Intra-arterial lipo-prostaglandin E1 infusion for arterial spasm in liver transplantation:A case report

BACKGROUND Hepatic artery obstruction is a critical consideration in graft outcomes after living donor liver transplantation.We report a case of diffuse arterial vasospasm that developed immediately after anastomosis and was managed with an intra-arterial infusion of lipo-prostaglandin E1(PGE1).CASE SUMMARY A 57-year-old male with hepatitis B virus-related liver cirrhosis and hepatocellular carcinoma underwent ABO-incompatible living donor liver transplant.The grafted hepatic artery was first anastomosed to the recipient’s right hepatic artery stump.However,the arterial pulse immediately weakened.Although a new anastomosis was performed using the right gastroepiploic artery,the patient’s arterial pulse rate remained poor.We attempted angiographic intervention immediately after the operation;it showed diffuse arterial vasospasms like‘beads on a string’.We attempted continuous infusion of lipo-PGE1 overnight via an intra-arterial catheter.The next day,arterial flow improved without any spasms or strictures.The patient had no additional arterial complications or related sequelae at the time of writing,1-year post-liver transplantation.CONCLUSION Angiographic evaluation is helpful in cases of repetitive arterial obstruction,and intra-arterial infusion of lipo-PGE1 may be effective in treating diffuse arterial spasms.

Effects of prostaglandin E combined with continuous renal replacement therapy on septic acute kidney injury

BACKGROUND The effects of prostaglandin E(PGE)combined with continuous renal replacement therapy(CRRT)on renal function and inflammatory responses in patients with septic acute kidney injury(SAKI)remain unclear.AIM To investigate the effects of PGE combined with CRRT on urinary augmenter of liver regeneration(ALR),urinary Na+/H+exchanger 3(NHE3),and serum inflammatory cytokines in patients with SAKI.METHODS The clinical data of 114 patients with SAKI admitted to Yichang Second People's Hospital from May 2017 to January 2019 were collected.Fifty-three cases treated by CRRT alone were included in a control group,while the other 61 cases treated with PGE combined with CRRT were included in an experimental group.Their urinary ALR,urinary NHE3,serum inflammatory cytokines,renal function indices,and immune function indices were detected.Changes in disease recovery and the incidence of adverse reactions were observed.The 28-d survival curve was plotted.RESULTS Before treatment,urinary ALR,urinary NHE3,blood urea nitrogen(BUN),serum creatinine(SCr),CD3+T lymphocytes,CD4+T lymphocytes,and CD4+/CD8+T lymphocyte ratio in the control and experimental groups were approximately the same.After treatment,urinary ALR and NHE3 decreased,while BUN,SCr,CD3+T lymphocytes,CD4+T lymphocytes,and CD4+/CD8+T lymphocyte ratio increased in all subjects.Urinary ALR,urinary NHE3,BUN,and SCr in the experimental group were significantly lower than those in the control group,while CD3+T lymphocytes,CD4+T lymphocytes,and CD4+/CD8+T lymphocyte ratio were significantly higher than those in the control group(P<0.05).After treatment,the levels of tumor necrosis factor-α,interleukin-18,and high sensitivity C-reactive protein in the experimental group were significantly lower than those in the control group(P<0.05).The time for urine volume recovery and intensive care unit treatment in the experimental group was significantly shorter than that in the control group(P<0.05),although there was no statistically significant difference in hospital stays between the two groups.The total incidence of adverse reactions did not differ statistically between the two groups.The 28-d survival rate in the experimental group(80.33%)was significantly higher than that in the control group(66.04%).CONCLUSION PGE combined with CRRT is clinically effective for treating SAKI,and the combination therapy can significantly improve renal function and reduce inflammatory responses.

