Blocking postsynaptic density-93 binding to C-X3-C motif chemokine ligand 1 promotes microglial phenotypic transformation during acute ischemic stroke

We previously reported that postsynaptic density-93 mediates neuron-microglia crosstalk by interacting with amino acids 357–395 of C-X3-C motif chemokine ligand 1(CX3 CL1) to induce microglia polarization. More importantly, the peptide Tat-CX3 CL1(comprising amino acids 357–395 of CX3 CL1) disrupts the interaction between postsynaptic density-93 and CX3 CL1, reducing neurological impairment and exerting a protective effect in the context of acute ischemic stroke. However, the mechanism underlying these effects remains unclear. In the current study, we found that the pro-inflammatory M1 phenotype increased and the anti-inflammatory M2 phenotype decreased at different time points. The M1 phenotype increased at 6 hours after stroke and peaked at 24 hours after perfusion, whereas the M2 phenotype decreased at 6 and 24 hours following reperfusion. We found that the peptide Tat-CX3 CL1(357–395 aa) facilitates microglial polarization from M1 to M2 by reducing the production of soluble CX3 CL1. Furthermore, the a disintegrin and metalloprotease domain 17(ADAM17) inhibitor GW280264 x, which inhibits metalloprotease activity and prevents CX3 CL1 from being sheared into its soluble form, facilitated microglial polarization from M1 to M2 by inhibiting soluble CX3 CL1 formation. Additionally, Tat-CX3 CL1(357–395 aa) attenuated long-term cognitive deficits and improved white matter integrity as determined by the Morris water maze test at 31–34 days following surgery and immunofluorescence staining at 35 days after stroke, respectively. In conclusion, Tat-CX3 CL1(357–395 aa) facilitates functional recovery after ischemic stroke by promoting microglial polarization from M1 to M2. Therefore, the Tat-CX3 CL1(357–395 aa) is a potential therapeutic agent for ischemic stroke.

C–C motif chemokine ligand 16 inhibits the progression of liver cirrhosis via inactivating hepatic stellate cells

Background:Liver cirrhosis results from many forms of chronic damage,characterized by accumulation of extracellular matrix.The present study aimed to explore a potential non-invasive biomarker and its mechanism in the progression of liver cirrhosis.Methods:Gene Expression Omnibus(GEO)dataset(GSE15654,n=216)was analyzed to screen genes associated with progression of liver cirrhosis.A total of 181 plasma samples,including healthy control(HC,n=20),chronic hepatitis B(CHB,n=77)and HBV-related liver cirrhosis(LC,n=84),were enrolled for validation.In vitro and in vivo experiments were employed for the mechanistic investigation.Results:GEO dataset analysis showed that relatively low mRNA-expression of C–C motif chemokine ligand 16(CCL16)was associated with elevated Child-Pugh score(P=0.034)and worse prognosis(P=0.025).Plasma CCL16 level decreased in a stepwise pattern,with a median concentration of 10.29,6.57 and 4.47 ng/mL in the HC,CHB and LC groups,respectively(P<0.001).Low plasma CCL16 was significantly related to hepatic dysfunction both in the CHB and LC groups(P<0.05).Combination of CCL16 and ALT showed improved distinguishing capability for LC compared to either alone.In vitro,CCL16 expression was downregulated by lipopolysaccharide and hypoxia.Overexpression of CCL16 from human normal liver cell line(LO2)reduced the extracellular matrix associated proteins(Col1 and Col4)in human hepatic stellate cell line(LX-2).In vivo,the pathological feature of cirrhosis was alleviated by the hepatocytespecific expression of CCL16.Conclusions:CCL16 could be a feasible plasma marker to predict the occurrence and progression of liver cirrhosis.CCL16 might impact liver cirrhosis through inactivating hepatic stellate cells.

Macrophage migration inhibitory factor facilitates astrocytic production of the CCL2 chemokine following spinal cord injury

Spinal cord injury causes accumulation of a large number of leukocytes at the lesion site where they contribute to excessive inflammation.Overproduced chemokines are responsible for the migratory process of the leukocytes,but the regulatory mechanism underlying the production of chemokines from resident cells of the spinal cord has not been fully elucidated.We examined the protein levels of macrophage migration inhibitory factor and chemokine C-C motif chemokine ligand 2 in a spinal cord contusion model at different time points following spinal cord injury.The elevation of macrophage migration inhibitory factor at the lesion site coincided with the increase of chemokine C-C motif chemokine ligand 2 abundance in astrocytes.Stimulation of primary cultured astrocytes with different concentrations of macrophage migration inhibitory factor recombinant protein induced chemokine C-C motif chemokine ligand 2 production from the cells,and the macrophage migration inhibitory factor inhibitor 4-iodo-6-phenylpyrimidine attenuated the stimulatory effect.Further investigation into the underlying mechanism on macrophage migration inhibitory factor-mediated astrocytic production of chemokine C-C motif chemokine ligand 2 revealed that macrophage migration inhibitory factor activated intracellular JNK signaling through binding with CD74 receptor.Administration of the macrophage migration inhibitory factor inhibitor 4-iodo-6-phenylpyrimidine following spinal cord injury resulted in the reduction of chemokine C-C motif chemokine ligand 2-recruited microglia/macrophages at the lesion site and remarkably improved the hindlimb locomotor function of rats.Our results have provided insights into the functions of astrocyte-activated chemokines in the recruitment of leukocytes and may be beneficial to develop interventions targeting chemokine C-C motif chemokine ligand 2 for neuroinflammation after spinal cord injury.

