Predictive value of angiopoietin-like protein 8 in metabolic dysfunction-associated fatty liver disease and its progression:A case-control study

BACKGROUND The prevalence of metabolic dysfunction-associated fatty liver disease(MAFLD)is rapidly increasing,currently affecting approximately 25%of the global population.Liver fibrosis represents a crucial stage in the development of MAFLD,with advanced liver fibrosis elevating the risks of cirrhosis and hepatocellular carcinoma.Simple serum markers are less effective in diagnosing liver fibrosis compared to more complex markers.However,imaging techniques like transient elastography face limitations in clinical application due to equipment and technical constraints.Consequently,it is imperative to identify a straightforward yet effective method for assessing MAFLD-associated liver fibrosis.AIM To investigate the predictive value of angiopoietin-like protein 8(ANGPTL8)in MAFLD and its progression.METHODS We analyzed 160 patients who underwent abdominal ultrasonography in the Endocrinology Department,Xiaogan Central Hospital affiliated to Wuhan University of Science and Technology,during September 2021-July 2022.Using abdominal ultrasonography and MAFLD diagnostic criteria,among the 160 patients,80 patients(50%)were diagnosed with MAFLD.The MAFLD group was divided into the liver fibrosis group(n=23)and non-liver fibrosis group(n=57)by using a cut-off fibrosis-4 index≥1.45.Logistical regression was used to analyze the risk of MAFLD and the risk factors for its progression.Receiver operating characteristic curves were used to evaluate the predictive value of serum ANGPTL8 in MAFLD and its progression.RESULTS Compared with non-MAFLD patients,MAFLD patients had higher serum ANGPTL8 and triglyceride-glucose(TyG)index(both P<0.05).Serum ANGPTL8(r=0.576,P<0.001)and TyG index(r=0.473,P<0.001)were positively correlated with MAFLD.Serum ANGPTL8 was a risk factor for MAFLD[odds ratio(OR):1.123,95%confidence interval(CI):1.066-1.184,P<0.001).Serum ANGPTL8 and ANGPTL8+TyG index predicted MAFLD[area under the curve(AUC):0.832 and 0.886,respectively;both P<0.05].Compared with MAFLD patients without fibrosis,those with fibrosis had higher serum ANGPTL8 and TyG index(both P<0.05),and both parameters were positively correlated with MAFLD-associated fibrosis.Elevated serum ANGPTL8(OR:1.093,95%CI:1.044-1.144,P<0.001)and TyG index(OR:2.383,95%CI:1.199-4.736,P<0.013)were risk factors for MAFLD-associated fibrosis.Serum ANGPTL8 and ANGPTL8+TyG index predicted MAFLD-associated fibrosis(AUC:0.812 and 0.835,respectively;both P<0.05).CONCLUSION The serum levels of ANGPTL8 are elevated and positively correlated with MAFLD.They can serve as predictors for the risk of MAFLD and liver fibrosis,with the ANGPTL8+TyG index potentially exhibiting even higher predictive value.

Clinical and experimental study on angiopoietin-like protein 8 associated with proliferative diabetic retinopathy

AIM：To confirm the role of angiopoietin-like protein 8（Angptl 8） in proliferative diabetic retinopathy（PDR）.METHODS：The sera and aqueous humor of 10 PDR patients and 10 non-diabetic retinopathy（NDR） patients（idiopathic macular hole patients） were collected and the expression of Angptl 8 was detected by enzyme linked immune-sorbent assay（ELISA）.Experimental diabetes mice model was induced with streptozotocin.The expression of glycosylated hemoglobin and Angptl 8 in sera was detected.Recombinant Angptl 8 was re-infused into wild type（WT） diabetic mice and spatial frequency threshold and contrast sensitivity were measured.In vitro retinal pigment epithelium（RPE） were stimulated by recombinant Angptl 8 for 24 h.MMT assay were used to detect cell proliferation.At the same time,q RT-PCR and Western blot was used to measure the expression of proliferation-related factors in PRE cells.RESULTS：The expression of Angptl 8 was markedly increased in the sera and aqueous humor of PDR patients（F=99.02,P〈0.0001 in sera;t=10.42,P〈0.0001 in aqueous）.After successfully establishing the diabetic mice model,we found that glycosylated hemoglobin and Angptl 8 expression levels were increased.Re-infusion of recombinant Angptl 8 into WT diabetic mice could further decrease spatial frequency threshold and contrast sensitivity（P〈0.01）.In vitro,RPE cells stimulated by recombinant Angptl 8could increase the relative absorbance of MMT assay（1.486±0.042 vs 1.000±0.104,P〈0.05） and proliferating cell nuclear antigen（PCNA） expression（0.55±0.01 vs 0.29±0.03,P〈0.05）.The proliferative effect of Angptl 8 is mainly mediated by increasing the expression of proliferation-activating factors cyclin A1（4.973±0.205 vs 2.720±0.197,P〈0.05）,cyclin F（5.690±0.219 vs 4.297±0.292,P〈0.05） and E2 F2（2.297±0.102 vs 1.750±0.146,P〈0.05）,and reducing the expression of proliferation-inhibiting factors cdkn1（2.370±0.074 vs 3.317±0.135,P〈0.05） and cdkn2（4.793±0.065 vs 5.387±0.149,P〈0.05）.CONCLUSION：The expression of Angptl 8 is increased in PDR,and the increased Angptl 8 can promote proliferation and increase proliferation-related factors.

