Maternal supplementation with n-3 fatty acids affects placental lipid metabolism, inflammation, oxidative stress, the endocannabinoid system, and the neonate cytokine concentrations in dairy cows

Background The placenta plays a crucial role in supporting and influencing fetal development.We compared the effects of prepartum supplementation with omega-3(n-3)fatty acid(FA)sources,flaxseed oil(FLX)and fish oil(FO),on the expression of genes and proteins related to lipid metabolism,inflammation,oxidative stress,and the endocannabinoid system(ECS)in the expelled placenta,as well as on FA profile and inflammatory response of neonates.Late-pregnant Holstein dairy cows were supplemented with saturated fat(CTL),FLX,or FO.Placental cotyledons(n=5)were collected immediately after expulsion,and extracted RNA and proteins were analyzed by RTPCR and proteomic analysis.Neonatal blood was assessed for FA composition and concentrations of inflammatory markers.Results FO increased the gene expression of fatty acid binding protein 4(FABP4),interleukin 10(IL-10),catalase(CAT),cannabinoid receptor 1(CNR1),and cannabinoid receptor 2(CNR2)compared with CTL placenta.Gene expression of ECS-enzyme FA-amide hydrolase(FAAH)was lower in FLX and FO than in CTL.Proteomic analysis identified 3,974 proteins;of these,51–59 were differentially abundant between treatments(P≤0.05,|fold change|≥1.5).Top canonical pathways enriched in FLX vs.CTL and in FO vs.CTL were triglyceride metabolism and inflammatory processes.Both n-3 FA increased the placental abundance of FA binding proteins(FABPs)3 and 7.The abundance of CNR1 cannabinoid-receptor-interacting-protein-1(CNRIP1)was reduced in FO vs.FLX.In silico modeling affirmed that bovine FABPs bind to endocannabinoids.The FLX increased the abundance of inflammatory CD44-antigen and secreted-phosphoprotein-1,whereas prostaglandin-endoperoxide synthase 2 was decreased in FO vs.CTL placenta.Maternal FO enriched neonatal plasma with n-3 FAs,and both FLX and FO reduced interleukin-6 concentrations compared with CTL.Conclusion Maternal n-3 FA from FLX and FO differentially affected the bovine placenta;both enhanced lipid metabolism and modulated oxidative stress,however,FO increased some transcriptional ECS components,possibly related to the increased FABPs.Maternal FO induced a unique balance of pro-and anti-inflammatory components in the placenta.Taken together,different sources of n-3 FA during late pregnancy enhanced placental immune and metabolic processes,which may affect the neonatal immune system.

Traditional Chinese medicine Pien-Tze-Huang ameliorates LPS-induced sepsis through bile acid-mediated activation of TGR5-STAT3-A20 signalling

Pien Tze Huang(PZH),a class-1 nationally protected traditional Chinese medicine(TCM),has been used to treat liver diseases such as hepatitis;however,the effect of PZH on the progression of sepsis is unknown.Here,we reported that PZH attenuated lipopolysaccharide(LPS)-induced sepsis in mice and reduced LPS-induced production of proinflammatory cytokines in macrophages by inhibiting the activation of mitogen-activated protein kinase(MAPK)and nuclear factor-kappa B(NF-κB)signalling.Mechanistically,PZH stimulated signal transducer and activator of transcription 3(STAT3)phosphorylation to induce the expression of A20,which could inhibit the activation of NF-κB and MAPK signalling.Knockdown of the bile acid(BA)receptor G protein-coupled bile acid receptor 1(TGR5)in macrophages abolished the effects of PZH on STAT3 phosphorylation and A20 induction,as well as the LPS-induced inflammatory response,suggesting that BAs in PZH may mediate its anti-inflammatory effects by activating TGR5.Consistently,deprivation of BAs in PZH by cholestyramine resin reduced the effects of PZH on the expression of phosphorylated-STAT3 and A20,the activation of NF-κB and MAPK signalling,and the production of proinflammatory cytokines,whereas the addition of BAs to cholestyramine resin-treated PZH partially restored the inhibitory effects on the production of proinflammatory cytokines.Overall,our study identifies BAs as the effective components in PZH that activate TGR5-STAT3-A20 signalling to ameliorate LPS-induced sepsis.