Protective effects of prostaglandin E1 perfusion againstspinal cord ischemia-reperfusion injury in a rabbit model

BACKGROUND： Prostaglandin E1 （PGE1） is known to be protective in ischemia-reperfusion of heart, lung,renal, and liver tissue. It still remains to be determined whether PGE1 exhibits similar protection against spinal cord ischemia-reperfusion injury in a rabbit model. OBJECTIVE： To observe the large, ventral horn, motor neurons of the spinal cord, as well as limb function, and to investigate whether perfusion of PGE1 exhibits protective effects against spinal cord ischemia-reperfusion injury in a rabbit model. DESIGN, TIME AND SETTING： Controlled observation. The experiment was performed at the Department of Orthopedics, First Affiliated Hospital of Liaoning Medical University between June and October 2007. MATERIALS： Twenty male, New Zealand white rabbits, weighing 2.0 kg and of mixed gender, were used in the present study. The following chemicals and compounds were used： prostaglandin E1 injectable powder,as well as malondialdehyde and ATPase kits. Animal intervention was in accordance with animal ethical standards. METHODS： We separated rabbits into control and experimental groups randomly, with 10 rabbits in each group. Rabbits were used as spinal cord ischemia models by segmentally cross-clamping the infrarenal aorta. The control group was subsequently perfused for five minutes with blood and saline solution, and the experimental group was perfused for 5 minutes with blood and saline solution containing PGE1 （100 ng/kg/min）. MAIN OUTCOME MEASURES： The neurological function of the hind limbs was assessed 12, 24, and 48 hours after model establishment. All animals were sacrificed and spinal cords were harvested for histological analyses. The large motor neurons in the ventral horn of L1-7 were observed by inverted microscope. RESULTS： All 20 rabbits were included in the final analysis, without any loss. In the ventral horn of the L5-7 segments, there were more large motor neurons that appeared viable in the experimental group than the control group （P 〈 0.05）. The scores of hind limb functions were greater in the experimental group after 12, 24, and 48 hours （P 〈 0.01）. CONCLUSION： Perfusion of PGE1 reduced the amount of neuronal damage in the spinal cord ischemia-reperfusion injury rabbit model. These results correlated with increased numbers large motor neurons in the ventral horn of the spinal cord, as well as improved hind limb function.

Expression of prostaglandin E_2 receptors in rat hippocampus

BACKGROUND: Prostaglandin E2 (PGE2) can directly regulate toxic injury of hippocampal neurons through participation by its receptor. Increase of excitability of hippocampal membrane and long-term synaptic elasticity are closely related to PGE2, PGD2 and PGF2A. This suggests that PGE2 may be a key molecule of neuronal signal passage and regulate the existence of neurons through its receptor. However, which isoforms of PGE2 receptor expressing in hippocampal neurons is still unclear. OBJECTIVE: To research the subtype expression of PGE2 receptor in hippocampus of rats through mRNA transcription and protein interpretation. DESIGN: Animal studies with random, control and operator and designer double-blind methods. SETTING: University of South Carolina, Animal Center. MATERIALS: Sprague-Dawley rats, aged 12 weeks, weighing 200 g, females 48 and males 48, were selected from Animal Center in South Carolina University. Tri ReagentTM kit was provided by Molecular Research Center, USA. METHODS: The experiment was carried out in Animal Center in South Carolina University from January to December 2005. The expression of the PGE2 receptors was profiled and compared in rat hippocampus using real-time RT-PCR and Western-blot techniques. MAIN OUTCOME MEASURES: Expression of PGE2 receptors in various isoforms of hippocampal neurons of rats. RESULTS: mRNAs of all four EP1-4 subtypes were detected in the hippocampus. Western-blot data showed consistently detectable bands at approximately Mr 50 000 of EP1, Mr 40 000 and Mr 52 000 of EP2, Mr 45 000, Mr 57 000 and Mr 105 000 of EP3, and Mr 46 000 of EP4. CONCLUSION: Identifying four subtypes of EPs heterogeneously expresses in the hippocampus.

The effect of prostaglandin E_1 on recovery of early renal graft functions after transplantation

Objective To investigate the effect of prostaglandin E1 (PGE1) on recovery of early renal graft functions after transplantation. Methods One hundred and seven patients after renal transplantation were allocated in the treated group, and treated by conventional treatment with injection of 10 μg prostaglandin E1 additionally twice a day for 14 days. And eighty-eight patients who received conventional treatment alone after renal transplantation at the corresponding period were allocated in the control group. Indexes of the two groups, including incidence of delayed graft function and acute rejection reaction, volume of urine, serum certaintie (SCr), endogenous certainties clearance rate (CCr), the blood flow resistance in graft as well as blood viscosity (BV), and platelet aggregation rate (PAR), were determined. Results The urinary volume and endogenous certainties clearance rate of the treated group were significantly higher, but the level of SCr, incidence of renal function recovery retardation, BV, PAR and blood flow resistance in graft were significantly lower than those of the control group (P<0.05). The difference of incidence of acute rejection reaction between the two groups was insignificant (P>0.05). Conclusion Prostaglandin E1 can improve blood microcirculation and decrease the incidence of renal function recovery retardation. These effects are helpful for recovery of renal function after renal transplantation.

