Gga-miRNA-181-5p family facilitates chicken myogenesis via targeting TGFBR1 to block TGF-βsignaling

MicroRNAs(miRNAs)have been demonstrated to control chicken skeletal muscle growth,however,the potential function of the miR-181-5p family in chicken myogenesis remains largely unknown.Here,our study identified the two chicken(Gallus gallus;Gga)miR-181-5p family members widely expressed in various tissues,specifically miR-181a-5p and miR-181b-5p.Besides,the breast muscles of fast-growing broilers expressed higher levels of miR-181a-5p and miR-181b-5p than those of slow-growing layers.Functionally,miR-181a-5p and miR-181b-5p both promote the expression level of myogenic factors including myogenin(MyoG),myogenic differentiation 1(MyoD1),and myosin heavy chain(MyHC),meanwhile accelerating the myotube formation of skeletal muscle satellite cells(SMSCs).Mechanistically,miR-181a-5p and miR-181b-5p directly bind to the 3′untranslated region(UTR)of the transforming growth factor beta receptor 1(TGFBR1)mRNA,further reducing the expression of TGFBR1.TGFBR1 is a key Transforming growth factor beta(TGF-β)signaling transduction receptor and had a negative function in muscle cell differentiation.Furthermore,knockdown of TGFBR1 facilitated the expression of chicken myogenic factors,boosted myotube formation,and decreased the SMAD family member 2/3(SMAD2/3)phosphorylation in chicken SMSCs.SMAD2/3 are downstream of TGF-βsignaling,and miR-181a-5p and miR-181b-5p could reduce the expression of TGFBR1 to further diminish the SMAD2/3 phosphorylation.Our findings revealed that the miR-181-5p family targets TGFBR1 to break the TGF-βsignaling transduction,which resulted in promoting chicken skeletal muscle development.

Argatroban promotes recovery of spinal cord injury by inhibiting the PAR1/JAK2/STAT3 signaling pathway

Argatroban is a synthetic thrombin inhibitor approved by U.S.Food and Drug Administration for the treatment of thrombosis.However,whether it plays a role in the repair of spinal cord injury is unknown.In this study,we established a rat model of T10 moderate spinal cord injury using an NYU Impactor ModerⅢand performed intraperitoneal injection of argatroban for 3 consecutive days.Our results showed that argatroban effectively promoted neurological function recovery after spinal cord injury and decreased thrombin expression and activity in the local injured spinal cord.RNA sequencing transcriptomic analysis revealed that the differentially expressed genes in the argatroban-treated group were enriched in the JAK2/STAT3 pathway,which is involved in astrogliosis and glial scar formation.Western blotting and immunofluorescence results showed that argatroban downregulated the expression of the thrombin receptor PAR1 in the injured spinal cord and the JAK2/STAT3 signal pathway.Argatroban also inhibited the activation and proliferation of astrocytes and reduced glial scar formation in the spinal cord.Taken together,these findings suggest that argatroban may inhibit astrogliosis by inhibiting the thrombin-mediated PAR1/JAK2/STAT3 signal pathway,thereby promoting the recovery of neurological function after spinal cord injury.

Enhancement of porcine in vitro embryonic development through luteolin‑mediated activation of the Nrf2/Keap1 signaling pathway

Background Oxidative stress,caused by an imbalance in the production and elimination of intracellular reactive oxygen species(ROS),has been recognized for its detrimental effects on mammalian embryonic development.Luteolin(Lut)has been documented for its protective effects against oxidative stress in various studies.However,its specific role in embryonic development remains unexplored.This study aims to investigate the influence of Lut on porcine embryonic development and to elucidate the underlying mechanism.Results After undergoing parthenogenetic activation(PA)or in vitro fertilization,embryos supplemented with 0.5μmol/L Lut displayed a significant enhancement in cleavage and blastocyst formation rates,with an increase in total cell numbers and a decrease in the apoptosis rate compared to the control.Measurements on D2 and D6 revealed that embryos with Lut supplementation had lower ROS levels and higher glutathione levels compared to the control.Moreover,Lut supplementation significantly augmented mitochondrial content and membrane potential.Intriguingly,activation of the Nrf2/Keap1 signaling pathway was observed in embryos supplemented with Lut,leading to the upregulation of antioxidant-related gene transcription levels.To further validate the relationship between the Nrf2/Keap1 signaling pathway and effects of Lut in porcine embryonic development,we cultured PA embryos in a medium supplemented with brusatol,with or without the inclusion of Lut.The positive effects of Lut on developmental competence were negated by brusatol treatment.Conclusions Our findings indicate that Lut-mediated activation of the Nrf2/Keap1 signaling pathway contributes to the enhanced production of porcine embryos with high developmental competence,and offers insight into the mechanisms regulating early embryonic development.

