Melanocortin 3,5 receptors immunohistochemical expression in colonic mucosa of inflammatory bowel disease patients:A matter of disease activity?

BACKGROUND Melanocortin 3 and 5 receptors(i.e.,MC3R and MC5R)belong to the melanocortin family.However,data regarding their role in inflammatory bowel diseases(IBD)are currently unavailable.AIM This study aims to ascertain their expression profiles in the colonic mucosa of Crohn’s disease(CD)and ulcerative colitis(UC),aligning them with IBD disease endoscopic and histologic activity.METHODS Colonic mucosal biopsies from CD/UC patients were sampled,and immunohisto-chemical analyses were conducted to evaluate the expression of MC3R and MC5R.Colonic sampling was performed on both traits with endoscopic scores(Mayo endoscopic score and CD endoscopic index of severity)consistent with inflamed mucosa and not consistent with disease activity(i.e.,normal appearing mucosa).RESULTS In both CD and UC inflamed mucosa,MC3R(CD:+7.7 fold vs normal mucosa,P<0.01;UC:+12 fold vs normal mucosa,P<0.01)and MC5R(CD:+5.5 fold vs normal mucosa,P<0.01;UC:+8.1 fold vs normal mucosa,P<0.01)were significantly more expressed compared to normal mucosa.CONCLUSION MC3R and MC5R are expressed in the colon of IBD patients.Furthermore,expression may differ according to disease endoscopic activity,with a higher degree of expression in the traits affected by disease activity in both CD and UC,suggesting a potential use of these receptors in IBD pharmacology.

METTL5 promotes cell proliferation,invasion,and migration by up-regulating Toll-like receptor 8 expression in colorectal cancer

BACKGROUND N6-methyladenosine(m6A)modification represents the predominant alteration found in eukaryotic messenger RNA and plays a crucial role in the progression of various tumors.However,despite its significance,the comprehensive investigation of METTL5,a key m6A methyltransferase,in colorectal cancer(CRC)remains limited.AIM To investigate the role of METTL5 in CRC.METHODS We assessed METTL5 expression levels in clinical samples obtained from CRC patients as well as in CRC cell lines.To elucidate the downstream targets of METTL5,we performed RNA-sequencing analysis coupled with correlation analysis,leading us to identify Toll-like receptor 8(TLR8)as a potential downstream target.In vitro functional assessments of METTL5 and TLR8 were conducted using CCK-8 assays,scratch assays,as well as assays measuring cell migration and invasion.RESULTS Our findings reveal a pronounced upregulation of METTL5 expression in both CRC cells and tissues,which correlated significantly with an unfavorable prognosis.In vitro experiments unequivocally demonstrated the oncogenic role of METTL5,as evidenced by its promotion of CRC cell proliferation,invasion,and migration.Notably,we identified TLR8 as a downstream target of METTL5,and subsequent down-regulation of TLR8 led to a significant inhibition of CRC cell proliferation,invasion,and tumor growth.CONCLUSION The heightened expression of METTL5 in CRC is strongly associated with clinicopathological features and a poor prognosis,thereby underscoring its potential utility as a critical marker for facilitating early diagnosis and prognostication in CRC.

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。

拮抗CC趋化因子受体5信号诱导肿瘤细胞凋亡并调节肿瘤微环境抑制肿瘤生长

目的探索拮抗CC趋化因子受体5(CCR5)信号通路对肿瘤生长和肿瘤微环境的影响。方法采用CCK8细胞毒试验研究CCR5选择性拮抗剂Maraviroc体外对小鼠Lewis肺腺癌细胞增殖的影响,并运用流式细胞术和RT-PCR检测肿瘤细胞凋亡和Caspase 8基因的表达。然后采用免疫荧光组织化学染色法研究了Maraviroc对小鼠体内肿瘤生长和肿瘤微环境中CD4^(+)和CD8^(+)以及Foxp3^(+)细胞比例的影响。结果拮抗CCR5信号在体内外均能够抑制癌细胞的生长。体外研究发现:CCR5拮抗剂可通过增强凋亡基因Caspase 8表达而诱导肿瘤细胞凋亡。在小鼠体内,CCR5拮抗剂可明显增加肿瘤微环境中CD4^(+)和CD8^(+)细胞的浸润而减少Foxp3^(+)细胞的浸润。结论拮抗CCR5信号可能通过诱导肿瘤细胞凋亡,逆转免疫抑制性肿瘤微环境而抑制肿瘤生长。

