with TMP inhibited microglial activation, migration and regeneration, especially in ganglion cell layer (GCL). In addition to the inhibition of TLR4, TMP significantly inhibited the translocation of NF-κB p-65 to the nucleus in vivo. The downstream genes of NF-κB, such as the pro-inflammatory cytokines interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α) and interleukin-1ß (IL-1ß), were significantly downregulated by TMP pretreatment in the retina. Accordingly, the increased expression of cleaved caspase-3 and the decreased ratio of B-cell lymphoma-2 (Bcl-2) to Bcl-2 associated X Protein (Bax) were significantly attenuated by TMP. TUNEL assay also demonstrated that TMP exerted neuroprotective effects in the retina. Therefore, this study elucidated a novel mechanism that TMP inhibits retinal inflammation by inhibiting microglial activation via a TLR4/NF-κB signalling pathway.

Network-based analysis with primary cells reveals drug response landscape of acute myeloid leukemia.

Acute myeloid leukemia (AML) is one of the most common, complex, and heterogeneous hematological malignancies in adults. Despite progresses in understanding the pathology of AML, the 5-year survival rates still remain low compared with CML, CLL, etc. The relationship between genomic features and drug responses is critical for precision medication. Herein, we depicted a picture for response of 145 drugs against 33 primary cell samples derived from AML patients with full spectrum of genomic features assessed by whole exon sequencing and RNA sequencing. In general, most of the samples were much more sensitive to the combinatorial chemotherapy regimens than the single chemotherapy drugs. Overall, these samples were moderately sensitive to the Traditional Chinese Medicine (TCM) and the targeted drugs. In the weighted gene coexpression network analysis (WGCNA), the TCM and targeted therapies displayed similar genetic signatures in the gene module correlation. Meanwhile, the expression of miRNAs, lncRNAs, and mRNAs did not display apparent gene module correlations among those different types of therapies. In addition, the combinatorial chemotherapy bear more module correlations than the single drugs. Interestingly, we found that the gene mutations and drug response were not enriched in any WGCNA module analysis. Most of the sensitive drug response biomarkers were enriched in the ribosome, endocytosis, cell cycle, and p53 associated signaling pathways. This study showed that gene expression modules might show better correlation than gene mutations for drug efficacy predictions.

Efficacy and safety of moxibustion for chronic low back pain: A systematic review and meta-analysis of randomized controlled trials.

INTRODUCTION: To systematically review and meta-analyze the efficacy of moxibustion in treating patients with chronic low back pain (CLBP). METHODS: A systematic search of the Cochrane Library, Web of Science, PubMed, Embase, EBSCO, CBM, Wanfang, CNKI and VIP (until November, 2019) was used to identify studies reporting pain intensity (VAS or NRS), disability (ODI or RMDQ), JOA score, and quality of life (SF-36) in patients with CLBP. Study selection, data extraction was performed critically and independently by two reviewers. Cochrane criteria for risk of bias was used to assess the methodological quality of the trials. The Grading of Recommendations Assessment, Development, and Evaluation Methodology (GRADE) was applied to test the quality of evidence from the quantitative analysis. RESULTS: Ten RCTs, including 987 patients, met the inclusion criteria. Moxibustion had a superior effect on VAS score when compared with western medicine [RR = -1.69, 95%CI(-2.40, -0.98), p < 0.00001] and acupuncture [RR = -0.47, 95%CI(-0.92, -0.02), p=0.04], but it failed to do so when compared with core stability training [RR = -0.41, 95%CI(-0.87, 0.05), p=0.08]. The result showed that moxibustion plus other active treatments (including western medicine, massage, acupuncture and core stability training) had better effects on low back pain relief compared with active treatments alone. Moxibustion showed favourable effects on disability [SMD = -3.80, 95%CI (-5.49, -2.11), p < 0.0001], JOA score [MD = 4.10, 95%CI(2.30, 5.90), p < 0.00001], and SF-36 score [MD = 13.41, 95%CI(9.68, 17.14), p < 0.00001]. The evidence level of the results from the ten studies was determined to be very low to low. CONCLUSIONS: It is difficult to draw firm conclusions that moxibustion is an effective intervention for treating CLBP due to the small sample size of eligible trails and the high risk of bias among the available articles. Rigorously designed large-scale RCTs are required to further confirm the results in this review.