Spi1 regulates the microglial/macrophage inflammatory response via the PI3K/AKT/mTOR signaling pathway after intracerebral hemorrhage

Preclinical and clinical studies have shown that microglia and macrophages participate in a multiphasic brain damage repair process following intracerebral hemorrhage.The E26 transformation-specific sequence-related transcription factor Spi1 regulates microglial/macrophage commitment and maturation.However,the effect of Spi1 on intracerebral hemorrhage remains unclear.In this study,we found that Spi1 may regulate recovery from the neuroinflammation and neurofunctional damage caused by intracerebral hemorrhage by modulating the microglial/macrophage transcriptome.We showed that high Spi1expression in microglia/macrophages after intracerebral hemorrhage is associated with the activation of many pathways that promote phagocytosis,glycolysis,and autophagy,as well as debris clearance and sustained remyelination.Notably,microglia with higher levels of Soil expression were chara cterized by activation of pathways associated with a variety of hemorrhage-related cellular processes,such as complement activation,angiogenesis,and coagulation.In conclusion,our results suggest that Spi1 plays a vital role in the microglial/macrophage inflammatory response following intracerebral hemorrhage.This new insight into the regulation of Spi1 and its target genes may advance our understanding of neuroinflammation in intracerebral hemorrhage and provide therapeutic targets for patients with intracerebral hemorrhage.

Exploring the effect of Bushen Bitong recipe-containing serum on IL-1β-induced chondrocyte apoptosis based on SOX9/NF-κB/MMP-13 signaling pathway

Objective:To observe the effect and possible mechanism of action of Bushen Bitong recipe(BSBT)containing serum on IL-1β-induced chondrocyte apoptosis.Methods:Generation 3 rat chondrocytes were randomized into Control,IL-1β,IL-1β+BSBT(L),IL-1β+BSBT(M),and IL-1β+BSBT(H)groups(5%,10%and 15%BSBT-containing serum),and then 24h after intervention respectively,the cell proliferation and Apoptosis rate;Western blot detected the expression levels of Bcl-2,BAX,Caspase-3,SOX9,NF-κB p65,MMP-13 proteins in chondrocytes.ELISA detected the levels of TNF-α,IL-6,and bFGF in the supernatants of chondrocyte culture.Results:Compared with Control group,cell proliferation activity decreased,apoptosis rate increased,NF-κB p65,MMP-13 protein level and TNF-α,IL-6 level increased,and SOX9 protein level and bFGF level decreased in IL-1βgroup;compared with IL-1βgroup,different concentrations of BSBT-containing serum group,cell proliferation activity increased,and apoptosis rate decreased.NF-κB p65,MMP-13 protein level and TNF-α,IL-6 level decreased,SOX9 protein level and bFGF level increased;compared with IL-1β+BSBT(L)group,cell proliferation activity increased,apoptosis rate decreased in IL-1β+BSBT(M)and IL-1β+BSBT(H)groups,and NF-κB p65,MMP-13 protein level and TNF-αlevel decreased.13 protein levels and TNF-αand IL-6 levels decreased,and SOX9 protein levels and bFGF levels increased.Conclusion:BSBT-containing serum may promote IL-1β-induced proliferation of chondrocytes,reduce apoptosis,improve the microenvironment of chondrocytes,and promote cartilage repair through the SOX9/NF-κB/MMP-13 signaling pathway.

IKIP downregulates THBS1/FAK signaling to suppress migration and invasion by glioblastoma cells

Background:Inhibitor of NF-κB kinase-interacting protein(IKIP)is known to promote proliferation of glioblastoma(GBM)cells,but how it affects migration and invasion by those cells is unclear.Methods:We compared levels of IKIP between glioma tissues and normal brain tissue in clinical samples and public databases.We examined the effects of IKIP overexpression and knockdown on the migration and invasion of GBM using transwell and wound healing assays,and we compared the transcriptomes under these different conditions to identify the molecular mechanisms involved.Results:Based on data from our clinical samples and from public databases,IKIP was overexpressed in GBM tumors,and its expression level correlated inversely with survival.IKIP overexpression in GBM cells inhibited migration and invasion in transwell and wound healing assays,whereas IKIP knockdown exerted the opposite effects.IKIP overexpression in GBM cells that were injected into mouse brain promoted tumor growth but inhibited tumor invasion of surrounding tissue.The effects of IKIP were associated with downregulation of THBS1 mRNA and concomitant inhibition of THBS1/FAK signaling.Conclusions:IKIP inhibits THBS1/FAK signaling to suppress migration and invasion of GBM cells.