基于5-HT及其受体的中医药干预消化系统疾病研究进展

5羟色胺(5-hydroxytryptamine,5-HT)是一种重要的单胺类神经递质,具有广泛的生物学效应。血清和胃肠道中的5-HT含量可占人体5-HT总量的95%以上,血液循环中的5-HT也主要来源于肠道。5-HT在消化系统疾病发生发展中的作用历来受到关注。此文从5-HT特征、5-HT受体以及基于5-HT及其受体探讨中医药对消化系统疾病的干预作用进行了概述,以期进一步为消化系统疾病的防治、临床与基础研究提供新思路。

橘皮竹茹汤对化疗性异食癖大鼠模型5-羟色胺3受体蛋白和mRNA表达的影响

目的观察橘皮竹茹汤对化疗性异食癖大鼠模型止呕作用及对5-羟色胺3受体(5-HT3R)蛋白和mRNA的影响。方法将36只Wistar雄性大鼠随机分为正常对照组、中药对照组(橘皮竹茹汤10.6 g/kg,不造模)、模型组、昂丹司琼组(2.6 mg/kg)、橘皮竹茹汤低剂量组(5.3 g/kg)和橘皮竹茹汤高剂量组(10.6 g/kg),每组6只。各组大鼠给予相应干预措施6 d(每日2次)后,采用腹腔注射顺铂(6 mg/kg)的方式诱导建立化疗性异食癖大鼠模型,观察并记录造模后24 h内大鼠体质量、摄食量和高岭土摄入量的变化。继续干预1 d后,苏木精-伊红(HE)染色观察各组大鼠胃窦、回肠组织形态学变化;实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法检测延髓、回肠组织中5-HT3R、色氨酸羟化酶(TPH)、单胺氧化酶A(MAOA)mRNA表达;Western blot法检测延髓组织中5-HT3R蛋白的表达。结果①与正常对照组比较,模型组大鼠体质量、摄食量降低(P<0.05),高岭土摄入量显著增加(P<0.05);与模型组比较,昂丹司琼组和橘皮竹茹汤低、高剂量组大鼠高岭土摄入量显著降低(P<0.05)。②HE染色显示,与正常对照组比较,模型组胃窦组织上皮细胞受损,固有层腺体排列疏松、紊乱,回肠上皮部分缺失,腺体排列紊乱;与模型组比较,昂丹司琼组和橘皮竹茹汤低、高剂组胃窦和回肠组织病理改变减轻。③RT-qPCR结果显示,与正常对照组比较,模型组大鼠延髓、回肠5-HT3R mRNA表达水平显著升高(P<0.05),回肠MAOA mRNA表达水平显著降低(P<0.05);与模型组比较,橘皮竹茹汤高剂量组大鼠延髓5-HT3R mRNA表达水平明显降低(P<0.05),橘皮竹茹汤低剂量组回肠TPH1 mRNA、橘皮竹茹汤高剂量组延髓TPH2 mRNA表达水平明显降低(P<0.05),昂丹司琼组、橘皮竹茹汤低剂量组、橘皮竹茹汤高剂量组回肠MAOA mRNA表达水平明显升高(P<0.05)。④Western blot结果显示,与正常对照组比较,模型组大鼠延髓5⁃HT3R蛋白表达水平显著升高(P<0.05);与模型组比较,橘皮竹茹汤高剂量组大鼠延髓5-HT3R蛋白表达水平显著降低(P<0.05)。结论橘皮竹茹汤可以通过抑制延髓5-HT3R蛋白和mRNA的表达,调控5-羟色胺合成与代谢相关酶,进而改善化疗导致的恶心呕吐。