Positive effects of dietary supplementation of three probiotics on milk yield, milk composition and intestinal flora in Sannan dairy goats varied in kind of probiotics.

In this study, we investigated the effects of Saccharomyces cerevisiae (SC), Bacillus subtilis (BS) and Enterococcus faecalis (EF), singly and in combination, on the dry matter intake (DMI), milk production and composition, and faecal microflora of Saanen dairy goats. Fifty goats were randomly divided into five groups: (a) basal diet (control); (b) basal diet + SC; (c) basal diet + BS; (d) basal diet + EF; and (e) basal diet + mixed probiotics. Each treated animal received 5 g/d of probiotics for a total administration of 5 × 1,011 CFU/goat per day. The inclusion of B. subtilis and E. faecalis in the diet of lactating Saanen goats increased DMI (p < .05). Enhanced milk yield was observed with BS and EF. Milk fat percentage was significantly increased by feeding mixed probiotics compared with the control (p < .05); supplying SC, BS and mixed probiotics enhanced the protein percentage (p < .05). The milk lactose percentage in the SC and BS groups was higher than in the control (p < .05). The amount of milk total solids was higher after feeding EF or mixed probiotics than in the control group (p < .05). Non-fat solids showed no notable differences among groups (p > .05). There was no significant influence on gut bacterial abundance and diversity from adding these three probiotics, singly or in combination. Bacteroidales, Escherichia-Shigella and Christensenellaceae abundances were decreased by supplying these probiotics but Succinivibrionaceae increased. In conclusion, there were positive influences of probiotic feed supplementation on intake, milk performance and intestinal microecology.

Tetramethylpyrazine downregulates transcription of the CXC receptor 4 (CXCR4) via nuclear respiratory factor­1 (Nrf­1) in WERI­Rb1 retinoblastoma cells.

Tetramethylpyrazine (TMP; an extract of the Chinese herbal medicine, Chuanxiong) has been shown to exert remarkable antiretinoblastoma (RB) effects. Based on our previous study, the target gene was found to be C­X­C chemokine receptor type 4 (CXCR4). CXCR4 is a prognostic marker in various types of cancer, but the exact mechanisms underlying the regulation of CXCR4 expression by TMP in WERI­Rb1 cells have yet to be fully elucidated. In the present study, it was revealed that TMP significantly downregulated CXCR4 expression and inhibited CXCR4 promoter activity in WERI­Rb1 cells, indicating that TMP inhibits CXCR4 expression in WERI­Rb1 cells through transcriptional regulatory mechanisms. Among the numerous transcription factors involved in CXCR4 function, including Yin Yang 1 (YY1), nuclear respiratory factor­1 (Nrf­1), Krüppel­like Factor 2 (KLF2), specificity protein 1 (SP1), and nuclear factor­κB subunit 1 (NF­κB1), only TMP led to a significant downregulation of Nrf­1 expression. Chromatin immunoprecipitation assays further indicated that Nrf­1 directly binds to the promoter region of CXCR4, and silencing Nrf­1 via siRNA transfection notably inhibited CXCR4 expression in WERI­Rb1 cells. In addition, the expression levels of both Nrf­1 and CXCR4 increased concomitantly with WERI­Rb1 cell density. Furthermore, the downregulation of Nrf­1 and CXCR4 expression in RB by TMP was confirmed in vivo. Taken together, the results of the present study have uncovered a novel mechanism in which CXCR4 expression is regulated by Nrf­1 in WERI­Rb1 cells, thereby identifying novel potential targets for the treatment of RB, and providing evidence for the clinical application of TMP in adjuvant retinoblastoma therapy.

One-month IOP in mitomycin C-augmented trabeculectomy can predict long-term IOP control in chronic primary angle-closure glaucoma.