Acoustic emission signal identification of different rocks based on SE-1DCNN-BLSTM network model

In order to study fracture mechanism of rocks in different brittle mineral contents,this study pro-poses a method to identify the acoustic emission signal released by rock fracture under different brittle miner-al content(BMC),and then determine the content of brittle matter in rock.To understand related interference such as the noises in the acoustic emission signals released by the rock mass rupture,a 1DCNN-BLSTM network model with SE module is constructed in this study.The signal data is processed through the 1DCNN and BLSTM networks to fully extract the time-series correlation features of the signals,the non-correlated features of the local space and the weak periodicity law.Furthermore,the processed signals data is input into the fully connected layers.Finally,softmax function is used to accurately identify the acoustic emission signals released by different rocks,and then determine the content of brittle minerals contained in rocks.Through experimental comparison and analysis,1DCNN-BLSTM model embedded with SE module has good anti-noise performance,and the recognition accuracy can reach more than 90 percent,which is better than the traditional deep network models and provides a new way of thinking for rock acoustic emission re-search.

Oleuropein alleviates sepsis-induced acute lung injury via the AMPK/Nrf-2/HO-1 signaling

Objective:To explore the effect of oleuropein on sepsis-induced acute lung injury(ALI)in vitro and in vivo and investigate the underlying mechanism.Methods:In an lipopolysaccharide(LPS)-mediated cell model of sepsis-induced ALI and a cecal ligation and puncture-induced mouse model of septic ALI,CCK-8 assay and flow cytometry analysis were used to detect cell activity and apoptosis.ELISA and relevant assay kits were used to measure the levels of inflammatory cytokines and oxidative stress,respectively.Western blot was applied to determine the expression of apoptosis-and AMP-activated protein kinase(AMPK)/nuclear factor erythroid 2-related factor-2(Nrf-2)/heme oxygenase-1(HO-1)signaling-associated proteins.JC-1 staining,adenosine triphosphate(ATP)assay kit,and MitoSOX Red assays were performed to detect mitochondrial membrane potential,ATP content,and mitochondrial ROS formation,respectively.Moreover,lung injury was evaluated by measuring lung morphological alternations,lung wet-to-dry ratio,myeloperoxidase content,and total protein concentration.Results:Oleuropein reduced inflammatory reaction,oxidative damage,and apoptosis,and ameliorated mitochondrial dysfunction in LPS-exposed BEAS-2B cells and mice with septic ALI.Besides,oleuropein activated the AMPK/Nrf-2/HO-1 signaling pathway.However,these effects of oleuropein were abrogated by an AMPK inhibitor compound C.Conclusions:Oleuropein can protect against sepsis-induced ALI in vitro and in vivo by activating the AMPK/Nrf-2/HO-1 signaling,which might be a potential therapeutic agent for the treatment of sepsis-induced ALI.

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.

Electroacupuncture improves myocardial fibrosis in heart failure rats by attenuating ECM collagen deposition through modulation of TGF-β1/Smads signaling pathway

