QSAR modeling of benzoquinone derivatives as 5-lipoxygenase inhibitors

The inhibitors of 5-LOX control the overproduction of pro-inflammatory mediators known as leukotrienes(LTs)and thus have therapeutic relevance in the treatment of various diseases like asthma,rheumatoid arthritis,inflammatory bowel disease and certain types of cancers.This has increased the search for efficient therapeutic agents for protein 5-LOX and this process is now primarily based on QSAR.In this study,we have developed four different quantitative structure and 5-LOX inhibition activity relationship models of benzoquinone derivative by exploiting CoMFA,RF,SVM,and MLR chemometric methods.Performance of the QSAR models was measured by using cross-validation technique as well as through the external test set prediction.RF model outperforms all other models.SVM and MLR models failed due to the poor performance of the external test set prediction.CoMFA model,which shows relatively good performance was used to explore the essential structural regions where the modification was necessary to design a novel scaffold with improved activity.Moreover,molecular docking of all the derivatives to the binding site of 5-LOX was done to show their binding mode and to identify critical interacting residues inside the active site of 5-LOX.The docking result confirms the stability and rationality of the CoMFA model.

Variants of the arachidonate 5-lipoxygenase-activating protein (ALOX5AP) gene and risk of ischemic stroke in Han Chinese of eastern China

Variants of the arachidonate 5-1ipoxygenase-activating protein （ALOX5AP） gene have been suggested to play an important role in the pathogenesis of atherosclerosis and ischemic stroke. This study was aimed to explore the association of ALOX5AP variants with ischemic stroke risk in Han Chinese of eastern China. A total of 690 ischemic stroke cases and 767 controls were recruited. The subjects were further subtyped according to the Trial of Org 10172 in Acute Stroke Treatment （TOAST） criteria. On the basis of that, two polymorphisms of the ALOX5AP gene （rs10507391 and rs12429692） were determined by TaqMan genotyping assay. In addition, plasma leukotriene B4 （LTB4） levels were analyzed in these subjects. There was no evidence of association between the two variants of ALOX5AP and the risk of ischemic stroke or its TOAST-subtypes. Haplotype analysis and stratification analysis according to sex, age, body mass index, hypertension, and diabetes also showed negative association. Analysis of LTB4 levels in a subset of cases and controls revealed that LTB4 levels were significantly higher in ischemic stroke cases than in the controls （70.06± 14.75 ng/L vs 57.34±10.93 ng/L; P = 0.000） and carriers of the T allele of the rs10507391 variant were associated with higher plasma LTB4 levels （P = 0.000）. The present study suggests there is no association of the two polymorphisms in the ALOX5AP gene with ischemic stroke risk in Han Chinese of eastern China.

15-Lipoxygenase inhibition of Commelina benghalensis,Tradescantia fluminensis,Tradescantia zebrina

Objective:To evaluate the 15-lipoxygenase inhibitory activity of the methanol leaf extracts of Commelina benghalensis,Tradescantia fluminensis(T.fluminensis)and Tradescantia zebrina.Method:The inhibitory activity was evaluated using a spectrophotometric assay by observing the increase in absorbance at 234 nm due to the formation of the product 13-hydroperoxyoctadecadienoic acid.The extracts were also tested for the presence of terpenoids,saponins,tannins,flavonoids,steroids,phenolic compounds,alkaloids and cardiac glycosides.Results:All the extracts inhibited the action of 15-lipoxygenase at a concentration of 0.2μg/mL.T.fluminensis and Tradescantia zebrina exhibited higher than 50%inhibition with T.fluminensis at 87.2%.T.fluminensis was partitioned with ethyl acetate and hexane and their IC_(50)values were determined at 8.72μg/mL and 98.04μg/mL,respectively.Conclusions:T.fluminensis is a potentially good source of 15-lipoxygenase inhibitors.

