LPA对牦牛卵丘细胞扩张因子HAS2、PTGS2和PTX3表达的影响

本研究以溶血磷脂酸(lysophosphatidic acid,LPA)在卵丘细胞扩张中的作用为切入点,旨在探讨不同浓度LPA对牦牛卵丘细胞(yak cumulus cells,YCCs)中卵丘扩张因子(透明质酸合成酶2(hyaluronate synthase 2,HAS2)、前列腺素内过氧化物合酶2(prostaglandin-endoperoxide synthase 2,PTGS2)和正五聚蛋白3(pentraxin 3,PTX3))表达的影响。本研究以健康成年(3~4岁)雌牦牛的YCCs为研究对象,取对数生长期的YCCs,将不同浓度的LPA(空白对照、阴性对照、5、15、30和50μmol·L^(-1))作用于体外培养的YCCs,分别培养12、24、36、48 h后,CCK-8检测YCCs的细胞活性,采用RT-qPCR和Western-blot法检测YCCs中HAS2、PTGS2和PTX3 mRNA和蛋白相对表达量,细胞免疫荧光染色法检测卵丘扩张因子HAS2、PTGS2和PTX3在YCCs中的分布。试验中每个处理组3个重复。结果显示,LPA孵育12、24、36和48 h,对YCCs的活性有着明显的促进作用。当孵育时间为24 h时,LPA对YCCs活性的促进作用最明显。相比较于对照组而言,当LPA浓度为15μmol·L^(-1)时,不同孵育时间中YCCs的活性提升最为显著(P<0.05)。当LPA浓度为15μmol·L^(-1)时,与空白对照组相比,HAS2、PTGS2和PTX3的mRNA和蛋白相对表达量最高(P<0.05),且YCCs中HAS2、PTGS2和PTX3的荧光强度明显增强。当LPA浓度大于15μmol·L^(-1)时,HAS2、PTGS2和PTX3的mRNA和蛋白相对表达量逐渐下降。本研究表明,LPA对YCCs活性具有促进作用。当孵育时间为24 h,并且LPA浓度为15μmol·L^(-1)时,活性提升最为显著(P<0.05)。LPA可以增强YCCs中卵丘扩张因子HAS2、PTGS2、PTX3的表达,且其作用浓度具有剂量依赖性,最佳浓度为15μmol·L^(-1)。研究结果为阐明LPA促进牦牛卵丘细胞扩张的分子机制提供了理论依据,为进一步提高牦牛卵母细胞的质量和体外受精(in vitro fertilization,IVF)成功率提供理论基础。

production of extracellular lysophosphatidic acid in the regulation of adipocyte functions and liver fibrosis

Lysophosphatidic acid(LPA), a glycerophospholipid, consists of a glycerol backbone connected to a phosphate head group and an acyl chain linked to sn-1 or sn-2 position. In the circulation, LPA is in submillimolar range and mainly derived from hydrolysis of lysophosphatidylcholine, a process mediated by lysophospholipase D activity in proteins such as autotaxin(ATX). Intracellular and extracellular LPAs act as bioactive lipid mediators with diverse functions in almost every mammalian cell type. The binding of LPA to its receptors LPA1-6 activates multiple cellular processes such as migration, proliferation and survival. The production of LPA and activation of LPA receptor signaling pathways in the events of physiology and pathophysiology have attracted the interest of researchers. Results from studies using transgenic and gene knockout animals with alterations of ATX and LPA receptors genes, have revealed the roles of LPA signaling pathways in metabolic active tissues and organs. The present review was aimed to summarize recent progresses in the studies of extracellular and intracellular LPA production pathways. This includes the functional, structural and biochemical properties of ATX and LPA receptors. The potential roles of LPA production and LPA receptor signaling pathways in obesity, insulin resistance and liver fibrosis are also discussed.