Background: To explore the effects of electroacupuncture on cardiac function and myocardial fibrosis in rat models of heart failure, and to elucidate the underlying mechanism of electroacupuncture in heart failure treatment. Methods: Healthy male Sprague-Dawley rats were allocated into three groups: Sham group, Model group, and electroacupuncture (Model + EA) group, with each group comprising 8 rats. The model underwent a procedure involving the ligation of the left anterior descending coronary artery to induce a model of heart failure. The Model + EA group was used for 7 consecutive days for electroacupuncture of bilateral Shenmen (HT7) and Tongli (HT5), once a day for 30 min each time. Left ventricular parameters in rats were assessed using a small-animal ultrasound machine to analyze changes in left ventricular end-diastolic volume, left ventricular end-systolic volume, left ventricular ejection fraction, and left ventricular fractional shortening. Serum interleukin-1β (IL-1β), cardiac troponin (cTn), and N-terminal brain natriuretic peptide precursor levels were measured using ELISA. Histopathological changes in rat myocardium were observed through HE staining, while collagen deposition in rat myocardial tissue was assessed using the Masson staining method. Picro sirius red staining, immunohistochemical staining, and RT-qPCR were utilized to distinguish between the various types of collagen deposition. The expression level of TGF-β1 and SMAD2/3/4/7 mRNA in rat myocardial tissues was determined using RT-qPCR. Additionally, western blot analysis was conducted to assess the protein expression levels of TGF-β1, SMAD3/7, and p-SMAD3 in rat myocardial tissues. Results: Compared with the Sham group, the left ventricular ejection fraction and left ventricular fractional shortening values of the Model group were significantly decreased (P < 0.01);the left ventricular end-diastolic volume and left ventricular end-systolic volume values were remarkably increased (P < 0.01);serum N-terminal brain natriuretic peptide precursor content was increased (P < 0.01);serum IL-1β and cTn levels were increased (P < 0.01);myocardial collagen volume fraction were increased (P < 0.01);and those of the expression of TGF-β1 and SMAD2/3/4 mRNA was increased (P < 0.01);the expression of SMAD7 mRNA was decreased (P < 0.01);the protein expression levels of TGF-β1, SMAD3, and p-Smad3 were increased (P < 0.01);the protein expression level of SMAD7 was decreased (P < 0.01) in the Model group. Compared to the Model group, the expression levels of the proteins TGF-β1, SMAD3, and p-Smad3 in myocardial tissue were found to be decreased (P < 0.01), and the expression level of the protein SMAD7 was found to be increased (P < 0.01) in the Model + EA group;the collagen volume fraction and deposition of type Ⅰ /Ⅲ collagen were decreased (P < 0.01) in the Model + EA group. Conclusion: Electroacupuncture alleviates myocardial fibrosis in rats with heart failure, and this effect is likely due to attributed to the modulation of the TGF-β1/Smads signaling pathway, which helps reduce collagen deposition in the extracellular matrix.

Exploring the mechanism of electroacupuncture at different acupoints on acute colitis rats based on JAK2/STAT3/SOCS1 signaling pathway

Objective:To investigate the mechanism of JAK2/STAT3/SOCS1 signaling pathway in electroacupuncture of different acupoints on acute colitis rats.Methods:36 SPF SD rats were randomly divided into 6 groups,with 6 rats in each group.The rat model of acute colitis was prepared by enema with glacial acetic acid solution.After the model was established,electroacupuncture was given to each acupoint group,with density wave,frequency 2Hz-50 Hz,intensity 2 mA,muscle tremor as the degree 20 min/time,1 time/day,for 3 consecutive days.Observe the general condition of rats;the pathological changes of colonic mucosa in rats were observed by HE method.The contents of serum interleukin-4(IL-4)and interleukin-8(IL-8)were detected by ELISA.Western blot and RT-PCR were used to detect the expression of JAK2,STAT3,SOCS1 protein and mRNA in rat colon tissue.Results:In contrast to the normal group,the overall condition of the model group was worse,the colonic mucosa was severely damaged,even necrotic,and the ulcer surface was obvious.The content of IL-4 in serum was obviously reduced,and the content of IL-8 was obviously go up(P<0.01).The protein content of JAK2,STAT3 and the expression of JAK2,STAT3 mRNA in colon tissue of rats were obviously go up,while the protein content of SOCS1 and the expression of SOCS1 mRNA were obviously reduced(P<0.01).In contrast to the model group,the general condition of rats in each acupoint group was significantly improved,the damage and necrosis of colonic mucosa and ulcer surface were obviously alleviated,the content of IL-4 in serum was obviously go up,and the content of IL-8 was significantly decreased(P<0.01).The protein content of JAK2,STAT3 and the expression of JAK2,STAT3 mRNA in colon tissue of rats were obviously reduced,while the protein content of SOCS1 and the expression of SOCS1 mRNA were obviously go up(P<0.05,P<0.01).Comparison of different acupoint groups,the colonic mucosal injury in the Zusanli group was significantly reduced,the content of serum IL-4 was significantly increased,and the content of IL-8 was significantly decreased(P<0.05,P<0.01).The protein content and mRNA expression of JAK2 and STAT3 in colon tissue were significantly down-regulated,while the protein content and mRNA expression of SOCS1 were significantly go up(P<0.05,P<0.01).Conclusion:Electroacupuncture at each acupoint can improve the damage of colonic mucosa and reduce the inflammatory response.The therapeutic effect of Zusanli(ST36)is better than that of Tianshu(ST25),Dachangshu(BL25)and Shangjuxu(ST37).The mechanism may be related to the regulation of JAK2/STAT3/SOCS1 signaling pathway related proteins and inflammatory cytokines IL-4 and IL-8.