Caffeic acid hinders the proliferation and migration through inhibition of IL-6 mediated JAK-STAT-3 signaling axis in human prostate cancer

Background:Caffeic acid(CA)is considered a promising phytochemical that has inhibited numerous cancer cell proliferation.Therefore,it is gaining increasing attention due to its safe and pharmacological applications.In this study,we investigated the role of CA in inhibiting the Interleukin-6(IL-6)/Janus kinase(JAK)/Signal transducer and activator of transcription-3(STAT-3)mediated suppression of the proliferation signaling in human prostate cancer cells.Materials and Methods:The role of CA in proliferation and colony formation abilities was studied using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide(MTT)assay and colony formation assays.Tumour cell death and cell cycle arrest were identified usingflow cytometry techniques.CA treatment-associated protein expression of mitogen-activated protein kinase(MAPK)families,IL-6/JAK/STAT-3,proliferation,and apoptosis protein expressions in PC-3 and LNCaP cell lines were measured using Western blot investigation.Results:We have obtained that treatment with CA inhibits prostate cancer cells(PC-3 and LNCaP)proliferation and induces reactive oxygen species(ROS),cell cycle arrest,and apoptosis cell death in a concentration-dependent manner.Moreover,CA treatment alleviates the expression phosphorylated form of MAPK families,i.e.,extracellular signal-regulated kinase 1(ERK1),c-Jun N-terminal kinase(JNK),and p38 in PC-3 cells.IL-6 mediated JAK/STAT3 expressions regulate the proliferation and antiapoptosis that leads to prostate cancer metastasis and migration.Therefore,to mitigate the expression of IL-6/JAK/STAT-3 is considered an important target for the treatment of prostate cancer.In this study,we have observed that CA inhibits the expression of IL-6,JAK1,and phosphorylated STAT-3 in both PC-3 and LNCaP cells.Due to the inhibitory effect of IL-6/JAK/STAT-3,it resulted in decreased expression of cyclin-D1,cyclin-D2,and CDK1 in both PC-3 cells.In addition,CA induces apoptosis by enhancing the expression of Bax and caspase-3;and decreased expression of Bcl-2 in prostate cancer cells.Conclusions:Thus,CA might act as a therapeutical application against prostate cancer by targeting the IL-6/JAK/STAT3 signaling axis.

Landscape of Sequence Variations in Homologous Copies of FAD2 and FAD3 in Rapeseed(Brassica napus L.)Germplasm with High/Low Linolenic Acid Trait

Genetic manipulation(either restraint or enhancement)of the biosynthesis pathway ofα-linolenic acid(ALA)in seed oil is an important goal in Brassica napus breeding.B.napus is a tetraploid plant whose genome often har-bors four and six homologous copies,respectively,of the two fatty acid desaturases FAD2 and FAD3,which con-trol the last two steps of ALA biosynthesis during seed oil accumulation.In this study,we compared their promoters,coding sequences,and expression levels in three high-ALA inbred lines 2006L,R8Q10,and YH25005,a low-ALA line A28,a low-ALA/high-oleic-acid accession SW,and the wildtype ZS11.The expression levels of most FAD2 and FAD3 homologs in the three high-ALA accessions were higher than those in ZS11 and much higher than those in A28 and SW.The three high-ALA accessions shared similar sequences with the pro-moters and CDSs of BnFAD3.C4 and BnFAD3.A3.In A28 and SW,substitution of three amino acid residues in BnFAD2.A5 and BnFAD2.C5,an absence of BnFAD2.C1 locus,and a 549 bp long deletion on the BnFAD3.A3 promoter were detected.The profile of BnFAD2 mutation in the two low-ALA accessions A28 and SW is different from that reported in previous studies.The mutations in BnFAD3 in the high-ALA accessions are reported for thefirst time.In identifying the sites of these mutations,we provide detailed information to aid the design of mole-cular markers for accelerated breeding schemes.

4-Hydroxycinnamic acid attenuates neuronal cell death by inducing expression of plasma membrane redox enzymes and improving mitochondrial functions

Many approaches to neurodegenerative diseases that focus on amyloid-βclearance and gene therapy have not been successful.Some therapeutic applications focus on enhancing neuronal cell survival during the pathogenesis of neurodegenerative diseases,including mitochondrial dysfunction.Plasma membrane(PM)redox enzymes are crucial in maintaining cellular physiology and redox homeostasis in response to mitochondrial dysfunction.Neurohormetic phytochemicals are known to induce the expression of detoxifying enzymes under stress conditions.In this study,mechanisms of neuroprotective effects of 4-hydroxycinnamic acid(HCA)were examined by analyzing cell survival,levels of abnormal proteins,and mitochondrial functions in two different neuronal cells.HCA protected two neuronal cells exhibited high expression of PM redox enzymes and the consequent increase in the NAD^(+)/NADH ratio.Cells cultured with HCA showed delayed apoptosis and decreased oxidative/nitrative damage accompanied by decreased ROS production in the mitochondria.HCA increased the mitochondrial complexes I and II activities and ATP production.Also,HCA increased mitochondrial fusion and decreased mitochondrial fission.Overall,HCA maintains redox homeostasis and energy metabolism under oxidative/metabolic stress conditions.These findings suggest that HCA could be a promising therapeutic approach for neurodegenerative diseases.

