Indian food habit & food ingredients may have a role in lowering the severity & high death rate from COVID-19 in Indians: findings from the first nutrigenomic analysis

Background & objectives: During the COVID-19 pandemic, the death rate was reportedly 5-8 fold lower in India which is densely populated as compared to less populated western countries. The aim of this study was to investigate whether dietary habits were associated with the variations in COVID-19 severity and deaths between western and Indian population at the nutrigenomics level. Methods: In this study nutrigenomics approach was applied. Blood transcriptome of severe COVID-19 patients from three western countries (showing high fatality) and two datasets from Indian patients were used. Gene set enrichment analyses were performed for pathways, metabolites, nutrients, etc., and compared for western and Indian samples to identify the food- and nutrient-related factors, which may be associated with COVID-19 severity. Data on the daily consumption of twelve key food componentsacross four countries were collected and a correlation between nutrigenomics analyses and per capita daily dietary intake was investigated. Results: Distinct dietary habits of Indians were observed, which may be associated with low death rate from COVID-19. Increased consumption of red meat, dairy products and processed foods by western populations may increase the severity and death rate by activating cytokine storm-related pathways, intussusceptive angiogenesis, hypercapnia and enhancing blood glucose levels due to high contents of sphingolipids, palmitic acid and byproducts such as CO2 and lipopolysaccharide (LPS). Palmitic acid also induces ACE2 expression and increases the infection rate. Coffee and alcohol that are highly consumed in western countries may increase the severity and death rates from COVID-19 by deregulating blood iron, zinc and triglyceride levels. The components of Indian diets maintain high iron and zinc concentrations in blood and rich fibre in their foods may prevent CO2 and LPS-mediated COVID-19 severity. Regular consumption of tea by Indians maintains high high-density lipoprotein (HDL) and low triglyceride in blood as catechins in tea act as natural atorvastatin. Importantly, regular consumption of turmeric in daily food by Indians maintains strong immunity and curcumin in turmeric may prevent pathways and mechanisms associated with SARS-CoV-2 infection and COVID-19 severity and lowered the death rate. Interpretation & conclusions: Our results suggest that Indian food components suppress cytokine storm and various other severity related pathways of COVID-19 and may have a role in lowering severity and death rates from COVID-19 in India as compared to western populations. However, large multi-centered case?control studies are required to support our current findings.

Effects of Chemically Interesterified Palm Olein on Lipid Profiles in Hamsters

@#Palm olein (POo) has been perceived as atherogenic due to its high proportion of palmitic acid (41.2%) content. It is interesting that most of the palmitic acid of POo is located at stereospecific numbering sn-1 and sn-3 positions of the triacylglycerol (TAG) backbone. The present study aims to investigate the effects of positional distribution of fatty acids on the lipid profiles of POo or chemically interesterified palm olein (CIE POo) fed hamsters in comparison to high oleic sunflower oil (HOSO) fed hamsters. Male weanling Syrian golden hamsters (n=10 for each group), were fed diets formulated with the above oils for 12 weeks. There was no significant difference between CIE POo and HOSO groups for total cholesterol (TC). CIE POo with increased amount of palmitic acid (43.2%) at sn-2 position did not cause significant increases in TC levels compared to the HOSO group. In addition, the POo group has significantly higher high-density lipoprotein cholesterol (HDL-C) than that of the HOSO group, P = 0.011 (< 0.05) while the HOSO group has significantly lower total cholesterol (TC) levels than that of the POo group, P = 0.012 (< 0.05).

Regulation of palmitic acid and lipopolysaccharide-induced macrophage polarization by adipokine WNT1-inducible signaling pathway protein 2 / 中华老年医学杂志