X-Paste improves wound healing in diabetes via NF-E2-related factor/HO-1 signaling pathway

BACKGROUND Diabetic foot ulcers(DFU),as severe complications of diabetes mellitus(DM),significantly compromise patient health and carry risks of amputation and mortality.AIM To offer new insights into the occurrence and development of DFU,focusing on the therapeutic mechanisms of X-Paste(XP)of wound healing in diabetic mice.METHODS Employing traditional Chinese medicine ointment preparation methods,XP combines various medicinal ingredients.High-performance liquid chromatography(HPLC)identified XP’s main components.Using streptozotocin(STZ)-induced diabetic,we aimed to investigate whether XP participated in the process of diabetic wound healing.RNA-sequencing analyzed gene expression differences between XP-treated and control groups.Molecular docking clarified XP’s treatment mechanisms for diabetic wound healing.Human umbilical vein endothelial cells(HUVECs)were used to investigate the effects of Andrographolide(Andro)on cell viability,reactive oxygen species generation,apoptosis,proliferation,and metastasis in vitro following exposure to high glucose(HG),while NF-E2-related factor-2(Nrf2)knockdown elucidated Andro’s molecular mechanisms.RESULTS XP notably enhanced wound healing in mice,expediting the healing process.RNA-sequencing revealed Nrf2 upregulation in DM tissues following XP treatment.HPLC identified 21 primary XP components,with Andro exhibiting strong Nrf2 binding.Andro mitigated HG-induced HUVECs proliferation,metastasis,angiogenic injury,and inflammation inhibition.Andro alleviates HG-induced HUVECs damage through Nrf2/HO-1 pathway activation,with Nrf2 knockdown reducing Andro’s proliferative and endothelial protective effects.CONCLUSION XP significantly promotes wound healing in STZ-induced diabetic models.As XP’s key component,Andro activates the Nrf2/HO-1 signaling pathway,enhancing cell proliferation,tubule formation,and inflammation reduction.

The flavonoid glycoside vaccarin inhibits adipogenesis and stimulates lipolysis via Hedgehog signaling in 3T3-L1 adipocytes

Vaccarin,a flavonoid glycoside isolated from Vaccaria segetalis,is non-toxic to 3T3-L1 cells up to concentrations of 200μM.Accordingly,we investigated the effects of this natural product on adipogenesis and lipolysis in 3T3-L1 adipocytes.Our results revealed that vaccarin significantly inhibited lipid accumulation by suppressing the adipogenesis-related transcription factors peroxisome proliferator-activated receptorγ(PPARγ)and the CCAAT/enhancer-binding proteinα(C/EBPα).Specifically,lipid accumulation decreased by up to 27.7±2.7%when 3T3-L1 adipocytes were treated with a 10μM concentration of vaccarin.Mechanistic studies showed that the compound inhibited adipogenesis through activation of the Hedgehog(Hh)signaling pathway and so restoring Smo and Gli1 expression at an early stage of differentiation.In mature 3T3-L1 cells,vaccarin significantly increased the secretion of glycerol into the surrounding medium and thus indicating that it accelerated the degradation of triglycerides.In addition,vaccarin,was shown to enhance lipolysis through stimulation of the transcription levels of lipoprotein lipase,monoglycerides lipase,adipose triacylglyceride lipase,hormone-sensitive lipase and adipose differentiated-related protein.All told,vaccarin suppressed lipid accumulation and enhanced lipolysis during adipocyte differentiation by restoring Hh signaling.As such,it is a phytochemical capable of halting adipocyte hyperplasia and,thereby,ameliorating the effects of obesity.