Sphingosine kinase 1 is upregulated with lysophosphatidic acid receptor 2 in human colorectal cancer

AIM: To examine the expression of SphK1, an oncogenic kinase that produces sphingosine 1-phosphate (S1P), and its correlation with the expression of LPAR2, a major lysophosphatidic acid (LPA) receptor overexpressed in various cancers, in human colorectal cancer.METHODS: Real-time reverse-transcription polymerase chain reaction was used to measure the mRNA expression of SphK1, LPAR2, and the three major S1P receptors in 27 colorectal cancer samples and corresponding normal tissue samples. We also examined the correlation between the expression of SphK1 and LPAR2.RESULTS: Colorectal cancer tissue in 22 of 27 patients had higher levels of SphK1 mRNA than in normal tissue. In two-thirds of the samples, SphK1 mRNA expression was more than two-fold higher than in normal tissue. Consistent with previous reports, LPAR2 mRNA expression in 20 of 27 colorectal cancer tissue samples was higher compared to normal tissue samples. Expression profiles of all three major S1P receptors, S1PR1, S1PR2, and S1PR3, varied without any trend, with no significant difference in expression between cancer and normal tissues. A highly significant positive correlation was found between SphK1 and LPAR2 expression [Pearson&#x02019;s correlation coefficient (r) = 0.784 and P &#x0003c; 0.01]. The mRNA levels of SphK1 and LPAR2 did not correlate with TNM stage.CONCLUSION: Our findings suggest that S1P and LPA may play important roles in the development of colorectal cancer via the upregulation of SphK1 and LPAR2, both of which could serve as new therapeutic targets in the treatment of colorectal cancer.

Effects of lysophosphatidic acid on human periodontal ligament stem cells from teeth extracted from dental patients

Despite their potential applications in future regenerative medicine, periodontal ligament stem cells(PDLSCs) are difficult to obtain in large amounts from patients. Therefore, maintaining sternness while expanding the cell numbers for medical use is the key to transitioning PDLSCs from the bench to the clinic. Lysophosphatidic acid(LPA), which is present in the human body and saliva, is a signaling molecule derived from phospholipids. In this study, we examined the effects of LPA on sternness maintenance in human PDLSCs. Several spindle-shaped and fibroblast-like periodontal ligament stem-like cell lines were established from PDLSC isolation. Among these cell lines, the most morphologically appropriate cell line was characterized. The expression levels of OCT4, NANOG(a stem cell marker), and CD90(a mesenchymal stem cell marker) were high. However, CD73(a negative marker of mesenchymal stem cells) expression was not observed. Notably, immunofluorescence analysis identified the expression of STRO-1, CD146(a mesenchymal stem cell marker), and sex determining region Y-box 2 at the protein level. In addition, lipid droplets were stained by Oil red O after the induction of adipogenesis for 21 days, and mineralized nodules were stained by Alizarin Red S after the induction of osteogenesis for 14 days. Alkaline phosphate staining also demonstrated the occurrence of osteogenesis. In summary, we established a human PDLSC line, which could be applied as a cell source for tissue regeneration in dental patients. However, further studies are needed to determine the detailed effects of LPA on PDLSCs.

Effects of lysophosphatidic acid on human colon cancer cells and its mechanisms of action