The prognostic value of C-X-C motif chemokine receptor 4 in patients with sporadic malignant peripheral nerve sheath tumors

Background: Recent studies indicate that C-X-C motif chemokine receptor 4(CXCR4) and its ligand, C-X-C motif chemokine ligand 12(CXCL12), stimulate expression of the cell cycle regulatory protein Cyclin D1 in neurofibromatosis 1-associated malignant peripheral nerve sheath tumor(MPNST) cells and promote their proliferation. In this study, we measured the expression of CXCR4, CXCL12, and Cyclin D1 proteins in sporadic MPNST tissues from Chinese patients and investigated their prognostic values.Methods: CXCR4, CXCL12, and Cyclin D1 protein expression in samples from 58 Chinese patients with sporadic MPNST was assessed with immunohistochemical staining.Their prognostic values were evaluated with Kaplan-Meier analysis and a log-rank test. Multivariate Cox regression analysis was used to identify independent prognostic factors.Results: High expression of CXCR4, CXCL12, and Cyclin D1 was observed in 19(32.8%), 32(55.2%), and 16(27.6%)samples, respectively. CXCR4 expression was positively correlated with CXCL12 expression(r = 0.334, P = 0.010) and Cyclin D1 expression(r = 0.309, P = 0.018). Patients with high CXCR4 expression showed longer overall survival than those with low CXCR4 expression(χ~2 = 4.642, P = 0.031).Conclusion: High CXCR4 expression may define a specific subtype of sporadic MPNST with favorable prognosis.

C-C基序趋化因子配体14的研究进展

C-C基序趋化因子配体14(CCL14)是一类趋化细胞定向移动的小分子蛋白质,可能参与多种疾病(包括肿瘤、免疫相关疾病、脓毒症、急性肾损伤)的发生发展。本文对近几年CCL14的研究进展进行了综述。

血清CX3CL1、CCL17与类风湿关节炎相关间质性肺疾病患者肺功能及预后的关系研究

目的分析血清C-X3-C基序趋化因子配体1(CX3CL1)、C-C基序趋化因子配体17(CCL17)与类风湿关节炎相关间质性肺疾病(RA-ILD)患者肺功能及预后的关系。方法选取2017年1月—2018年1月上海中医药大学附属龙华医院风湿科收治RA患者295例,根据是否合并ILD分为ILD组115例和非ILD组180例,根据预后情况将RA-ILD患者分为预后不良亚组和预后良好亚组。检测血清CX3CL1、CCL17水平及肺功能指标[第1秒用力呼气容积(FEV_(1))、用力肺活量(FVC)、FEV_(1)/FVC和肺一氧化碳弥散量(DLCO)]。采用Pearson相关性分析RA-ILD患者血清CX3CL1、CCL17水平与肺功能指标的相关性,多因素Logistic回归分析RA-ILD患者预后不良的影响因素,受试者工作特征(ROC)曲线分析血清CX3CL1、CCL17水平预测RA-ILD患者预后不良的价值。结果与非ILD组比较,ILD组血清CX3CL1、CCL17水平升高,FEV_(1)、FVC、FEV_(1)/FVC、DLCO降低(t/P=14.359/<0.001、13.855/<0.001、12.015/<0.001、2.732/0.007、14.749/<0.001、14.010/<0.001)。Pearson相关性分析显示,RA-ILD患者血清CX3CL1、CCL17水平与FEV_(1)、FVC、FEV_(1)/FVC、DLCO呈负相关(r=-0.762、-0.711、-0.577、-0.534、-0.707、-0.692、-0.735、-0.672,P均<0.001)。随访5年,115例RA-ILD患者预后不良发生率为46.96%。多因素Logistic回归分析显示,普通型间质性肺炎和高分辨率电子计算机断层扫描(HRCT)评分、CX3CL1、CCL17升高为RA-ILD患者预后不良的独立危险因素[OR(95%CI)=3.745(1.245~11.264)、1.051(1.008~1.095)、1.008(1.003~1.012)、1.037(1.012~1.062)],FEV_(1)、FEV_(1)/FVC、DLCO升高为独立保护因素[OR(95%CI)=0.952(0.896~0.988)、0.892(0.867~0.981)、0.857(0.789~0.949)]。ROC曲线分析显示,血清CX3CL1、CCL17水平联合预测RA-ILD患者预后不良的曲线下面积为0.875,大于CX3CL1、CCL17单独预测的0.783、0.788(Z=2.807、2.698,P=0.005、0.007)。结论RA-ILD患者血清CX3CL1、CCL17水平升高,与肺功能降低和预后不良有关,血清CX3CL1、CCL17水平联合检测对RA-ILD患者预后具有较高的预测价值。