POU1F1基因SNP位点与尼罗罗非鱼体质量和形态性状的相关性

【目的】研究POU1F1基因的单核苷酸多态性(SNP),评估多态性与尼罗罗非鱼(Oreochromis niloticus)的体质量和形态性状的相关性,为罗非鱼以生长性状为目的的选育提供参考。【方法】利用PCR产物测序方法,从POU1F1中共筛查到28个多态性较高的位点,分析尼罗罗非鱼高要亲代群体的这些位点与其体质量及全长、体长、头长、体高、体宽等6个形态性状的相关性,并在尼罗罗非鱼高要子代群体和番禺群体中验证,将获得的体质量和形态相关位点进一步在尼罗罗非鱼海南群体中验证。【结果与结论】高要亲代群体和子代群体中,分别有6个位点[S3(A-400G)、S4(A-469T)、S5(I-539D)、S6(A-881G)、S7(A-888G)和S12(C-1365T)]和5个位点[S3、S5、S11(I-1358D)、S13(C-1511T)、S14(A-1539T)]与体质量、形态性状相关。POU1F1基因11个SNP位点中,未发现与番禺群体体质量和形态性状相关联的位点。POU1F1基因6个SNP位点与海南雌雄群体关联分析表明,S4位点与海南雌性群体体质量相关,S3和S5位点与雄性群体体质量相关。双倍型与各群体体质量、各形态性状关联分析表明,在高要亲代群体中获得体宽相关双倍型2个;在高要子代群体、番禺群体及海南雄性群体中未获得与生长性状相关双倍型;在海南雌性群体中获得与体质量相关的双倍型1个。

人参皂苷Rg3通过调控E2F1对人胃癌SGC-7901细胞生物行为学的影响

目的探究人参皂苷Rg3通过调控E2F1对人胃癌SGC-7901细胞生物行为学的影响。方法MTT测定不同浓度人参皂苷Rg3(0、80、160、320μmol·L^(-1))对细胞增殖影响;流式细胞术测定不同浓度人参皂苷Rg3对细胞凋亡的影响;划痕愈合实验和Transwell实验测定不同浓度人参皂苷Rg3对细胞迁移及侵袭的影响;Western blot测定不同浓度人参皂苷Rg3对E2F1、MMP-2、MMP-9、BCL-2、Bax表达的影响。结果80、160、320μmol·L^(-1)人参皂苷Rg3组细胞存活率与空白对照组比较明显降低,且呈浓度依赖性(P<0.05)。80、160、320μmol·L^(-1)人参皂苷Rg3组细胞凋亡率与空白对照组比较明显增加,且呈浓度依赖性(P<0.05)。80、160、320μmol·L^(-1)人参皂苷Rg3组细胞迁移数目与空白对照组比较明显降低,且呈浓度依赖性(P<0.05)。80、160、320μmol·L^(-1)人参皂苷Rg3组细胞侵袭数目与空白对照组比较明显降低,且呈浓度依赖性(P<0.05)。80、160、320μmol·L^(-1)人参皂苷Rg3组E2F1 mRNA与E2F1蛋白表达量相较空白对照组明显减少且,呈浓度依赖性(P<0.05)。80、160、320μmol·L^(-1)人参皂苷Rg3组细胞中MMP-2、MMP-9、BCL-2蛋白表达量与空白对照组比较明显降低,BCL-2与空白对照组比较明显升高,且呈浓度依赖性(P<0.05)。结论人参皂苷Rg3能降低胃癌SGC-7901细胞增殖能力,抑制细胞迁移及侵袭能力,同时还促进SGC-7901细胞凋亡,且具有浓度依赖性,其作用机制可能通过E2F1因子下调MMP-2、MMP-9、BCL-2表达,上调Bax表达有关。

咪达唑仑通过调控LncRNA NR2F2-AS1对肾癌786-O细胞增殖、迁移及凋亡的影响

目的 探讨咪达唑仑对肾癌786-O细胞增殖、迁移及凋亡的影响及其可能作用机制。方法 体外培养人肾癌细胞786-O,随机分组:咪达唑仑0、20、40、60μmol/L组、si-NC组、si-核受体亚家族2组F成员反义RNA(NR2F2-AS)1组、咪达唑仑+pcDNA组、咪达唑仑+pcDNA-NR2F2-AS1组;CCK-8法、流式细胞术与Transwell实验分别检测细胞增殖、凋亡及迁移;实时荧光定量-聚合酶链反应(RT-qPCR)检测LncRNA NR2F2-AS1表达量;Western印迹检测B细胞淋巴瘤(Bcl)-2、Bcl-2相关蛋白(Bax)表达量。结果 与咪达唑仑0μmol/L组比较,咪达唑仑20、40、60μmol/L组细胞增殖抑制率、细胞凋亡率和Bax蛋白水平明显升高,迁移细胞数和Bcl-2蛋白水平明显降低,长链非编码(Lnc)RNA NR2F2-AS1表达量明显降低,且呈剂量依赖性(P<0.05);转染si-NR2F2-AS1可明显抑制细胞增殖及迁移,并可明显提高细胞凋亡率(P<0.05);与咪达唑仑+pcDNA组比较,咪达唑仑+pcDNA-NR2F2-AS1组细胞增殖抑制率、细胞凋亡率和Bax蛋白水平明显降低,迁移细胞数和Bcl-2蛋白水平明显升高(P<0.05)。结论 咪达唑仑可通过抑制LncRNA NR2F2-AS1表达而减弱肾癌细胞增殖及迁移能力,并可诱导肾癌细胞凋亡。