Ferulic acid reduces inflammatory response induced by radiation through Sirt1-NLRP3 pathway

Objective:A model of inflammatory damage was induced by radiation to investigate whether ferulic acid(FA)can reduce the inflammatory response through the Sirt1-NLRP3 inflammatory pathway.This will help discover radiation-protective drugs and elucidate the molecular mechanisms related to radiation-induced inflammatory damage.Methods:A mouse model of radiation-induced immunoinflammatory injury was established to verify the anti-inflammatory effects of FA in vivo.C57BL/6J mice were randomly divided into six groups,and 5 Gy whole-body irradiation was used for modeling.Mice were administered a gastric solvent,amifostine,or 25,50,or 100 mg/kg FA daily for 12 days,consecutively,before irradiation.The serum of mice was collected 24 hour after irradiation to observe the content of inflammatory factors interleukin(IL)-1β,IL-18,IL-6,and tumor necrosis factor(TNF)-α.The spleen and thymus tissues of mice were weighed and the organ index was calculated for pathological testing and immunofluorescence detection.Results:FA reduced the radiation-induced decrease in the spleen and thymus indices.FA significantly reduced the secretion of inflammatory factors in the serum and reversed the radiation-induced reduction in lymphocytes in the spleen and thymus of mice.FA activated Sirt1 and inhibited the expression of the NLRP3 inflammasome to alleviate the inflammatory response.Conclusions:FA reduced radiation-induced inflammation in animals,possibly by activating Sirt1 and reducing nucleotide oligomerization domain(NOD)-like receptor thermal protein domain associated protein 3(NLRP3)inflammasome expression,thereby reducing the secretion of inflammatory factors.

Acetic acid additive in NaNO_(3)aqueous electrolyte for long-lifespan Mg-air batteries

Mg-air batteries have attracted tremendous attention as a potential next-generation power source for portable electronics and e-transportation due to their remarkable high theoretical volumetric energy density,environmental sustainability,and cost-effectiveness.However,the fast hydrogen evolution reaction(HER)in NaCl-based aqueous electrolytes impairs the performance of Mg-air batteries and leads to poor specific capacity,low energy density,and low utilization.Thus,the conventionally used NaCl solute was proposed to be replaced by NaNO_(3)and acetic acid additive as a corrosion inhibitor,therefore an electrolyte engineering for long-life time Mg-air batteries is reported.The resulting Mg-air batteries based on this optimized electrolyte demonstrate an improved discharge voltage reaching~1.8 V for initial 5 h at a current density of 0.5 mA/cm^(2) and significantly prolonged cells'operational lifetime to over 360 h,in contrast to only~17 h observed in NaCl electrolyte.X-ray photoelectron spectroscopy and time-of-flight secondary ion mass spectrometry were employed to analyse the composition of surface film and scanning electron microscopy combined with transmission electron microscopy to clarify the morphology changes of the surface layer as a function of acetic acid addition.The thorough studies of chemical composition and morphology of corrosion products have allowed us to elucidate the working mechanism of Mg anode in this optimized electrolyte for Mg-air batteries.

Understanding the roles of Brønsted/Lewis acid sites on manganese oxide-zeolite hybrid catalysts for low-temperature NH_(3)-SCR

Although metal oxide-zeolite hybrid materials have long been known to achieve enhanced catalytic activity and selectivity in NO_(x)removal reactions through the inter-particle diffusion of intermediate species,their subsequent reaction mechanism on acid sites is still unclear and requires investigation.In this study,the distribution of Brønsted/Lewis acid sites in the hybrid materials was precisely adjusted by introducing potassium ions,which not only selectively bind to Brønsted acid sites but also potentially affect the formation and diffusion of activated NO species.Systematic in situ diffuse reflectance infrared Fourier transform spectroscopy analyses coupled with selective catalytic reduction of NO_(x)with NH_(3)(NH_(3)-SCR)reaction demonstrate that the Lewis acid sites over MnO_(x)are more active for NO reduction but have lower selectivity to N_(2)than Brønsted acids sites.Brønsted acid sites primarily produce N_(2),whereas Lewis acid sites primarily produce N_(2)O,contributing to unfavorable N_(2)selectivity.The Brønsted acid sites present in Y zeolite,which are stronger than those on MnO_(x),accelerate the NH_(3)-SCR reaction in which the nitrite/nitrate species diffused from the MnO_(x)particles rapidly convert into the N_(2).Therefore,it is important to design the catalyst so that the activated NO species formed in MnO_(x)diffuse to and are selectively decomposed on the Brønsted acid sites of H-Y zeolite rather than that of MnO_(x)particle.For the physically mixed H-MnO_(x)+H-Y sample,the abundant Brønsted/Lewis acid sites in H-MnO_(x)give rise to significant consumption of activated NO species before their inter-particle diffusion,thereby hindering the enhancement of the synergistic effects.Furthermore,we found that the intercalated K+in K-MnO_(x)has an unexpected favorable role in the NO reduction rate,probably owing to faster diffusion of the activated NO species on K-MnO_(x)than H-MnO_(x).This study will help to design promising metal oxide-zeolite hybrid catalysts by identifying the role of the acid sites in two different constituents.