Casticin-induced apoptosis involves death receptor 5 upregulation in hepatocellular carcinoma cells

AIM:To investigate the apoptotic activities of casticin in hepatocellular carcinoma(HCC) cells and its molecular mechanisms.METHODS:PLC/PRF/5 and Hep G2 cell lines were cultured in vitro and the inhibitory effect of casticin on the growth of cells was detected by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolim bromide(MTT) assay.The apoptotic cell death was examined using the cell apoptosis enzyme linked immunosorbent assay(ELISA) detection kit,flow cytometry(FCM) after propidium iodide(PI) staining and DNA agarose gel electrophoresis.The caspase activities were measured using ELISA.Reactive oxygen species(ROS) production was evaluated by FCM after dichlorodihydrofluorescein diacetate(DCFH-DA) probe labeling.Intracellular glutathione(GSH) content was measured using a glutathione assay kit.The expression of death receptor(DR)4 and DR5 proteins was analyzed by Western blotting and FCM.RESULTS:Casticin significantly inhibited the growth of human HCC(PLC/PRF/5 and Hep G2) cells in a dosedependent manner(P < 0.05).Casticin increased the percentage of the sub-G1 population in HCC cells in a concentration-dependent manner.The potency of casticin to PLC/PRF/5 cells was higher than that of 5-flurouracil(26.8% ± 4.8% vs 17.4% ± 5.1%) at 10 μmol/L for 24 h.Casticin increased the levels of Histone/DNA fragmentation and the levels of active caspase-3,-8 and-9 in a concentration-dependent manner(P < 0.05).Treatment with 30 μmol/L casticin for 24 h resulted in the formation of a DNA ladder.Casticin reduced the GSH content(P < 0.05),but did not affect the level of intracellular ROS in PLC/PRF/5 and Hep G2 cells.The thiol antioxidants,acetylcysteine(NAC) and GSH restored GSH content and attenuated casticin-induced apoptosis.In contrast,the nonthiol antioxidants,butylated hydroxyanisole and mannitol failed to do so.In the HCC cells treated with casticin for 24 h,DR5 protein level was increased.The expression of DR5 protein induced by casticin was inhibited by NAC.Pretreatment with DR5/Fc chimera protein,a blocking antibody,effectively attenuated the induction of apoptosis by casticin.CONCLUSION:Casticin-induced apoptosis of HCC cells is involved in GSH depletion and DR5 upregulation.

LRP5基因突变导致骨质疏松症-假性胶质瘤综合征一例报告

报告一例常染色体隐性遗传发病的骨质疏松症-假性胶质瘤综合征。先证者女性,23岁,父母非近亲结婚,出生后发现双目失明,婴儿期因发现右眼视网膜母细胞瘤行右眼球摘除术,9岁开始反复发生轻微外力骨折,诊断为成骨不全。查体发现脊柱侧凸畸形、胸廓畸形、双上肢肘外翻、四肢关节韧带松弛。双能X线吸收检测仪(dual energy X-ray absorptiometry,DXA)骨密度明显低于同龄人,腰椎1-4骨密度Z值-5,左髋骨密度Z值-1.8。X线摄片示全身骨小梁稀疏。Sanger测序显示低密度脂蛋白受体相关蛋白-5(lowdensity lipoprotein receptor-related protein 5,LRP5)基因的6号外显子和23号外显子发生复合杂合突变,导致p.Pro382Leu+p.Cys1611LeufsX33。本文通过文献复习对该病的临床表现和诊疗特点进行讨论及总结,以期帮助临床医生提高对这一疾病的认识。