AIM： To study the effects of lysophosphatidic acid （LPA） on proliferation, adhesion, migration, and apoptosis in the human colon cancer cell line, SW480, and its mechanisms of action. METHODS： Methyl tetrazolium assay was used to assess cell proliferation. Flow cytometry was employed to detect cell apoptosis. Cell migration was measured by using a Boyden transweU migration chamber. Cell adhesion assay was performed in 96-well plates according to protocol. RESULTS： LPA significantly stimulated SW480 cell proliferation in a dose-dependent and timeependent manner compared with the control group （P 〈 0.05） while the mitogen-activated protein kinase （MAPK） inhibitor, PD98059, significantly blocked the LPA stimulation effect on proliferation. LPA also significantly stimulated adhesion and migration of SW480 cells in a dosedependent manner （P 〈 0.05）. Rho kinase inhibitor, Y-27632, significantly inhibited the upegulatory effect of LPA on adhesion and migration （P 〈 0.05）. LPA significantly protected cells from apoptosis induced by the chemotherapeutic drugs, cisplatin and 5-FU （P 〈 0.05）, but the phosphoinositide 3-kinase （PI3K） inhibitor, LY294002, significantly blocked the protective effect of LPA on apoptosis. CONCLUSION： LPA stimulated proliferation, adhesion,migration of 5W480 cells, and protected from apoptosis. The Ras/Raf-MAPK, G12/13-Rho-RhoA and PI3K- AKT/PKB signal pathways may be involved.

Yangxueqingnao particles inhibit rat vascular smooth muscle cell proliferation induced by lysophosphatidic acid

Objective: To observe the effect of Yangxueqingnao particles on rat vascular smooth muscle cell (VSMC) prolif- eration induced by lysophosphatidic acid (LPA). Methods: The amount of 3H-TdR (3H-thymidine) admixed in cultured rat VSMC was measured and mitogen-activated protein kinase (MAPK) activity and lipid peroxidation end product malondialdehyde (MDA) content of the VSMC were assayed. Results: 1×10?9, 1×10?8, 1×10?7 mol/L LPA in a concentration dependent manner, induced the amount of 3H-TdR admixed, MAP kinase activity, and MDA content of the cultured rat VSMC to increase. However, 5%, 10%, and 15% Yangxueqingnao serum preincubation resulted in a decrease of 23.0%, 42.0%, and 52.0% (P<0.01) respectively in the amount of 3H-TdR admixed, a decline in VSMC MAP kinase activity of 13.9% (P<0.05), 29.6% (P<0.01), and 48.9% (P<0.01) respectively, and also, a decrease in MDA content of VSMC of 19.4%, 24.7%, and 43.2% (P<0.01) respectively, in the 1×10?7 mol/L LPA-treated VSMC. Conclusions: LPA activates the proliferation and lipid peroxidation of VSMC in a concentration dependent manner. The LPA-induced VSMC proliferation is related to the activity of MAP kinases, enzymes involved in an intracellular signalling pathway. The results of the present study showed that Yangxueqingnao particles can effectively inhibit LPA-induced VSMC proliferation, MAP kinase activation, and reduce lipid peroxidative lesion.

Lysophosphatidic acid transactivates both c-Met and epidermal growth factor receptor, and induces cyclooxygenase-2 expression in human colon cancer LoVo cells

AIM： To examine whether lysophosphatidic acid （LPA） induces phosphorylation of c-Met and epidermal growth factor receptor （EGFR）, both of which have been proposed as prognostic markers of colorectal cancer, and whether LPA induces cyclooxygenase-2 （COX-2） expression in human colon cancer cells. METHODS： Using a human colon cancer cell line, LoVo cells, we performed immunoprecipitation analysis, followed by Western blot analysis. We also examined whether LPA induced COX-2 expression, by Western blot analysis. RESULTS： Immunoprecipitation analysis revealed that 10 μmol/L LPA induced tyrosine phosphorylation of c-Met and EGFR in LoVo cells within a few minutes. We found that c-Met tyrosine phosphorylation induced by LPA was not attenuated by pertussis toxin or a matrix metalloproteinase inhibitor, in marked contrast to the results for EGFR. In addition, 0.2-40 IJmol/L LPA induced COX-2 expression in a dose-dependent manner. CONCLUSION： Our results suggest that LPA acts upstream of various receptor tyrosine kinases （RTKs） and COX-2, and thus may act as a potent stimulator of colorectal cancer. 2005 The WJG Press and Elsevier Inc. All rights reserved.