Catalytic Performance of Aquathermolysis and Viscosity Reduction of Heavy Oil over a WO_(3)/ZrO_(2) Solid Acid

Tungstated zirconia(WO_(3)/ZrO_(2))solid acid catalysts with different WO_(3) contents were prepared by a hydrothermal method and then used in the catalytic aquathermolysis of heavy oil from Xinjiang.The WO_(3)/ZrO_(2) solid acid catalyst was characterized by a range of characterization methods,including X-ray diffraction,NH3-temperature programmed desorption,and pyridine infrared spectroscopy.The WO_(3) content of the WO_(3)/ZrO_(2) catalysts had an important impact on the structure and property of the catalysts.When the WO_(3) mass fraction was 20%,it facilitated the formation of tetragonal zirconia,thereby enhancing the creation of robust acidic sites.Acidity is considered to have a strong impact on the catalytic performance of the aquathermolysis of heavy oil.When the catalyst containing 20%WO_(3) was used to catalyze the aquathermolysis of heavy oil under conditions of 14.5 MPa,340℃,and 24 h,the viscosity of heavy oil decreased from 47266 to 5398 mPa·s and the viscosity reduction rate reached 88.6%.The physicochemical properties of heavy oil before and after the aquathermolysis were analyzed using a saturates,aromatics,resins,and asphaltenes analysis,gas chromatography,elemental analysis,densimeter etc.After the aquathermolysis,the saturate and aromatic contents significantly increased from 43.3%to 48.35%and 19.47%to 21.88%,respectively,with large reductions in the content of resin and asphaltene from 28.22%to 25.06%and 5.36%to 2.03%,respectively.The sulfur and nitrogen contents,and the density of the oil were significantly decreased.These factors were likely the main reasons for promoting the viscosity reduction of heavy oil during the aquathermolysis over the WO_(3)/ZrO_(2) solid acid catalysts.

Branched-chain fatty acids from goat milk alleviate ulcerative colitis via the TLR4/NF-κB/NLRP3 pathway

Branched-chain fatty acids(BCFAs)are new bioactive fatty acids with anti-inflammatory properties.However,the role of BCFAs in alleviating ulcerative colitis has not been clarified.Herein,we evaluated the protective effect of BCFAs from goat milk in mice with colitis induced using dextran sodium sulfate(DSS)and explored the corresponding mechanism.These results show that BCFAs extracted from goat milk can significantly alleviate weight loss in mice,and reduce the disease activity index and the activity of myeloperoxidase while increasing the content of antioxidant enzymes in colon tissue and reducing the oxidation stress response.These data also show that BCFAs can down-regulate the gene and protein expression of the toll-like receptor 4(TLR4)/nuclear factorκB p65(NF-κB p65)/NOD-like receptor thermal protein domain associated protein 3(NLRP3)signaling pathway,and at the same time significantly reduce the expression of pro-inflammatory factors tumor necrosis factorα(TNF-α),interleukin 1β(IL-1β),and IL-18 in colon tissue,and significantly increase the expression of the anti-inflammatory factor IL-10.In conclusion,these results demonstrated that BCFAs in goat milk exerted effects on colitis-related inflammatory cytokines and inhibited inflammation by inducing the TLR4/NF-κB/NLRP3 pathway to alleviate DSS-induced ulcerative colitis.This study provides evidence for the potential of BCFAs as bioactive fatty acids in food products and to ameliorate ulcerative colitis development in mice.

Elaidic acid-induced intestinal barrier damage led to gut-liver axis derangement and triggered NLRP3 inflammasome in the liver of SD rats

Previous studies have shown that trans fatty acids(TFA) are associated with several chronic diseases,the gut microbiota is directly influenced by dietary components and linked to chronic diseases.Our research investigated the effects of elaidic acid(EA),a typical TFA,on the gut microbiota to understand the underlying mechanisms of TFA-related chronic diseases.16S rDNA gene sequencing on faecal samples from Sprague-Dawley rats were performed to explore the composition change of the gut microbiota by EA gavage for 4 weeks.The results showed that the intake of EA increased the abundance of well-documented harmful bacteria,such as Proteobacteria,Anaerotruncus,Oscillibacter and Desulfovibrionaceae.Plus,EA induced translocation of lipopolysaccharides(LPS) and the above pathogenic bacteria,disrupted the intestinal barrier,led to gut-liver axis derangement and TLR4 pathway activation in the liver.Overall,EA induced intestinal barrier damage and regulated TLR4-MyD88-NF-κB/MAPK pathways in the liver of SD rats,leading to the activation of NLRP3 inflammasome and inflammatory liver damage.