华法林相关基因CYP2C9、VKORC1、CYP4F2的多态性分布

目的研究CYP2C9、VKORC1、CYP4F2基因多态性在陕西省宝鸡地区汉族人群中的分布情况,为华法林个体化用药提供遗传依据。方法选取2022年1月至2023年5月在宝鸡市中医医院心内科接受华法林治疗并进行相关基因检测的住院患者475例,采用飞行时间质谱仪对华法林药物相关基因CYP2C9、VKORC1、CYP4F2分别进行检测,然后对检测结果的基因多态性进行分析。结果华法林药物相关各基因分布频率符合Hardy-Weinberg equilibrium规律,475例患者VKORC1基因型分别是AA型、AG型、GG型,基因频率分别为83.79%,15.58%,0.63%;CYP2C9基因型分别是*1/*1型、*1/*3型、*1/*2型,基因频率分别为95.17%,4.74%,0.11%,未检出*2/*2、*2/*3、*3/*3基因型;CYP4F2基因型分别是*1/*1型、*1/*3型、*3/*3型,基因频率分别为52.40%,39.37%,8.21%。各基因型分布在患者不同性别、年龄之间无统计学差异。结论陕西宝鸡地区汉族人群华法林药物基因VKORC1(c.-1639G>A)以变异型为主,包括纯合变异和杂合变异,而CYP2C9(*1、*2、*3)和CYP4F2(*1,*3)以野生型为主。

益肾填精凉血汤联合西药治疗慢性过敏性紫癜性肾炎的疗效及外周血ET-1、TXB2、6-Keto-PGF1α水平的影响

目的:探讨益肾填精凉血汤联合西药治疗慢性过敏性紫癜性肾炎(Henoch-schonlein purpura nephritis,HSPN)患者的疗效及对肾功能、外周血内皮素-1(ET-1)、血栓素B2(TXB2)、6-酮-前列腺素F1α(6-Keto-PGF1α)水平的影响。方法:将97例HSPN患者,采用随机数字表法分为2组,研究组(49例)和对照组(48例)。对照组中脱落3例,剩余45例患者进行常规治疗,包括口服贝那普利、泼尼松以及双嘧达莫。研究组中脱落4例,剩余45例患者在对照组基础上,给予益肾填精凉血汤口服。2组均治疗12 w。观察2组治疗后的临床疗效,治疗前后肾外症状(皮肤紫癜疹消退时间、关节肿痛消失时间、腹痛消失时间、血尿消失时间)、中医证候积分、24 h尿蛋白定量(24 hUpro)、血肌酐(Scr)、血清尿素氮(BUN)、血β2-微球蛋白(β2-MG)、尿红细胞计数(U-RBC)、免疫T细胞亚群(CD4^(+)、CD8^(+)阳性率及CD4^(+)/CD8^(+)比值)水平、血清细胞因子[白介素-1(IL-1)、白介素-17(IL-17)、肿瘤坏死因子-α(TNF-α)]水平、肾脏微循环指标(ET-1、TXB2、6-Keto-PGF1α)及不良反应发生情况。结果:研究组总有效率(86.67%)高于对照组(62.22%)(P<0.05)。研究组皮肤紫癜疹消退时间、关节肿痛消失时间、腹痛消失时间、血尿消失时间均缩短;中医证候积分,24 hUpro、Scr、BUN、β2-MG、U-RBC,CD8^(+)水平,IL-1、IL-17、TNF-α,ET-1、TXB2均降低;CD4^(+)、CD4^(+)/CD8^(+)水平,6-Keto-PGF1α均升高(P<0.05),且研究组与对照组比较差异有统计学意义(P<0.05)。治疗期间,肝、肾功能等均未有严重不良反应发生。结论:益肾填精凉血汤联合西药治疗慢性HSPN的疗效确切,能缩短肾外症状消失时间,调节肾功能,提高机体免疫功能,抑制炎症反应,改善肾脏微循环,减轻单纯服用西药的不良反应。