Mating behaviors in ovoviviparous black rockfish(Sebastes schlegelii):molecular function of prostaglandin E2 as both a hormone and pheromone

Prostaglandins(PGs)are profound hormones in teleost sexual behavior,especially in mating.PGs act as pheromones that affect the olfactory sensory neurons of males,inducing the initiation of a series of mating behaviors.However,the molecular mechanism by which PGs trigger mating behavior in ovoviviparous teleosts is still unclear.In the present study,we employed the ovoviviparous black rockfish(Sebastes schlegelii),an economically important marine species whose reproductive production is limited by incomplete fertilization,as a model species.The results showed that when the dose of PGE2 was higher than 10 nmol/L,a significant(P<0.05)increase in mating behaviors was observed.Dual-fluorescence in situ hybridization indicated that PGE2 could fire specific neurons in different brain regions and receptor cells in the olfactory sac.After combining with specific neurons in the central nervous system(CNS),a series of genes related to reproduction are activated.The intracerebroventricular administration of PGE_(2) significantly increased lhb levels(P<0.05)in both sexes.Moreover,steroidogenesis in gonads was also affected,inducing an increase(P<0.05)in E_(2) levels in males and T levels in females.PGE_(2) levels were also increased significantly(P<0.05)in both sexes.The present study revealed that PGE2 can activate mating behavior in black rockfish in both hormone and pheromone pathways,leading to variations in sex steroid levels and activation of reproductive behaviors.Our results provide not only novel insight into the onset of mating behaviors in ovoviviparous teleosts but also solutions for the incomplete fertilization caused by natural mating in cage aquaculture.

Pathogenesis of chronic enteropathy associated with the SLCO2A1 gene:Hypotheses and conundrums

Chronic enteropathy associated with the SLCO2A1 gene(CEAS)is a complex gastroenterological condition characterized by multiple ulcers in the small intestine with chronic bleeding and protein loss.This review explores the potential mechanisms underlying the pathogenesis of CEAS,focusing on the role of SLCO2A1-encoded prostaglandin transporter OATP2A1 and its impact on prostaglandin E2(PGE2)levels.Studies have suggested that elevated PGE2 levels contribute to mucosal damage,inflammation,and disruption of the intestinal barrier.The effects of PGE2 on macrophage activation and Maxi-Cl channel functionality,as well as its interaction with nonsteroidal anti-inflammatory drugs play crucial roles in the progression of CEAS.Understanding the balance between its protective and pro-inflammatory effects and the complex interactions within the gastrointestinal tract can shed light on potential therapeutic targets for CEAS and guide the development of novel,targeted therapies.

Effect of Intravenous Administration of Liposomal Prostaglandin E1 on Microcirculation in Patients with ST Elevation Myocardial Infarction Undergoing Primary Percutaneous Intervention

Background：Several studies have demonstrated that primary percutaneous coronary intervention （PCI） can result in reperfusion injury.This study aims to investigate the effectiveness of liposomal prostaglandin E l （Lipo-PGE1,Alprostadil,Beijing Tide Pharmaceutical Co.,Ltd.） for enhancing microcirculation in reperfusion injury.In addition,this study determined the optimal administration method for acute ST elevation myocardial infarction （STEMI） patients undergoing primary PCI.Methods：Totally,68 patients with STEMI were randomly assigned to two groups：intravenous administration ofLipo-PGE 1 （Group A）,and no Lipo-PGE1 administration （Group B）.The corrected thrombolysis in myocardial infarction （TIMI） frame count （cTFC） and myocardial blush grade （MBG） were calculated.Patients were followed up for 6 months.Major adverse cardiac events （MACE） were also measured.Results：There was no significant difference in the baseline characteristics between the two groups.The cTFC parameter in Group A was significantly lower than Group B （18.06 ± 2.06 vs.25.31 ± 2.59,P ＜ 0.01）.The ratio of final MBG grade-3 was significantly higher （P ＜ 0.05） in Group A （87.9％） relative to Group B （65.7％）.There was no significant difference between the two groups in final TIMI-3 flow and no-reflow.Patients were followed up for 6 months,and the occurrence of MACE in Group A was significantly lower than that in Group B （6.1％ vs.25.9％ respectively,P ＜ 0.05）.Conclusions：Myocardial microcirculation of reperfusion injury in patients with STEMI,after primary PCI,can be improved by administering Lipo-PGE1.