Oleanolic acid inhibits colon cancer cell stemness and reverses chemoresistance by suppressing JAK2/STAT3 signaling pathway

Background:Oleanolic acid(OA),a pentacyclic triterpenoid exhibiting specific anti-cancer properties and highly effective antioxidant activity,was isolated from traditional Chinese medicinal herbs.Conversely,the OA that impacts colon cancer(CC)cells and its underlying mechanisms remain poorly understood.Methods:The cytotoxic effect of OA alone or OA-5-Fluorouracil(5-FU)combination on normal and CC cells was analyzed by methyl thiazolyl diphenyl-tetrazolium bromide(MTT).Then,the impact of OA on CC cell lines(LoVo and HT-29)proliferation and stemness were measured using colon formation and tumorsphere formation assays.Octamer-binding transcription factor 4(Oct4),Prominin-1(CD133),Nanog,and transcription factor SOX-2(SOX2)are cell stemness-related indicators whose expression was assessed usingfluorescence qPCR assay,Western blotting,and immunohistochemistry.The effect of OA on the proliferative potency of CC cells was evaluated using an in vivo model.Results:The stem-like characteristics and clone production of colon cancer cells were markedly reduced by OA alone or in combination with OA-5-FU.Moreover,OA increases the susceptibility of CC cells to 5-FU by blocking the cell stemness-related markers(CD133,Nanog,SOX2,and Oct4)expression levels both in vitro and in vivo,as well as by inactivating the activator of transcription 3(STAT3 signaling)and Janus kinase 2/signal transducer(JAK2).Conclusion:Thesefindings imply that oleanolic acid,both in vitro and in vivo,suppresses the JAK2/STAT3 pathway,which in turn reverses chemoresistance and decreases colon cancer cell stemness.Therefore,by reducing the recommended amount of 5-FU,this strategy may improve chemotherapeutic effectiveness and minimize undesired side effects.

Gossypol acetic acid regulates leukemia stem cells by degrading LRPPRC via inhibiting IL-6/JAK1/STAT3 signaling or resulting mitochondrial dysfunction

BACKGROUND Leukemia stem cells(LSCs)are found to be one of the main factors contributing to poor therapeutic effects in acute myeloid leukemia(AML),as they are protected by the bone marrow microenvironment(BMM)against conventional therapies.Gossypol acetic acid(GAA),which is extracted from the seeds of cotton plants,exerts anti-tumor roles in several types of cancer and has been reported to induce apoptosis of LSCs by inhibiting Bcl2.AIM To investigate the exact roles of GAA in regulating LSCs under different microenvironments and the exact mechanism.METHODS In this study,LSCs were magnetically sorted from AML cell lines and the CD34+CD38-population was obtained.The expression of leucine-rich pentatricopeptide repeat-containing protein(LRPPRC)and forkhead box M1(FOXM1)was evaluated in LSCs,and the effects of GAA on malignancies and mitochondrial RESULTS LRPPRC was found to be upregulated,and GAA inhibited cell proliferation by degrading LRPPRC.GAA induced LRPPRC degradation and inhibited the activation of interleukin 6(IL-6)/janus kinase(JAK)1/signal transducer and activator of transcription(STAT)3 signaling,enhancing chemosensitivity in LSCs against conventional chemotherapies,including L-Asparaginase,Dexamethasone,and cytarabine.GAA was also found to downregulate FOXM1 indirectly by regulating LRPPRC.Furthermore,GAA induced reactive oxygen species accumulation,disturbed mitochondrial homeostasis,and caused mitochondrial dysfunction.By inhibiting IL-6/JAK1/STAT3 signaling via degrading LRPPRC,GAA resulted in the elimination of LSCs.Meanwhile,GAA induced oxidative stress and subsequent cell damage by causing mitochondrial damage.CONCLUSION Taken together,the results indicate that GAA might overcome the BMM protective effect and be considered as a novel and effective combination therapy for AML.

超声波辅助合成色酮-3-甲酸

以苯酚为原料,经傅克酰基化、甲酰化、水解三步反应制备了色酮-3-甲酸,总收率57.4%,产物纯度98.6%。甲酰化反应以3-(2-羟基苯基)-3-氧代丙酸甲酯0.05 mol为模型,考察了不同溶剂、甲酸钠用量、超声频率、超声功率、反应温度、反应时间的影响。研究结果表明最优化的反应条件为四氢呋喃作溶剂、甲酸钠与3-(2-羟基苯基)-3-氧代丙酸甲酯的用量摩尔比为1.5:1、超声频率50 kHz、超声功率550 W、温度60℃、反应11 h,该步反应收率为80.0%。