宣肺合剂联合西药治疗慢性支气管炎急性发作的疗效

目的 观察宣肺合剂联合西药治疗慢性支气管炎急性发作的疗效,及其对血清CXC趋化因子受体4(CXCR4)、半乳糖凝集素-9(Gal-9)和趋化因子3(CCL3)水平的影响。方法 选择2020年1月-2021年12月在本院诊治的慢性支气管炎急性发作的患者96例,分为观察组和对照组各48例。对照组予以常规西药治疗,观察组在对照组的基础上予以宣肺合剂治疗。比较两组治疗后的疗效和不良反应,比较治疗前后喘息、咳痰、咳嗽积分、第一秒用力呼气量(FEV1)、用力肺活量(FVC)、最大通气量(MVV)、CXCR4、Gal-9和CCL3水平的变化。结果 观察组的总有效率为95.83%,对照组的总有效率为79.17%,观察组显著优于对照组(P<0.05);两组的不良反应率差异无统计学意义(P>0.05)。两组治疗前喘息、咳痰、咳嗽积分、FEV1、FVC、MVV、CXCR4、Gal-9和CCL3水平差异无统计学意义(P>0.05);治疗后两组的喘息、咳痰、咳嗽积分、CXCR4、Gal-9和CCL3水平均较治疗前显著降低,与对照组比较观察组出现明显降低(均P<0.01);治疗后两组的FEV1、FVC和MVV水平均较治疗前显著升高,与对照组比较观察组显著升高(均P<0.01)。结论 宣肺合剂能够提高西药对慢性支气管炎急性发作的疗效,缓解症状和改善肺功能,其机制可能与降低机体CXCR4、Gal-9和CCL3的水平有关。

Chemokine signaling involving chemokine (C-C motif) ligand 2 plays a role in descending pain facilitation

Objective Despite accumulating evidence on a role of immune cells and their associated chemicals in mecha- nisms of pain, few studies have addressed the potential role of chemokines in the descending facilitation of persistent pain. The present study was undertaken to test the hypothesis that the chemokine （C-C motif） ligand 2 （CCL2） （commonly known as monocyte chemoattractant protein-1） signaling in the rostral ventromedial medulla （RVM）, a pivotal structure in brainstem pain modulatory circuitry, is involved in descending pain facilitation in rats. Methods An L5 spinal nerve ligation （SNL） was produced in rats under pentobarbital anesthesia. Western blot and immunohistochemistry were used to detect the expression levels of CCL2 and CCL2 receptor （CCR2）, and examine their distributions compared with the neuronal marker NeuN as well as glial markers glial fibrillary acidic protein （GFAP, astroglial） and CD 11 b （microglial）, respectively. Results SNL induced an increase in CCL2 expression in the RVM, and this returned to the control level at 4 weeks after injury. The induced CCL2 colocalized with NeuN, but not with GFAP and CD1 lb. CCR2 was also upregu- lated by SNL in the RVM, and this increase lasted for at least 4 weeks. CCR2 was colocalized with CD1 lb but not GFAP. Few RVM neurons also exhibited CCR2 staining. Neutralizing CCL2 with an anti-CCL2 antibody （0.2-20 ng） or injecting RS-102895 （0.1-10 pmol）, a CCR2b chemokine receptor antagonist, into the RVM on day 1 after SNL, significantly at- tenuated the established thermal and mechanical hypersensitivity. In addition, injection of recombinant rat CCL2 （0.03-3 pmol） into the RVM induced dose-dependent hyperalgesia, which was prevented by pretreatment with RS-102895 （10 pmol）. Interleukin-β （IL-1]3）, a potent inducer of neuronal CCL2, was also selectively upregulated in RVM reactive as- trocytes. Injection of IL-1 ]3 （120 fmol） into the RVM induced behavioral hyperalgesia, which was blocked by RS-102895 （10 pmol）. However, an IL-1 receptor antagonist （3 pmol） did not prevent CCL2 （3 pmol）-induced hyperalgesia. These results suggest that the effect of CCL2 is downstream to IL-113 signaling. Conclusion The IL-1 β and CCL2-CCR2 signaling cascades play a role in neuron-glia-cytokine interactions and the descending facilitation of neuropathic pain.

Intestinal epithelial chemokine (C-C motif) ligand 7 overexpression protects against high fat diet-induced obesity and hepatic steatosis in mice