Host Factors Alter Effects of Angiopoietin-Like Protein 8 on Glucose Homeostasis in Diabetic Mice

Recovery of functional beta cell mass offers a biological cure for type 1 diabetes. However, beta cell mass is difficult to regain once lost since the proliferation rate of beta cells after youth is very low. Angiopoietin like-protein 8 (ANGPTL8), a peptide that has a role in the regulation of lipoprotein lipase activity, was reported to increase beta cell proliferation in mice in 2013. Subsequent studies of human ANGPTL8 for short term (3 to 8 days) in non-diabetic mice showed little or no increase in beta cell proliferation. Here, we examined the effect of ANGPTL8 on glucose homeostasis in models that have not been examined previously. We expressed mouse ANGPTL8 using adenovirus in 2 mouse models of diabetes (streptozotocin and Non-Obese Diabetic (NOD) mice) over 2 weeks. Also, we tested ANGPTL8 in NOD mice deficient in leukocyte 12-lipoxygenase (12LO), an enzyme that contributes to insulitis and loss of beta cell function in NOD, in an effort to determine whether 12LO deficiency alters the response to ANGPTL8. Adenovirus-mediated expression of ANGPTL8 lowered blood glucose levels in streptozotocin treated mice without an increase in beta cell proliferation or serum insulin concentration. While ANGPTL8 did not reverse hyperglycemia in overtly hyperglycemic NOD mice or alter glucose homeostasis of non-diabetic NOD mice, ANGPTL8 reduced blood glucose levels in 12LOKO NOD mice. However, the lower glucose levels in 12LOKO NOD were not associated with higher serum insulin levels or beta cell proliferation. In summary, while mouse ANGPTL8 does not increase beta cell proliferation in NOD mice or streptozotocin treated mice in agreement with studies in non-diabetic mice, it lowers blood glucose levels in multiple low-dose streptozotocin induced diabetes and 12LO deficiency indicating that host factors influence the impact of ANGPTL8 on glucose homeostasis.

急性心肌梗死患者血清ANGPTL8、KLF2表达与冠脉病变程度及主要心脏不良事件发生的关系

目的探讨急性心肌梗死(AMI)患者血清血管生成素样蛋白8(ANGPTL8)、Kruppel样因子2(KLF2)表达与冠脉病变程度及主要心脏不良事件(MACE)发生的关系。方法选取106例AMI患者为研究对象,根据冠脉病变程度将患者分为轻度组(52例)和重度组(54例),根据MACE发生情况将患者分为MACE组(18例)和非MACE组(88例)。收集患者一般资料,酶联免疫吸附试验(ELISA)法检测血清ANGPTL8、KLF2水平,Spearman相关分析AMI患者血清ANGPTL8、KLF2水平与Gensini评分的相关性,多因素Logistic回归分析AMI患者冠脉病变程度的影响因素;绘制受试者工作特征(ROC)曲线分析血清ANGPTL8、KLF2水平预测AMI患者发生MACE的价值。结果重度组AMI患者高血压史、高血脂史患者比例,收缩压、舒张压、三酰甘油(TG)、N-末端B型利钠肽原(NT-proBNP)、心肌肌钙蛋白I(cTnI)水平,Gensini评分以及血清ANGPTL8水平高于轻度组(P<0.05),高密度脂蛋白胆固醇(HDL-C)水平以及血清KLF2水平低于轻度组(P<0.05);且轻度组和重度组AMI患者病变支数比较,差异有统计学意义(P<0.05)。AMI患者血清ANGPTL8水平与Gensini评分呈正相关(r=0.638,P<0.05),血清KLF2水平与Gensini评分呈负相关(r=-0.612,P<0.05)。高血压史、高血脂史、cTnI、ANGPTL8是AMI患者冠脉病变进展为重度的危险因素(P<0.05),而HDL-C、KLF2是保护因素(P<0.05)。MACE组血清ANGPTL8水平高于非MACE组(P<0.05),而血清KLF2水平低于非MACE组(P<0.05)。血清ANGPTL8、KLF2水平及二者联合预测AMI患者发生MACE的曲线下面积分别为0.740(95%CI:0.646~0.820)、0.799(95%CI:0.710~0.870)、0.806(95%CI:0.717~0.876)。结论血清ANGPTL8、KLF2表达均与AMI患者冠脉病变程度密切相关,且对MACE发生具有一定预测价值。