5-HT受体在瘙痒中的作用研究进展

由组胺介导的经典瘙痒信号通路以及非组胺介导的其他瘙痒信号通路对于瘙痒的产生和感知具有重要作用。5-羟色胺(5-hydroxytrptamine,5-HT)又称血清素,是一种神经递质和血管活性胺类物质,可以引起明显的瘙痒行为。大量研究揭示了5-HT及其多种受体在急性和慢性瘙痒发生中的重要性,然而介导5-HT能瘙痒的信号机制尚不完全清楚。本文对5-HT及其受体家族在瘙痒中的作用及其相关信号机制进行简要梳理总结,为瘙痒相关疾病的临床诊疗或药物开发提供一定的参考。

瞬时受体电位通道5对间歇性低氧致心肌焦亡的影响

目的探讨瞬时受体电位通道5(TRPC5)对间歇性低氧致心肌焦亡的影响。方法TRPC5^(-/-)大鼠及SD大鼠各12只,两种大鼠分别随机分为慢性间歇性低氧组(CIH组)和常氧组(Control组),即WT-Control组、WT-CIH组、TRPC5^(-/-)-Control组、TRPC5^(-/-)-CIH组,每组6只。通过Masson染色观察大鼠心肌纤维化情况,ELISA检测血清炎症因子水平,Western blot检测TRPC5及焦亡相关蛋白相对表达水平。结果Masson染色显示,TRPC5^(-/-)-CIH组胶原容积分数高于TRPC5^(-/-)-Control组,低于WT-CIH组。ELISA结果显示,TRPC5^(-/-)-CIH组血清IL-1、IL-6、TGF-β水平高于TRPC5^(-/-)-Control组,低于WT-CIH组。Western Blot结果显示,TRPC5^(-/-)-CIH组焦亡相关蛋白Caspase-1、NLRP3、GSDMD、GSDMD-N相对表达量高于TRPC5^(-/-)-Control组、低于WT-CIH组。结论TRPC5缺乏减轻间歇性低氧导致的心肌焦亡,并改善心肌纤维化。

脑源性神经营养因子和神经营养因子-5在女性生殖内分泌领域的研究进展

脑源性神经营养因子(BDNF)和神经营养因子-5(NT-5)属于神经营养因子家族,是一类由神经支配的靶组织分泌的分泌型多肽,两者通过作用于相同的受体发挥作用。BDNF和NT-5在卵巢表达广泛,发挥着促进卵泡组装和发育、卵母细胞成熟、排卵、调节颗粒细胞和膜细胞类固醇激素分泌等多种生理作用。不孕症是由多种病因导致的一种生育障碍状态,不孕症患者卵巢局部的BDNF和NT-5存在异常分泌情况。因此,了解BDNF和NT-5在女性生殖内分泌领域的研究进展,可能为不孕症治疗提供新方向,为辅助生殖结局的预测提供新指标。

Detachment of esophageal carcinoma cells from extracellular matrix causes relocalization of death receptor 5 and apoptosis

AIM:To investigate the effect of detachment of esophageal cancer cells from extracellular matrix on the localization of death receptor 5(DR5) and apoptosis.METHODS:Anchorage-dependent EC9706 cells of esophageal squamous cell carcinoma were pretreated or not treated with brefeldin A.Detached cells were harvested by ethylenediaminetetraacetic acid digestion.Expression and localization of DR5 in these cells were determined by immunocytochemical and immunofluorescence assays,as well as flow cytometry analysis.Apoptosis of EC9706 cells was detected by flow cytometry after stained with fluorescein isothiocyanate-labeled annexin V/propidium iodide.Activation of caspase 8 was detected by Western blot analysis.RESULTS:Immunocytochemical assay indicated that DR5 was predominantly perinuclear in adherent cells but was mainly localized in cell membrane in detached cells.In addition,immunofluorescence assay also confirmed the above-mentioned results,and further demonstrated that DR5 was present in the form of coarse granules in detached cells,but in the form of fine granules in adherent cells.Cytometry analysis revealed higher levels of DR5 expression on the surfaces of brefeldin-A-untreated cells than on the surfaces of brefeldin-A-treated cells,but brefeldin A treatment did not affect the total DR5 expression levels.Moreover,nocodazole did not influence the extracelluar DR5 expression levels in EC9706 cells.Apoptosis assay revealed that detached cells were more sensitive to DR5 antibody-induced apoptosis than adherent cells.Western blotting showed that caspase 8 was activated in temporarily detached cells 4 h earlier than in adherent cells.CONCLUSION:Progress from adhesion to detachment of EC9706 cells causes DR5 relocalization,and promotes cytoplasmic translocation of DR5 to cell surfaces via a Golgi-dependent pathway.Moreover,it might also result in DR5 aggregation to render apoptosis of detached cells.