Lysophosphatidic acid induced nuclear translocation of nuclear factor-κB in Panc-1 cells by mobilizing cytosolic free calcium

AIM: To clarify whether Lysophosphatidic acid (LPA) activates the nuclear translocation of nuclear factor-κB (NF-κB) in pancreatic cancer. METHODS: Panc-1, a human pancreatic cancer cell line, was used throughout the study. The expression of LPA receptors was confirmed by reverse-transcript polymerase chain reaction (RT-PCR). Cytosolic free calcium was measured by fluorescent calcium indicator fura-2, and the localization of NF-κB was visualized by immunofluorescent method with or without various agents, which effect cell signaling. RESULTS: Panc-1 expressed LPA receptors, LPA1, LPA2 and LPA3. LPA caused the elevation of cytosolic free calcium dose-dependently. LPA also caused the nuclear translocation of NF-κB. Cytosolic free calcium was attenuated by pertussis toxin (PTX) and U73122, an inhibitor of phospholipase C. The translocation of NF-κB was similarly attenuated by PTX and U73122, but phorbol ester, an activator of protein kinase C, alone did not translocate NF-κB. Furthermore, the translocation of NF-κB was completely blocked by Ca2+ chelator BAPTA-AM. Thapsigargin, an endoplasmic- reticulum Ca2+-ATPase pump inhibitor, also promoted the translocation of NF-κB. Staurosporine, a proteinkinase C inhibitor, attenuated translocation of NF-κB induced by LPA. CONCLUSION: These findings suggest that protein kinase C is activated endogenously in Panc-1, and protein kinase C is essential for activating NF-κB with cytosolic calcium and that LPA induces the nuclear translocation of NF-κB in Panc-1 by mobilizing cytosolic free calcium.

KAI1/CD82 gene and autotaxin-lysophosphatidic acid axis in gastrointestinal cancers

The KAI1/CD82 gene inhibits the metastasis of most tumors and is remarkably correlated with tumor invasion and prognosis.Cell metabolism dysregulation is an important cause of tumor occurrence,development,and metastasis.As one of the important characteristics of tumors,cell metabolism dysregulation is attracting increasing research attention.Phospholipids are an indispensable substance in the metabolism in various tumor cells.Phospholipid metabolites have become important cell signaling molecules.The pathological role of lysophosphatidic acid(LPA)in tumors was identified in the early 1990s.Currently,LPA inhibitors have entered clinical trials but are not yet used in clinical treatment.Autotaxin(ATX)has lysophospholipase D(lysoPLD)activity and can regulate LPA levels in vivo.The LPA receptor family and ATX/lysoPLD are abnormally expressed in various gastrointestinal tumors.According to our recent pre-experimental results,KAI1/CD82 might inhibit the migration and metastasis of cancer cells by regulating the ATX-LPA axis.However,no relevant research has been reported.Clarifying the mechanism of ATX-LPA in the inhibition of cancer metastasis by KAI1/CD82 will provide an important theoretical basis for targeted cancer therapy.In this paper,the molecular compositions of the KAI1/CD82 gene and the ATX-LPA axis,their physiological functions in tumors,and their roles in gastrointestinal cancers and target therapy are reviewed.

Autotaxin and lysophosphatidic acid signalling in lung pathophysiology

Autotaxin(ATX or ENPP2) is a secreted glycoprotein widely present in biological fluids. ATX primarily functions as a plasma lysophospholipase D and is largely responsible for the bulk of lysophosphatidic acid(LPA) production in the plasma and at inflamed and/or malignant sites. LPA is a phospholipid mediator produced in various conditions both in cells and in biological fluids, and it evokes growth-factor-like responses, including cell growth, survival, differentiation and motility, in almost all cell types. The large variety of LPA effector functions is attributed to at least six G-protein coupled LPA receptors(LPARs) with overlapping specificities and widespread distribution. Increased ATX/LPA/LPAR levels have been detected in a large variety of cancers and transformed cell lines, as well as in non-malignant inflamed tissues, suggesting a possible involvement of ATX in chronic inflammatory disorders and cancer. In this review, we focus exclusively on the role of the ATX/LPA axis in pulmonary pathophysiology, analysing the effects of ATX/LPA on pulmonary cells and leukocytes in vitro and in the context of pulmonary pathophysi-ological situations in vivo and in human diseases.