Background:We previously found that the intestinal epithelial chemokine(C-C motif)ligand 7(CCL7)plays an important role in the development of toxin-induced acute liver damage.The detailed effects of intestinal epithelial CCL7 on chronic diseases;however,are still unclear.Here,we aimed to investigate the impact of intestinal epithelial CCL7 overexpression on high-fat diet(HFD)-induced obesity and steatohepatitis in mice.Methods:Intestinal epithelial CCL7 overexpression(CCL7tgIEC)mice and their wild-type(WT)littermates were fed with normal chow or HFD for 16 weeks to induce obesity and non-alcoholic fatty liver disease.Body weight gain,as well as adipose tissue index were assessed.Liver injury was monitored by histological analysis and real time polymerase chain reaction.Gut microbial composition was analyzed by 16S rRNA gene sequencing.Results:We found that the CCL7tgIEC mice on a HFD had markedly decreased weight gain(8.9 vs.17.0 g,P<0.05)and a lower adipose tissue index that include mesenteric fat(1.0%vs.1.76%,P<0.05),gonadal fat(2.1%vs.6.1%,P<0.05),subcutaneous fat(1.0%vs.2.8%,P<0.05)compared to WT animals.HFD-induced glucose intolerance and insulin resistance were also significantly improved in CCL7tgIEC mice compared to WT.Furthermore,HFD-fed CCL7tgIEC mice displayed less hepatic lipid accumulation and lower expression of inflammatory factors than WT mice.16S rRNA gene sequencing demonstrated that CCL7 overexpression in intestinal epithelial cells improved HFD-induced gut microbial dysbiosis.Conclusions:Our study revealed that CCL7 overexpression in the intestinal epithelium protects mice against the progression of diet-induced obesity,hepatic steatosis,and enteric dysbiosis.

The miR-9-5p/CXCL11 pathway is a key target of hydrogen sulfide-mediated inhibition of neuroinflammation in hypoxic ischemic brain injury

We previously showed that hydrogen sulfide(H2S)has a neuroprotective effect in the context of hypoxic ischemic brain injury in neonatal mice.However,the precise mechanism underlying the role of H2S in this situation remains unclear.In this study,we used a neonatal mouse model of hypoxic ischemic brain injury and a lipopolysaccharide-stimulated BV2 cell model and found that treatment with L-cysteine,a H2S precursor,attenuated the cerebral infarction and cerebral atrophy induced by hypoxia and ischemia and increased the expression of miR-9-5p and cystathionineβsynthase(a major H2S synthetase in the brain)in the prefrontal cortex.We also found that an miR-9-5p inhibitor blocked the expression of cystathionineβsynthase in the prefrontal cortex in mice with brain injury caused by hypoxia and ischemia.Furthermore,miR-9-5p overexpression increased cystathionine-β-synthase and H2S expression in the injured prefrontal cortex of mice with hypoxic ischemic brain injury.L-cysteine decreased the expression of CXCL11,an miR-9-5p target gene,in the prefrontal cortex of the mouse model and in lipopolysaccharide-stimulated BV-2 cells and increased the levels of proinflammatory cytokines BNIP3,FSTL1,SOCS2 and SOCS5,while treatment with an miR-9-5p inhibitor reversed these changes.These findings suggest that H2S can reduce neuroinflammation in a neonatal mouse model of hypoxic ischemic brain injury through regulating the miR-9-5p/CXCL11 axis and restoringβ-synthase expression,thereby playing a role in reducing neuroinflammation in hypoxic ischemic brain injury.

新生儿持续肺动脉高压患儿血清CXCL8、CXCL12与一氧化氮吸入治疗临床转归的关系

目的探讨新生儿持续肺动脉高压(persistent pulmonary hypertension of newborn,PPHN)患儿血清人CXC型趋化因子配体8(C-X-C motif chemokine ligand 8,CXCL8)、CXCL12与一氧化氮吸入治疗临床转归的关系。方法选择2021-08/2023-05月作者医院收治并给予一氧化氮吸入治疗的PPHN患儿135例为研究对象。根据患儿出院时临床转归结局分为死亡组(n=32)和存活组(n=103)。比较两组PPHN患儿血清CXCL8、CXCL12水平。单因素及多因素Logistic回归模型分析接受一氧化氮吸入治疗PPHN患儿临床转归的影响因素。受试者工作特征(receiver operating characteristic,ROC)曲线分析血清CXCL8、CXCL12对接受一氧化氮吸入治疗PPHN患儿临床转归的预测价值。结果死亡组患儿血清CXCL8、CXCL12水平显著高于存活组(P均<0.05)。多因素Logsitic回归分析结果显示,血清CXCL8水平升高、血清CXCL12水平升高、早产、出生时Apgar评分0~3分、合并并发症是接受一氧化氮吸入治疗的PPHN患儿死亡的危险因素,肺表面活性物质应用、吸入一氧化氮早期反应则是保护因素(P<0.05)。ROC曲线分析结果显示,血清CXCL8、CXCL12联合检测对接受一氧化氮吸入治疗的PPHN患儿死亡预测的曲线下面积(area under the curve,AUC)为0.828,大于血清CXCL8、CXCL12单独检测(AUC分别为0.762、0.714)。结论PPHN患儿血清CXCL8、CXCL12水平升高与接受一氧化氮治疗的不良临床转归有关,且CXCL8、CXCL12水平升高是PPHN患儿死亡的危险因素。CXCL8、CXCL12联合检测对接受一氧化氮治疗PPHN患儿死亡具有较高的预测价值。