母体血清GDF-15、ANGPTL8对胎儿先天性心脏病产前诊断的临床意义

目的探讨血清生长分化因子-15(GDF-15)、血管生成素样蛋白8(ANGPTL8)在先天性心脏病(CHD)胎儿母体中的表达及其诊断胎儿CHD的价值。方法收集2020年1月—2023年3月甘肃省妇幼保健院产前诊断中心产前筛查疑似胎儿CHD的120例孕妇作为研究对象,根据产前彩色多普勒超声心动图诊断结果分为CHD组(n=86)和非CHD组(n=34)。比较2组血清GDF-15、ANGPTL8水平;多因素Logistic回归分析胎儿CHD发病的影响因素;ROC曲线分析血清GDF-15、ANGPTL8对胎儿CHD的诊断效能。结果与非CHD组比较,CHD组血清GDF-15、ANGPTL8水平显著升高(t/P=7.860/<0.001、7.334/<0.001);多因素Logistic回归分析结果显示,孕次≥3次、CHD家族史及血清GDF-15、ANGPTL8升高均是胎儿CHD发病的危险因素[OR(95%CI)=1.383(1.082~1.767),1.596(1.148~2.218),3.596(1.694~7.633),3.175(1.571~6.417)];血清GDF-15、ANGPTL8及二者联合预测胎儿CHD的AUC分别为0.805、0.835、0.903,二者联合诊断胎儿CHD优于血清GDF-15、ANGPTL8各自单独诊断(Z=2.052、2.219,P=0.040、0.027)。结论CHD胎儿的母体血清GDF-15、ANGPTL8水平显著升高,二者联合对胎儿CHD具有较高的辅助诊断潜能。

FZD7、GAL-8在肝内胆管癌中的表达及其与临床病理特征和预后的关系

目的分析肝内胆管癌(ICC)中卷曲同源蛋白7(FZD7)、半乳糖凝集素8(GAL-8)的表达及其与临床病理特征和预后的关系。方法选取2017年4月—2020年4月榆林市第一医院肿瘤科诊治的ICC患者84例作为研究对象,患者均进行根治性手术。采用免疫组织化学法检测癌组织和癌旁组织FZD7、GAL-8蛋白表达;Spearman秩相关分析FZD7与GAL-8的相关性;Kaplan-Meier法分析FZD7、GAL-8表达对ICC患者生存预后的影响;Cox回归分析ICC的预后影响因素。结果ICC癌组织中FZD7、GAL-8阳性率分别为73.81%(62/84)、71.43%(60/84),高于癌旁组织的4.76%(4/84)、7.14%(6/84),差异有统计学意义(χ^(2)=83.950,72.770,P均<0.001);ICC中FZD7与GAL-8表达呈正相关(r s=0.745,P<0.001);低分化程度、有淋巴结转移、TNM分期Ⅲ期的ICC癌组织中FZD7、GAL-8阳性率高于高中分化、无淋巴结转移、TNM分期Ⅰ~Ⅱ期(FZD7:χ^(2)/P=8.221/<0.001,6.097/0.014,13.014/<0.001;GAL-8:χ^(2)/P=7.207/0.007,5.555/0.018,11.760/0.001)。FZD7阳性组3年总生存率为32.26%(20/62),低于阴性组的68.19%(15/22)(Log rankχ^(2)=8.723,P=0.003);GAL-8阳性组3年总生存率为28.33%(17/60),低于GAL-8阴性组的75.30%(18/24)(Log rankχ^(2)=24.310,P<0.001)。TNM分期Ⅲ期、低分化程度、淋巴结转移、FZD7阳性、GAL-8阳性是影响ICC患者预后的独立危险因素[HR(95%CI)=1.614(1.215~2.145),1.516(1.219~1.884),1.916(1.315~2.791),1.826(1.222~2.729),1.737(1.237~2.438)]。结论ICC癌组织中FZD7、GAL-8表达上调,两者均参与ICC肿瘤进展,是评估ICC患者预后的肿瘤标志物。

NapA蛋白诱导巨噬细胞分泌趋化因子单核细胞趋化蛋白1和白细胞介素8的机制

目的:研究幽门螺杆菌NapA蛋白诱导巨噬细胞分泌趋化因子单核细胞趋化蛋白1(monocyte chemoattractant protein-1,MCP-1)和白细胞介素8(interleukin-8,IL-8)的机制。方法:利用NapA蛋白处理巨噬细胞,然后使用ELISA法检测上清中MCP-1和IL-8的表达量。使用C29、ST2825、SB203580、SP600125以及PDTC和巨噬细胞预先共孵育1 h,分别抑制Toll样受体2(toll-like receptors 2,TLR2)、髓样分化因子(myeloid differentiation factor 88,MyD88)、核糖核酸酶p蛋白亚基p38(ribonuclease p-protein subunit p38,p38)、应激活化蛋白激酶(c-Jun N-terminal kinase,c-Jun)和核因子κB(nuclear factor kappa B,NF-κB)的活性,然后再加入NapA蛋白孵育4 h,收集上清并检查其中MCP-1和IL-8的表达量。结果:NapA蛋白刺激巨噬细胞后,MCP-1和IL-8的表达量明显高于未处理组。利用抑制剂C29抑制TLR2的活性,NapA蛋白诱导的MCP-1和IL-8表达量降低。使用ST2825、PDTC以及SB203580分别抑制MyD88、NF-κB以及p38的活性,能够降低NapA蛋白诱导巨噬细胞分泌MCP-1和IL-8的能力,但是抑制c-Jun不影响NapA蛋白诱导巨噬细胞分泌MCP-1和IL-8。结论:幽门螺杆菌NapA蛋白通过TLR2/MyD88/NF-κB通路诱导巨噬细胞分泌MCP-1和IL-8。