PURPOSE: To investigate the predictors of long-term intraocular pressure (IOP) in chronic primary angle-closure glaucoma (CPACG) treated with primary trabeculectomy. METHODS: This study systematically reviewed cases of CPACG treated with primary trabeculectomy. The scleral flaps in all cases were sutured with two stitches in situ and two releasable sutures to ensure watertight under normal IOP conditions during surgery. Mitomycin C was used in all eyes. All patients were followed for 2 years. Digital massage of the bulbus and removal of the releasable suture were performed according to the IOP and shape of the filtering bleb. Demographic data and clinical outcomes were recorded. Factors predicting long-term IOP were identified. RESULTS: A total of 72 patients (88 eyes) with a mean age of 58.51 ± 10.60 years were included in this study. The complete success rate was 89.77% after 2 years. The IOP began to stabilize after 7 days and reached its lowest point at the 1-month follow-up. The preoperative and early postoperative high or low IOP does not affect long-term effects (P > 0.05). There was a positive correlation between postoperative IOP at the 1-month and 2-year follow-ups (r = 0.64, P < 0.001). CONCLUSION: In CPACG patients undergoing primary trabeculectomy, scleral flaps sutured watertightly with two stitches in situ and two releasable sutures under normal IOP conditions can ensure controllable, effective and safe treatment of CPACG. The preoperative and early postoperative high or low IOP does not affect long-term effects. One-month postoperative IOP can be used as a predictor of long-term IOP control.

Discovery of N-(4-(6-Acetamidopyrimidin-4-yloxy)phenyl)-2-(2-(trifluoromethyl)phenyl)acetamide (CHMFL-FLT3-335) as a Potent FMS-like Tyrosine Kinase 3 Internal Tandem Duplication (FLT3-ITD) Mutant Selective Inhibitor for Acute Myeloid Leukemia.

Most of the current FMS-like tyrosine kinase 3 (FLT3) inhibitors lack selectivity between FLT3 kinase and cKIT kinase as well as the FLT3 wt and internal tandem duplication (ITD) mutants. We report a new compound 27, which displays GI50 values of 30-80 nM against different ITD mutants and achieves selectivity over both FLT3 wt (8-fold) and cKIT kinase in the transformed BaF3 cells (>300-fold). 27 potently inhibits the proliferation of the FLT3-ITD-positive acute myeloid leukemia cancer lines through suppression of the phosphorylation of FLT3 kinase and downstream signaling pathways, induction of apoptosis, and arresting the cell cycle into the G0/G1 phase. 27 also displays potent antiproliferative effect against FLT3-ITD-positive patient primary cells, whereas it does not apparently affect FLT3 wt primary cells. In addition, it also exhibits a good therapeutic window to PBMC compared to PKC412. In the in vivo studies, 27 demonstrates favorable PK profiles and suppresses the tumor growth in the MV4-11 cell inoculated mouse xenograft model.

[Chemical constituents from roots of Viburnum setigerum].

This experiment was to study the constituents of the roots of Viburnum setigerum through various column chromatographic techniques. Thirteen compounds were obtained and their structures were identified using chemical and spectroscopic methods as (7αH, 8'αH)-4, 4', 8α-trihydroxy-3, 3', 9-trimethoxy-7, 9'-epoxylignan (1), (7αH, 8'αH)-4, 4', 8α, 9-tetrahydroxy-3, 3'-dimethoxy-7, 9'-epoxylignan (2), alashinol G (3), alashinol F (4), (-)-secoisolariciresinol (5), (7R, 7'R, 8R, 8'S)-3, 3'-dimethoxy-7, 7'-epoxylignane -4, 4', 9, 9'-tetraol (6), (7αH, 8αH, 8'ßH)-4, 4', 7'α, 9-tetrahydroxy-3, 3'-dimethoxy-7, 9'-epoxylignan (7), loganin (8), dihydroquercetin (9), protocatechuic acid (10), 4-hydroxy-3-methoxy-benzoic acid (11), adoxoside (12), and catechin (13). Compound 1 was a new compound. Compounds 3-7 and 11 were reported from the genus Viburnum for the first time. All compounds were separated from this plant for the first time.