老年肌少症患者血清硬骨素和n-3脂肪酸表达水平及临床价值研究

目的探讨血清硬骨素、n-3脂肪酸在老年肌少症患者中的表达水平及临床意义。方法收集2021年5月—2023年5月新疆医科大学第五附属医院老年病科收治的老年肌少症患者118例为病例组,根据病情程度分为肌少症前期(n=39)、肌少症期(n=46)和重度肌少症期(n=33),选取同期医院健康体检者60例为健康对照组。酶联免疫吸附法检测血清硬骨素,气相色谱法检测n-3脂肪酸水平;人体成分分析仪和生物电阻抗体法检测体脂百分比、上臂围、内脏脂肪面积、蛋白质质量;6M步行法测量步速,Jamar握力计测量双手握力,双能X线吸收仪测量全身及四肢骨骼肌质量,计算相对四肢骨骼肌质量指数(RSMI);硬骨素和n-3脂肪酸与病情严重程度及上述指标的相关性用Pearson积矩相关或Spearman秩相关分析;采用Logistic回归分析老年肌少症的影响因素,ROC曲线评估血清硬骨素和n-3脂肪酸对老年肌少症的诊断价值。结果病例组患者血清硬骨素、n-3脂肪酸水平低于健康对照组(t/P=13.342/<0.001、13.116/<0.001);血清硬骨素、n-3脂肪酸水平比较,肌少症前期>肌少症期>重度肌少症期(F/P=59.138/<0.001、79.217/<0.001);病例组患者体脂百分比、内脏脂肪面积大于健康对照组(t/P=8.732/<0.001、5.124/<0.001),上臂围、蛋白质质量、步速、握力、全身骨骼肌质量、四肢骨骼肌质量、RSMI水平低于健康对照组(t/P=3.859/<0.001、8.459/<0.001、5.758/<0.001、12.492/<0.001、7.006/<0.001、10.334/<0.001、11.813/<0.001);老年肌少症患者血清硬骨素、n-3脂肪酸与体脂百分比、内脏脂肪面积呈负相关(硬骨素:r/P=-0.537/<0.001、-0.612/<0.001;n-3脂肪酸:r/P=-0.498/<0.001、-0.523/<0.001),与上臂围、蛋白质质量、步速、握力、全身骨骼肌质量、四肢骨骼肌质量、RSMI呈正相关(硬骨素:r/P=0.593/<0.001、0.624/<0.001、0.639/<0.001、0.597/<0.001、0.601/<0.001、0.607/<0.001、0.638/<0.001;n-3脂肪酸:r/P=0.569/<0.001、0.611/<0.001、0.570/<0.001、0.592/<0.001、0.549/<0.001、0.534/<0.001、0.587/<0.001);年龄、体脂百分比、内脏脂肪面积升高是老年肌少症危险因素[OR(95%CI)=1.702(1.115~2.600)、1.551(1.052~2.287)、1.387(1.006~1.913)],BMI、上臂围、蛋白质质量、步速、握力、全身骨骼肌质量、四肢骨骼肌质量、RSMI、硬骨素、n-3脂肪酸升高是老年肌少症的保护因素[OR(95%CI)=0.728(0.539~0.982)、0.768(0.593~0.995)、0.845(0.723~0.986)、0.815(0.668~0.995)、0.585(0.382~0.897)、0.746(0.573~0.972)、0.733(0.559~0.964)、0.713(0.541~0.940)、0.822(0.695~0.973)、0.803(0.664~0.971)];血清硬骨素、n-3脂肪酸及二者联合诊断老年肌少症的AUC分别为0.822、0.818、0.894,二者联合诊断老年肌少症的AUC大于其各自单独诊断(Z=2.205、2.328,P=0.002、0.001)。结论老年肌少症患者血清硬骨素和n-3脂肪酸降低,两指标与病情严重程度密切相关,早期联合检测可辅助临床诊断老年肌少症。

基于PI3K/AKT/GSK-3β信号通路探讨EA改善APP/PS1双转基因小鼠认知功能障碍的内在机制

目的探讨鞣花酸(ellagicacid,EA)对APP/PS1双转基因小鼠认知功能的影响,并基于磷脂酰肌醇3-激酶/蛋白激酶B/糖原合成酶激酶-3(PI3K/AKT/GSK-3β)信号通路探讨鞣花酸对双转基因小鼠海马氧化应激水平的调节机制。方法将32只SPF级6月龄APP/PS1双转基因小鼠随机分为4组,即APP/PS1组、APP/PS1+EA组、APP/PS1+LY294002组、APP/PS1+EA+LY294002组,每组8只,另外选取8只SPF级C57BL/6J野生型小鼠(Wildtype)作为空白对照组,即WT组。APP/PS1+EA组给予50mg·kg^(-1)·d^(-1)灌胃EA;APP/PS1+LY294002组予以1.5mg·kg^(-1)·d^(-1)腹腔注射PI3K抑制剂LY294002;APP/PS1+EA+LY294002组予以50mg·kg^(-1)·d^(-1)灌胃EA,同时按1.5mg·kg^(-1)·d^(-1)腹腔注射LY294002;WT组和APP/PS1组于相同时间点灌胃等体积10%二甲基亚砜(DMSO)。每日给药1次,连续给药60天。Morris水迷宫检测小鼠学习和记忆能力,免疫组化、蛋白免疫印迹法检测PI3K、AKT、GSK-3β相关蛋白的表达,透射电镜观察小鼠海马组织超微结构变化。结果与WT组相比,其他四组的逃避潜伏期均增长(P<0.05),穿越平台次数明显减少(P<0.01);APP/PS1组、APP/PS1+LY294002组和APP/PS1+EA+LY294002组中的PI3K、AKT蛋白表达量显著降低(P<0.01),GSK-3β表达量显著升高(P<0.01);APP/PS1+EA组的PI3K表达量降低(P<0.05),AKT表达量显著降低(P<0.01),GSK-3β表达量升高(P<0.05);与WT组相比,APP/PS1组海马神经元细胞数目较少,线粒体结构破坏,大部分线粒体出现肿胀,线粒体的内膜和外模不完整,部分线粒体嵴消失,微管、微丝缠结,排列紊乱,而APP/PS1+EA组神经元细胞数较APP/PS1组增多,线粒体结构较清晰,可见清楚的线粒体嵴,线粒体轻度水肿。微管、微丝排列较整齐有序。结论鞣花酸改善AD模型小鼠的学习和记忆能力、减少海马神经元细胞损伤和凋亡,其作用机制可能是通过调节PI3K、AKT、GSK-3β等相关蛋白降低AD模型小鼠海马氧化应激水平。