支气管镜介入治疗对RMPP患儿的疗效及其对外周血CRP、SF、IL-8水平及CD4^(+)/CD8^(+)比值的影响

目的探讨支气管镜介入治疗对难治性肺炎支原体肺炎(RMPP)患儿的疗效及其对外周血C反应蛋白(CRP)、铁蛋白(SF)、白细胞介素(IL)-8水平及CD4^(+)/CD8^(+)比值的影响。方法选取2020年12月至2022年12月河北省儿童医院收治的195例确诊为RMPP患儿作为研究对象,采用随机数字表法分为对照组和介入组(病程≥14 d归为晚期介入组、病程<14 d归为早期介入组)。根据临床疗效将130例接受支气管镜介入治疗的患儿分为有效组和无效组。对照组进行常规治疗,晚期介入组和早期介入组在对照组的基础上实施支气管镜介入治疗。比较对照组、晚期介入组和早期介入组临床疗效、临床症状与体征改善情况;检测并比较对照组、晚期介入组和早期介入组外周血CRP、SF、IL-8水平及CD4^(+)/CD8^(+)比值;比较对照组、晚期介入组和早期介入组不良反应发生率;比较有效组和无效组外周血CRP、SF、IL-8水平及CD4^(+)/CD8^(+)比值。采用受试者工作特征(ROC)曲线分析外周血CRP、SF、IL-8、CD4^(+)/CD8^(+)比值对支气管镜介入治疗RMPP疗效的预测价值。结果早期介入组总有效率高于对照组和晚期介入组,差异均有统计学意义(P<0.05);对照组和晚期介入组总有效率比较,差异无统计学意义(P>0.05)。早期介入组咳嗽持续时间、发热持续时间、住院时间均短于对照组和晚期介入组,差异均有统计学意义(P<0.05)。对照组、晚期介入组和早期介入组治疗后外周血CD4^(+)/CD8^(+)比值均高于治疗前,CRP、SF、IL-8水平均低于治疗前,差异均有统计学意义(P<0.05);早期介入组治疗后外周血CD4^(+)/CD8^(+)比值均高于对照组和晚期介入组,CRP、SF、IL-8水平均低于对照组和晚期介入组,差异均有统计学意义(P<0.05)。早期介入组出现不良反应8例,晚期介入组出现不良反应10例,均暂停灌洗操作后很快恢复。对照组未出现上述不良反应。无效组外周血CRP、SF、IL-8水平均高于有效组,CD4^(+)/CD8^(+)比值低于有效组,差异均有统计学意义(P<0.05)。ROC曲线分析结果显示,CRP、SF、IL-8、CD4^(+)/CD8^(+)比值联合检测预测支气管镜介入治疗RMPP无效的曲线下面积、灵敏度、特异度分别为0.871、81.81%、83.80%。结论早期应用支气管镜介入治疗对RMPP的疗效较好,可改善患儿外周血CRP、SF、IL-8水平及CD4^(+)/CD8^(+)比值,且外周血CRP、SF、IL-8、CD4^(+)/CD8^(+)比值联合检测有助于预测支气管镜介入治疗RMPP的疗效。

LncRNA MEG8通过靶向miR-15a-5p调控MICA/B介导结直肠癌细胞免疫逃逸

目的:探究长链非编码RNA母系表达基因8(LncRNA MEG8)通过靶向miR-15a-5p调控MHCⅠ类相关蛋白A/B(MICA/B)介导的结直肠癌(CRC)细胞免疫逃逸机制。方法:RT-qPCR、Western blot检测CRC组织和细胞系MEG8、miR-15a-5p、MICA、MICB、NKG2D蛋白水平;双荧光素酶实验验证MEG8对miR-15a-5p的调控关系;采用脂质体转染法将NC、MEG8、miR-NC、miR-15a-5p分别或共转染至SW480、SW620细胞,记为NC组、MEG8组、MEG8+miR-NC组、MEG8+miR-15a-5p组;将NK细胞分别与SW480、SW620细胞共培养;ELISA检测共培养液中TNF-α、IFN-γ水平;CCK-8、EdU染色、Transwell实验检测细胞增殖、迁移和侵袭能力;RT-qPCR、Western blot检测细胞MEG8、miR-15a-5p、MICA、MICB、NKG2D蛋白水平。结果:与癌旁组织或正常结肠上皮细胞相比,CRC组织和细胞系中MEG8、MICA、MICB、NKG2D mRNA和蛋白水平降低,miR-15a-5p水平升高(P<0.05)。MEG8靶向调控miR-15a-5p。共培养体系中,与NC组相比,MEG8组细胞MICA、MICB、NKG2D蛋白水平、共培养上清液中TNF-α、IFN-γ水平明显升高,培养1 d、2 d、3 d后,OD值、EdU阳性率、迁移和侵袭细胞数明显降低(P<0.05);过表达miR-15a-5p能部分逆转过表达MEG8对细胞MICA、MICB、NKG2D、TNF-α、IFN-γ水平和增殖、侵袭转移能力的影响(P<0.05)。结论:MEG8通过靶向miR-15a-5p调控MICA、MICB表达,促进NK细胞活性,抑制CRC细胞免疫逃逸。