Adiponectin receptor 1 mediates the difference in adiponectininduced prostaglandin E2 production in rheumatoid arthritis and osteoarthritis synovial fibroblasts

The synovial fluid concentrations of adiponectin are significantly higher in patients with rheumatoid arthritis （RA） than in patients with osteoarthritis （OA）. Accumulating evidence suggests that adiponectin may be an inducer of inflammation in arthritis, but the mechanism remains unclear. The objectives of this study were to compare the expression levels of adiponectin receptors in rheumatoid arthritis synovial fibroblasts （RASF） and osteoarthritis synovial fibroblasts （OASF）, evaluate the roles of adiponectin receptors in adiponectin-induced prostaglandin E2 （PGE2） production, and then investigate the effects of a nonsteroidal anti-inflammatory drug （NSAID） and a cyclooxygenase （COX）-2-selective inhibitor on adiponectin-induced PGE2 release. Methods The expressions of adiponectin receptor 1 （AdipoR1） and AdipoR2 mRNA and protein in synovial fibroblasts from seven patients with RA and eight patients with OA undergoing total knee replacement were evaluated by real-time polymerase chain reaction, immunofluorescence microscopy and Western blotting analysis. Adiponectin-induced PGE2 production was detected by enzyme-linked immunosorbent assay. RNA interference against the AdipoR1 and AdipoR2 genes was performed to investigate the effects of the adiponectin receptors on adiponectin-induced PGE2 production in both RASF and OASE Results AdipoR1 and AdipoR2 mRNA and protein were expressed by both RASF and OASF. Compared with OASF, RASF exhibited higher levels of AdipoR1, but there was no significant difference for AdipoR2. Adiponectin induced the production of PGE2 by the synovial fibroblasts in a concentration-dependent manner, and this was more obvious in RASF. RNA interference showed that the difference may be mediated by the diverse distribution of AdipoRl. The adiponectin-induced PGE2 production was efficiently relieved by the NSAID and COX-2-selective inhibitor. Conclusion The present findings suggest that AdipoR1 may mediate the difference in adiponectin-induced PGE2 production in RASF and OASE

Follicular fluid levels of prostaglandin E2 and the effect of prostaglandin E2 on steroidogenesis in granulosa-lutein cells in women with moderate and severe endometriosis undergoing in vitro fertilization and embryo transfer

Background The mechanisms of endometriosis with infertility have not been fully studied. The present study aimed to assess the follicular fluid （FF） levels of prostaglandin E2 （PGE2）, which plays a critical role within the ovary, and to investigate the effect of PGE2 on steroidogenesis in granulosa-lutein cells （GLCs） from women with and without endometriosis. Methods Thirty-three women with laparoscopically documented endometriosis and 40 controls undergoing in vitro fertilization （IVF） were studied. We assayed the concentrations of PGE2 in FF, the production of E2 and progesterone in FF and in culture medium, and the expression of steroidogenic acute regulatory protein （STAR） and CYP19A1 in GLCs with the intervention of PGE2. Results PGE2 and progesterone concentrations were increased and displayed positive correlation in endometriotic FE PGE2 induced the expression of STAR and the production of progesterone in GLCs from women with endometriosis, and the expression of STAR and the production of progesterone were increased in GLCs from women with endometriosis. However, there were no significant effects of PGE2 on promoting the production of E2 or the expression of CYP19A1 in GLCs. Moreover, the production of E2 and the expression of CYP19A1 in GLCs from women with endometriosis were significantly decreased compared to the controls. Conclusions PGE2 concentrations are increased in endometriotic FF, along with concomitant increases in progesterone and STAR. In contrast, the E2 and CYP19A1 are decreased in GLCs, which may delay the development of the follicles and cause an imbalance in the follicular steroid hormone levels. These changes may have close relationship with endometriosis-associated infertility.