Objective:To investigate the regulatory effect of WNT1-inducible signaling pathway protein 2(WISP2)on macrophage polarization in palmitic acid(PA)and lipopolysaccharide(LPS)-induced inflammation.Methods:The macrophage cell line RAW264.7 was treated with different concentrations of WISP2 protein, and cell viability was determined by means of luminescence assay using Cell-Titer Glo to determine the concentration of WISP2.The cells were divided into control group, palmitic acid group, palmitic acid combined with different concentrations of WISP2 group(10 μg/L and 100 μg/L)and lipopolysaccharide group, lipopolysaccharide combined with different concentrations of WISP2 group(10 μg/L and 100 μg/L). mRNA expression of M1 and M2 macrophages phenotype of each group were detected by real-time quantitative polymerase chain reaction.The protein expression of important inflammatory factors, TNF-α and IL-6, were evaluated by ELISA.Results:Compared with the control group, both 10 μg/L and 100 μg/L WISP2 groups had no effect on the activity of RAW264.7 cells, but significantly up-regulated the expression of various inflammatory factors, including Tnfα(1.877±0.039, 2.202±0.034, F=309.7, P<0.001), Il6(1.418±0.056, 1.506±0.059, F=81.39, P<0.001), Mcp1(1.620±0.014, 1.982±0.125, F=71.45, P<0.001), Ccl3(1.892±0.118, 1.942±0.132, F=32.93, P<0.001), and iNos(1.691±0.201, 1.548±0.090, F=13.60, P<0.05). mRNA in macrophages, and significantly down-regulated the expression of anti-inflammatory factors, including Tgfβ(1.376±0.025, 2.152±0.107, F=1.846, P<0.05), CD206(2.123±0.031, 3.139±1.663, F=8.037, P<0.05), Il4(2.098±0.464, 2.494±0.141, F=48.68, P<0.01), and Il10(1.303±0.216, 1.574±0.274, F=5.774, P<0.05)mRNA, causing M1 type macrophage polarization.Compared with the control group, 100 μmol/L palmitic acid could mildly but significantly increase the expression of inflammatory factors such as TNF-α and IL-6 at the transcriptional and protein levels.Compared with palmitic acid stimulation alone, the combination of palmitic acid and WISP2 further promoted the protein expression of macrophage inflammatory factors TNF-α[(589.4±17.0)ng/L, (692.6±83.4)ng/L, F=56.38, P<0.05], IL-6[(15.13±1.14)ng/L, (13.33±1.22)ng/L, F=23.32, P<0.001]and the mRNA expression of chemokines Mcp1(160±9.796, 140±18.91, F=141.1, P<0.0001)and C cl3(17.76±1.92, 14.41±1.27, F=125.2, P<0.0001). Compared with the control group, 100 μg/L lipopolysaccharide strongly stimulated the expression of inflammatory factors such as TNF-α[(3444±423)ng/L, F=71.20, P<0.0001]and IL-6[(497.0±41.2)ng/L, F=63.50, P<0.0001]in macrophages at the protein level.Compared with lipopolysaccharide stimulation alone, the combination of lipopolysaccharide and WISP2 further significantly up-regulated the mRNA expression of chemokines Mcp1(106.8±8.7, 118.7±4.6, F=251.5, P<0.0001)and Ccl3(35.3±12.5, 116.4±4.5, F=160.1, P<0.0001). Conclusions:The adipokine WISP2 can promote M1 macrophage polarization in palmitic acid and lipopolysaccharide-induced inflammation, and it had distinct regulation in macrophage polarization under different inflammatory response conditions.

Palmitic acid suppresses autophagy in neonatal rat cardiomyocytes via the cGAS-STING-IRF3 pathway / 南方医科大学学报

OBJECTIVE@#To investigate the effect of palmitic acid (PA) on autophagy in neonatal rat cardiomyocytes (NRCMs) and explore the underlying mechanism.@*METHODS@#NRCMs were isolated and cultured for 24 h before exposure to 10% BSA and 0.1, 0.3, 0.5, or 0.7 mmol/L PA for 24 h. After the treatments, the expressions of Parkin, PINK1, p62, LC3Ⅱ/ LC3Ⅰ, cGAS, STING and p-IRF3/IRF3 were detected using Western blotting and the cell viability was assessed with CCK8 assay, based on which 0.7 mmol/L was selected as the optimal concentration in subsequent experiments. The effects of cGAS knockdown mediated by cGAS siRNA in the presence of PA on autophagy-related proteins in the NRCMs were determined using Western blotting, and the expressions of P62 and LC3 in the treated cells were examined using immunofluorescence assay.@*RESULTS@#PA at different concentrations significantly lowered the expressions of Parkin, PINK1, LC3 Ⅱ/LC3 Ⅰ and LC3 Ⅱ/LC3 Ⅰ+Ⅱ (P < 0.05), increased the expression of p62 (P < 0.05), and inhibited the viability of NRCMs (P < 0.05). Knockdown of cGAS obviously blocked the autophagy-suppressing effect of PA and improved the viability of NRCMs (P < 0.05).@*CONCLUSION@#PA inhibits autophagy by activating the cGAS-STING-IRF3 pathway to reduce the viability of NRCMs.

Effects of oleic (18: 1n-9) and palmitic (16: 0) fatty acids on the metabolic state of adipocytes / Efectos de los ácidos grasos oleico (18: 1n-9) y palmítico (16: 0) en el estado metabólico del adipocito

Abstract Background: Elevated serum-free fatty acid (FFA) levels induce insulin resistance (IR) or a protective mechanism to IR development in humans; it depends on FFA type. Objetive: This study explores the effects of oleic (OLA - unsatured) and palmitic (PAM - saturated) fatty acids on insulin action in mature adipocytes effect. Methods: Cells were incubated 18 h with or without OLA and PAM at 250 μM, and 500 μM. After the culture period, were measured: adipocyte viability, size, fatty acids mobilisation, insulin signalling proteins, and glucose uptake. Results: Adipocytes exhibited optimal viability tolerances regardless of the kinds of fatty acids used for treatment. However, adipocytes were hypertrophic after OLA and PAM stimuli. Additionally, lipogenesis (lipid synthesis), and lipolysis (lipid breakdown) were significantly increased by treatment with OLA, or PAM (500 μM) compared to control. Moreover, OLA results showed that there was no significant reduction in signalling cascades, except for a downstream proinflammatory response. Instead, PAM hypertrophic adipocytes were insulin resistant with alteration of proinflammatory and stress markers. Conclusions: Current findings suggest that PAM induces insulin resistance, mitochondrial and reticulum stress on fat cells compared to those treated with OLA that, protects adipocytes to all those alterations.