CXCL10、LPA蛋白在卵巢癌组织中的表达及与预后的关系

目的 分析CXC趋化因子配体10(CXCL10)、溶血磷脂酸(LPA)蛋白在卵巢癌组织中的表达及与预后的关系。方法 选取2017年1月—2019年12月行卵巢切除术的103例原发性卵巢癌以及同期60例卵巢良性肿瘤为研究对象。所有患者术后取卵巢组织标本,采用免疫组化法检测CXCL10、LPA蛋白表达情况。比较不同卵巢组织CXCL10、LPA蛋白表达情况,分析CXCL10、LPA蛋白表达与卵巢癌患者临床特征及预后的关系。结果 卵巢癌组织CXCL10、LPA蛋白阳性表达率分别为68.93%(71/103)、75.73%(78/103),高于卵巢良性肿瘤组织的26.67%(16/60)、33.33%(20/60)(P<0.05);CXCL10、LPA蛋白表达与卵巢癌分化程度、国际妇产科联合会分期、腹水、淋巴结转移、肿瘤直径有关(P<0.05,P<0.01),而与年龄、组织学类型无关(P>0.05)。Kaplan-Meier生存曲线分析结果显示,CXCL10、LPA蛋白阳性表达组3年无进展生存率分别为23.94%、20.51%,低于阴性表达组的40.63%、56.00%(P<0.05)。结论 卵巢癌组织中CXCL10与LPA蛋白呈高表达状态,不同临床特征卵巢癌患者CXCL10与LPA蛋白表达存在差异,二者与预后有关。

原发性肝癌患者血清FGF19、GP73、LPA的水平及临床意义

目的探讨血清成纤维细胞生长因子19(FGF19)、高尔基体跨膜糖蛋白73(GP73)和溶血磷脂酸(LPA)在原发性肝癌患者中的水平及临床意义。方法将2020年5月至2023年5月诊治的106例原发性肝癌患者设为观察组,选取同期53例健康体检者设为对照组,对比两组的血清FGF19、GP73、LPA水平变化;采用Kendall′s tau-b法分析FGF19、GP73、LPA水平与原发性肝癌患者临床病理特征的相关性;Logistic回归模型和受试者工作特征(ROC)曲线分析FGF19、GP73、LPA联合诊断原发性肝癌的效能。结果观察组的FGF19、GP73、LPA水平均高于对照组(P<0.05)。不同TNM分期、分化程度、淋巴结转移患者的血清FGF19、GP73、LPA水平比较,差异有统计学意义(P<0.05)。FGF19、GP73、LPA水平与TNM分期、分化程度、淋巴结转移呈正相关(P<0.05)。ROC曲线分析显示,FGF19、GP73、LPA及三项联合诊断原发性肝癌的曲线下面积分别为0.833、0.846、0.864和0.945,敏感度分别为77.40%、82.10%、94.30%和92.50%,特异度分别为83.00%、79.20%、81.10%和83.00%。结论血清FGF19、GP73、LPA在原发性肝癌患者中呈异常表达,监测其水平变化有助于为及时发现原发性肝癌提供重要依据。