血必净注射液调控HMGB1/TLR4/NF-κB通路对脓毒症小鼠肺损伤的保护作用

目的 研究血必净注射液调控高迁移率族蛋白1(HMGB1)/Toll样受体4(TLR4)/核因子-κB(NF-κB)通路对脓毒症小鼠肺损伤的保护作用。方法 雄性C57BL/6小鼠随机分为对照组,模型组和低、中、高剂量血必净组,阴性对照(NC)组,NC+模型组,NC+高剂量血必净组,HMGB1+高剂量血必净组。采用盲肠结扎穿孔术建立脓毒症肺损伤模型,造模前给予NC慢病毒或HMGB1慢病毒尾静脉注射,造模当天给予血必净注射液(剂量5、10、15mL/kg)腹腔注射,2次/d,连续3 d,末次给药后24 h进行取材和检测。比较各组间肺组织病理改变,湿重(W)/干重(D)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、丙二醛(MDA)、超氧化物歧化酶(SOD)含量,裂解型Caspase-3、HMGB1、TLR4、NF-κB表达水平的差异。结果 模型组小鼠肺组织出现肺损伤的病理改变,W/D、TNF-α、IL-1β、IL-6、MDA的含量及裂解型Caspase-3、HMGB1、TLR4、NF-κB的表达水平均高于对照组,SOD的含量低于对照组(P<0.05)。不同剂量血必净组小鼠肺损伤的病理改变减轻,W/D、TNF-α、IL-1β、IL-6、MDA的含量及裂解型Caspase-3、HMGB1、TLR4、NF-κB的表达水平低于模型组,SOD的含量高于模型组(P<0.05)。HMGB1+高剂量血必净组小鼠肺损伤的病理改变加重,W/D、TNF-α、IL-1β、IL-6、MDA的含量及裂解型Caspase-3、HMGB1、TLR4、NF-κB的表达水平均高于NC+高剂量血必净组,SOD的含量低于NC+高剂量血必净组(P<0.05)。结论 血必净注射液对脓毒症小鼠肺损伤具有保护作用,并减轻炎症反应、氧化应激、细胞凋亡,其相关的分子机制为抑制HMGB1/TLR4/NF-κB通路。

金雀异黄酮通过Nrf2/HO-1信号通路减轻皮质神经元低氧/复氧损伤

目的研究金雀异黄酮(GEN)对皮质神经元低氧/复氧(H/R)损伤的影响,并基于核因子E2相关因子2/血红素加氧酶1(Nrf2/HO-1)信号通路探讨其机制。方法分离并体外培养C57BL/6J小鼠胎鼠(妊娠15 d)皮质神经元,设正常对照(Normal)组、模型(H/R)组、GEN(12.5μmol·L^(-1))组、TBHQ(Nrf2激动剂,10μmol·L^(-1))组、GEN(12.5μmol·L^(-1))+TBHQ(10μmol·L^(-1))组。除正常对照组外,其他组采用低氧(5%CO_(2)+95%N_(2))4 h后复氧(5%CO_(2)+95%空气)24 h的方法制备H/R损伤皮质神经元模型,各组分别于造模前2h给药干预。采用CCK-8法、流式细胞术检测神经元活力和凋亡率,DCFH-DA荧光探针法检测神经元活性氧(ROS)含量,分光光度法检测神经元中丙二醛(MDA)含量和抗氧化酶[超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)]活性,RT-PCR法检测神经元Nrf2、HO-1 mRNA表达,Western blot法检测神经元Nrf2、HO-1、B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)、激活型Caspase-3(cleaved Caspase-3)蛋白表达。结果与H/R组比较,GEN组、TBHQ组和GEN+TBHQ组皮质神经元活力明显升高、凋亡率明显降低(P<0.05);神经元中ROS、MDA含量明显降低,SOD、GSH-Px活性明显升高(P<0.05);Nrf2、HO-1 mRNA表达量明显升高(P<0.05);Nrf2、HO-1、Bcl-2蛋白表达量及Bcl-2/Bax比值明显升高,Bax、cleaved Caspase-3蛋白表达量明显降低(P<0.05)。GEN+TBHQ组对H/R损伤皮质神经元活力、凋亡率、氧化应激指标、Nrf2/HO-1信号通路相关mRNA和蛋白表达的调控作用均明显优于GEN组和TBHQ组(P<0.05)。结论GEN可通过促进Nrf2/HO-1信号通路活化抑制氧化应激损伤和神经元凋亡,对皮质神经元H/R损伤起到保护作用。