Resumen Introducción: Los niveles elevados de ácidos grasos libres (AGL) en suero inducen resistencia a insulina (RI) o un mecanismo de protección del desarrollo de RI en humanos, esto depende del tipo de AGL. Objetivo: Este estudio explora los efectos de los ácidos grasos oleico (insaturados - OLA) y palmítico (saturados - PAM) sobre la insulina en adipocitos maduros. Métodos: Las células se incubaron 18 h con o sin OLA y PAM a 250 μM y 500 μM. Después del período de cultivo, se evaluó en adipocitos: viabilidad, tamaño, movilización de ácidos grasos, proteínas de señalización de insulina y absorción de glucosa. Resultados: Los adipocitos mostraron viabilidad óptima independientemente de los tipos de ácidos grasos utilizados en el tratamiento. Los adipocitos eran hipertróficos tras estimulo con OLA y PAM. La lipogénesis (síntesis de lípidos) y la lipólisis (degradación de lípidos) aumentaron significativamente con el tratamiento con OLA o PAM (500 μM) en comparación con el control. En los resultados de OLA no se evidenció una reducción significativa en las cascadas de señalización de insulina, a excepción de una respuesta proinflamatoria posterior. En cambio, los adipocitos hipertróficos tratados con PAM presentaron resistencia a la insulina y alteración de los marcadores proinflamatorios y de estrés. Conclusiones: Nuestros hallazgos sugieren que PAM induce resistencia a la insulina, estrés mitocondrial y del retículo en las células grasas en comparación con aquellos tratados con OLA, AGL que, en cambio, protegen a los adipocitos de todas esas alteraciones.

Low-glucose combined with palmitic acid enhances the radiosensitivity of colorectal cancer through inducing reactive oxygen species generation and DNA damage / 中华放射肿瘤学杂志

Objective:To investigate the mechanism underlying the inhibiting effect of low-glucose combined with palmitic acid on human colon cancer cells and its influence on the radiosensitivity.Methods:Under the treatment of low-glucose, palmitic acid and low-glucose combined with palmitic acid, the treatment condition that significantly inhibited the proliferation of SW480 was screened by CCK-8 assay. The reactive oxygen species (ROS) level, mitochondrial membrane potential and apoptosis rate were detected by flow cytometry. The changes in the radiosensitivity were detected by immunofluorescence-based γ-H 2AX quantification and colony formation assay. The protein expression level was detected by Western blot. Results:Compared with the control group, the condition of low-glucose combined with 120μmol/L palmitic acid significantly inhibited the proliferation of SW480 cells ( P<0.01). The expression levels of CPT1a, PFKFB3 and PKM were significantly up-regulated, the expression levels of NDUFV1, NDUFV2 and NDUFS1 were remarkably down-regulated, the ROS level was significantly increased and the ATP level was considerably reduced in the cells under metabolic stress (all P<0.01). After irradiation, the number of γ-H 2AX foci was significantly increased ( P<0.05), and the D 0 value was significantly reduced ( P<0.01), the ROS level was considerably increased ( P<0.001), the apoptosis rate was significantly increased ( P<0.001) and the expression level of γ-H 2AX protein was remarkably up-regulated ( P<0.01) in the low-glucose combined with 120μmol/L palmitic acid group. Pretreatment with NAC could reverse the changes of ROS, apoptosis and γ-H 2AX protein expression. Conclusions:The combination of low-glucose and palmitic acid can induce metabolic stress in SW480 cells, inhibit tumor proliferation and increase the radiosensitization when combined with radiotherapy by inducing the generation of ROS and DNA damage.

Period circadian clock 3 inhibits palmitic acid-induced oxidative stress and inflammatory factor secretion in podocytes / 中南大学学报(医学版)

