Replication of interleukin 23 receptor and autophagy-related 16-like 1 association in adult-and pediatric-onset inflammatory bowel disease in Italy

AIM: To investigate gene variants in a large Italian inflammatory bowel disease (IBD) cohort, and to analyze the correlation of sub-phenotypes (including age at diagnosis) and epistatic interaction with other IBD genes. METHODS: Total of 763 patients with Crohn's disease (CD, 189 diagnosed at age < 19 years), 843 with ulcerative colitis (UC, 179 diagnosed <19 years), 749 healthy controls, and 546 healthy parents (273 trios) were included in the study. The rs2241880 [autophagy-related 16-like 1 (ATG16L1)], rs11209026 and rs7517847 [interleukin 23 receptor (IL23R)], rs2066844, rs2066845, rs2066847 (CARD15), rs1050152 (OCTN1), and rs2631367 (OCTN2) gene variants were genotyped. RESULTS: The frequency of G allele of ATG16L1 SNP (Ala197Thr) was increased in patients with CD compared with controls (59% vs 54% respectively) (OR = 1.25, CI = 1.08-1.45, P = 0.003), but not in UC (55%). The frequency of A and G (minor) alleles of Arg381Gln, rs11209026 and rs7517847 variants of IL23R were reduced significantly in CD (4%, OR = 0.62, CI = 0.45-0.87, P = 0.005; 28%, OR = 0.64, CI = 0.55-0.75, P < 0.01), compared with controls (6% and 38%, respectively). The A allele (but not G) was also reduced signifi cantly in UC (4%, OR = 0.69, CI = 0.5-0.94, P = 0.019). No association was demonstrated with sub-phenotypes and interaction with CARD15 , and OCTN1/2 genes, although both gene variants were associated with pediatric-onset disease. CONCLUSION: The present study confirms the association of IL23R polymorphisms with IBD, and ATG16L1 with CD, in both adult- and pediatric-onset subsets in our study population.

自噬相关16样蛋白1、半乳凝集素9在炎症性肠病患者中的表达及相关性

目的探讨自噬相关16样蛋白1(ATG16L1)、半乳凝集素9(Galectin-9)在炎症性肠病患者中的表达及相关性。方法选取炎症性肠病患者50例作为观察组,另选取因肠胃不适需进行体检者40例作为对照组。分别采用ELISA、Western blotting及RT-PCR检测两组血清和肠上皮组织中ATG16L1、Galectin-9的表达水平,并分析其相关性。结果观察组患者血清ATG16L1、Galectin-9表达,肠上皮组织ATG16L1、Galectin-9 mRNA及其蛋白表达均低于对照组(P均<0.05)。ATG16L1、Galectin-9表达与炎症性肠病的发生均呈负相关(r分别为-0.381、-0.401,P均<0.05);ATG16L1与Galectin-9在炎症性肠病肠组织中表达呈正相关(r=0.411,P<0.05);ATG16L1与Galectin-9在血清中的表达呈正相关(r=0.281,P<0.05);血清中ATG16L1与组织中ATG16L1表达呈正相关(r=0.382,P<0.05);血清中Galectin-9与组织中Galectin-9表达呈正相关(r=0.403,P<0.05)。结论ATG16L1、Galectin-9在炎症性肠病患者血清及肠上皮组织中的表达均显著下调,二者在血清和肠组织中的表达呈正相关。

ATG4A及ATG16L1基因多态性与肺结核易感性的关系

目的:探讨自噬相关基因4A(autophagy related 4A,ATG4A)及自噬相关基因16L1(autophagy related 16 like 1,ATG16L1)的多态性位点与中国西南地区人群肺结核易感性的关系。方法:采用聚合酶链反应-限制性片段长度多态性技术检测218例肺结核患者及248例健康对照者ATG4A基因rs807185位点及ATG16L1基因rs2241880位点的基因型,用logistic回归分析上述2个位点与肺结核病易感的相关性。结果:2组人群中rs807185位点的A等位基因、AT基因型及显性模型AA+AT vs.TT的频率分布存在统计学差异(P<0.05),且在女性中差异更突出。而rs2241880位点的各指标频率分布差异均不明显(P>0.05)。结论:ATG4A基因rs807185位点多态性与中国西南地区人群肺结核易感性呈负相关,而ATG16L1基因rs2241880位点多态性则与该地区人群肺结核易感性无明显相关。