Prostaglandin E2-EP4 signaling persistently amplifies CD40-mediated induction of IL-23 p19 expression through canonical and non-canonical NF-κB pathways

While there is mounting evidence that interleukin （IL）-23-IL-17 axis plays a critical role in the pathogenesis of various autoimmune diseases, much remains to be elucidated on how IL-23 is induced in the pathological processes. IL-23 is a heterodimer composed of p19 and p40, the latter being shared with IL-12. We previously reported that prostaglandin （PG） E2 promotes CD40-mediated induction of 1123a （p19） expression through its E receptor subtype 4 （EP4） receptor in splenic dendritic cells （DCs）. Here, we have analyzed signaling pathways regulating 1123a induction in the cross talk between EP4 and CD40 in bone marrow-derived DCs. We found that PGE2 synergistically induced 1123a transcription with CD40 signaling. An EP4 agonist, but not agonists of EP1, EP2, or EP3, reproduced this action. Stimulation of CD40 with an agonist antibody evoked biphasic induction of 1123a expression, with the early phase peaking at 1 h and the late phase peaking at 12 h and lasting up to 36 h after stimulation, whereas induction by lipopolysaccharide or tumor necrosis factor-α was transient. The early phase induction by CD40 stimulation was absent in DCs derived from Nfkbl-deficient mice, and the late phase induction was eliminated by RNA interference of nuclear factor-kappa B （NF-κB） p100 subunit. Further, cAMP response element-binding protein （CREB） depletion completely eliminated the induction of 1123a by CD40 stimulation. The addition of the EP4 agonist amplified the induction in both phases through the cAMP-protein kinase A （PKA） pathway. These results suggest that 1123a expression in DCs is synergistically triggered by the PG E2-EP4-cAMP-PKA pathway and canonical/non-canonical NF-KB pathways and CREB activated by CD40 stimulation.

咬合重建固定义齿修复术对牙周病伴牙列缺损患者颞下颌功能、脑血流速度及龈沟液IL-23、PGE_(2)的影响

目的 探讨咬合重建固定义齿修复术对牙周病(PD)伴牙列缺损患者颞下颌功能、脑血流速度及龈沟液白介素23(IL-23)、前列腺素E_(2)(PGE_(2))的影响。方法 选取2016年7月至2019年8月郑州大学第五附属医院收治的130例PD伴牙列缺损患者,根据治疗方案分为对照组和试验组,各65例。对照组接受垫式可摘局部义齿修复,试验组接受咬合重建固定义齿修复。治疗后观察6个月,统计两组修复效果、咀嚼功能、肌肉压痛指数、颞下颌关节紊乱指数、颞下颌关节功能障碍指数、龈沟液IL-23、PGE_(2)、舒张期末峰流速(Vd)、右侧大脑中动脉的收缩期峰流速(Vs)、平均峰流速值(Vm)、治疗满意度。结果 试验组总有效率高于对照组(P<0.05),咀嚼功能优良率高于对照组(P<0.05);修复6个月后试验组颞下颌关节功能障碍指数、肌肉压痛指数、颞下颌关节紊乱指数低于对照组,咀嚼状态下大脑中动脉Vs、Vd、Vm大于对照组(P<0.05),龈沟液IL-23水平低于对照组,PGE_(2)水平高于对照组(P<0.05),试验组语音功能、固位功能、咀嚼功能满意度高于对照组(P<0.05)。结论 咬合重建固定义齿修复术可改善PD伴牙列缺损患者咀嚼功能、颞下颌功能及脑血流速度,调节龈沟液IL-23、PGE_(2)水平,提高治疗满意度。