OBJECTIVES@#High fat-induced podocyte injury is one of the important factors leading to obesity related nephropathy (ORG), but the mechanism is not clear. This study aims to explore the mechanism of period circadian clock 3 (PER3) in the oxidative stress and inflammation induced by palmitic acid (PA) in podocytes.@*METHODS@#The C57BL/6J mice were fed with chow and high-fat diet for 16 weeks. The PER3 expression in kidney tissues were detected in the normal body weight group and the obesity group. The PER3 mRNA and protein expression were detected after the podocytes were induced with different concentrations (0, 50, 150 and 300 μmol/L) of PA for 48 h. The PER3 mRNA and protein expression were detected after the podocytes were induced with 150 μmol/L PA for 0, 24, 36, and 48 h. Triglyceride (TG) levels were examined in the PA group, the adenovirus (ad)-PER3+PA group, and the siRNA-PER+PA group after the podocytes were transfected by Ad-PER3 or small interfering RNA (siRNA)-PER3 for 48 h and subsequently were induced with 150 μmol/L PA for 48 h. The differential gene expression was detected using RNA sequencing (RNA-seq) after podocytes were transfected by siRNA-PER3 (siRNA-PER3 group) and siRNA-control (siRNA-control group), respectively. The mRNA levels of nephrin, podocin, podocalyxin, podoplanin, superoxide dismutase 1 (SOD1), glutathione peroxidase 1 (GPX1), catalase (CAT), and the levels of malondialdehyde (MDA), glutathione (GSH), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β) and interleukin-2 (IL-2) were detected after podocytes were transfected with Ad-PER3 or Ad-control for 48 h and then they were induced by 150 μmol/L PA for 48 h.@*RESULTS@#The PER3 was down-regulated in the obesity group compared with the normal body weight group (@*CONCLUSIONS@#PER3 can decrease the PA-induced oxidative stress and inflammatory factor secretion via inhibiting the lipogenesis in podocytes.

Guanxin Zhitong Capsules attenuate human endothelial cell damage induced by palmitic acid via MAPK signaling pathway / 中国中药杂志

The present study observed the effect of Guanxin Zhitong Capsules(GXZT) on the lipotoxicity of vascular endothelial cells and investigated the mechanism of GXZT in atherosclerosis treatment. The lipotoxicity model in human umbilical vein endothelial cells(HUVECs) was induced by palmitic acid(PA) stimulation. These cells were divided into a normal control group(NC, 15% normal serum), a model group(PA, 0.6 mmol·L~(-1) PA+15% normal serum), a high-dose GXZT group(GXZT-H, 0.6 mmol·L~(-1) PA+15% GXZT-medicated serum), a medium-dose GXZT group(GXZT-M, 0.6 mmol·L~(-1) PA+10% GXZT-medicated serum+5% normal serum) and a low-dose GXZT group(GXZT-L, 0.6 mmol·L~(-1) PA+5% GXZT-medicated serum+10% normal serum). HUVECs were detected for cell viability by cell counting kit-8(CCK-8) assay, apoptosis by flow cytometry, mitochondrial membrane potential(MMP) by JC-1 labeled laser scanning confocal microscopy, and total and phosphorylated proteins of p38, ERK1/2, and JNK1/2 in the mitogen-activated protein kinases(MAPK) signaling pathway by Western blot. The phosphorylated level was calcula-ted. Compared with the NC group, the PA group showed decreased cell viability and MMP(P<0.01, P<0.01), elevated apoptosis(P<0.01), and up-regulated phosphorylated levels of p38, ERK1/2, and JNK1/2(P<0.01, P<0.01, P<0.01). Compared with the PA group, the GXZT-H, GXZT-M, and GXZT-L groups showed increased cell viability and MMP(P<0.01, P<0.01, P<0.01), reduced apoptosis(P<0.01), and down-regulated protein expression and phosphorylated levels of p38, ERK1/2 and JNK1/2 in the MAPK signaling pathway(P<0.01, P<0.01, P<0.01). In conclusion, the results suggest that GXZT functions via blocking MAPK signaling pathway to relieve the damage of HUVECs induced by PA.

Effect and mechanism of plasminogen activator inhibitor-1 on cardiomyocytes apoptosis stimulated by palmitate / 解放军医学杂志

Objective To investigate the effect and mechanism of plasminogen activator inhibitor-1(PAI-1) on cardiomyocytes apoptosis stimulated by palmitate. Methods Mouse primary cardiomyocytes were isolated and cultured, and then divided into control group and palmitic acid (PA) group, the expressions of PAI-1, apoptotic protein cleaved-caspase 3 and BCL2-associated X protein (Bax) were examined using Western blotting. PAI-1 low expression cells and over expression cells were established, and the expressions of PAI-1, cleaved-caspase 3 and Bax were examined by real-time PCR and Western blotting. After PAI-1 siRNA conditions were determined, cells were stimulated with palmitate, and then divided into four groups including control, PAI-1 low expression group, palmitate group and PAI-1 low expression+palmitate group. Western blotting was performed to examine the expressions of cleaved-caspase 3, Bax and phosphorylated-nuclear factor kappa-B (p-NF-κB). At last, the cells in palmitate group were given with NF-κB inhibitor Bay11-7082, and then divided into palmitate group, PAI-1 low expression + palmitate group, Bay11- 7082 + palmitate group, and PAI-1 low expression + Bay11-7082 + palmitate group, then the expressions of cleaved-caspase 3 and Bax were detected by Western blotting. Results The expressive levels of cleaved-caspase 3, Bax and PAI-1 were significantly higher in palmitate group than those in control group (P<0.01). The expressions of cleaved-caspase 3, Bax and p-NF-κB were obviously lower in PAI-1 low expression+palmitate group than those in palmitate group (P<0.01). Besides, the expressions of cleaved-caspase 3 and Bax were obviously lower in Bay11-7082+palmitate group than those in palmitate group (P<0.01), and the expressions of cleaved-caspase 3 and Bax were markedly lower in PAI-1 low expression+Bay11-7082+palmitate group than those in Bay11- 7082+palmitate group (P<0.01). Conclusion By affecting the NF-?B pathway, PAI-1 could involve in the apoptosis regulation of palmitate stimulated cardiomyocytes.