Generation of Retinal Organoids with Mature Rods and Cones from Urine-Derived Human Induced Pluripotent Stem Cells.

Urine cells, a body trash, have been successfully reprogrammed into human induced pluripotent stem cells (U-hiPSCs) which hold a huge promise in regenerative medicine. However, it is unknown whether or to what extent U-hiPSCs can generate retinal cells so far. With a modified retinal differentiation protocol without addition of retinoic acid (RA), our study revealed that U-hiPSCs were able to differentiate towards retinal fates and form 3D retinal organoids containing laminated neural retina with all retinal cell types located in proper layer as in vivo. More importantly, U-hiPSCs generated highly mature photoreceptors with all subtypes, even red/green cone-rich photoreceptors. Our data indicated that a supplement of RA to culture medium was not necessary for maturation and specification of U-hiPSC-derived photoreceptors at least in the niche of retinal organoids. The success of retinal differentiation with U-hiPSCs provides many opportunities in cell therapy, disease modeling, and drug screening, especially in personalized medicine of retinal diseases since urine cells can be noninvasively collected from patients and their relatives.

Tetramethylpyrazine in a Murine Alkali-Burn Model Blocks NFκB/NRF-1/CXCR4-Signaling-Induced Corneal Neovascularization.

Purpose: Tetramethylpyrazine (TMP) is the active ingredient extracted from the Chinese herb Chuanxiong. The purpose of our study was to identify the mechanism of therapeutic TMP suppression of pathologic chemokine receptor 4 (CXCR4) transcription. Methods: C57BL/6J mice with alkali-burned corneas were treated with either TMP eye drops (1.5 mg/mL) or PBS. Corneal neovascularization (CNV) was measured and a clinical assessment was made by slit lamp microscopy. Expression of CXCR4 and the transcription factors nuclear respiratory factor-1 (NRF-1), nuclear factor kappa B (NFκB), forkhead box C1, and yin yang 1 were tracked by real-time RT-PCR and immunofluorescence staining of murine corneas. Western blot, real-time PCR, and immunofluorescence evaluated expression of related genes in human umbilical vein endothelial cells (HUVECs) after 200-µmol/L TMP treatment. In addition, plasmid transfection and chromatin immunoprecipitation assays elucidated the relationship among NRF-1, NFκB, and CXCR4. Results: Corneas treated with TMP had smaller areas of neovascularization and scored better in clinical assessments. Injured corneas showed significantly elevated expressions of NRF-1, NFκB, and CXCR4 that were normalized in vivo by TMP treatment. Similarly, in HUVECs in vitro, TMP decreased expression of NRF-1, NFκB, and CXCR4. Overexpression of NFκB or NRF-1 raised the expression of CXCR4 in HUVECs, but not synergistically. Chromatin immunoprecipitation assays detected only NRF-1 bound to the CXCR4 promoter region, suggesting NFκB controls CXCR4 expression by upregulating NRF-1. Together, our data suggest TMP downregulates CXCR4 by repressing NRF-1 expression in CNV, likely indirectly by downregulating NFκB. Conclusions: Our results implicate a novel mechanism wherein TMP inhibits neovascularization via an NFκB/NRF-1/CXCR4 circuit.

[Protective effects of LBP on cerebral ischemia reperfusion injury in mice and mechanism of inhibiting NF-κB, TNF-α, IL-6 and IL-1ß].

To observe the protective effects of Lycium barbarum polysaccharides (LBP) on cerebral ischemia reperfusion injury in mice and explore its mechanism. Common carotid artery thread was used to cause middle cerebral artery ischemia, and the thread was taken out after 2 h ischemia to achieve cerebral ischemia reperfusion injury in mice. Therefore, the transient middle cerebral artery occlusion (tMCAO) models were established to observe the effects of LBP (25,50, 100 mg•kg⁻¹) on neurological outcome, infarct size and water contents. HE staining was used to observe its effects on neurocytes of cerebral tissues in mice. Western blotting was used to evaluate the protein expression levels of NF-κB p65. ELISA was used to evaluate the levels of TNF-α, IL-6 and IL-1ß in the serum. According to the results, LBP markedly improved neurologic deficits, and decreased infarct size and water contents at 24 h after reperfusion in mice. Pathological section of brain tissues also proved its protective effects on neurocytes. Western blot analysis indicated that LBP markedly down-regulated the protein level of NF-κB p65. ELISA indicated that LBP decreased the levels of TNF-α, IL-6 and IL-1ß in the serum 24 h after reperfusion.In conclusion, LBP has protective effects on cerebral ischemia reperfusion injury in mice, and this effect may be associated with inhibiting NF-κB and inflammatory reactions.