信号转导和转录激活因子3/CC趋化因子配体2信号通路对乳腺癌细胞活力、迁移和侵袭的影响实验研究

目的:探讨信号转导和转录激活因子3(STAT3)/CC趋化因子配体2(CCL2)信号通路对乳腺癌细胞活力、迁移和侵袭的影响。方法:体外培养人乳腺癌细胞株(HCC1937、MCF-7、MDA-MB-231、ZR-75-1)和人正常乳腺上皮细胞(MCF-10A)。采用蛋白印迹法检测HCC1937、MCF-7、MDA-MB-231、ZR-75-1和MCF-10A细胞磷酸化STAT3(p-STAT3)、STAT3、CCL2蛋白表达。选取p-STAT3/STAT3蛋白比值、CCL2蛋白表达最高的人乳腺癌细胞进行后续实验。将MCF-7细胞分为对照组、STAT3抑制剂(WP1066)Ⅰ组(2μmol/L WP1066)、Ⅱ组(4μmol/L WP1066)、Ⅲ组(8μmol/L WP1066)。分别采用细胞计数试剂盒-8、划痕实验、Transwell实验检测MCF-7细胞活力、迁移率及侵袭能力。采用蛋白印迹法检测MCF-7细胞p-STAT3、STAT3、CCL2、基质金属蛋白酶(MMP)-2、MMP-9蛋白表达。结果:与MCF-10A细胞比较,HCC1937、MCF-7、MDA-MB-231、ZR-75-1细胞p-STAT3/STAT3蛋白比值、CCL2蛋白表达升高(均P<0.05)。其中MCF-7细胞p-STAT3/STAT3蛋白比值、CCL2蛋白表达最高,因此选用MCF-7细胞进行后续实验。与对照组比较,STAT3抑制剂Ⅰ、Ⅱ、Ⅲ组MCF-7细胞活力、迁移率、侵袭数目依次降低(均P<0.05)。与对照组比较,STAT3抑制剂Ⅰ、Ⅱ、Ⅲ组MCF-7细胞p-STAT3/STAT3蛋白比值、CCL2、MMP-2及MMP-9蛋白表达依次降低(均P<0.05)。结论:乳腺癌细胞中STAT3/CCL2呈高表达,抑制STAT3/CCL2信号通路能够降低乳腺癌细胞活力,减少迁移和侵袭。

十溴联苯醚灌胃后小鼠宫颈癌皮下移植瘤组织肿瘤相关成纤维细胞标志基因表达变化及其意义

目的观察十溴联苯醚(BDE-209)灌胃后小鼠宫颈癌皮下移植瘤组织肿瘤相关成纤维细胞(CAFs)标志基因表达变化,并探讨其参与宫颈癌发生发展的相关机制。方法将40只雌性C57BL/6小鼠随机分为对照组、BDE-209低剂量组、BDE-209中剂量组、BDE-209高剂量组,每组10只。各组小鼠均采用右前腋下接种宫颈癌U14细胞悬液的方法建立宫颈癌皮下移植瘤模型,BDE-209低剂量组、BDE-209中剂量组、BDE-209高剂量组经口灌胃给予20、100、500 mg/kg BDE-209,对照组给予等体积玉米油,连续21 d。取各组小鼠宫颈癌皮下移植瘤组织,进行转录组测序后筛选差异表达基因,与查阅文献得到的小鼠CAFs标志基因取交集后获得CAFs标志差异表达基因。对CAFs标志差异表达基因进行基因本体论(GO)功能和基因组百科全书(KEGG)通路富集分析,建立蛋白质相互作用网络(PPI)图后采用Betweenness算法确定排名前十的CAFs标志差异表达基因,即为CAFs标志差异核心基因。取BDE-209高剂量组与对照组小鼠部分宫颈癌皮下移植瘤组织,采用实时荧光定量PCR法检测其10个CAFs标志差异核心基因表达。结果共筛选到CAFs标志差异表达基因30个,其中BDE-209干预后上调29个、下调1个。GO功能富集分析结果显示,CAFs标志差异基因主要富集在细胞外基质(ECM)、生殖结构发育、PI3K/Akt信号转导等;KEGG通路富集分析结果显示,CAFs标志差异基因主要富集在补体和凝血级联反应、肿瘤发病途径等信号通路。PPI图和Betweenness算法筛选得到排名前十的CAFs标志差异核心基因分别为核心蛋白聚糖(Dcn)、细胞间黏附分子1(Icam1)、C-X-C基序趋化因子配体12(Cxcl12)、基质金属蛋白酶2(MMP-2)、血小板衍生生长因子受体(Pdgfrb)、Ⅴ型胶原蛋白α2(Col5a2)、补体成分1的子成分1(C1s1)、肝细胞生长因子(Hgf)、补体成分3(C3)、纤溶酶原激活物(Plat)。与对照组比较,BDE-209高剂量组小鼠移植瘤组织Dcn、Icam1、Cxcl12、MMP-2、Pdgfrb、Col5a2、C1s1、Hgf、C3、Plat mRNA相对表达量均升高(P均<0.05),Pdgfrb mRNA相对表达量无明显变化(P>0.05)。结论BDE-209灌胃会导致小鼠宫颈癌皮下移植瘤组织CAFs相关标志基因Dcn、Icam1、Cxcl12、MMP-2、Col5a2、C1s1、Hgf、C3、Plat表达升高,上述基因表达改变可能通过诱导CAFs的ECM变化和激活PI3K/Akt信号通路参与宫颈癌的发生发展。