绝经后2型糖尿病LRP5基因多态性与骨量异常的研究

目的分析低密度脂蛋白受体相关蛋白5(low density lipoprotein receptor related protein 5,LRP5)rs556442、rs312778位点基因多态性及突变在绝经后女性2型糖尿病(type 2 diabetes mellitus,T2DM)患者中骨量异常的意义。方法收集2021年5月至2023年5月新疆石河子地区的142例绝经后女性资料进行回顾性分析,分为正常对照组(A组,n=29)、T2DM组(B组,n=30)、骨量降低组(C组,n=28)、骨量降低+T2DM组(D组,n=55)。收集记录相关基线资料,运用全自动生化测定仪测定受试者血糖、血脂及骨代谢指标。通过双能X线骨密度仪测定骨密度(bone mineral density BMD),LRP5基因位点多态性采用飞行时间质谱法(MALDI-TOF-MS)测定,采用SNPscan技术对上述SNP位点进行基因分型。结果①四组基线资料比较,绝经年限、年龄及腰臀比(waist hip ratio,WHR)比较差异具有统计学意义(P<0.05);②与A组相比,B组的空腹血糖(FPG)、糖化血红蛋白(HbA1c)升高(P<0.05);D组的FPG、HbA1c的血清学水平升高,甘油三酯(TG)血清学水平降低(P<0.05);③rs556442位点:与A组相比,D组基因型分布(AA/AG/GG基因型)有统计学意义(P<0.05);rs312778位点组间基因型(CC/CT/TT基因型)及等位基因分布频率均无统计学意义(P>0.05);④rs556442位点:与AA基因型相比,B、C组AG/GG基因型的股骨颈BMD水平降低;D组AG/GG型的HDL血清学水平降低(P<0.05)。rs312778位点:与CC基因型相比,A组HbA1C、FPG血清学水平较CT/TT基因型低(P<0.05);D组CT/TT基因型的低密度脂蛋白(LDL)血清学水平升高(P<0.05);⑤多元线性回归分析:rs556442位点,TG增加是腰L_(1~4)BMD降低的危险因素(P<0.05);rs312778位点,体质量指数(body mass index,BMI)降低是腰L_(1~4)及股骨颈BMD降低的危险因素,TG增加是腰L_(1~4)BMD降低的危险因素(P<0.05);⑥在rs556442、rs312778位点骨量异常与基因型分布均无统计学意义(P>0.05)。结论新疆石河子地区绝经后T2DM女性患者LRP5基因rs556442、rs312778基因位点的突变可能与骨量降低有关。

