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 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.

CCL11、MK对分化型甲状腺癌患者预后的评估价值

目的探讨CC修饰趋化因子11(CCL11)、中期因子(MK)对分化型甲状腺癌(DTC)患者预后的评估价值。方法纳入2015年1月-2017年1月河南省人民医院收治的150例DTC患者作为DTC组,另选取同期150例良性甲状腺病变患者为良性组,150名健康志愿者为对照组。比较三组血清CCL11、MK水平。DTC患者按预后情况分为生存、死亡亚组,比较生存与死亡患者的临床资料,采用Cox回归分析DTC预后的影响因素;采用受试者工作特征(ROC)曲线分析血清CCL11、MK水平对DTC预后的评估价值。结果与对照组比较,DTC组、良性组确诊时血清CCL11、MK水平升高,且DTC组高于良性组(P<0.05);DTC组、良性组术后3 d血清CCL11、MK水平均低于确诊时,且DTC组高于良性组(P<0.05);DTC患者生存135例,死亡15例;DTC生存与死亡患者的年龄、肿瘤直径、TNM分期、淋巴结转移、包膜侵犯、分化程度及血清甲状腺球蛋白(Tg)、CCL11、MK水平比较,差异有统计学意义(P<0.05);Cox回归分析结果显示,TNM分期、淋巴结转移及血清Tg、CCL11、MK水平为DTC预后的影响因素(P<0.05);ROC曲线分析结果显示,血清CCL11、MK水平对DTC预后的评估价值较高,二者联合时预测效能更高(敏感度为93.33%,特异度为73.33%,ROC曲线下面积为0.835)。结论DTC患者血清CCL11、MK水平异常升高,二者联合检测对DTC预后的评估价值较高。

不同程度活动期溃疡性结肠炎患者血清IFABP、MUC1、CCL11水平变化及对预后的影响

目的探讨不同程度活动期溃疡性结肠炎(UC)患者血清肠型脂肪酸结合蛋白(IFABP)、黏蛋白1(MUC1)、C-C基序趋化因子配体11(CCL11)水平变化及对预后的影响。方法选择2018年6月至2020年1月陕西省西安市高陵区医院收治的活动期UC患者80例作为UC组,根据病情严重程度将UC患者分为轻度组(36例)、中度组(28例)和重度组(16例),根据患者随访2年预后情况分为预后不良组(29例)和预后良好组(51例);另选择同期在陕西省西安市高陵区医院健康体检者60例作为对照组。采用酶联免疫吸附试验检测各组研究对象血清IFABP、MUC1、CCL11水平,分析活动期UC患者预后不良的影响因素,采用受试者工作特征(ROC)曲线分析血清IFABP、MUC1、CCL11水平对活动期UC患者预后不良的预测价值。结果UC组患者血清IFABP、MUC1、CCL11水平均高于对照组,差异均有统计学意义(P<0.05)。轻度组、中度组、重度组患者血清IFABP、MUC1、CCL11水平依次升高,差异有统计学意义(P<0.05)。80例活动期UC患者预后不良发生率为36.25%(29/80)。病情为重度、IFABP≥161.43 pg/mL、MUC1≥17.62 U/mL、CCL11≥161.13 pg/mL为活动期UC患者预后不良的独立危险因素(P<0.05)。血清IFABP、MUC1、CCL11水平联合检测预测活动期UC患者预后不良的ROC曲线下面积大于各项指标单独检测,差异有统计学意义(P<0.05)。结论活动期UC患者血清IFABP、MUC1、CCL11水平升高与病情加重和预后不良有关,可作为活动期UC患者预后辅助预测指标。

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.

血清CCL11、GPX3和Gal-1水平在分化型甲状腺癌诊断中的临床价值

目的探讨血清嗜酸性粒细胞趋化因子(CCL11)、谷胱甘肽过氧化物酶3(GPX3)和半乳糖凝集素-1(Gal-1)水平在分化型甲状腺癌诊断中的临床价值。方法选择2019年1月至2020年12月在该院经手术病理确诊的分化型甲状腺癌患者97例为分化型甲状腺癌组;选择同期在该院就诊的良性甲状腺疾病患者65例和体检健康者45例,分别为甲状腺良性疾病组和健康对照组。采用酶联免疫吸附试验检测各组血清CCL11、GPX3和Gal-1水平。结果分化型甲状腺癌组血清CCL11和Gal-1水平高于甲状腺良性疾病组和健康对照组(P<0.05),甲状腺良性疾病组高于健康对照组(P<0.05),分化型甲状腺癌组术后血清CCL11和Gal-1水平较术前出现明显降低(P<0.05);分化型甲状腺癌组血清GPX3水平低于甲状腺良性疾病组和健康对照组(P<0.05),甲状腺良性疾病组低于健康对照组(P<0.05),分化型甲状腺癌组术后血清GPX3水平较术前出现明显升高(P<0.05)。分化型甲状腺癌患者血清CCL11、GPX3和Gal-1水平与淋巴结转移情况、浸润深度和TNM分期有关(P<0.05),而与性别、年龄、肿瘤最大径、组织学类型和分化程度无关(P>0.05)。血清CCL11、GPX3和Gal-1对分化型甲状腺癌具有较高的诊断效能,3项联合检测的灵敏度为91.8%,特异度为92.3%,曲线下面积为0.962,明显高于各指标单独检测(P<0.05)。分化型甲状腺癌患者血清CCL11(r=-0.625,P<0.05)和Gal-1(r=-0.716,P<0.05)水平与GPX3水平呈负相关,而血清CCL11水平与Gal-1水平呈正相关(r=0.647,P<0.05)。结论血清CCL11、GPX3和Gal-1参与了分化型甲状腺癌的发生发展过程,联合检测有助于提高对分化型甲状腺癌的诊断效能。