5-Lipoxygenase and cysteinyl leukotriene receptors in neuroinflammation and neuronal injury

Brian ischemic injury and central neurodegenerative diseases as leading contributors to disability and death have become a majorclinical and public health concern worldwide.Neuroinflammation plays a pivotal role in the pathological progression of cerebral ischemia and neurodegenerative diseases including Parkinson disease(PD).Therefore,it is important to find effective therapeutic targets to attenuate inflammation and delay the progression of brain injury.Cysteinyl leukotrienes(CysLTs) are potent inflammatory mediators synthesized from arachidonic acid by 5-lipoxygenase(5-LOX) in the central nervous system.Two distinct G-protein-coupled receptors,CysLT1 R and CysLT2 R,mediate most of the known CysLTs biological responses.Accumulating evidence has demonstrated that postischemic inflammation and neuronal loss are mediated by 5-LOX and CysLTRs fol owing focal cerebral ischemia.We recently reported that the expression of 5-LOX,CysLT1R and inflammatory vascular cell adhesion molecule-1(VCAM-1) was upregulated in the hippocampus of rats with transient global cerebral ischemia,which was closely associated with delayed neuronal death in the hippocampal CA1 area.5-LOX inhibitor zileuton,CysLT1R antagonist ONO-1078 and montelukast dose-dependently reduced hippocampal CA1 neuronal death and inhibited the increased expression of 5-LOX and VCAM-1.In vitro ischemia-like injury in 5-LOXtransfected PC12 cells,oxygen-glucose deprivation(OGD) induced cell death mediated by5-LOX via ROS/P38 MAPK pathway.The nonselective 5-LOX inhibitor caffeic acid inhibited OGDstimulated activation of 5-LOX and ROS/P38 MAPK signaling and improved neuronal survival.In PD model,high concentrations of rotenone caused directly PC12 neurotoxicity,which was modulated by 5-LOX and abolished by suppression of 5-LOX.It is well known that microglia is major modulators of inflammatory response after brain injury.Overactivated microglia and production of proinflammatory cytokine IL-1β,IL-6 and TNF-α contribute to the neuroinflammation and brain injury.5-LOX,CysLT1R and CysLT2R are involved in microglial activation and resultant neurotoxic responses.It has been found that low concentrations of rotenone can activate 5-LOX and CysLT1R on microglial cells to enhance microglial inflammation and microglia-dependent neuronal death in vitro.5-LOX inhibitor zileuton and CysLT1R antagonist montelukast protected neurons from microglia-dependent rotenone neurotoxicity.Furthermore,lipopolysaccharide(LPS)induced microglial activation and microglial neurotoxicity mediated by CysLT2R in vitro.Both pharmacological blockade(CysLT2R antagonist HAMI3379) and RNA interference(specific short hairpin RNA) of CysLT2 R significantly attenuated LPS-triggered microglial inflammation and subsequent neuronal death.Collectively,the present results indicate the role of 5-LOX and CysLTRs in neuroinflammation and brain injury.Modulation of 5-LOX and CysLTRs may be potential therapeutic approaches for inflammation-related brain disorders such as cerebral ischemia and PD.However,further research is needed to clarify the mechanisms underlying the regulation of neuinflammatory processes by 5-LOX and CysLTRs.

The effects of 5-lipoxygenase inhibitor and cysteinyl leukotriene receptor 1 antagonist on 1-methyl-4-phenylpyridine-induced neurotoxicity