血清IL-8、ESM-1、VAP-1在慢性阻塞性肺疾病临床诊断及病情评估中的应用

目的探讨慢性阻塞性肺疾病(COPD)患者血清白细胞介素-8(IL-8)、内皮特异性分子-1(ESM-1)、血管黏附蛋白-1(VAP-1)表达水平,分析其与病情程度相关性及其对COPD的诊断价值。方法选取2021年11月至2022年8月郑州大学第一附属医院收治的153例COPD患者为研究对象,其中COPD稳定期42例(设为SCOPD组)、COPD急性加重期111例(设为AECOPD组),同时选取同期于医院体检的健康志愿者51例为对照组。比较两组临床资料及血清IL-8、ESM-1、VAP-1水平。对比不同病情程度患者血清IL-8、ESM-1、VAP-1水平。分析血清各指标与肺功能、生化指标、病情程度相关性。评价血清各指标对SCOPD、AECOPD的诊断价值。结果AECOPD组、SCOPD组血清IL-8、ESM-1、VAP-1水平高于对照组,且AECOPD组高于SCOPD组(P<0.05);IL-8、ESM-1、VAP-1与生化指标、COPD评估测试表(CAT)、病情程度呈正相关,与肺功能呈负相关(P<0.05);血清各指标联合诊断SCOPD与AECOPD的曲线下面积(AUC)大于单独指标诊断(P<0.05)。结论血清IL-8、ESM-1、VAP-1与COPD病情严重程度相关,联合检测其水平对COPD具有一定诊断价值,可能作为疾病诊断、病情评估的重要指标。

血管生成素样蛋白8在心血管疾病发病中作用的研究进展

血管生成素样蛋白8(ANGPTL8)是由肝脏和脂肪组织合成、分泌的一种糖蛋白,能够促进血管重塑、调节炎性反应,并通过这些机制参与冠心病、高血压、主动脉瘤、病理性心肌肥厚等多种心血管疾病的发生与发展,有望成为心血管疾病新的预防和治疗靶点。

Angiopoietin-Like Protein 3 Expression is Down-Regulated in Experimentally Pregnant Toxemic Goats

Pregnancy toxemia is a metabolic disorder of lipid and glucose. Recent investigations have found that angiopoietin-like protein 3 （ANGPTL3） can contribute to disorder of carbohydrate and lipid metabolism. The present study was conducted to investigate the change of ANGPTL3 expression during pregnancy toxemia, We firstly cloned the full-length cDNA of ANGPTL3 in Liuyang Black goats, revealing that goat ANGPTL3 had the typical structure of the angiopoietin-like family, and its mRNA was exclusively expressed in liver. Pregnancy toxemia of pregnant goat does with twins during late gestation was induced by being fasted for 72 h, and then they were recovered after 5 d ofrefeeding. Hepatic ANGPTL3 gene expression was significantly down-regulated concomitantly with decreased serum glucose concentration, elevated serum β-hydroxybutyrate and free fatty acid levels in pregnant toxemic goats, and these changes were reversed after refeeding. These results suggest ANGPTL3 may play a certain role in the development of pregnancy toxemia in goats.

Recombinant angiopoietin-like protein 4 attenuates intestinal barrier structure and function injury after ischemia/reperfusion

BACKGROUND Intestinal barrier breakdown,a frequent complication of intestinal ischemiareperfusion(I/R)including dysfunction and the structure changes of the intestine,is characterized by a loss of tight junction and enhanced permeability of the intestinal barrier and increased mortality.To develop effective and novel therapeutics is important for the improvement of outcome of patients with intestinal barrier deterioration.Recombinant human angiopoietin-like protein 4(rhANGPTL4)is reported to protect the blood-brain barrier when administered exogenously,and endogenous ANGPTL4 deficiency deteriorates radiationinduced intestinal injury.AIM To identify whether rhANGPTL4 may protect intestinal barrier breakdown induced by I/R.METHODS Intestinal I/R injury was elicited through clamping the superior mesenteric artery for 60 min followed by 240 min reperfusion.Intestinal epithelial(Caco-2)cells and human umbilical vein endothelial cells were challenged by hypoxia/reoxygenation to mimic I/R in vitro.RESULTS Indicators including fluorescein isothiocyanate-conjugated dextran(4 kilodaltons;FD-4)clearance,ratio of phosphorylated myosin light chain/total myosin light chain,myosin light chain kinase and loss of zonula occludens-1,claudin-2 and VE-cadherin were significantly increased after intestinal I/R or cell hypoxia/reoxygenation.rhANGPTL4 treatment significantly reversed these indicators,which were associated with inhibiting the inflammatory and oxidative cascade,excessive activation of cellular autophagy and apoptosis and improvement of survival rate.Similar results were observed in vitro when cells were challenged by hypoxia/reoxygenation,whereas rhANGPTL4 reversed the indicators close to normal level in Caco-2 cells and human umbilical vein endothelial cells significantly.CONCLUSION rhANGPTL4 can function as a protective agent against intestinal injury induced by intestinal I/R and improve survival via maintenance of intestinal barrier structure and functions.