Neuroinflammation in mild respiratory COVID‑19:insights into cognitive impairment in milder cases

Severe acute respiratory syndrome coronavirus 2(SARSCoV-2)infection has been extensively shown to cause many neurological sequelae,and cognitive deficits(known as“brain fog”)may particularly and widely occur even in individuals with mild symptoms[1].Peripheral hyperinflammation as well as central nervous system(CNS)local immune responses may synergistically contribute to brain autoimmune injury.In addition to the direct neuroinvasion of SARS-CoV-2 and nonimmune effects such as severe systemic hypoxemia and vascular thrombosis,the central mechanism by which SARSCoV-2 accelerates cognitive-related symptoms may be related to immune effects[2].However,the precise neuroinflammatory mechanisms of SARS-CoV-2 infection have not been fully established.Fernández-Casta-da et al.[3]provided direct evidence and unique insights into the potential mechanism of cognitive impairment in mild respiratory coronavirus disease 2019(COVID-19)cases.

Transforming growth factor-β and toll-like receptor-4 polymorphisms are not associated with fibrosis in haemochromatosis

AIM:To investigate the role of genetic polymorphisms in the progression of hepatic fibrosis in hereditary haemochromatosis.METHODS:A cohort of 245 well-characterised C282Y homozygous patients with haemochromatosis was studied,with all subjects having liver biopsy data and DNA available for testing.This study assessed the association of eight single nucleotide polymorphisms(SNPs)in a total of six genes including toll-like receptor 4(TLR4),transforming growth factor-beta(TGF-β),oxoguanine DNA glycosylase,monocyte chemoattractant protein 1,chemokine C-C motif receptor 2 and interleukin-10 with liver disease severity.Genotyping was performed using high resolution melt analysis and sequencing.The results were analysed in relation to the stage of hepatic fibrosis in multivariate analysis incorporating other cofactors including alcohol consumption and hepatic iron concentration.RESULTS:There were significant associations between the cofactors of male gender(P=0.0001),increasing age(P=0.006),alcohol consumption(P=0.0001),steatosis(P=0.03),hepatic iron concentration(P<0.0001)and the presence of hepatic fibrosis.Of the candidate gene polymorphisms studied,none showed a significant association with hepatic fibrosis in univariate or multivariate analysis incorporating cofactors.We also specifically studied patients with hepatic iron loading above threshold levels for cirrhosis and compared the genetic polymorphisms between those with no fibrosis vs cirrhosis however there was no significant effect from any of the candidate genes studied.Importantly,in this large,well characterised cohort of patients there was no association between SNPs for TGF-βor TLR4and the presence of fibrosis,cirrhosis or increasing fibrosis stage in multivariate analysis.CONCLUSION:In our large,well characterised group of haemochromatosis subjects we did not demonstrate any relationship between candidate gene polymorphisms and hepatic fibrosis or cirrhosis.

Expression Changes of Serum IL-1α,CCL2,and CXCL2 in Patients With Pemphigus

Objective:This study was performed to explore the possible changes of the serum levels of the cytokines including interleukin 1α(IL-1α),chemokine monocyte chemotactic protein 1(also known as chemokine[C-C motif]ligand 2[CCL2]),and C-X-C motif chemokine ligand 2(CXCL2)in patients with pemphigus.Methods:The expression levels of IL-1α,CCL2,and CXCL2 in the serum of 57 patients with pemphigus PV(including 42 patients in progressive stage and 15 patients in remission stage)and 31 healthy controls were examined by enzyme-linked immunosorbent assay.The indepent-samples t-test was used to compare the two groups.Oneway analysis of variance was used for multiple-group comparisons,and the post-hoc least significant difference test was used to detect differences among multiple groups.Results:The serum expression levels of CCL2 and IL-1a were all significantly higher in the patients in progressive stage than in the controls([2.69±0.23]ng/mL vs.[2.55±0.28]ng/mL,P=0.043;[0.62±0.27]ng/mL vs.[0.48±0.23]ng/mL,P=0.038,respectively).In addition,the serum expression level of CXCL2 was significantly higher in patients in progressive stage than in in the remission stage([61.70±46.38]ng/mL vs.[24.97±18.46]ng/mL,P=0.037).Sex,disease classification,disease severity,treatment,and mucosal involvement had no significant influence on the expression of IL-1α,CCL2,or CXCL2 in the serum of patients groups and controls(all P>0.05).Conclusion:IL-1α,CCL2,and CXCL2 are heavily involved in the occurrence and development of pemphigus and may be related to the activity of the disease.