OBJECTIVE Previously we demonstrated the neuroprotective effect of 5-lipoxygenase(5-LOX)inhibitor as well as cysteinyl leukotriene receptor 1(Cys LT1)antagoniston rotenone-induced microglial activation and neuronal death.In this study,we determined the effects of 5-LOX inhibitor zileuton and Cys LT1 antagonist montelukast on neurotoxicity induced by 1-methyl-4-phenylpyridine(MPP+)in an in vitro model of Parkinson disease(PD).METHODS The neurotoxicity of MPP+,a neurotoxin relevant to PD,on the PC12 cells was measured by MTT assay,lactate dehydrogenase(LDH)release and double fluorescence staining with Hoechst/propidiumiodide(PI).The protective effects of 5-LOX inhibitor zileuton and Cys LT1 antagonist montelukast were investigated by the above methods.RESULTS We found that exposure of PC12 cells to MPP+led to a reduced cell viability and an increased level of LDH in a concentration-dependent manner.Pretreatment with zileuton and montelukast significantly attenuated viability loss and LDH release in MPP+-treated PC12 cells.Furthermore,MPP+increasednecrotic cell death in PC12 cells.Administration of montelukast significantly decreased MPP+-induced cell necrosis in PC12 cells.CONCLUSION The 5-LOX inhibitor zileuton and Cys LT1 antagonist montelukast have a neuroprotective effects on MPP+-induced neurotoxicity in PC12 cells.The 5-LOX inhibitor and Cys LT1 antagonist might raise a possibility as potential therapeutic agent for PD and other inflammation-related the central nervous system disorders.

Involvement of eicosanoids in the pathogenesis of pancreatic cancer: the roles of cyclooxygenase-2 and 5-lipoxygenase

The interplay between inflammation and cancer progression is a growing area of research. A combination of clinical, epidemiological, and basic science investigations indicate that there is a relationship between inflammatory changes in the pancreas and neoplastic progression. Diets high in ω-6 polyunsaturated fatty acids provide increased substrate for arachidonic acid metabolism by cyclooxygenase-2(COX-2) and 5-lipoxygenase(5-LOX) to form eicosanoids. These eicosanoids directly contribute to pancreatic cancer cell proliferation. Both COX-2 and 5-LOX are upregulated in multiple cancer types, including pancreatic cancer. In vitro studies using pancreatic cancer cell lines have demonstrated upregulation of COX-2 and 5-LOX at both the mRNA and protein levels. When COX-2 and 5-LOX are blocked via a variety of mechanisms, cancer cell proliferation is abrogated both in vitro and in vivo.The mechanism of COX-2 has been shown to include effects on apoptosis as well as angiogenesis. 5-LOX has been implicated in apoptosis. The use of COX-2 and 5-LOX inhibitors in clinical studies in patients with pancreatic cancer has been limited. Patient enrollment has been restricted to those with advanced disease which makes evaluation of these drugs as chemopreventive agents difficult. COX-2 and 5-LOX expression have been shown to be present during the early neoplastic changes of pancreatic cancer, well before progression to invasive disease. This indicates that the ideal role for these interventions is early in the disease process as preventive agents, perhaps in patients with chronic pancreatitis or hereditary pancreatitis.

Expression of 5-Lipoxygenase in human colorectal cancer

AIM: To evaluate the 5-lipoxygenases (Loxs) expression level in human colorectal cancer specimens in order to determine its clinicopathologic significance in human tumorigenesis. METHODS: The relative quantity of 5-Lox mRNA in paired 91 colorectal tumor and adjacent normal mucosa samples was determined by real time quantitative PCR. Additionally, the expression of 5-Lox and cyclooxygenase (Cox)-2 proteins was also examined using immunohistochemical staining methods. RESULTS: There was a marked increase in 5-Lox mRNA levels in the tumor compared with paired normal mucosa samples (P < 0.0001). Sixty six (72.5%) tumors showed high 5-Lox mRNA levels. The positivity rate of 5-Lox and Cox-2 protein expression was 68.7% and 79.1% respectively. There was a significant association between tumoral 5-Lox mRNA level and tumor size (Rho = 0.392, P = 0.0002), depth or vessel invasion. CONCLUSION: These results suggest that 5-Lox is up-regulated in colorectal cancer and that inhibition of its expression might be valuable in the prevention and treatment of colorectal cancer.