Angiopoietin-like protein 3 (ANGPTL3) deficiency and familial combined hypolipidemia

Three members of the angiopoietin-like(ANGPTL) protein family-ANGPTL3, ANGPTL4 and ANGPTL8-are important regulators of plasma lipoproteins. They inhibit the enzyme lipoprotein lipase, which plays a key role in the intravascular lipolysis of triglycerides present in some lipoprotein classes. This review focuses on the role of ANGPTL3 as emerged from the study of genetic variants of Angptl3 gene in mice and humans. Both loss of function genetic variants and inactivation of Angptl3 gene in mice are associated with a marked reduction of plasma levels of triglyceride and cholesterol and an increased activity of lipoprotein lipase and endothelial lipase. In humans with ANGPTL3 deficiency, caused by homozygous loss of function(LOF) variants of Angptl3 gene, the levels of all plasma lipoproteins are greatly reduced. This plasma lipid disorder referred to as familial combined hypolipidemia(FHBL2) does not appear to be associated with distinct pathological manifestations. Heterozygous carriers of LOF variants have reduced plasma levels of total cholesterol and triglycerides and are at lower risk of developing atherosclerotic cardiovascular disease, as compared to non-carriers. These observations have paved the way to the development of strategies to reduce the plasma level of atherogenic lipoproteins in man by the inactivation of ANGPTL3, using either a specific monoclonal antibody or anti-sense oligonucleotides.

MicroRNA-370-5p inhibits pigmentation and cell proliferation by downregulating mitogen-activated protein kinase kinase kinase 8 expression in sheep melanocytes

In mammals,microRNAs(miRNAs)play key roles in multiple biological processes by regulating the expression of target genes.Studies have found that the levels of miR-370-5p expression differ significantly in the skins of sheep with different hair colors;however,its function remains unclear.In this study,we investigated the roles of miR-370-5p in sheep melanocytes and found that the overexpression of miR-370-5p significantly inhibited cell proliferation(P<0.01),tyrosinase activity(P=0.001)and significantly reduced(P<0.001)melanin production.Functional prediction revealed that the 3′-untranslated region(UTR)of MAP3K8 has a putative miR-370-5p binding site,and the interaction between these two molecules was confirmed using luciferase reporter assays.In situ hybridization assays revealed that MAP3K8 is expressed in the cytoplasm of melanocytes.The results of quantitative RT-PCR and Western blotting analyses revealed that overexpression of miR-370-5p in melanocytes significantly inhibits(P<0.01)MAP3K8 expression via direct targeting of its 3′UTR.Inhibition of MAP3K8 expression by siRNA-MAP3K8 transfection induced a significant inhibition(P<0.01)of melanocyte proliferation and significant reduction(P<0.001)in melanin production,which is consistent with our observations for miR-370-5p.Target gene rescue experiments indicated that the expression of MAP3K8 in melanocytes co-transfected with miR-370-5p and MAP3K8-cDNA(containing sites for the targeted binding to miR-370-5p)was significantly rescued(P≤0.001),which subsequently promoted significant increases in cell proliferation(P<0.001)and melanin production(P<0.01).Collectively,these findings indicate that miR-370-5p plays a functional role in inhibiting sheep melanocyte proliferation and melanogenesis by downregulating the expression of MAP3K8.

血清SAA、HBP、IL-8水平与脓毒症患者病情严重程度及28天预后的关系

目的:探讨血清淀粉样蛋白A(SAA)、肝素结合蛋白(HBP)、白细胞介素8(IL-8)水平与脓毒症患者病情严重程度及28 d预后的相关性。方法:将106例脓毒症患者按照病情严重程度分为非休克组(n=82)及休克组(n=24);按照28 d预后情况将患者分死亡组(n=32)与存活组(n=74)。比较不同病情严重程度患者血清SAA、HBP、IL-8水平的差异,采用Logistic回归分析与脓毒症患者28 d预后相关的风险因素;绘制受试者工作特征(ROC)曲线验证各指标的诊断效能。结果:休克组患者血清SAA、HBP及IL-8水平均高于非休克组(P<0.05)。死亡组患者APACHEⅡ评分、NEWS评分、SOFA评分、WBC、CRP、PCT、SAA、HBP及IL-8水平均高于存活组(P<0.05)。Logistic回归分析结果显示,高APACHEⅡ评分、高SAA、HBP及IL-8水平是脓毒症患者28 d死亡的风险因素(P<0.05)。ROC曲线结果显示,SAA、HBP及IL-8预测脓毒症患者28 d死亡的曲线下面积(AUC)分别为0.871、0.828、0.769,APACHEⅡ评分预测的AUC为0.801;各项指标联合诊断的AUC为0.982,敏感度和特异度分别为93.75%和97.30%。结论:血清SAA、HBP、IL-8水平均能较好评估脓毒症患者病情严重程度及28 d预后,且联合APACHEⅡ评分诊断时效能更高。