Spinal CCL2 Promotes Central Sensitization, Long-Term Potentiation, and Inflammatory Pain via CCR2： Further Insights into Molecular, Synaptic, and Cellular Mechanisms

Mounting evidence supports an important role of chemokines, produced by spinal cord astrocytes, in promoting central sensitization and chronic pain. In particular, CCL2 （C-C motif chemokine ligand 2） has been shown to enhance N-methyl-D-aspartate （NMDA）-induced currents in spinal outer lamina II （Iio） neurons. However, the exact molecular, synaptic, and cellular mechanisms by which CCL2 modulates central sensitization are still unclear. We found that spinal injection of the CCR2 antagonist RS504393 attenuated CCL2- and inflammation-induced hyperalgesia. Single-cell RT-PCR revealed CCR2 expres- sion in excitatory vesicular glutamate transporter subtype 2-positive （VGLUT2＋） neurons. CCL2 increased NMDA- induced currents in CCR2＋/VGLUT2＋ neurons in lamina IIo; it also enhanced the synaptic NMDA currents evoked by dorsal root stimulation; and furthermore, it increased the total and synaptic NMDA currents in somatostatin- expressing excitatory neurons. Finally, intrathecal RS504393 reversed the long-term potentiation evoked in the spinal cord by C-fiber stimulation. Our findings suggest that CCL2 directly modulates synaptic plasticity in CCR2- expressing excitatory neurons in spinal lamina Iio, and this underlies the generation of central sensitization in patho- logical pain.

Spinal CCL2 Promotes Pain Sensitization by Rapid Enhancement of NMDA-Induced Currents Through the ERK-GluN2B Pathway in Mouse Lamina Ⅱ Neurons

Previous studies have shown that CCL2(C-C motif chemokine ligand 2)induces chronic pain,but the exact mechanisms are still unknown.Here,we established models to explore the potential mechanisms.Behavioral experiments revealed that an antagonist of extracellular signal-regulated kinase(ERK)inhibited not only CCL2-induced inflammatory pain,but also pain responses induced by complete Freund’s adjuvant.We posed the question of the intracellular signaling cascade involved.Subsequent experiments showed that CCL2 up-regulated the expression of phosphorylated ERK(pERK)and N-methyl D-aspartate receptor[NMDAR]subtype 2B(GluN2B);meanwhile,antagonists of CCR2 and ERK effectively reversed these phenomena.Whole-cell patchclamp recordings revealed that CCL2 enhanced the NMDAR-induced currents via activating the pERK pathway,which was blocked by antagonists of GluN2B and ERK.In summary,we demonstrate that CCL2 directly interacts with CCR2 to enhance NMDAR-induced currents,eventually leading to inflammatory pain mainly through the CCL2-CCR2-pERK-GluN2B pathway.

Functional roles of CCL5/RANTES in liver disease

Inflammation,which is mediated by leukocyte trafficking and activation,plays a prominent role in the pathogenesis of acute and chronic liver injury.Chemokines are critical mediators involved in the migration of leukocytes into the diseased liver via binding to their G protein-coupled receptors.CeC motif ligand 5(CCL5)belongs to the CC-chemokine family and is secreted by several hepatic cell pop-ulations including hepatocytes,macrophages,hepatic stellate cells,and endothelial cells upon activation.CCL5 regulates the recruitment and migration of T cells(via CCR5)and NK cells(via CCR1).Moreover,CCL5 activates and stimulates T cell proliferation and cytokine production,sequentially regulating in-flammatory responses.Accumulating studies have identified crucial effects of CCL5 both in liver-disease patients and in experimental models,in which CCL5 is elevated and displays distinct effects according to pathological conditions.In this review,we discussed the crucial functions of CCL5 in liver diseases,including acute liver failure,hepatic ischemia-reperfusion injury,acute liver failure,acute and viral hepatitis,alcoholic liver disease,non-alcoholic fatty liver disease,fibrosis,and hepatocellular carcinoma.Continued understanding the roles of CCL5 in liver disease and their mechanisms of activation are indispensable for the development of effective clinical therapeutics.