水稻溶血磷脂酸酰基转移酶基因(LPAT)家族生物信息学分析及在籽粒油脂合成中的作用

【目的】挖掘水稻溶血磷脂酸酰基转移酶(Lysophosphatidic acid acyltransferase,LPAT)基因家族成员信息,分析其生物信息学特征及在水稻籽粒油脂合成中的作用。【方法】通过生物信息学分析,对OsLPAT基因家族进行基因结构、系统进化树、组织表达谱以及激素和逆境表达谱等分析。利用比较代谢组和转录组学的方法,分析OsLPAT家族各成员在水稻籽粒油脂合成中的作用。【结果】水稻基因组中LPAT家族包含5个成员,分别命名为OsLPAT1~OsLPAT5。除OsLPAT1外,其他成员均含有4~6个外显子,且各成员均包含Acyltransferase C-terminus(PF01553)结构域;进化分析显示,LPAT基因在单子叶植物中较为保守;OsLPAT2的表达受多种逆境条件及激素诱导,OsLPAT3和OsLPAT5的表达受到脱落酸(Abscisic acid,ABA)的强烈诱导,而OsLPAT4则特异性地受到渗透胁迫的诱导;比较代谢组和转录组学分析显示,OsLPAT家族各成员分别在籽粒发育的不同阶段发挥功能,从而促进油脂(Triacylglycerol,TAG)合成。【结论】水稻基因组中存在5个LPAT基因,其表达受到不同激素的诱导,可能在水稻激素响应过程中发挥作用。该家族成员参与了不同发育阶段籽粒中的油脂前体磷脂酸(Phosphatidic acid,PA)的合成,从而正调控籽粒中的油脂合成。

miR-129-3p与LPAR3在肝癌细胞中的表达水平及其靶向关系研究

目的探究微小RNA(miR)-129-3p与溶血磷脂酸受体3(LPAR3)在肝癌细胞中的表达水平及其靶向调控机制。方法细胞学实验:选取3种肝癌细胞系HepG2、BEL-7402、SMMC-7721及正常肝细胞系HL-7702作为研究对象,通过实时荧光定量聚合酶链式反应(qPCR)检测miR-129-3p与LPAR3的表达水平,生物信息学软件DBmiR预测miR-129-3p与LPAR3的靶向关系,并使用双荧光报告实验探究miR-129-3p与LPAR3在4种细胞系HepG2、BEL-7402、SMMC-7721和HL-7702的靶向关系。裸鼠荷瘤实验:将20只裸鼠分为野生组(n=10)和miR-129-3p组(n=10),分别接种野生型HepG2细胞和转染miR-129-3p的HepG2细胞,绘制肿瘤生长曲线并通过小动物活体成像检测肿瘤体积,qPCR和western blot检测肿瘤组织miR-129-3p,LPAR3的表达水平及PI3K/AKT信号通路分子。结果细胞实验表明,与对照细胞相比,3种肝癌细胞中miR-129-3p显著低表达,LPAR3显著高表达,差异有统计学意义(P<0.01),生物信息学软件和双荧光报告实验证实miR-129-3p与LPAR3具有靶向调控关系(P<0.01)。裸鼠荷瘤实验表明:与对照组相比,miR-129-3p组肿瘤生长显著受到抑制,肿瘤体积显著降低,qPCR和Western blot实验表明肿瘤组织中miR-129-3p显著升高,LPAR3、PI3K、AKT表达水平显著降低,差异有统计学意义(P<0.01)。结论miR-129-3p在肝癌细胞中显著低表达,与LPAR3具有靶向调控关系;miR-129-3p的高表达水平可以显著抑制肿瘤生长,其机制可能与PI3K/AKT信号通路的调控相关。