Effect of CHI3L1 on the oxidative stress of human umbilical vein endothelial cells treated with palmitic acid / 解放军医学杂志

Objective To observe the effect of chitinase-3-like protein 1 (CHI3L1) on the oxidative stress of human umbilical vein endothelial cells (HUVECs) after treatment with palmitic acid, and preliminarily explore the molecular mechanism. Methods HUVECs were cultured in vitro, treated with different concentrations of CHI3L1 for 24 hours, and the MTT assay was used to determine the proliferation of HUVECs. The cells were divided into the control group (untreated group), palmitic acid group and CHI3L1+palmitic acid group. Cells in palmitic acid group were treated with 100 μmol/L palmitic acid for 24 hours, and in CHI3L1+palmitic acid group were pretreated with 100 ng/ml CHI3L1 for 2 hours, then 100 μmol/L palmitic acid were added and co-treating for 24 hours. Western blotting was used to detect the contents of nuclear and cytoplasmic p65, nuclear Lamin B, Actin, heme oxygenase (HMOX), reduced coenzyme /Ⅱ dependent quinone oxidoreductase (NQO1), phosphorylated protein kinase B (p-Akt), endoplasmic reticulum redox 1-like protein (Ero1-Lα), Calnexin, protein disulfide isomerase (PDI), chaperonin/ glucose regulatory protein (Bip1/Grp) 78, endoplasmic reticulum nuclear signal transduction protein (IRE)-1α and phosphorylated eukaryotic translation initiation factor (p-eIF) 2α. The contents of IL-6 and TNF-α were measured by ELISA, the activity of NF-κB was measured by DNA binding assay, and the nuclear translocation of Nrf2 was detected by immunofluorescence. Results MTT results showed that compared with the control, treatment with 100 ng/ml CHI3L1 for 24 hours did not decrease the activity of HUVECs (94.17±6.13 vs. 100.00±0.00). Western blotting results showed that the p65 level in nucleus of CHI3L1+palmitic acid group was lower than that in palmitic acid group (0.77±0.04 vs. 0.92±0.09, P<0.05), but in cytoplasm was higher than that in palmitic acid group (0.45±0.04 vs. 0.27±0.05, P<0.05). The DNA binding activity of NF-κB in CHI3L1+palmitic acid group was lower than that in palmitic acid group (0.26±0.04 vs. 0.43±0.07, P<0.05). ELISA results showed that the contents of TNF-α and IL-6 were lower in CHI3L1+palmitic acid group than those in palmitic acid group [(85.91±21.16) pg/ml vs. (221.12±18.71) pg/ml; (71.43±10.56) pg/ml vs. (95.03±11.2) pg/ml, P<0.05]. Western blotting results showed that the levels of HMOX and NQO1 were significantly higher in CHI3L1+palmitic acid group than those in palmitic acid group (0.58±0.07 vs. 0.32±0.09; 1.08±0.04 vs. 0.62±0.09, P<0.05). Immunofluorescence results showed that compared with palmitic acid group, the content of Nrf2 increased significantly in CHI3L1+palmitic acid group (10.26%±4.53% vs. 78.64%±3.16%, P<0.05). Western blotting results showed that the phosphorylation level of PI3K/Akt was significantly higher in CHI3L1+palmitic acid group than those in palmitic acid group (0.51±0.04 vs. 0.15±0.09, P<0.05), and the expression levels of Ero1-Lα, Calnexin, PDI, BIP1/GRP78, IRE-1α and p-eIF2α in endoplasmic reticulum were significantly reduced in CHI3L1+palmitic acid group than those in palmitic acid group (0.32±0.02 vs. 0.39±0.09; 0.42±0.04 vs. 0.54±0.07; 0.19±0.02 vs. 0.24±0.05; 0.11±0.01 vs. 0.17±0.03; 0.19±0.03 vs. 0.33±0.04; and 0.22±0.07 vs. 0.67±0.09. P<0.05). Conclusion CHI3L1 can inhibit the cytokine secretion of HUVECs treated with palmitic acid, and reduce the stress level of endoplasmic reticulum.