白藜芦醇激活细胞外信号调节激酶5信号蛋白促进小鼠MC3T3-E1细胞增殖

背景:细胞外信号调节激酶5信号蛋白对生物体的存活不可或缺,白藜芦醇能通过多种途径促进成骨细胞增殖,但其是否能通过细胞外信号调节激酶5信号蛋白调控成骨细胞功能还需进一步验证。目的:探究细胞外信号调节激酶5对MC3T3-E1细胞增殖以及相关分泌蛋白的调控作用,进一步验证白藜芦醇通过激活细胞外信号调节激酶5完成上述过程。方法:小鼠MC3T3-E1前成骨细胞分别用完全培养基、XMD8-92(细胞外信号调节激酶5抑制剂)、表皮生长因子(细胞外信号调节激酶5激活剂)和白藜芦醇单独干预及XMD8-92+表皮生长因子、白藜芦醇+XMD8-92干预后,通过Western blot检测各组细胞内细胞外信号调节激酶5、磷酸化细胞外信号调节激酶5蛋白,增殖相关蛋白Cyclin D1、CDK4、PCNA,以及成骨细胞分泌蛋白骨保护素、核因子κB受体活化因子配体的表达情况,使用细胞免疫荧光染色检测各组细胞外信号调节激酶5、骨保护素和核因子κB受体活化因子配体荧光强度,使用EdU染色检测各组细胞增殖情况。白藜芦醇干预MC3T3-E1细胞的适宜浓度及时间由细胞形态学观察和CCK-8实验确定。结果与结论:①细胞外信号调节激酶5信号蛋白的激活能有效促进MC3T3-E1细胞增殖、上调骨保护素/核因子κB受体活化因子配体比值;②白藜芦醇干预MC3T3-E1细胞的适宜浓度及时间为5μmol/L,24 h;③白藜芦醇可以激活细胞外信号调节激酶5信号蛋白,进而促进成骨细胞增殖,并上调骨保护素/核因子κB受体活化因子配体比值;④研究结果表明,白藜芦醇可以通过激活细胞外信号调节激酶5信号蛋白促进MC3T3-E1细胞增殖,并通过激活细胞外信号调节激酶5信号蛋白上调骨保护素/核因子κB受体活化因子配体比值。

5-HT_(2A)受体拮抗剂治疗神经精神疾病幻觉症状研究进展

帕金森病、阿尔茨海默病和精神分裂症等神经精神疾病在发展过程中会出现幻觉、妄想等精神病症状,这些症状发病率高、治愈难,严重影响患者生活质量。尽管经典抗精神病药物氯丙嗪、舒必利和奋乃静等通过拮抗多巴胺2型(D_(2))受体可治疗相关症状,但也会引发不可控的锥体外系反应和高泌乳素症等不良反应。近年来研究发现,奥氮平、氯氮平、利培酮和匹莫范色林等非经典抗精神病药物通过拮抗5-羟色胺2A(5-HT_(2A))受体或同时拮抗5-HT_(2A)受体(强)和D_(2)受体(弱)治疗神经精神疾病的幻觉症状。非临床研究结果表明,在多种因素诱导的幻觉模型上,非经典抗精神病药物均表现出良好的治疗作用;临床研究进一步证实,该类药物显著改善精神病症状(以幻觉和妄想为主),尤其是匹莫范色林,其对氯氮平、利培酮不敏感或耐受的患者仍能表现出良好的治疗效果。同时,非经典抗精神病药物不良反应的发生率和严重程度较低,总体耐受性较好。本文综述了5-HT_(2A)受体拮抗剂改善神经精神疾病伴随的幻觉症状的研究进展,为设计开发新型神经精神疾病治疗药物提供参考。

TGR5在心血管疾病中的作用研究进展

武田G蛋白偶联受体5(TGR5)是一种位于细胞膜表面的胆汁酸受体,广泛分布于体内许多组织细胞中,能被体内绝大多数胆汁酸直接激活。TGR5在多种生理和病理生理过程中发挥着重要作用,包括细胞Ca^(2+)转运、氧化应激、细胞增殖、炎症反应和线粒体代谢等,从而维持线粒体稳态和血管内膜功能,抑制动脉粥样硬化、心肌肥大、心肌梗死后心肌重塑等心血管疾病的进展。目前,随着许多胆汁酸及胆汁酸衍生物相关药物逐步投入临床应用,有必要进一步深入研究以明确TGR5在心血管系统中的作用。本文针对TGR5与心血管系统的相关性,从TGR5参与调节巨噬细胞、血管内膜功能、血管平滑肌、心肌细胞和线粒体代谢5个方面进行阐述,总结梳理最新的研究进展,以期为TGR5作为心血管疾病治疗靶点提供理论依据。