miR-525-5p、围脂滴蛋白3的表达与胶质瘤预后的关系研究

目的 探讨微核糖核酸-525-5p(mi R-525-5p)、围脂滴蛋白3(PLIN3)的表达与胶质瘤预后的关系。方法选取2018年2月至2020年7月青岛市市立医院收治的102例胶质瘤患者,收集术中部分瘤组织和瘤旁组织采用实时荧光定量聚合酶链式反应检测mi R-525-5p、PLIN3 m RNA表达,免疫组织化学染色法检测PLIN3表达。通过Target Scan数据库预测mi R-525-5p与PLIN3的结合位点,分析二者在胶质瘤组织中表达的相关性。根据胶质瘤组织中PLIN3 m RNA表达均值分为高表达组和低表达组,采用Kaplan-Meier法绘制高/低PLIN3 m RNA表达胶质瘤患者生存曲线,多因素Cox回归分析胶质瘤患者预后影响因素。结果 与瘤旁组织比较,胶质瘤组织中mi R-525-5p表达降低,PLIN3 m RNA表达升高(P<0.05)。与瘤旁组织比较,胶质瘤组织中PLIN3阳性率升高(P<0.05)。mi R-525-5p与PLIN3存在结合位点。mi R-525-5p与PLIN3 m RNA表达在胶质瘤组织中呈负相关(P<0.05)。PLIN3 m RNA高表达组3年总生存率低于低表达组(P<0.05)。多因素Cox回归分析显示,低分化、世界卫生组织中枢神经系统肿瘤分级Ⅲ~Ⅳ级、PLIN3 m RNA≥1.86为胶质瘤患者死亡的独立危险因素(HR>1,P<0.05)。结论 胶质瘤组织中mi R-525-5p低表达和PLIN3 m RNA高表达,二者表达呈负相关,PLIN3 m RNA高表达与胶质瘤患者预后不良有关。

入院时血清TGF-β1、Smad2、Smad3、HA、LN、PCⅢ、CⅣ水平与CHB肝纤维化严重程度的相关性及对疾病预后的预测价值

目的探讨入院时血清转化生长因子-β1(TGF-β1)、Smad同源蛋白2(Smad2)、Smad同源蛋白3(Smad3)及透明质酸(HA)、Ⅲ型前胶原(PCⅢ)、层黏连蛋白(LN)、Ⅳ型胶原(CⅣ)水平与慢性乙型肝炎(CHB)肝纤维化严重程度的相关性及联合检测对疾病预后的预测价值。方法选取河南省中医院2021年3月至2022年3月收治的78例CHB肝纤维化患者作为研究组,选择同期78名健康体检者作为对照组。比较研究组和对照组及不同肝纤维化分期、不同炎症活动分级CHB肝纤维化患者入院时血清TGF-β1、Smad2、Smad3、HA、PCⅢ、LN、CⅣ水平;分析入院时血清TGF-β1、Smad2、Smad3、HA、PCⅢ、LN、CⅣ水平与肝纤维化分期、炎症活动分级的相关性。CHB肝纤维化患者治疗3个月后,根据患者预后分为预后良好和预后不良亚组,比较预后良好和预后不良患者入院时血清TGF-β1、Smad2、Smad3、HA、PCⅢ、LN、CⅣ水平;分析入院时血清TGF-β1、Smad2、Smad3、HA、PCⅢ、LN、CⅣ水平联合检测对CHB肝纤维化患者预后不良的预测价值。结果研究组入院时血清TGF-β1、Smad2、Smad3、HA、LN、PCⅢ、CⅣ高于对照组(P<0.05);不同肝纤维化分期、炎症活动分级CHB肝纤维化患者入院时血清TGF-β1、Smad2、Smad3、HA、LN、PCⅢ、CⅣ比较:S1<S2<S3<S4、G1<G2<G3<G4,差异有统计学意义(P<0.05);入院时血清TGF-β1、Smad2、Smad3、HA、LN、PCⅢ、CⅣ水平与肝纤维化分期、炎症活动分级均呈正相关(P<0.05)。预后良好患者入院时血清TGF-β1、Smad2、Smad3、HA、LN、PCⅢ、CⅣ水平均低于预后不良患者(P<0.05);入院时血清TGF-β1、Smad2、Smad3、HA、LN、PCⅢ、CⅣ水平联合预测肝纤维化患者预后不良的曲线下面积(AUC)优于各指标单一检测(P<0.05)。结论CHB肝纤维化患者入院时血清TGF-β1、Smad2、Smad3、HA、PCⅢ、LN、CⅣ水平均呈现高表达,且与肝纤维化分期、炎症活动分级密切相关,其联合检测对CHB肝纤维化患者预后有较高的预测价值,可用于评估CHB肝纤维化患者病情严重程度和预后,为制定针对性治疗措施提供参考。