5-lipoxygenase expression in a brain damage model induced by chronic oral administration of aluminum

A preliminary study has found that the 5-lipoxygenase inhibitor, caffeic acid, has a marked protective effect on acute brain injury induced by intracerebroventricular microinjection of aluminum. In this experiment, chronic brain injury and neuronal degeneration model was established in rats by chronic oral administration of aluminum, and then intervened using caffeic acid. Results showed that caffeic acid can downregulate chronic aluminum overload-induced 5-1ipoxygenase mRNA and protein expression, and repair the aluminum overload-induced hippocampal neuronal damage and spatial orientation impairment. It is suggested that direct intervention of 5-lipoxygenase expression has a neuroprotective role in the degeneration induced by chronic aluminum overload brain injury model.

A Facile Total Synthesis of 2-Geranyl-2',3,4',4-tetrahydroxydihydrochalcone with Highly 5-Lipoxygenase Inhibiting Activity

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C1q/TNF-related protein 5 promotes atherogenesis by enhancing transcytosis and oxidative modification of low-density lipoprotein through increasing 12/15-lipoxygenase

Objective Increased transcytosis of low-density lipoprotein (LDL)across the endothelium and oxidation of LDL deposited within the subendothelial space are crucial early events in atherogenesis. C1q/TNF-related protein (CTRP) 5 is a novel secreted glycoprotein and its biological functions are largely undefined.

Further evidence for the roles of 5-lipoxygenase and cysteinyl leukotriene receptors in cerebral ischemic injury

Arachidonic acid-metabolizing enzyme 5-lipoxygenase (5-LOX) produces pro-inflammatory mediators:leukotrienes (including cysteinyl leukotrienes, CysLTs). 5-LOX and CysLTs are involved in the pathophysiological process after brain injury, and the actions of CysLTs are mediated via activation of their receptors, CysLT1 and CysLT2. We have recently reported the expressions of 5-LOX, CysLT1 and CysLT2 in human brains with traumatic injury and tumors, and short-term neuroprotective effects of Cys-LT1 antagonists in stroke models of rats and mice.

Expression manner of 5-lipoxygenase in human traumatic brain injury and astrocytoma

5-lipoxygenase (5-LO) is a key enzyme of arachidonic acid metabolism. The metabolites, leuklotrienes, are important mediators in asthma, inflammatory and allergic disorders. The activation of 5-LO includes the gathering of 5-LO,adhering onto the membranes of nuclei and entrance to the nuclei. In the central nervous system, 5-LO is widely

Short-term intensive atorvastatin therapy improves endothelial function partly via attenuating perivascular adipose tissue inflammation through 5-lipoxygenase pathway in hyperlipidemic rabbits

Background Atherosclerosis is a kind of disease with multiple risk factors,of which hyperlipidemia is a major classical risk factor resulting in its pathogenesis and development.The aim of this study was to determine the effects of short-term intensive atorvastatin （IA） therapy on vascular endothelial function and explore the possible mechanisms that may help to explain the clinical benefits from short-term intensive statin therapy.Methods After exposure to high-fat diet （HFD） for 8 weeks,the animals were,respectively,treated with IA or low-dose atorvastatin （LA） for 5 days.Blood lipids,C-reactive protein （CRP）,tumor necrosis factor-α （TNF-α）,interleukin-6 （IL-6）,nitric oxide （NO）,endothelin-1 （ET-1）,and endothelium-dependent vasorelaxation function were,respectively,measured.mRNA and protein expression of CRP,TNF-α,IL-6,macrophage chemoattractant protein-1 （MCP-1）,and 5-lipoxygenase （5-LO） were also evaluated in pericarotid adipose tissue （PCAT） and cultured adipocytes.Results HFD increased serum inflammatory factor levels; induced significant hyperlipidemia and endothelial dysfunction,including imbalance between NO and ET-1; enhanced inflammatory factors and 5-LO expression; and promoted macrophage infiltration into adipose tissue.Five-day IA therapy could significantly decrease serum inflammatory factor levels and their expression in PCAT; restore the balance between NO and ET-1; and improve endothelial function and macrophage infiltration without significant changes in blood lipids.However,all of the above were not observed in LA therapy.In vitro experiment found that lipopolysaccharide （LPS） enhanced the expression of inflammatory factors and 5-LO in cultured adipocytes,which could be attenuated by short-time （6 hours） treatment of high-dose （5 pmol/L） but not low-dose （0.5 μmol/L） atorvastatin.In addition,inhibiting 5-LO by Cinnamyl-3,4-dihydroxy-α-cyanocinnamate （CDC,a potent and direct 5-LO inhibitor） could significantly downregulate the above-mentioned gene expression in LPS-treated adipocytes.Conclusion Short-term IA therapy could significantly ameliorate endothelial dysfunction induced by HFD,which may be partly due to attenuating inflammation of PCAT through inhibiting 5-LO pathway.

