儿童慢性非萎缩性胃炎中核苷酸结合寡聚化结构域样受体含pyrin结构域蛋白6的表达研究

Study on the expression of NLR family,pyrin domain containing 6 in children with chronic non-atrophic gastritis

目的探讨儿童慢性非萎缩性胃炎(chronic non-atrophic gastritis,CNG)患儿胃组织及胃液中核苷酸结合寡聚化结构域样受体含pyrin结构域蛋白6(NLR family,pyrin domain containing6,NLRP6)的表达,并分析幽门螺杆菌(Helicobacter pylori,Hp)感染对NLRP6表达的影响。方法选取2020年10月至2021年7月于徐州医科大学附属医院儿科就诊的CNG患者120例作为观察对象,采用病例对照研究方法。根据病理学诊断、内镜下胃黏膜损伤程度及Hp感染情况分为4组:轻度CNG组Hp阴性;中重度CNG组Hp阴性;轻度CNG组Hp阳性;中重度CNG组Hp阳性。采用酶联免疫吸附测定法(enzyme-linked immune sorbent assay,ELISA)检测4组胃组织及胃液中NLRP6的表达;免疫印迹法检测4组胃组织中NLRP6的表达,并分析在儿童CNG中表达的意义。呈正态分布的计量资料两组间比较采用独立样本t检验;多组间比较采用单因素方差分析,两两比较采用LSD-t检验。计数资料组间比较采用χ^(2)检验。结果中重度CNG组中Hp阳性率62.96%(34/54)高于轻度CNG组37.04%(20/54),差异有统计学意义(χ^(2)=18.32,P<0.001)。在同等Hp条件下,轻度CNG组中NLRP6表达[Hp阴性轻度CNG:胃组织(653.73±37.71)ng/L、胃液(471.75±38.47)ng/L;Hp阳性轻度CNG:胃组织(616.69±43.33)ng/L、胃液(445.29±36.39)ng/L]高于中重度CNG组[Hp阴性中重度CNG:胃组织(623.82±52.99)ng/L、胃液(446.48±47.49)ng/L;Hp阳性中重度CNG:胃组织(580.43±62.75)ng/L、胃液(406.88±51.85)ng/L],差异均有统计学意义(Hp阴性轻度与中重度CNG比较:胃组织P=0.035,胃液P=0.046;Hp阳性轻度与中重度CNG比较:胃组织P=0.010,胃液P=0.002);同等程度胃黏膜损伤中,Hp阴性组NLRP6表达[Hp阴性轻度CNG:胃组织(653.73±37.71)ng/L,胃液(471.75±38.47)ng/L;Hp阴性中重度CNG:胃组织(623.82±52.99)ng/L,胃液(446.48±47.49)ng/L高于阳性组(Hp阳性轻度CNG:胃组织(616.69±43.33)ng/L,胃液(445.29±36.39)ng/L;Hp阳性中重度CNG:胃组织(580.43±62.75)ng/L,胃液(406.88±51.85)ng/L],差异均有统计学意义(轻度CNGHp阴性与阳性比较:胃组织P=0.005,胃液P=0.023;中重度CNG阴性与阳性相比:胃组织P=0.004,胃液P=0.003)。结论在同等Hp条件下,胃黏膜损伤程度越重,NLRP6越低;在同等黏膜损伤程度的情况下,Hp阴性组的NLRP6表达水平明显高于Hp阳性组。提示NLRP6在慢性胃炎中起到抑制炎症的作用,维持上皮细胞的完整性,且Hp能够抑制NLRP6的表达。

Objective To explore the expression of the nucleotide-binding oligomerization domain-like receptor containing pyrin domain protein 6(NLR family,pyrin domain containing 6,NLRP6)in the gastric tissue and gastric juice of children with chronic non-atrophic gastritis(CNG),and to analyze the influence of Helicobacter pylori(Hp)infection on the expression of NLRP6.Methods A case-control study was conducted to select 120 CNG patients in pediatrics of Department of Pediatrics,The Affiliated Hospital of Xuzhou Medical University from October 2020 to July 2021.According to pathological diagnosis,endoscopic gastric mucosal damage and Hp infection,they were divided into 4 groups:mild CNG group Hp negative,Moderate to severe CNG group Hp negative,Mild CNG group Hp positive,Moderate to severe CNG group Hp positive.The enzyme-linked immunosorbent assay(ELISA)was used to detect the expression level of NLRP6 in the four groups of gastric tissue and gastric juice,and Western blot was used to detect the expression of NLRP6 in the gastric tissue of the 4 groups,and the significance of expression in CNG of children is analyzed.Independent sample t-test was used to compare the mean between the two groups.One way ANOVA was used to compare the mean of multiple groups of samples,and LSD t-test was used for pairwise comparison.Comparison between count data groupsχ^(2) inspection.Results The positive rate of Hp in the moderate to severe chronic non-atrophic gastritis group was 62.96%(34/54)higher than that in the mild chronic non-atrophic gastritis group 37.04%(20/54),and the difference was statistically significant(χ^(2)=18.32,P<0.001).Under the same Hp conditions,the expression of NLRP6 in the mild chronic non-atrophic gastritis group(Hp negative mild CNG:gastric tissue(653.73±37.71)ng/L,gastric juice(471.75±38.47)ng/L;Hp positive mild CNG:Gastric tissue(616.69±43.33)ng/L,gastric juice(445.29±36.39)ng/L was higher than the moderate to severe chronic non-atrophic gastritis group(Hp negative moderate to severe CNG:gastric tissue(623.82±52.99)ng/L,gastric juice(446.48±47.49)ng/L;Hp positive Moderate to severe CNG:gastric tissue(580.43±62.75)ng/L,gastric juice(406.88±51.85)ng/L,the difference is statistically significant(under Hp negative,mild compared with moderate to severe CNG:gastric tissue P=0.035;gastric juice P=0.046;Under Hp positive,mild compared with moderate to severe CNG:gastric tissue P=0.010;gastric juice P=0.002);in the same degree of gastric mucosal injury,NLRP6 expression in Hp-negative group(Hp-negative mild CNG:gastric tissue(653.73±37.71)ng/L,gastric juice(471.75±38.47)ng/L;Hp negative moderate to severe CNG:gastric tissue(623.82±52.99)ng/L,gastric juice(446.48±47.49)ng/L higher than the positive group(Hp positive mild CNG:gastric tissue(616.69±43.33)ng/L,gastric juice(445.29±36.39)ng/L;Hp positive moderate to severe CNG:gastric tissue(580.43±62.75)ng/L,gastric juice(406.88±51.85)ng/L,the difference is statistically significant(under mild CNG,Hp negative is compared with positive:Gastric tissue P=0.005;gastric juice P=0.023;under moderate to severe CNG,negative versus positive:gastric tissue P=0.004;gastric juice P=0.003).Conclusion Under the same Hp conditions,the more severe the gastric mucosal damage,the lower the NLRP6;under the same degree of mucosal damage,the expression level of NLRP6 in the Hp-negative group was significantly higher than that of the Hp-positive group.It is suggested that NLRP6 plays a role in inhibiting inflammation in chronic gastritis,maintaining the integrity of epithelial cells,and Hp can inhibit the expression of NLRP6.