Analysis of chemical constituents of fatty acids and their esters of Coicis Semen by UPLC-Triple-TOF-MS / 中草药

Objective: To establish a rapid qualitative analysis method for fatty acids and esters in Coicis Semen by ultra-performance liquid chromatography triple quadrupole time-of-flight mass spectrometry (UPLC-Triple-TOF-MS). Methods: Agilent ZORBAX-SB C18 (250 mm × 4.6 mm, 5.0 μm) column was used. The mobile phase was acetonitrile-isopropanol (1:1) elution, the flow rate was 1 mL/min, the detection wavelength was 210 nm, the column temperature was 30 ℃, and the injection quantity was 10 μL. Electrospray ion source positive ion mode was adopted, and the scanning range was m/z 100-1 500. The sample data were collected by full scanning mode, and the fatty acid chemical composition of Coicis Semen was quickly identified according to the information obtained by high-resolution mass spectrometry combined with secondary mass spectrometry. Results: A total of 29 kinds of fatty acids and esters in Coicis Semen were detected, and the cracking rules of the compounds were analyzed. Through the mass-to-charge ratio of molecular ion peaks and fragment ions, the Scifinder and Reaxy network databases, and the literature, these compounds were different under the action of ion source by losing the structure of oleic acid, linoleic acid, palmitic acid, oxidizing oleic acid and the like. The mass-to-charge ratio of the fragment ions, and the name and structural formula of the 29 fatty acids and their ester compounds were inferred. Conclusion: The method of qualitative analysis of the fatty acids and esters of Coicis Semen established in this study is accurate, rapid and sensitive, which provides experimental basis for improving the quality control level of Coicis Semen and further elucidating the pharmacodynamic substance basis of Coicis Semen.

Mechanism of Shaoyao Decoction in treatment of ulcerative colitis based on network pharmacology and molecular docking technology / 中草药

Objective: Network pharmacology and molecular docking technology were used to study the material basis and possible mechanism of Shaoyao Decoction in treatment of ulcerative colitis (UC). Methods: TCMSP and TCMID were used to obtain the potential active components and drug targets of Shaoyao Decoction. GeneCard and OMIM were used to search disease targets. The common targets obtained by matching drug targets and disease targets were imported into String to construct a PPI network, and Cyto NCA plug-in was used to screen key targets. The network diagram was drawn by connecting the key targets with the corresponding components, so as to screen the key components. GO and KEGG enrichment analyses were performed on key targets. SYBYL-X2.0 software was used to dock the molecules of the key components with the key targets. The rat UC model was replicated in vivo. After the intervention of Shaoyao Decoction, the disease activity index was observed, the colonic pathological damage was evaluated, and the levels of TNF-α, IL-4 and CXCR4 were detected by ELISA. Results: A total of 424 potential active components were found in Shaoyao Decoction. The key components included quercetin, palmitic acid, catechin, and procyanidins, etc. Its 41 key targets for UC were mainly related to the positive regulation of transcription, the negative regulation of apoptosis process, the signal transduction, and other biological processes. The key targets played a role in treating UC through signaling pathways such as TNF, HIF-1, cancer pathway, TLR, PI3K-Akt, et al. Molecular docking results showed that key components had good binding activity with corresponding targets. Shaoyao Decoction improved colon pathological damage, down-regulated the level of TNF-α and CXCR4, and up-regulated the level of IL-4 in vivo. Conclusion: Shaoyao Decoction has the characteristics of "multi-components, multi-targets, multi-pathways" in treatment of UC, which lays a foundation for further study of its mechanism.

GC-MS analysis of volatile oil from traditional She medicine Clematis Florida var. plena flower / 中草药

Objective: Clematis florida var. plena is a traditional She medicine. In this paper, we aim to study the main chemical components of essential oil of C. florida var. plena flower. The study will provide reference for large-scale reasonable cultivation and quality standard establishment of C. florida var. plena. Methods: C. florida var. plena flower's essential oil was extracted with steam distillation and it was identified by GC-MS. Area normalization method was used to measure the percentage of each components. Results: In the experiment, 32 element were detected in essential oil of C. florida var. plena flower, among which 20 chemical components were identified. There are 13 chemical components over 1%, and the top five compounds are palmitic acid (26.94%), phytol (10.58%), linoleic acid (6.13%), pentadecane (4.54%) and n-tricosane (3.84%). Conclusion: GC-MS was first used to analyze the main chemical components of C. florida var. plena's flower essential oil. The study provides scientific basis for cultivation, production, development, utilization of C. florida var. plena.

Associação entre Staphylococcus epidermidis e o nível de ácido palmítico em pacientes com acne vulgar / The association between Staphylococcus epidermidis and palmitic acid level in patients with acne vulgaris

INTRODUÇÃO: A acne vulgar é um distúrbio inflamatório da glândula pilossebácea. O ácido palmítico é um dos principais tipos de ácidos graxos livres e pode desempenhar um papel na patogênese da acne. Além disso, estudos recentes sugeriram que o Staphylococcus epidermidis pode estar envolvido na acne. OBJETIVO: Explorar a associação entre a Staphylococcus epidermidis e o ácido palmítico na acne vulgar. MÉTODOS: 43 estudantes do ensino médio de uma área urbana do sul de Sulawesi, na Indonésia, foram incluídos. O nível de ácido palmítico foi medido utilizando cromatografia gasosa e os comedões foram cultivados para detectar o perfil do microbioma. O teste de Mann-Whitney foi utilizado para analisar a diferença do nível palmítico médio entre os grupos com diferentes graus de gravidade da acne vulgar. RESULTADOS: 14 pacientes (32,6%) apresentavam acne vulgar leve, enquanto 14 e 15 pacientes apresentavam acne vulgar moderada e grave, respectivamente. O grupo grave e moderado apresentou nível de ácido palmítico significativamente maior em comparação ao grupo leve. A análise de subgrupo de pacientes com acne vulgar moderada e grave, positiva para S. epidermidis, mostrou um nível significativamente maior de ácido palmítico comparado ao grupo leve. CONCLUSÕES: Esses resultados sugerem que S. epidermidis pode estar associado ao nível de ácido palmítico e pode contribuir na patogênese da acne.