5-HT1A受体拮抗剂对七氟烷致老年认知功能障碍模型大鼠突触可塑性的作用及其机制

目的探讨5-HT1A受体拮抗剂对七氟烷致老年认知功能障碍模型大鼠突触可塑性的作用及其机制。方法将30只18月龄Sprague-Dawley大鼠随机分为对照组、模型组以及治疗组。模型组吸入50%空气、氧气混合物(2 L/min)和2%七氟烷4 h后左侧脑室给予生理盐水(5μL),治疗组在吸入50%空气、氧气混合物(2 L/min)和2%七氟烷4 h后左侧脑室给予5-HT1A受体拮抗剂(3μg),对照组仅吸入50%空气、氧气混合物(2 L/min)4 h。水迷宫法检测各组大鼠的学习记忆能力;HE染色法观察各组大鼠海马组织病理变化情况;尼氏染色和高尔基染色观察海马区神经元和突触的病理形态变化;免疫荧光法检测MeCP2、p250GAP、PSD-95、GAP-43与Syn蛋白表达;实时荧光定量PCR检测PKA、CREB1、BDNF mRNA表达水平;Western blotting检测PKA、CREB1、p-CREB1、BDNF蛋白表达水平。结果与对照组比较,模型组大鼠逃避潜伏期显著延长,穿越平台次数显著减少(P<0.05);与模型组比较,治疗组大鼠逃避潜伏期显著缩短,穿越平台次数明显增多(P<0.05)。HE、尼氏和高尔基染色显示,与对照组比较,模型组大鼠海马神经元形态不规则,排列松散,周围组织间隙扩大,细胞核固缩深染,部分神经元内尼氏体减少,树突分支数量以及树突棘密度明显减少;与模型组比较,治疗组大鼠海马神经元形态规则,结构相对完整,排列均匀,神经元内尼氏体数量增多,树突分支数量及树突棘密度显著增加。与对照组比较,模型组大鼠脑组织MeCP2、PSD-95、GAP-43、Syn蛋白,PKA、CREB1、BDNF mRNA和蛋白表达明显减少(P<0.05),p250GAP蛋白表达明显增加(P<0.05)。与模型组比较,治疗组MeCP2、PSD-95、GAP-43、Syn蛋白,PKA、CREB1、BDNF mRNA和蛋白表达明显增加(P<0.05),p250GAP蛋白表达明显减少(P<0.05)。结论5-HT1A受体拮抗剂通过激活CREB/BDNF通路,增强PSD-95、GAP-43、Syn表达,促进突触重塑,保护大鼠海马神经元细胞,从而改善七氟烷致老年认知功能障碍模型大鼠的学习记忆能力。

Inhibiting 5-hydroxytryptamine receptor 3 alleviates pathological changes of a mouse model of Alzheimer's disease

Extracellular amyloid beta(Aβ) plaques are main pathological feature of Alzheimer’s disease.However,the specific type of neuro ns that produce Aβ peptides in the initial stage of Alzheimer’s disease are unknown.In this study,we found that 5-hydroxytryptamin receptor 3A subunit(HTR3A) was highly expressed in the brain tissue of transgenic amyloid precursor protein and presenilin-1 mice(an Alzheimer’s disease model) and patients with Alzheimer’s disease.To investigate whether HTR3A-positive interneurons are associated with the production of Aβ plaques,we performed double immunostaining and found that HTR3A-positive interneurons were clustered around Aβ plaques in the mouse model.Some amyloid precursor protein-positive or β-site amyloid precursor protein cleaving enzyme-1-positive neurites near Aβ plaques were co-localized with HTR3A interneurons.These results suggest that HTR3A-positive interneurons may partially contribute to the generation of Aβ peptides.We treated 5.0-5.5-month-old model mice with tro pisetron,a HTR3 antagonist,for 8 consecutive weeks.We found that the cognitive deficit of mice was partially reversed,Aβ plaques and neuroinflammation we re remarkably reduced,the expression of HTR3 was remarkably decreased and the calcineurin/nuclear factor of activated T-cell 4 signaling pathway was inhibited in treated model mice.These findings suggest that HTR3A interneurons partly contribute to generation of Aβ peptide at the initial stage of Alzheimer’s disease and inhibiting HTR3 partly reve rses the pathological changes of Alzheimer’s disease.