血浆D-D、LPA、t-PA对髋关节置换患者并发下肢深静脉血栓的诊断价值

目的探讨血浆D-二聚体(D-D)、溶血磷脂酸(LPA)及组织型纤溶酶原激活剂(t-PA)对髋关节置换患者并发下肢深静脉血栓(DVT)的诊断价值。方法选取2017年1月至2019年3月我院实施髋关节置换手术的148例患者为研究对象,根据患者术后是否并发DVT分为DVT组23例和非DVT组125例;对比分析两组患者血浆中D-D、LPA、t-PA水平及凝血功能指标,采用受试者工作曲线(ROC)分析三项指标对DVT的诊断价值。结果DVT组患者血浆D-D、LPA水平高于非DVT组(P<0.05),t-PA水平低于非DVT组(P<0.05);DVT组患者血浆PT、APTT、INR水平低于非DVT组(P<0.05),PLT水平高于非DVT组(P<0.05);血浆D-D、LPA、t-PA单独检测诊断髋关节置换患者并发DVT的灵敏度、特异度、AUC分别为86.96%、72.00%和0.871,82.61%、75.20%和0.846,65.22%、67.20%和0.694;三指标联合检测能明显提高诊断效能,灵敏度、特异度、AUC分别为91.30%、88.41%和0.944。结论血浆D-D、LPA、t-PA单独检测对髋关节置换患者并发DVT均具有较高的临床诊断价值,而三者联合检测诊断价值更高。

Circular RNA LPAR3通过miR-143-5p/USP14通路调控食管癌细胞糖酵解的研究

目的探究调控食管癌细胞糖酵解的机制。方法逆转录实时定量PCR(quantitative real-time,qRT-PCR)法分别检测环状RNA溶血磷脂酸受体(circular RNA lysophosphatidic acid receptor 3,CircLPAR3),微小RNA-143-5(microRNA-143-5p,miR-143-5p)和泛素特异性蛋白酶14(ubiquitin-specific protease 14,USP14)的表达水平;使用Seahorse XF 24细胞外通量分析仪测量细胞外酸化率(extracellular acidification rate,ECAR)和耗氧率(oxygen comsumpition rate,OCR);Western印迹法检测糖酵解途径关键酶HK2的表达水平;使用Starbase数据库预测CircLPAR3与miR-143-5p以及miR-143-5p与USP14的靶向结合位点,双荧光素酶试验验证CircLPAR3和miR-143-5p以及miR-143-5p与USP14的靶向关系。结果与正常人食管上皮细胞(normal human esophageal epithelial cells,Het-1A)组比较,食管癌细胞系中CircLPAR3高表达,且KYSE450细胞系的表达最高,同时miR-143-5p低表达而USP14高表达。与Het-1A组比较,KYSE450组HK2的蛋白表达水平增加,同时细胞ECAR增加,整体糖酵解OCR减少;敲低CircLPAR3的表达导致细胞HK2的蛋白表达水平减少,ECAR减少,整体糖酵解OCR增加;在抑制CircLPAR3的表达情况下,抑制miR-143-5p的表达能够部分逆转细胞的糖酵解途径;双荧光素酶实验验证了CircLPAR3和miR-143-5p的靶向关系,以及miR-143-5p和USP14的靶向关系。结论CircLPAR3能够通过调控miR-143-5p/USP14通路调节食管癌细胞糖酵解途径。

LPAR1基因对脂多糖刺激下Ⅱ型肺泡上皮细胞中组织因子和纤溶酶原激活物抑制剂1表达的影响

目的 了解脂多糖(LPS)刺激下Ⅱ型肺泡上皮细胞(AEC Ⅱ)中差异表达基因溶血磷脂酸受体1(LPAR1)的动态表达特点,以及该基因对LPS刺激下组织因子(TF)和纤溶酶原激活物抑制剂1(PAI-1)表达的调节作用。方法 对LPS损伤后的RLE-6TN细胞进行RNA-seq测序,通过生物信息学分析,以NF-κB信号通路为核心,找到该信号通路富集到的的差异表达基因LPAR1。使用LPS刺激处于生长对数期的RLE-6TN细胞6、12、24、48及72 h,检测细胞中LPAR1基因的动态表达水平。在此基础上通过慢病毒转染敲低细胞中LPAR1基因的表达,观察该基因对细胞中TF和PAI-1表达的调节作用。结果 RLE-6TN细胞中LPAR1的表达在刺激6 h后开始升高,24 h时候达到顶峰,之后逐渐降低;利用慢病毒转染技术,成功低敲了细胞中LPAR1基因水平。LPS刺激24 h后,RLE-6TN细胞中TF和PAI-1的mRNA和蛋白的表达水平与正常对照组比较显著增加,差异有统计学意义(P <0.05);低敲LPAR1基因后,TF和PAI-1的表达水平显著降低(P <0.05),接近正常对照水平(P> 0.05)。结论 LPAR1基因对LPS刺激下RLE-6TN细胞TF和PAI-1的表达具有调节作用。该基因有望成为纠正ARDS肺泡内纤维蛋白沉积的新靶标。