Tetramethylpyrazine-mediated regulation of CXCR4 in retinoblastoma is sensitive to cell density.

Retinoblastoma is the most common ocular tumor in children, and it causes extensive damage. Current treatment options for retinoblastoma include surgery, chemotherapy, radiotherapy and cryotherapy. However, the majority of chemotherapy medicines cause complications and side effects that lead to severe impairment of patient health. Previous studies have reported that tetramethylpyrazine (TMP), which is an extract of the Chinese herbal medicine Chuanxiong, reduces the risk of multidrug resistance in chemotherapy and inhibits the proliferation and metastasis of various types of cancer cells. However, the underlying molecular mechanism of TMP in retinoblastoma remains unclear. The current study demonstrated that C-X-C chemokine receptor type 4 (CXCR4) was expressed in WERI­Rb1 cells and in retinoblastoma. Using reverse transcription­quantitative polymerase chain reaction and western blotting techniques, the current study demonstrated that TMP significantly downregulated the expression of CXCR4 in WERI­Rb1 cells cultured at high density, whereas it had a minor effect in low­density WERI­Rb1 cells; additionally, this effect occurred in a time­dependent manner. TMP inhibited the proliferation of WERI­Rb1 cells as effectively as a CXCR4 antagonist, AMD3100, consistent with a role of CXCR4 in cancer development. Notably, TMP did not affect the cell cycle of cells cultured at low density (1x105 cells/ml), whereas it induced G1­phase arrest in high­density cells (7.5x105 cells/ml; P<0.05). In addition, the expression of CXCR4 in primary rat retinal neurocytes was significantly downregulated by TMP treatment, and this treatment protected primary rat retinal neurocytes from H2O2­induced damage. Thus, the results of this study indicate that TMP is a potential candidate for use in treatment of retinoblastoma, and also provides novel insights into the mechanisms of the anti­cancer and neuroprotective effects of this extract.

4-O-Sulfation in sea cucumber fucodians contribute to reversing dyslipidiaemia caused by HFD.

Fucodians extracted from sea cucumbers (SC-FUCs) possess linear chains with uniform repeating units. Sulfation patterns endow SC-FUCs unique structures and bioactivity. The present study investigated the anti-hyperlipidemic activity of two fucodians isolated from sea cucumbers Pearsonothuria graeffei (fuc-Pg) and Isostichopus badionotus (fuc-Ib). The results indicate fuc-Pg dominated with a 4-O-sulfation pattern shows strong activity in reducing body weight, regulating lipid disorder (TC, TG, HLD-C, and LDL-C level), improving liver function (liver weight, GOP, GPT, and TBA concentrations) and increasing adiponectin level (adiponectin concentration) caused by HFD. However, fuc-Ib dominated with a 2-O-sulfation pattern has only moderate effects. These results suggest that different sulfation pattern may contribute to the differences in the hyperlipidemic activities. Further analysis by quantitative reverse transcription-polymerase chain reaction and Western blot analysis indicate that Fuc-Pg can suppress the expression of CD 36, increase the level of PPARα and decrease the level of CYP7A1, thus, the transportation of fatty acids into liver tissue and lipid metabolism, while fuc-Ib had only limited effects. Our results indicated the sulfation pattern may contribute to anti-hyperlipidemic activity of fucodian, and fuc-Pg dominated with 4-O-sulfation shows better effect and could be further developed as a potential anti-hyperlipidemic food supplement.