亚油酸诱导植物乳杆菌p-8产生共轭亚油酸的蛋白组和类组蛋白乙酰化分析

研究亚油酸(linoleic acid,LA)诱导下植物乳杆菌(Lactobacillus plantarum)p-8的蛋白组和类组蛋白乙酰化水平差异,探究共轭亚油酸(conjugated linoleic acid,CLA)产生的机制。结果表明,LA的最佳诱导质量浓度为1 mg/mL,在此质量浓度条件下,蛋白质组学分析发现,烯酰-酰基载体蛋白还原酶、酰基载体蛋白等脂肪酸合成的关键蛋白因子下调,脂肪酸合成下调,乙酰基转移酶水平上调。Western blot结合体外添加NaAc实验表明,乙酰辅酶A含量增加导致类组蛋白乙酰化水平提升。实时聚合酶链式反应检测显示,添加LA和NaAc使CLA相关酶、转录调控因子和Sigma因子的mRNA表达升高,CLA含量也提高。相关性分析得出,乙酰辅酶A与LA水合酶转录水平和CLA含量之间呈显著正相关,揭示乙酰辅酶A和乙酰基转移酶含量上调导致类组蛋白乙酰化加强而上调CLA合成。结果为分子改良或科学调控乳酸菌以提高CLA转化率奠定了基础。

激素敏感型原发性肾病综合征患儿血清ANGPTL8、新蝶呤水平及临床意义

目的 探究激素敏感型原发性肾病综合征(NS)患儿血清血管生成素样蛋白8(ANGPTL8)、新蝶呤水平及临床意义。方法 选取2018年1月至2021年1月该院诊治的159例激素敏感型原发性NS患儿为研究对象(NS组)。随访1年,根据是否发生频复发,分为非频复发亚组(n=93)和频复发亚组(n=66),另选取该院儿外科择期手术的60例腹股沟斜疝患儿为对照组。酶联免疫吸附试验(ELISA)检测血清ANGPTL8、新蝶呤水平。比较各组血清ANGPTL8、新蝶呤水平。多因素Logistic回归分析影响激素敏感型原发性NS患儿频复发的危险因素。受试者工作特征(ROC)曲线分析血清ANGPTL8、新蝶呤对激素敏感型原发性NS患儿频复发的预测价值。结果 NS组血清ANGPTL8、新蝶呤水平明显高于对照组,差异有统计学意义(t=20.948、44.288,P<0.001、0.001)。频复发亚组血清ANGPTL8、新蝶呤水平、CD8^(+)T细胞、24 h尿蛋白定量及尿蛋白转阴时间明显高于非频复发亚组,而血白蛋白、CD4^(+)T细胞、CD4^(+)/CD8^(+)T细胞比值低于非频复发亚组,差异有统计学意义(均P<0.05)。NS患儿血清ANGPTL8、新蝶呤水平与24 h尿蛋白量、CD8^(+)T细胞呈显著正相关,与血白蛋白、CD4^(+)T细胞、CD4^(+)/CD8^(+)呈显著负相关(均P<0.05)。血清ANGPTL8、新蝶呤水平是影响激素敏感型原发性NS患儿频复发的独立危险因素(P<0.05)。血清ANGPTL8、新蝶呤联合模型对激素敏感型原发性NS患儿频复发预测的曲线下面积(AUC)为0.852(95%CI:0.813~0.889),高于ANGPTL8、新蝶呤单一指标检测的0.764(95%CI:0.722~0.812),0.749(95%CI:0.711~0.790),差异具有统计学意义(Z=3.623、3.987,P=0.003、<0.001)。结论 血清ANGPTL8、新蝶呤水平升高是影响激素敏感型原发性NS患儿频复发的危险因素。血清ANGPTL8、新蝶呤联合检测对激素敏感型原发性NS患儿频复发具有较高的预测价值。