INTRODUCTION: Acne vulgaris is an inflammatory disorder of the pilosebaceous gland. Palmitic acid is one of the major types of free fatty acid and may play a role in acne pathogenesis. In addition, recent studies suggested that Staphylococcus epidermidis might be involved in acne. OBJECTIVE: To explore the association between Staphylococcus epidermidis and palmitic acid in acne vulgaris. METHODS: Forty-three high school students at an urban area in South Sulawesi, Indonesia, were included. The palmitic acid level was measured using gas chromatography and comedone was cultured to detect the microbiota profile. Mann-Whitney test was used to analyze the median palmitic level diference between groups with different acne vulgaris severity. RESULTS: Fourteen patients (32.6%) had mild acne vulgaris, while 14 and 15 patients had moderate and severe acne vulgaris, respectively The severe and moderate group showed significantly higher palmitic acid level compared with the mild group. Subgroup analysis of patients with moderate and severe acne vulgaris positive for S. epidermidis showed a significantly higher palmitic acid level compared with the mild group. CONCLUSIONS: This result suggests that S. epidermidis may be associated with the palmitic acid level and may contribute to acne pathogenesis.

Emodin exerts protective effect against palmitic acid-induced endoplasmic reticulum stress in HepG2 cells / 한국영양학회지

PURPOSE: Protein overloading in the endoplasmic reticulum (ER) leads to endoplasmic reticulum stress, which exacerbates various disease conditions. Emodin, an anthraquinone compound, is known to have several health benefits. The effect of emodin against palmitic acid (PA) - induced ER stress in HepG2 cells was investigated. METHODS: HepG2 cells were treated with varying concentrations of palmitic acid to determine the working concentration that induced ER stress. ER stress associated genes such as ATF4, XBP1s, CHOP and GRP78 were checked using RT- PCR. In addition, the expression levels of unfolded protein response (UPR) associated proteins such as IRE1α, eIF2α and CHOP were checked using immunoblotting to confirm the induction of ER stress. The effect of emodin on ER stress was analyzed by treating HepG2 cells with 750 µM palmitic acid and varying concentrations of emodin, then analyzing the expression of UPR associated genes. RESULTS: It was evident from the mRNA and protein expression results that palmitic acid significantly increased the expression of UPR associated genes and thereby induced ER stress. Subsequent treatment with emodin reduced the mRNA expression of ATF4, GRP78, and XBP1s. Furthermore, the protein levels of p-IRE1α, p-elF2α and CHOP were also reduced by the treatment of emodin. Analysis of sirtuin mRNA expression showed that emodin increased the levels of SIRT4 and SIRT7, indicating a possible role in decreasing the expression of UPR-related genes. CONCLUSION: Altogether, the results suggest that emodin could exert a protective effect against fatty acid-induced ER stress and could be an agent for the management of various ER stress related diseases.

Palmitic acid-induced lipotoxicity promotes a novel interplay between Akt-mTOR, IRS-1, and FFAR1 signaling in pancreatic ß-cells

BACKGROUND: Free fatty acid receptor 1 (FFAR1) is G-protein coupled receptor predominantly expressed in pancreatic ß-cells that is activated by a variety of free fatty acids (FFAs). Once activated, it promotes glucose-stimulated insulin secretion (GSIS). However, increased levels of FFAs lead to lipotoxicity, inducing loss of ß-cell function. FFAR1 plays a key role in the development of type 2 diabetes (T2D), and previous studies have indicated the importance of developing anti-diabetic therapies against FFAR1, although its role in the regulation of ß-cell function remains unclear. The present study investigated the role of FFAR1 under lipotoxic conditions using palmitic acid (PA). The rat insulinoma 1 clone 832/13 (INS-1 832/13) cell line was used as a model as it physiologically resembles native pancreatic ß-cells. Key players of the insulin signaling pathway, such as mTOR, Akt, IRS-1, and the insulin receptor (INSR1ß), were selected as candidates to be analyzed under lipotoxic conditions. RESULTS: We revealed that PA-induced lipotoxicity affected GSIS in INS-1 cells and negatively modulated the activity of both IRS-1 and Akt. Reduced phosphorylation of both IRS-1 S636/639 and Akt S473 was observed, in addition to decreased expression of both INSR1ß and FFAR1. Moreover, transient knockdown of FFAR1 led to a reduction in IRS-1 mRNA expression and an increase in INSR1ß; mRNA. Finally, PA affected localization of FFAR1 from the cytoplasm to the perinucleus. CONCLUSIONS: In conclusion, our study suggests a novel regulatory involvement of FFAR1 in crosstalk with mTOR-Akt and IRS-1 signaling in ß-cells under lipotoxic conditions.