APPLICABILITY OF PROSTAGLANDIN E_1 INDIRECT PORTAL VEIN ANGIOGRAPHY IN PATIENTS WITH PORTAL HYPERTENSION

The imaging quality of the portal vein was obviously improved with prostaglandin E1 (PGE1) indirect portal vein digital subtraction angiography (DSA) in 23 cases. The time-density curve showed that the occurrence rate of opposite hepatic blood flow of splenic vein (SV) was the highest (17.4%). The total visualization rate of the left gastric vein (LGV) was 78.3%, and the visualization rate of the short gastric vein (SGV) was 36.4%. 38.9% of the LGV and all the SGV were visualized with indirect portal vein DSA through SA. Indirect portal vein angiography through superior mesenteric artery and that through splenic artery were of equal importance. In portal hypertension patients with hemorrhage of the digestive tract, when LGV and SGV could not be visualized in PGE1 indirect portal vein DSA, the possibility of non-varices vein bleeding should be considered. When opposite hepatic blood flow with obvious dilation appeared in LGV and SGV, devascularization of the pericardial blood vessels would be justifiable.

Prostaglandin E2 promotes hematopoietic development from human embryonic stem cells

Recent studies have suggested that prostaglan-din(PG)E2(PGE2)and the prostaglandin pathway are essential for hematopoietic stem cell growth and develop-ment.However,similar studies on hematopoietic commit-ment from human embryonic stem cells(hESCs)are still limited.Here we report that the addition of PGE2 promotes hematopoietic differentiation of hESCs.The induced cells from hESCs/OP9 co-culture and in the presence of PGE2 were characterized by reverse transcription-PCR(RT-PCR),flow cytometry,colony-forming assays and Wright-Giemsa staining.Our results demonstrated that PGE2 exposure could alter the gene expression pattern and morphology of co-cultured hESCs and resulted in a robust hematopoietic differentiation with higher frequencies of CD34+and CD45+cells.Furthermore,the Smad signaling pathway may be involved in PGE2 and OP9 induced hematopoietic differentiation of hESCs.This research may improve our knowledge of stem cell regulation and hopefully lead to better stem cell-based therapeutic options.

Damage control:Harnessing prostaglandin E2 as a potential healing factor of tissue injuries

Increasing prostaglandin E2 by knocking out its inhibitor 15-hydroxyprostaglandin dehydrogenase(15-PDGH)or administering a compound that inhibits 15-PDGH was recently found to improve healing in hematopoietic stem cell transplants,colitis recovery,and hepatogenesis after transection in mice.These results are suggestive of pharmacologic therapies or even genetic therapy that could improve patient outcomes,especially since the excess PGE2 and the 15-PDGH inhibitor have proven to be non-toxic.However,elevated levels of PGE2 are associated with increased risk of cancer and blood clotting problems.It would be unacceptable to treat a cancer patient with chemotherapy and replenish the hematopoietic stem cells with the help of PGE2,only to have increased expression of PGE2 and induce another cancer.Therefore,to assess the most therapeutic aspects of PGE2,it is important to consider effects that could induce disease.

Measurement of prostaglandin metabolites is useful in diagnosis of small bowel ulcerations

BACKGROUND We recently reported on a hereditary enteropathy associated with a gene encoding a prostaglandin transporter and referred to as chronic enteropathy associated with SLCO2 A1 gene(CEAS). Crohn's disease(CD) is a major differential diagnosis of CEAS, because these diseases share some clinical features. Therefore, there is a need to develop a convenient screening test to distinguish CEAS from CD.AIM To examine whether prostaglandin E major urinary metabolites(PGE-MUM) can serve as a biomarker to distinguish CEAS from CD.METHODS This was a transactional study of 20 patients with CEAS and 98 patients with CD.CEAS was diagnosed by the confirmation of homozygous or compound heterozygous mutation of SLCO2 A1. We measured the concentration of PGEMUM in spot urine by radioimmunoassay, and the concentration was compared between the two groups of patients. We also determined the optimal cut-off value of PGE-MUM to distinguish CEAS from CD by receiver operating characteristic(ROC) curve analysis.RESULTS Twenty Japanese patients with CEAS and 98 patients with CD were enrolled.PGE-MUM concentration in patients with CEAS was significantly higher than that in patients with CD(median 102.7 vs 27.9 μg/g × Cre, P < 0.0001). One log unit increase in PGE-MUM contributed to 7.3 increase in the likelihood for the diagnosis of CEAS [95% confidence interval(CI) 3.2-16.7]. A logistic regression analysis revealed that the association was significant even after adjusting confounding factors(adjusted odds ratio 29.6, 95%CI 4.7-185.7). ROC curve analysis revealed the optimal PGE-MUM cut-off value for the distinction of CEAS from CD to be 48.9 μg/g × Cre with 95.0% sensitivity and 79.6% specificity.CONCLUSION PGE-MUM measurement is a convenient, non-invasive and useful test for the distinction of CEAS from CD.