Structural optimization and biological evaluation of 1,5-disubstituted pyrazole-3-carboxamines as potent inhibitors of human 5-lipoxygenase

Human 5-lipoxygenase(5-LOX)is a well-validated drug target and its inhibitors are potential drugs for treating leukotriene-related disorders.Our previous work on structural optimization of the hit compound 2 from our in-house collection identified two lead compounds,3a and 3b,exhibiting a potent inhibitory profile against 5-LOX with IC_(50) values less than 1 mmol/L in cell-based assays.Here,we further optimized these compounds to prepare a class of novel pyrazole derivatives by opening the fusedring system.Several new compounds exhibited more potent inhibitory activity than the lead compounds against 5-LOX.In particular,compound 4e not only suppressed lipopolysaccharide-induced inflammation in brain inflammatory cells and protected neurons from oxidative toxicity,but also significantly decreased infarct damage in a mouse model of cerebral ischemia.Molecular docking analysis further confirmed the consistency of our theoretical results and experimental data.In conclusion,the excellent in vitro andin vivo inhibitory activities of these compounds against 5-LOX suggested that these novel chemical structures have a promising therapeutic potential to treat leukotriene-related disorders.

非小细胞肺癌组织中lncRNA GAS5、LHPP表达与上皮间质化相关性及临床意义

目的探讨非小细胞肺癌(NSCLC)患者癌组织中长链非编码核糖核酸生长抑制特异性基因5(lncRNA GAS5)、磷酸赖氨酸磷酸组氨酸无机焦磷酸盐磷酸酶(LHPP)表达与上皮间质化(EMT)相关性及临床意义。方法收集2018年6月至2020年1月在河北北方学院附属第一医院行手术切除的NSCLC 90例患者的癌组织及癌旁组织,采用实时荧光定量聚合酶链反应检测lncRNA GAS5、LHPP和EMT相关蛋白[E-钙黏蛋白(E-Cad)、N-钙黏蛋白(N-Cad)和波形蛋白(VIM)]表达。分析NSCLC患者癌组织中lncRNA GAS5、LHPP mRNA与临床病理特征的关系,并通过Pearson相关性分析NSCLC患者癌组织中lncRNA GAS5、LHPP mRNA与EMT相关蛋白表达的相关性。采用Kaplan-Meier法绘制不同lncRNA GAS5、LHPP mRNA表达的NSCLC患者生存曲线,多因素Cox回归分析NSCLC患者预后的影响因素。结果NSCLC患者癌组织中lncRNA GAS5、LHPP mRNA、E-Cad mRNA表达低于癌旁组织,N-Cad mRNA、VIM mRNA表达高于癌旁组织,差异有统计学意义(P<0.05)。Pearson相关性分析显示,NSCLC患者癌组织中lncRNA GAS5与E-Cad mRNA表达呈正相关(r=0.724,P<0.001),与N-Cad mRNA、VIM mRNA表达呈负相关(r=-0.699、-0.689,P<0.001);lncRNA GAS5与LHPP mRNA表达呈正相关(r=0.651,P<0.001)。不同分化程度、肿瘤TNM分期、淋巴结转移的NSCLC患者癌组织中lncRNA GAS5、LHPP mRNA表达比较差异有统计学意义(P<0.05)。Kaplan-Meier生存曲线分析显示,lncRNA GAS5高表达组的3年总生存率[68.18%(30/44)]高于lncRNA GAS5低表达组的3年总生存率[36.96%(17/46)];LHPP mRNA高表达组的3年总生存率[67.39%(31/46)]高于LHPP mRNA低表达组的3年总生存率[36.36%(16/44)],差异有统计学意义(χ^(2)=10.274、10.322,P<0.05)。低分化、肿瘤TNM分期Ⅲ期、有淋巴结转移为NSCLC患者死亡的独立危险因素,lncRNA GAS5≥1.32、LHPP mRNA≥1.12为独立保护因素(P<0.05)。结论NSCLC患者癌组织中lncRNA GAS5、LHPP mRNA低表达,与EMT相关蛋白表达、分化程度、肿瘤TNM分期、淋巴结转移和预后有关,可能成为NSCLC诊治的新靶点。