High fatty acid induced H9c2 cardiomyocytes injury through mitochondrial pathways / 中国药理学通报

Aim: To explore the role of mitochondrial apoptosis pathway in high fatty acid induced injury in H9c2 cardiomyocytes. Methods: Cardiomyocytes were exposed to different concentrations of palmitic acid (PA) at 0 ∼0. 4 mmol · L-1 for 24 h and different time points (0 ∼48 h) of PA 0.2 mmol · L-1. Cell viability was measured by MTT; the intracellular reactive oxygen species (ROS) was detected by ROS kit; the cells were detected by apoptosis kit; the cell mitochondrial membrane potential changes were detected by mitochondrial membrane potential kit, and the protein expressions of mitochondrial apoptosis pathway such as Cyt-C, Cleaved caspase-3, Bax, and Bcl-2 were detected by Western blot. Results: When the cells were stimulated with PA for 24 h, the cell proliferation rates of 0. 2 and 0. 4 mmol · L-1 PA groups significantly decreased. The level of ROS increased gradually, the cell mitochondrial membrane potential decreased and the cell apoptosis increased. When the cells were stimulated with PA (0. 2 mmol · L-1) for 24 h, 36 h and 48 h, and all of the cell proliferation rates showed significant decline. Cardiomyocytes exposed to PA (0. 4 mmol · L-1) for 24 h showed an increase in the expression of mitochondrial related proteins (Cyt-C, Cleaved caspase-3 and Bax) (P < 0. 05), while Bcl-2 expression was significantly reduced (P < 0. 05). Conclusion: Mitochondrial apoptosis signaling pathway might play an important role in high fatty acid induced H9c2 myocardial injury.

Role of mitochondrial apoptosis pathway in palmitic acid-induced cell apoptosis in vascular endothelial cells / 中国药理学通报

Aim: To explore the role of mitochondrial apoptosis pathway in palmitic acid (PA) -induced cell apoptosis in human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were exposed to different concentrations of PA (0. 1, 0. 2, 0. 4, 0. 8 mmol · L-1) for 24 h and different time points of 0.4 mmol · L-1 PA (0, 12, 24, 48 h). Cell viability was measured by MTT. The expression of reactive oxygen species (ROS) in HUVECs was detected by immunofluorescence. The level of intracellular apoptosis was detected by TUNEL assay. The protein expressions of mitochondrial apoptosis pathway proteins such as AIF, Cyt-C, cleaved caspase-3, Bcl-2, Bax were determined by Western blot. Results: HUVECs exposed to PA at 0. 4 mmol · L-1 for 24 h showed a decrease in their viability and an increase in the level of AIF, Cyt- C, cleaved caspase-3, Bax/Bcl-2 (P < 0. 05). Cell apoptosis level was significantly up-regulated (P < 0. 05). The intracellular apoptosis levels in the vascular endothelial cells treatment with mitochondrial apoptosis pathway inhibitor ciclosporine A (CsA) were significantly lower than those of PA group (P < 0. 05). Conclusion: Activated mitochondrial apoptosis pathway might play an important role in PA-induced cell apoptosis in vascular endothelial cells.

Marein inhibited oxidative stress and inflammation of high glucose induced HBZY-1 cells via AMPK signaling pathway / 中国药理学通报

Aim To study the role of marein mediated AMPK signaling pathway in delaying oxidative stress, inflammation and fibrotic protein expression in diabetic nephropathy ( DN ). Methods In vitro diabetic nephropathy model was established by HG + PA induced rat mesangial cells ( HBZY-1 ) , and the cultured HBZY-1 cells were divided into normal control group (NG), HG + PA( GlucoselOO mmol • L'1 + Palmitic acid 250 [imo\ • L'1, HG +PA) model group, HG + PA + marein with different doses of 25 p,mol • L"1, 50 p,mol • L"1, 100 jimol • L"1, and 200 junol • L"1 groups. MTS was used to detect the effect of marein on HBZY-1 cell proliferation, and the optimal concentration was selected. Western blot was used to test the protein expression of NOX4, TGF-fU, MCP-1, a-SMA, FN, Collagen VI. Adenosine monophosphate activated( AMPK) protein kinase family of AMPK7I, p-AMPK a expression were measured. Results Marein inhibited high glucose palmitate-induced proliferation of HBZY-1 cells, down-regulated NOX4, TGF-(31, MCP-1, cx-SMA, FN and Collagen VI expression in model cells. Meanwhile, marein up-regulated both AMPK 7I and p-AMPKa expression. Conclusions Marein may inhibit the HBZY-1 cell proliferation, oxidative stress, inflammation and fibrosis factors expression in HG + PA induced HBZY-1 cell by activating of both AMPK 7I and AMPK signaling pathway, thus delaying renal injury in diabetic nephropathy.