5-HT_(2A)受体反向激动剂LPM6690061改善普拉克索致PD小鼠躁狂

为探讨5-HT_(2A)受体反向激动剂LPM6690061对普拉克索引起的帕金森病(PD)躁狂小鼠的症状改善作用及作用机制,采用1-甲基-4-苯基-1,2,3,6,-四氢吡啶(MPTP)建立小鼠PD模型,同时腹腔注射PPX建立小鼠PD躁狂模型,后灌胃给予2.0,6.0,18.0 mg/kg LPM6690061进行治疗。转棒、爬杆实验检测PD小鼠的运动障碍;悬尾、强迫游泳实验检测PD躁狂小鼠的情绪障碍;旷场实验检测PD躁狂小鼠的过度活动;Y迷宫实验检测PD躁狂小鼠的空间记忆;苏木精-伊红染色法检测海马体齿状回(DG)区细胞的损伤;蛋白免疫印迹法检测小鼠海马体糖原合酶激酶3β(GSK-3β)、磷酸化GSK-3β(P-GSK-3β)、丝氨酸/苏氨酸激酶(Akt)、磷酸化Akt(P-Akt)、细胞外调节蛋白激酶(Erk)、磷酸化Erk(P-Erk)、环磷腺苷效应元件结合蛋白(CREB)、磷酸化CREB(P-CREB)的表达。结果表明,5-HT_(2A)受体反向激动剂LPM6690061可改善普拉克索引起的PD小鼠的躁狂症状,抑制Akt/GSK-3β信号通路,降低海马体P-GSK-3β、P-Akt蛋白的表达;增强Erk/CREB通路,增加海马体P-Erk和P-CREB蛋白的表达,减少神经元凋亡,改善PD躁狂小鼠的空间记忆能力,减轻海马体DG区的颗粒细胞损伤。

2013—2022年房山区5岁以下儿童死亡情况分析

目的 分析房山区5岁以下儿童死亡率变化趋势及不同年龄儿童死亡原因的变化情况,针对5岁以下儿童死亡的主要原因,制定综合有效的管理措施,降低新生儿及婴儿死亡率。方法 采用北京市妇幼信息系统5岁以下儿童死亡监测网络收集2013—2022年5岁以下儿童死亡个案报告卡,统计儿童死亡率及不同年龄段死亡构成比,分析新生儿、婴儿根本死因及顺位情况。结果 2019—2022年早期新生儿、新生儿、婴儿死亡率大体上呈下降趋势,早期新生儿死亡率下降幅度最大。2019—2021年早期新生儿死亡的构成比大体上呈下降趋势。10年期间持续年份较长的根本死因顺位第一位是早产或低出生体重、出生窒息的新生儿,早产或低出生体重的婴儿;第二位是早产或低出生体重、出生窒息的新生儿,出生窒息、先天性心脏病的婴儿;第三位是其他先天异常的新生儿及婴儿。结论 加强孕妇围产期保健管理及高危孕妇孕期早期干预,督促定期随诊,提高高危孕妇分娩前的评估水平,提升产科医生孕期及分娩时的诊治技术、新生儿科医生危重新生儿安全转运评估质量及新生儿复苏技术成功水平,是降低新生儿及婴儿死亡的关键措施。