C-X3-C基序趋化因子配体1/C-X3-C基序趋化因子受体1通路参与失血性休克/复苏大鼠记忆功能恢复

目的探讨C-X3-C基序趋化因子配体1(CX3CL1)/C-X3-C基序趋化因子受体1(CX3CR1)通路调控的小胶质细胞活化对失血性休克/复苏大鼠记忆功能的影响。方法实验分为2部分。第1部分,将大鼠随机分为假手术组,模型-0.5 h组,模型-1.5 h组,模型-3 h组,每组10只,模型组之间失血性休克时间存在差异。第2部分,大鼠随机分为对照组与CX3CL1组,每组10只。CX3CL1组大鼠脑室注射CX3CL1蛋白,对照组大鼠注射生理盐水。所有大鼠在模型制作前开展Morris水迷宫训练,模型制作后4 d,开展水迷宫测试。完成后,取全脑组织进行HE染色与免疫组织化学染色,取脑脊液检测炎性细胞因子含量,取脑组织进行Real-time PCR检测与Western blotting检测。结果与假手术组相比,模型组大鼠逃避潜伏期增加,穿越平台次数与第Ⅲ象限停留时间减少,且HE染色中显示神经元状态受损。此外,与假手术组相比,模型组大鼠脑组织中离子钙结合衔接分子1(Iba1)表达升高,脑脊液中肿瘤细胞坏死因子α(TNF-α)与白细胞介素(IL)-6含量升高,M1型小胶质细胞标记CD16、TNF-α、IL-1β与诱导性一氧化氮合酶(iNOS)mRNA含量升高。与此同时,与假手术组相比,模型组大鼠脑组织中CX3CL1、CX3CR1蛋白表达降低,磷酸化核因子κB(p-NF-κB)与核苷酸结合寡聚结构域(NOD)样受体蛋白3(NLRP3)蛋白表达升高。然而,与模型组相比,CX3CL1组大鼠逃避潜伏期减少,穿越平台次数与第Ⅲ象限停留时间增加,且神经元状态恢复。此外,与对照组相比,CX3CL1组大鼠脑组织中Iba1表达降低,脑脊液中TNF-α与IL-6含量降低,M1型小胶质细胞标记CD16、TNF-α、IL-1β与iNOS mRNA含量降低,M2型小胶质细胞标记CD206、转化生长因子β(TGF-β)、精氨酸酶1(Arg1)、几丁质酶3样蛋白1(Ym1)mRNA含量升高。结论CX3CL1有助于抑制小胶质细胞过度激活,诱导小胶质细胞向M2型极化,抑制M1型极化,降低炎性细胞因子释放,减轻失血性休克/复苏诱发的记忆功能损伤。

细胞外基质刚度诱导CCL5合成以提高非小细胞肺癌免疫治疗响应

目的探讨非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞外基质刚度调控趋化因子配体5(C-C motif chemokine ligand 5,CCL5)的机制,揭示CCL5对NSCLC的免疫治疗响应的影响。方法通过TCGA数据库分析NSCLC细胞外基质刚度与CCL5表达的相关性;根据NSCLC细胞外基质刚度设计不同刚度的聚丙烯酰胺水凝胶,获得响应水凝胶基质刚度力学加载的H1299细胞并对其进行转录组测序,利用q-PCR验证高基质刚度对CCL5表达的影响;进一步利用NSCLC患者转录组数据探究CCL5与免疫治疗响应的相关性。结果高细胞外基质刚度可以上调CCL5的表达,并且干扰素-γ(interferonγ,IFN-γ)介导的信号通路可能参与了该过程;CCL5高表达的NSCLC患者,肿瘤组织中细胞毒性T细胞的丰度更高,与抗程序性死亡受体-1(programmed cell death protein 1,PD-1)治疗响应性有关。结论细胞外基质刚度增加可促进CCL5合成,CCL5可通过增加肿瘤组织中细胞毒性T细胞浸润提升NSCLC的免疫治疗响应性。