远缘杂交仁用杏新品种京仁5号的选育

京仁5号通过仁用杏品种龙王帽与扁桃品种意大利2号远缘杂交育成。果实卵圆形,果顶圆凸,在北京地区7月中下旬成熟;成熟时果皮底色黄色,阳面着片状红色,平均单果质量30.5 g;果核卵圆形,壳面较平滑,平均单核质量2.54 g,纵横侧径分别为3.28 cm、2.12 cm、1.23 cm;核仁饱满,离核,甜仁,平均单仁鲜质量1.05 g、干质量0.90 g,出仁率37.02%;杏仁脂肪含量(w,后同)435.63 g·kg^(-1),蛋白质含量269.84 g·kg^(-1),钙含量1.26 g·kg^(-1),铁含量24.08 mg·kg^(-1)。丰产稳产,适应性强,综合性状优良。

‘陇核5号’果实发育规律及疏果施肥对坚果品质的影响

【目的】明确核桃果实发育规律及影响坚果品质的因素,为核桃果园生产管理提供参考。【方法】以‘陇核5号’为研究对象,对其果实发育过程中外观形态变化进行拍照记录,测定果实三径、果实质量、果皮厚度、坚果三径、坚果质量、核仁质量、出仁率、壳厚等指标,利用Logistic方程、二次方程、三次方程等模型对各指标变化趋势进行拟合。通过疏果和不同时期喷施叶面肥,分析不同处理对核桃坚果经济性状的影响。【结果】在发育过程中,果实(坚果)大小和质量均表现为缓慢增长、迅速增长、缓慢增长的趋势,其中果实质量分别在花后50 d时和花后100 d时出现了增长高峰,坚果质量在花后110 d时出现了增长高峰。花后30~40 d时果实缓慢增长,随后迅速膨大,果实质量快速增加,花后50 d时开始出现水状核仁,花后70 d时硬核开始出现;花后70 d后,果实大小和质量增长变慢,果壳开始形成并逐渐硬化变厚;花后90~120 d时,坚果质量、果壳厚度、核仁质量及出仁率等指标均快速增加;花后120~140 d时,坚果大小增长速度变慢,坚果质量开始下降。方差分析结果表明,除坚果三径外,其他指标在不同处理间差异显著(P<0.05)。其中以7月初—8月初喷施叶面肥处理下核桃坚果经济指标值最高且与对照差异极显著(P<0.01);其次是5月底—6月底喷施叶面肥,与对照差异显著(P<0.05);疏果处理下核桃坚果经济指标与对照差异不显著(P>0.05)。【结论】‘陇核5号’果实生长发育总体为缓慢、快速、缓慢的趋势,呈单“S”形曲线。疏果、施肥均能改善核桃坚果的经济性状。