针灸配合枳术汤对ACI患者神经功能及LPA的影响

目的探讨针灸配合枳术汤对急性脑梗死(acute cerebral infarct,ACI)患者神经功能及溶血磷脂酸(lysophosphatidic acid,LPA)的影响。方法选择2019年4月至2022年4月86例急性脑梗死患者,随机分为两组各43例,对照组予以常规西医治疗,试验组在对照组基础上加针灸配合枳术汤治疗。治疗前后采用美国国立卫生院神经功能缺损评分(national institute of health stroke scale,NIHSS)、Fugl-Meyer评分(FMA)、巴塞尔指数(Barthel)对患者进行评估,并检测LPA水平。结果两组的NIHSS评分都有降低的趋势,且试验组更低,差异有统计学意义(P<0.05)。另两项评分均有明显提高,且试验组更高,差异有统计学意义(P<0.05)。两组LPA水平均明显降低,但差异无统计学意义(P>0.05)。试验组愈显率比对照组高一些,差异有统计学意义(P<0.05),但两组之间的总有效率比较差异无统计学意义(P>0.05)。结论针灸配合枳术汤可有效促进ACI患者的神经功能恢复,提高临床疗效,但对血浆LPA无明显影响。

脑梗死患者脑脊液溶血磷脂酸水平与神经功能缺损状态的相关性分析

目的探讨与分析脑梗死(cerebral infarction,CI)患者脑脊液溶血磷脂酸(plasma lysophosphatidic acid,LPA)水平与神经功能缺损状态的相关性分析。方法选择2020年3月—2022年11月在北京市昌平区中西医结合医院诊治的67例脑梗死患者作为研究对象,所有患者在入院时进行神经功能缺损状态美国国立卫生研究院卒中量表(National Institute of Health Stroke Scale,NIHSS)评分,检测脑脊液溶血磷脂酸水平并进行相关性分析。结果在67例患者中,平均神经功能缺损状态NIHSS评分为(18.32±2.15)分,其中>21分17例(重度组)。重度组性别、身体质量指数、受教育程度、合并疾病等与非重度组对比,差异无统计学意义(P>0.05),重度组与非重度组的年龄、发病到入院时间等对比,差异有统计学意义(P<0.05)。重度组与非重度组的血糖、同型半胱氨酸、总胆固醇、甘油三酯含量对比,差异无统计学意义(P>0.05),重度组的低密度脂蛋白胆固醇含量高于非重度组,差异有统计学意义(P<0.05)。重度组的脑脊液溶血磷脂酸水平显著高于非重度组,差异有统计学意义(P<0.05)。Pearson分析显示NIHSS评分与年龄、发病到入院时间、脑脊液溶血磷脂酸水平都存在正相关性(P<0.05)。logistic回归分析显示年龄、发病到入院时间、脑脊液溶血磷脂酸水平为影响脑梗死患者NIHSS评分的重要因素(P<0.05)。结论脑梗死患者的神经功能缺损状态比较严重,神经功能缺损状态越差的脑梗死患者,脑脊液溶血磷脂酸水平越高,脑梗死患者脑脊液溶血磷脂酸水平与神经功能缺损状态存在相关性。