良恶性肺疾病组织前列腺素E受体4基因甲基化水平变化及其诊断价值研究

目的探讨良恶性肺疾病组织中前列腺素E受体4(PTGER4)基因甲基化水平及其诊断肺恶性肿瘤价值。方法收集我院胸外科患者术后冰冻组织标本174例,根据病理结果分为肺腺癌组60例,鳞癌组38例,错构瘤组17例、良性疾病组31例和肺正常组织组28例,收集临床基础资料,分别提取组织DNA。PTGER4基因甲基化水平检测采用甲基化敏感限制性内切酶-荧光定量PCR方法,根据PTGER4基因与内参ACTB基因的循环阈值差异(ΔCt值)判断甲基化程度。结果肺腺癌组、肺鳞癌组、良性疾病组、错构瘤组和肺正常组织组ΔCt值的中位数(四分位数)[M(P 25,P 75)]分别为0.21(-0.67,1.20)、1.75(-0.04,4.10)、6.02(4.36,7.57)、7.17(5.72,8.78)和4.61(3.65,5.10),根据ΔCt比较,肺腺癌和肺鳞癌组PTGER4甲基化水平显著高于良性疾病组、错构瘤组和肺正常组织组(P<0.05)。肺腺癌甲基化水平显著高于肺鳞癌(P=0.044)。ROC曲线分析显示,PTGER4基因甲基化诊断总的肺癌的曲线下面积(AUC)、灵敏度和特异性分别为0.923、79.6%和98.7%,诊断肺腺癌为0.984、90.0%和98.7%,鳞癌为0.828、65.8%和96.1%,腺癌AUC显著高于鳞癌(P<0.05)。肺癌患者PTGER4基因甲基化水平与年龄、性别、吸烟、肿瘤标志物(CEA、NSE、CYFRA21-1、SCC)和肿瘤分期无显著性相关(P>0.05)。结论肺恶性肿瘤组织中PTGER4基因甲基化呈现特异性升高;PTGER4基因甲基化对肺腺癌的诊断效能更高。

胃食管反流病患者反流症状评分与血清PGE2、Ghrelin及GAS水平的相关性分析

目的探讨胃食管反流病(GERD)患者反流症状评分与血清前列腺素E_(2)(PGE_(2))、胃饥饿素(Ghrelin)及胃泌素(GAS)水平的相关性。方法选择2019年8月至2022年8月陕西航天医院收治的GERD患者150例(GERD组)和同期健康体检者120例(对照组)为研究对象,采用酶联免疫吸附试验检测血清PGE_(2)、Ghrelin和GAS水平。比较GERD组与对照组血清PGE_(2)、Ghrelin、GAS水平和胃食管反流病问卷(GERD-Q)评分,分析不同GERD-Q评分患者血清PGE_(2)、Ghrelin、GAS水平,采用Pearson相关分析GERD患者GERD-Q评分与血清PGE_(2)、Ghrelin、GAS水平的相关性。结果GERD组GERD-Q评分和血清PGE_(2)水平高于对照组,血清Ghrelin、GAS水平低于对照组,差异有统计学意义(P<0.05)。GERD-Q评分为7~10分者血清PGE_(2)水平低于GERD-Q评分为11~14分和15~18分者,血清Ghrelin、GAS水平高于GERD-Q评分为11~14分和15~18分者,差异有统计学意义(P<0.05);GERD-Q评分为11~14分者血清PGE_(2)水平低于GERD-Q评分为15~18分者,血清Ghrelin、GAS水平高于GERD-Q评分为15~18分者,差异有统计学意义(P<0.05)。Pearson相关分析显示,GERD患者GERD-Q评分与血清PGE_(2)水平呈正相关(r=0.510,P<0.05),与血清Ghrelin、GAS水平均呈负相关(r=-0.524、-0.568,P<0.05)。结论血清PGE_(2)、Ghrelin、GAS水平均与GERD患者GERD-Q评分相关,有望成为评估患者反流症状严重程度的血清学标志物。