脓毒症继发急性肺损伤患者血清CCL25和PARK7表达水平及其临床价值研究

目的探讨脓毒症患者血清C-C模体趋化因子配体25(C-C motif chemokine ligand 25,CCL25),人帕金森病蛋白7(Parkinson’s disease protein 7,PARK7)水平与急性肺损伤(acute lung injury,ALI)的关系及临床意义。方法选取2019年2月~2023年2月合肥京东方医院诊治的138例脓毒症患者为脓毒症组,根据是否继发ALI分为ALI组(n=40)和非ALI组(n=98),以同期体检的70例健康人为对照。酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测血清CCL25和PARK7水平。Pearson相关分析血清CCL25,PARK7与临床指标的相关性。多因素Logistic回归分析脓毒症继发ALI的危险因素。受试者工作特征曲线分析血清CCL25和PARK7水平对脓毒症继发ALI的预测价值。结果脓毒症组患者血清CCL25(367.52±46.87ng/L),PARK7(54.26±17.45μg/L)水平高于对照组(48.17±5.26ng/L,12.31±4.12μg/L),差异具有统计学意义(t=46.825,19.813,均P<0.05)。ALI组患者血清CCL25(434.65±52.87ng/L vs 340.12±42.64ng/L),PARK7(103.47±22.51μg/L vs 34.18±7.46μg/L),呼吸指数(1.58±0.48 vs 0.88±0.07)、动脉血二氧化碳分压(Pa CO_(2))(50.11±6.27mm Hg vs 40.42±5.20mm Hg)、急性生理学与慢性健康状况评价Ⅱ(APACHEⅡ)评分(23.37±3.82分vs 17.15±3.41分)、序贯器官衰竭(SOFA)评分(13.56±2.93分vs 10.18±2.81分)均高于非ALI组,而氧合指数(237.14±23.56分vs341.14±21.37分)、动脉血氧分压(Pa O_(2))(55.87±8.03mm Hg vs 63.11±7.14mm Hg)低于非ALI组,差异具有统计学意义(t=10.998,27.151,14.145,9.342,9.385,6.332,25.172,5.210,均P<0.05)。ALI组患者血清CCL25,PARK7水平与APACHEⅡ评分、SOFA评分、呼吸指数、Pa CO_(2)呈正相关(r=0.579~0.801,均P<0.05),与氧合指数、Pa O_(2)呈负相关(r=-0.687,-0.643;-0.654,-0.712,均P<0.05)。血清CCL25(OR=1.309,95%CI:1.040~1.646),PARK7(OR=1.288,95%CI:1.016~1.633),APACHEⅡ评分(OR=1.188,95%CI:1.019~1.384),SOFA评分(OR=1.197,95%CI:1.006~1.425)是影响脓毒症患者继发ALI的独立危险因素。血清CCL25,PARK7联合对脓毒症继发ALI预测的曲线下面积(95%CI)[0.833(0.784~0.872)]大于单项指标[0.770(0.725~0.835),0.741(0.691~0.790)],差异具有统计学意义(Z=4.602,4.318,均P<0.05)。结论脓毒症患者血清CCL25和PARK7水平升高,是影响脓毒症继发ALI发生的独立危险因素,两者联合对脓毒症继发ALI具有较高的预测价值。

血清miR-31、miR-155和CCL26在溃疡性结肠炎中预后不良的诊断价值

目的观察血清miR-31、miR-155和趋化因子配体26(CCL26)在溃疡性结肠炎中预后不良的诊断价值。方法选择2019年1月—2020年6月在我院收治的溃疡性结肠炎的患者119例,根据Mayo评分将其分为活动期组(65例)和缓解期组(54例);同时根据Mayo评分将活动期组分为轻度亚组(25例),中度亚组(22例)和重度亚组(18例);另选择同期在我院健康体检者45例为对照组。比较各组血清miR-31、miR-155和CCL26水平的变化;分析血清miR-31、miR-155和CCL26水平与溃疡性结肠炎严重程度的关系及影响溃疡性结肠炎预后不良的单因素和多因素分析;评估血清miR-31、miR-155和CCL26水平在2年内溃疡性结肠炎发生预后不良的诊断价值。结果溃疡性结肠炎活动期组血清miR-31、miR-155和CCL26水平明显高于缓解期组和对照组(P<0.05),而缓解期组明显高于对照组(P<0.05)。血清miR-31、miR-155和CCL26水平随着溃疡性结肠炎严重程度的增加而升高(P<0.05)。单因素和多因素分析发现UCEIS评分>6分,血清miR-31、miR-155和CCL26水平升高是发生溃疡性结肠炎预后不良的独立危险因素(P<0.05)。血清miR-31、miR-155和CCL26水平在诊断溃疡性结肠炎发生预后不良具有较高的诊断效能,联合检测的灵敏度为80%,特异度为99.4%,AUC为0.963明显高于单个指标miR-31(z=2.727,P=0.006)、miR-155(z=3.012,P=0.003)和CCL26(z=2.091,P=0.036)。结论miR-31、miR-155和CCL26参与了溃疡性结肠炎的发生发展,与溃疡性结肠炎的严重程度有关,联合检测有助于提高对溃疡性结肠炎2年内发生预后不良的诊断价值。