无有机模板法制备Cu-ZSM-5催化剂及其选择性催化氧化苯乙烯

不使用任何有机模板剂,硝酸铜和正硅酸四乙酯(TEOS)经过酸性水解、碱性陈化和水热晶化,制备得到四配位铜修饰ZSM-5沸石(Cu-ZSM-5)。考察凝胶的Si/Al摩尔比、pH值、晶化温度和晶化时间对Cu-ZSM-5的晶型和相对结晶度的影响。采用X射线衍射(XRD)、红外光谱(FT-IR)、紫外漫反射谱和X-射线光电子能谱等手段对Cu-ZSM-5进行分析表征。结果表明:Cu-ZSM-5沸石具有MFI结构,结晶度高,无其他杂晶;铜原子以四配位为主。四配位铜是催化氧化苯乙烯的活性位点;在m(Cu-ZSM-5-50)=0.1 g、n(TBHP)/n(Styrene)=1.5、T=60℃、t=5 h条件下,苯乙烯转化率和苯甲醛选择性分别为76.5%和69.4%,表明Cu-ZSM-5-50具有较高的催化活性和较好的重复利用性。该制备方法不使用有机模板剂,无需高温焙烧,Cu-ZSM-5催化剂的制备成本大大降低。

单纯型大疱性表皮松解症患儿的角蛋白5基因突变研究

目的:对1例单纯型大疱性表皮松解症(EBS)患儿及其家系进行基因突变检测和致病性分析。方法:应用高通量二代测序技术捕获目标序列,对患儿进行全外显子组测序。发现致病位点后,应用Sanger测序法进行家系验证,查阅人类基因突变数据库(HGMD),运用生物信息学蛋白功能预测软件,分析变异位点的致病性。结果:患儿角蛋白5(KRT5)基因的第一外显子检出杂合变异c.536T>C(p.F179S),该变异造成KRT5蛋白的第179位氨基酸改变,患儿父母均未检测到相同突变。结论:患儿KRT5基因的杂合变异c.536T>C(p.F179S)为新发致病性变异,导致患儿KRT5缺陷,进而引发疱疹样型EBS(DM-EBS)。该变异位点在国内未见报道,扩大了我国人群KRT5的基因突变谱,为家系的遗传咨询和产前诊断提供依据。

电针预处理对切口痛大鼠中脑导水管周围灰质5-HT_(7)受体表达的影响

目的:建立足趾部切口痛大鼠模型,探讨重复电针预处理对切口痛大鼠镇痛效果及其对中脑导水管周围灰质(periaqueductal gray,PAG)5-HT_(7)受体(5-HT_(7)R)表达的影响。方法:40只成年雄性SD大鼠按随机数字表法均等分为对照组(Control,Con组)、切口痛模型组(Incision pain,IP组)、正常+电针预处理组(Control+Electroacupuncture,Con+EA组)、模型+电针预处理组(Incision Pain+Electroacupuncture,IP+EA组)。IP组和IP+EA组大鼠右足趾部行疼痛造模,且造模前,Con+EA组和IP+EA组大鼠行右侧“足三里”穴和“环跳”穴电针刺激(2/10 Hz疏密波,刺激强度数值为1档,每日1次30 min),连续5天。于第1次电针预处理前2 h(T1)、术前2 h(T2)、术后4 h(T3)、术后24 h(T4)测定大鼠机械刺激缩足反射阈值(mechanical withdrawal threshold,MWT)和热缩足潜伏期(thermal withdrawal latency,TWL);酶联免疫吸附法检测脑脊液中5-HT浓度;免疫组化和免疫荧光方法分别检测大鼠PAG中c-Fos和5-HT_(7)R蛋白表达情况。结果:与Con组比较,IP组大鼠T3、T4时间点MWT和TWL均明显降低,脑脊液中5-HT浓度增加,PAG中c-Fos蛋白和5-HT_(7)R表达明显上调(P<0.05);Con+EA组和IP+EA组脑脊液中5-HT含量和PAG中5-HT_(7)R表达均上升(P<0.05)。与IP组相比,IP+EA组大鼠T3、T4时间点的MWT和TWL显著升高,PAG中c-Fos蛋白表达减少,5-HT_(7)R蛋白表达增加(P<0.05)。结论:电针预处理可能通过上调PAG中5-HT_(7)R蛋白表达发挥镇痛作用。