多不饱和脂肪酸对Δ15 Des转基因小鼠前列腺素E_(2)水平的影响

目的探讨多不饱和脂肪酸(PUFAs)对Δ15脂肪酸去饱和酶(Δ15 Des)转基因小鼠前列腺素E_(2)(PGE_(2))水平的影响。方法取C57BL/6野生型(WT)雄性小鼠和Δ15 Des转基因雄性小鼠作为对照组(n=5,WT组)和实验组(n=5,TG组),6%花生四烯酸(ARA)饲养8周,每周记录小鼠体质量变化;8周后麻醉处死2组小鼠,冰上分离小鼠肝脏、睾丸及肌肉组织,采用气相色谱(GC)检测2组小鼠各组织中脂肪酸(FAs)的组成及水平,采用酶联免疫吸附(ELISA)和实时荧光定量PCR(RT-qPCR)测定2组小鼠各组织中PGE_(2)的水平及前列腺素E合酶1(PTGES1)、前列腺素E合酶2(PTGES2)信使RNA(mRNA)的相对表达。结果第1~8周,2组小鼠体质量均在增长,2组间比较、差异无统计学意义(P>0.05);WT组小鼠各组织中PUFAs主要是欧米伽-6 PUFAs(n-6 PUFAs);与WT组相比,TG组小鼠各组织中ARA水平降低、二十碳五烯酸(EPA)和二十二碳六烯酸(DHA)水平升高(P<0.05或P<0.01),睾丸组织中亚油酸(LA)水平升高、二十二碳四烯酸(DTA)水平降低(P<0.05);TG组小鼠睾丸、肌肉组织中PGE_(2)水平低于WT组(P<0.05或P<0.01);TG组小鼠肝脏、睾丸组织中PTGES1、PTGES2 mRNA表达低于WT组(P<0.05或P<0.01)。结论PUFAs可减少Δ15 Des转基因小鼠ARA的合成,降低与PGE_(2)相关的合成酶表达,从而调节体内PGE_(2)的水平。

Endogenous prostaglandin D₂synthesis inhibits e-selectin generation in human umbilical vein endothelial cells

We examined the role of prostaglandin D2 (PGD2) in the formation of E-selectin following inter-leukin-1 (IL-1) stimulation in human umbilical vein endothelial cells (HUVEC) transfected with lipocaline-type PGD2 synthase (L-PGDS) genes. HUVEC were isolated from human umbilical vein and incubated with 20 U/mL IL-1 and various concentrations of authentic PGD2. The isolated HUVEC were also transfected with L-PGDS genes by electroporation. The L-PGDS-transfected HUVEC were used to investigate the role of endogenous PGD2 in IL-1-stimulated E-selectin biosynthesis. We also used an anti-PGD2 antibody to examine whether an intracrine mechanism was involved in E-selectin production. PGD2 and E-selectin levels were determined by radio-immunoassay and enzyme- immunoassay, respectively. E-selectin mRNA was assessed by real-time RT-PCR. IL-1-stimulated E-selectin production by HUVEC was dose-dependently inhibited by authentic PGD2 at concentrations greater than 10–6 mol/L. L-PGDS gene-transfected HUVEC produced more PGD2 than HUVEC transfected with the reporter gene alone. IL-1 induced increases in E-selectin production in HUVEC transfected with the reporter genes alone. However, this effect was significantly attenuated in the case of IL-1 stimulation of HUVEC trans-fected with L-PGDS genes, and accompanied by an apparent suppression of E-selectin mRNA expression. Neutralization of extracellular PGD2 by anti-PGD2- specific antibody influenced neither E-selectin mRNA expression nor E-selectin biosynthesis. HUVEC transfected with L-PGDS genes showed increased PGD2 synthesis. This increase was associated with attenuation of both E-selectin generation and E-selectin mRNA expression. The results suggest that endogenous PGD2 decreases E-selectin synthesis and E-selectin mRNA expression, probably through an intracrine mechanism.