血清CX3C基序趋化因子配体1及Ⅰ型纤溶酶原激活抑制物水平与慢性阻塞性肺疾病合并Ⅱ型呼吸衰竭患者病情严重程度及预后的关系研究

目的 研究慢性阻塞性肺疾病(COPD)合并Ⅱ型呼吸衰竭患者血清CX3C基序趋化因子配体1(CX3CL1)、Ⅰ型纤溶酶原激活抑制物(PAI-1)水平与病情严重程度及预后的关系。方法 选取2019年1月至2021年1月重庆医科大学附属永川医院收治的COPD合并Ⅱ型呼吸衰竭患者96例为研究对象(呼吸衰竭组)、同期诊治的单纯COPD患者60例为单纯COPD组、同期健康体检的健康人群60例为健康对照组。检测受试者血清CX3CL1、PAI-1水平。比较不同健康状况受试者、不同病情严重程度呼吸衰竭组患者血清CX3CL1、PAI-1水平差异。分析影响COPD合并Ⅱ型呼吸衰竭患者不良预后的因素以及血清CX3CL1、PAI-1对不良预后的预测价值。结果 呼吸衰竭组、单纯COPD组CX3CL1、PAI-1水平均高于健康对照组[(266±29)、(219±35)ng/L比(156±23)ng/L,(15.2±2.3)、(12.3±2.2)μg/L比(8.2±1.8)μg/L],且呼吸衰竭组高于单纯COPD组(均P<0.05)。呼吸衰竭组中轻度组、中度组、重度组血清CX3CL1、PAI-1水平随疾病严重程度加重呈上升趋势(均P<0.05)。COPD合并Ⅱ型呼吸衰竭患者血清CX3CL1、PAI-1水平与第1秒用力呼气容积占预计值百分比(FEV1%)、第1秒用力呼气容积与用力肺活量比值(FEV1/FVC比值)呈负相关(均P<0.001)。呼吸衰竭组中预后不良组血清CX3CL1、PAI-1水平均高于预后良好组(均P<0.05)。多因素Logistic回归分析结果显示,血清CX3CL1、PAI-1升高是COPD合并Ⅱ型呼吸衰竭患者不良预后的独立危险因素(均P<0.001)。血清CX3CL1、PAI-1联合检测对COPD合并Ⅱ型呼吸衰竭患者不良预后预测的曲线下面积大于CX3CL1、PAI-1单独诊断(均P<0.001)。结论 COPD合并Ⅱ型呼吸衰竭患者血清CX3CL1、PAI-1水平升高,二者与病情严重程度有关,联合检测有助于评估COPD合并Ⅱ型呼吸衰竭患者的临床预后。

急性髓系白血病病人免疫球蛋白κ链C区蛋白和趋化因子配体2表达及意义

目的:探讨急性髓系白血病病人免疫球蛋白κ链C区蛋白(IGKC)、趋化因子配体2(CCL2)表达水平及临床意义。方法:选取急性髓系白血病病人80例作为观察组,另选取同期体检健康者40名作为对照组。采用酶联免疫吸附法检测2组病人血清IGKC、CCL2水平,并比较观察组不同分型病人血清IGKC、CCL2水平;Pearson法分析观察组病人IGKC、CCL2水平与降钙素原(PCT)、C反应蛋白(CRP)的相关性;ROC曲线分析血清IGKC、CCL2、PCT及CRP水平对观察组不同分型病人的诊断价值。结果:观察组病人血清IGKC、CCL2、PCT、CRP水平均明显高于对照组(P<0.01);不同WHO分型、ELN分型病人血清IGKC、CCL2、PCT、CRP水平间差异均有统计学意义(P<0.01),其中,M3~M4型、M5~M6型病人血清IGKC、CCL2、PCT、CRP水平均高于M1~M2型(P<0.05);与良好型病人相比,中等型和不良型病人血清IGKC、CCL2、PCT、CRP水平均高于良好型病人(P<0.05)。Pearson相关分析显示,观察组病人血清IGKC与CCL2、PCT、CRP均呈正相关关系(r=0.393、0.302、0.487,P<0.05),CCL2与PCT、CRP均呈正相关关系(r=0.510、0.516,P<0.05)。ROC曲线分析结果显示,血清IGKC、CCL2诊断M1~M2型及M5~M6型急性髓系白血病水平较高,AUC均达到0.75以上。结论:急性髓系白血病病人血清IGKC、CCL2均呈高表达,二者对急性髓系白血病有一定的辅助诊断价值。

ZCCHC10通过激活P53促进急性髓系白血病细胞中CCL18基因的转录

肿瘤抑制因子P53是一种转录因子,可激活一系列靶基因的转录,从而调控细胞周期停滞、DNA修复、免疫、炎症和细胞凋亡等多种生理或病理过程。前期研究表明,CCHC型锌指蛋白10(zinc finger CCHC-type containing 10,ZCCHC10)通过激活P53在肺癌和急性髓系白血病中发挥抑癌作用。为进一步探讨ZCCHC10在急性髓系白血病中的作用机制,本文通过RNA测序(RNA sequencing,RNA-seq)技术对过表达ZCCHC10或空载体的ML2细胞进行了转录组分析,一共鉴定到1284个差异基因[|log2(fold change)|逸1,q值约0.05],包括778个上调基因和506个下调基因。其中,趋化因子CCL18在过表达ZCCHC10的ML2细胞中上调18倍。生物信息学分析显示,CCL18基因启动子上含有两个P53反应元件。生物素标记DNA亲和实验和染色质免疫共沉淀实验证实,P53可结合到CCL18基因启动子上。荧光素酶活性分析表明,P53可以增强CCL18基因启动子的活性。这些研究表明ZCCHC10通过激活P53促进CCL18基因的表达。