3-吲哚丙酸抑制腹膜间皮细胞EMT和纤维化的机制研究

目的 评估3-吲哚丙酸(3-indoleacetic acid, IPA)对脂多糖(lipolyaccharide, LPS)诱导的小鼠腹膜间皮细胞上皮-间质转化(epithelial-mesenchymal transition, EMT)和纤维化的影响。方法 CCK-8法检测不同浓度(0.1、1.0和10μmol/L)的IPA对LPS处理前后的小鼠腹膜间皮细胞增殖活性的影响,确定IPA最佳使用剂量。将小鼠腹膜间皮细胞分成Control组、LPS组、LPS+IPA组、LPS+LY364947(TGF-β1/Smad3通路抑制剂)组、LPS+IPA+LY364947组和LPS+IPA+SRI-011381(TGF-β1/Smad3通路激活剂)组;CCK-8法检测细胞增殖活力,Transwell法检测细胞侵袭,ELISA检测培养上清中间质表型α-平滑肌肌动蛋白(α-smooth muscle actin, α-SMA)的浓度,Western blot检测细胞中α-SMA,EMT相关因子E-cadherin, TGF-β1/Smad3通路相关因子Smad3、p-Smad3的蛋白表达。结果 IPA的最佳使用剂量为1.0μmol/L。与Control组相比,LPS组细胞增殖活力、E-cadherin表达水平下降(均P<0.01),侵袭能力、α-SMA表达水平和p-Smad3/Smad3升高(均P<0.01)。与LPS组相比,LPS+IPA组和LPS+LY364947组细胞增殖活力、E-cadherin表达水平上升(均P<0.01),侵袭能力、α-SMA表达和p-Smad3/Smad3下降(均P<0.01)。与LPS+IPA组相比,LPS+IPA+LY364947组所测指标趋势更加显著,LPS+IPA+SRI-011381组所测指标变化趋势均被逆转。结论 IPA通过抑制TGF-β1/Smad3通路中Smad3蛋白的磷酸化,从而抑制细胞EMT进展,减轻LPS诱导的腹膜间皮细胞损伤。

Gabra3通过AKT/mTOR途径促进膀胱癌T24细胞侵袭和迁移

目的:探究γ-氨基丁酸受体亚基α-3(Gamma-aminobutyric acid receptor subunit alpha-3,Gabra3)基因对膀胱癌T24细胞凋亡、迁移和侵袭的作用及其机制。方法:TCGA数据库分析Gabra3基因在膀胱癌中的表达以及转移和生存的相关性。建立Gabra3过表达和Gabra3突变的T24细胞株,根据转染质粒不同,分为空白T24细胞(Control)、空pDONR223载体转染T24细胞(NC)、野生型Gabra3过表达T24细胞株(wt-Gabra3)、突变型Gabra3 T24细胞株(mut-Gabra3)。在回补实验中,分为转染突变型Gabra3 T24组(mut-Gabra3)、转染空pDONR223载体mut-Gabra3组(mut-Gabra3-NC)、转染野生型Gabra3过表达组(mut-Gabra3+wt-Gabra3)。用TUNEL染色检测T24细胞凋亡,细胞划痕和Transwell分别检测T24细胞迁移和侵袭,Western blot检测AKT/mTOR通路相关分子生物表达水平。结果:Gabra3基因在膀胱癌中显著高表达(P<0.05),生存曲线证实远处转移存在的患者生存期明显较短(P<0.05)。成功构建Gabra3过表达和突变的T24细胞株。与Control组相比,wt-Gabra3组细胞凋亡能力显著降低,迁移、侵袭能力显著增强(P<0.05),而mut-Gabra3组细胞凋亡显著增加,侵袭能力显著减弱(P<0.05);wt-Gabra3组细胞p-AKT、p-mTOR、p-4E-BP1、p-S6K蛋白表达均显示上凋(P<0.05),而mut-Gabra3组细胞上述蛋白无差异。在回补实验中,过表达wt-Gabra3的mut-Gabra3突变T24细胞凋亡能力受到抑制(P<0.05),迁移和侵袭能力显著增强(P<0.05),并且该组细胞AKT/mTOR通路相关分子显著激活(P<0.05)。结论:外源性的未编辑的Gabra3可以促进膀胱癌T24细胞的迁移、侵袭能力,并且可能与激活AKT/mTOR途径通路密切相关。