FKBP8介导的线粒体自噬在重复性离心运动诱导骨骼肌损伤中的作用

目的:探讨重复性离心运动对大鼠骨骼肌线粒体结构、功能及自噬的影响,分析FK506结合蛋白8(FKBP8)介导的线粒体自噬在运动致骨骼肌线粒体损伤中的作用。方法:32只成年雄性Sprague-Dawley大鼠,随机分为2周安静对照组(2C组,n=8)、4周安静对照组(4C组,n=8)、2周运动组(2E组,n=8)和4周运动组(4E组,n=8)。2E组和4E组大鼠分别运动2周和4周,均采取持续性下坡跑,跑台坡度为-16°,速度为16 m/min,每天运动90 min,每周5天。在末次离心运动训练结束后休息24 h,对所有大鼠施加一次性力竭运动,分别在力竭运动之前及之后通过尾静脉测定其血乳酸值,并记录完成力竭运动的距离。采用透射电子显微镜观察比目鱼肌线粒体超微结构的变化;应用Western blot方法检测大鼠比目鱼肌线粒体琥珀酸脱氢酶亚基B(SDHB)、细胞色素C氧化酶亚基1(MTCO1)、FKBP8和微管相关蛋白1轻链3(LC3)的蛋白表达;使用免疫荧光双标技术检测比目鱼肌FKBP8与LC3、LC3与线粒体膜蛋白细胞色素C氧化酶亚基COXⅣ(COXⅣ)、COXⅣ与溶酶体相关膜蛋白2(LAMP2)共定位的量。结果:(1)2E组大鼠完成一次力竭运动的距离和运动之前、之后血乳酸值均明显高于2C组与4E组(P<0.05或P<0.01),4E组力竭运动距离亦显著高于4C组(P<0.01),但4E组大鼠力竭运动之前、之后血乳酸值与4C组相比差异不具有显著性(P>0.05)。(2)2E组与4E组大鼠比目鱼肌线粒体均出现明显肿胀、肌膜下积聚等超微结构异常变化,有自噬体和自噬溶酶体形成,同时线粒体数量明显减少(P<0.05),且4E组比目鱼肌线粒体损伤程度较2E组更为严重。(3)2E组与4E组大鼠比目鱼肌线粒体SDHB和MTCO1蛋白表达均明显低于同期2C组与4C组,且4E组显著低于2E组(P<0.05或P<0.01)。(4)2E组与4E组大鼠比目鱼肌线粒体FKBP8和LC3蛋白表达,以及FKBP8与LC3、LC3与COXⅣ、COXⅣ与LAMP2共定位的量均明显高于同期2C组与4C组,且4E组显著高于2E组(P<0.05或P<0.01)。结论:重复性离心运动后骨骼肌线粒体结构、数量和功能受损,同时FKBP8信号激活介导线粒体自噬发生,但仍可能不足以降解清除受损的线粒体,从而导致大鼠骨骼肌损伤及其运动能力先升高后下降。

血清ANGPTL8、ITLN-1对川崎病患儿冠状动脉损伤的预测价值

目的探讨血清血管生成素样蛋白8(ANGPTL8)、凝集蛋白-1(ITLN-1)对川崎病(KD)患儿冠状动脉损伤的预测价值。方法选取2020年5月至2023年5月在本院就诊的108例KD患儿作为研究组,根据是否发生冠状动脉损伤分为冠状动脉损伤组(n=37)与无冠状动脉损伤组(n=71);另选取同期体检健康儿童72例作为对照组。采用双抗体夹心-酶联免疫吸附法检测所有受试者血清炎症相关指标及ANGPTL8、ITLN-1水平;Pearson法分析冠状动脉损伤KD患儿血清ANGPTL8、ITLN-1水平与炎症指标的相关性;评估血清ANGPTL8、ITLN-1水平对KD患儿冠状动脉损伤的预测价值。结果冠状动脉损伤组KD患儿PLT、CRP、IL-6和TNF-α水平明显高于无冠状动脉损伤组及对照组(F/χ^(2)值分别为67.198、75.056、71.876及94.158,P<0.05);与对照组相比,研究组KD患儿血清ANGPTL8水平明显升高(t=17.490,P<0.05),ITLN-1水平明显降低(t=13.162,P<0.05);与无冠状动脉损伤组相比,冠状动脉损伤组患儿血清ANGPTL8水平明显升高(t=7.454,P<0.05),ITLN-1水平明显降低(t=7.631,P<0.05)。Pearson相关分析结果显示,冠状动脉损伤KD患儿血清ANGPTL8水平与PLT及CRP、IL-6、TNF-α水平均呈正相关(r=0.514、0.638、0.484、0.674,P<0.05),ITLN-1水平与PLT及CRP、IL-6、TNF-α水平均呈负相关(r=-0.603、-0.659、-0.439、-0.646,P<0.05)。ROC曲线分析结果显示,血清ANGPTL8、ITLN-1联合预测KD患儿冠状动脉损伤的曲线下面积(AUC)为0.950,敏感度为89.19%,特异度为85.91%。多因素Logistic回归分析结果显示,PLT、CRP、IL-6、TNF-α、ANGPTL8水平升高是KD患儿冠状动脉损伤的独立危险因素,ITLN-1水平升高是KD患儿冠状动脉损伤的保护因素。其OR值及95%CI分别为1.623(1.088~2.421)、1.722(1.266~2.342)、1.413(1.035~1.930)、1.725(1.210~2.460)、1.534(1.114~2.111)和0.685(0.485~0.967)。结论KD冠状动脉损伤患儿血清ANGPTL8水平明显升高,ITLN-1水平明显降低,且ANGPTL8水平升高、ITLN-1水平降低为KD患儿冠状动脉损伤的独立危险因素,联合检测二者血清水平对KD患儿冠状动脉损伤有较高的预测价值。