Interaction of the major inflammatory bowel disease susceptibility alleles in Crohn’s disease patients

AIM:To investigate the interaction of interleukin-23 receptor(IL23R)(rs1004819 and rs2201841),autophagy-related 16-like 1(ATG16L1)(rs2241880), caspase recruitment domain-containing protein 15 (CARD15)genes,and IBD5 locus in Crohn's disease(CD) patients. METHODS:A total of 315 unrelated subjects with CD and 314 healthy controls were genotyped.Interactions and specific genotype combinations of a total of eight variants were tested.The variants of IBD5locus(IGR2198a_1 rs11739135 and IGR2096a_1 rs12521868),CARD15(R702W rs2066845 and L1007fs rs2066847),ATG16L1(rs2241880)and IL23R (rs1004819,rs2201841)genes were genotyped by PCR-RFLP,the G908R(rs2066844)in CARD15 was determined by direct sequencing. RESULTS:The association of ATG16L1 T300A with CD was confirmed[P=0.004,odds ratio(OR)=1.69, 95%CI:1.19-2.41],and both IL23R variants were found to represent significant risk for the disease(P= 0.008,OR=2.05,95%CI:1.20-3.50 for rs1004819 AA;P<0.001,OR=2.97,95%CI:1.65-5.33 for rs2201841 CC).Logistic regression analysis of pairwise interaction of the inflammatory bowel disease (IBD)loci indicated that IL23R,ATG16L1,CARD15 and IBD5(IGR2198a_1)contribute independently to disease risk.We also analysed the specific combina- tions by pair of individual ATG16L1,IL23R rs1004819, rs2201841,IGR2198a_1,IGR2096a_1 and CARD15 genotypes for disease risk influence.In almost all cases,the combined risk of susceptibility pairs was higher in patients carrying two different risk-associated gene variants together than individuals with just one polymorphism.The highest OR was found for IL23R rs2201841 homozygous genotype with combination of positive CARD15 status(P<0.001,OR=9.15,95% CI:2.05-40.74). CONCLUSION:The present study suggests a cumulative effect of individual IBD susceptibility loci.

脂肪间充质干细胞外泌体调控心脏成纤维细胞自噬和NLRP3炎症小体平衡抑制心肌梗死后不良心室重塑

[目的]探讨脂肪间充质干细胞(ADMSC)外泌体(Exo)对心肌梗死(MI)后不良心室重塑的抑制作用和机制。[方法]观察心脏成纤维细胞经过氧化氢(H_(2)O_(2))处理后自噬和炎症表型的改变。MI大鼠经尾静脉注射等体积的生理盐水、ADMSC外泌体(MSC-Exo)、成纤维细胞外泌体(MEF-Exo),观察心脏成纤维细胞自噬相关16样蛋白1(ATG16L1)、自噬相关蛋白7(ATG7)和NOD样受体蛋白3(NLRP3)炎症小体的表达,炎症反应,心肌纤维化程度以及心功能。[结果]心脏成纤维细胞经H_(2)O_(2)处理后,自噬相关蛋白ATG16L1和ATG7表达显著降低(P<0.001),NLRP3表达显著升高(P<0.001),促炎细胞因子白细胞介素1β(IL-1β)和IL-18水平显著增加(P<0.001)。MI大鼠经MSC-Exo干预后,自噬相关蛋白ATG16L1和ATG7表达显著上调(P<0.001),NLRP3表达显著下调(P<0.001),血清IL-1β和IL-18水平显著降低(P<0.001),纤维化相关蛋白胶原蛋白Ⅰ和Ⅲ显著减少(P<0.001),心肌纤维化程度显著减轻(P<0.001),心功能明显改善(P<0.001)。[结论]脂肪MSC-Exo通过调控心脏成纤维细胞自噬和NLRP3炎症小体的平衡,发挥抑制MI后不良心室重塑的作用。