Toll样受体4在原发性肾病综合征患儿肾组织及外周血中表达的意义

目的探讨Toll样受体4(TLR4)在原发性肾病综合征(INS)患儿肾组织及外周血中的表达及意义。方法采集2015年10月至2018年6月在新乡医学院第一附属医院确诊为INS的78例患儿肾活检组织及21例儿童正常肾组织(对照组1),应用免疫组织化学方法检测标本中TLR4表达水平,比较INS不同肾脏病理类型、不同临床分型中TLR4表达水平的差异,分析其与24 h尿蛋白、血清清蛋白的相关性;应用酶联免疫吸附试验(ELISA)检测INS患儿治疗前(活动期)及治疗后(缓解期)和23例健康儿童(对照组2)外周血中TLR4表达水平,比较INS不同肾脏病理类型、不同临床分型患儿血清中TLR4表达水平的差异,分析其与24 h尿蛋白、血清清蛋白的相关性;对INS患儿肾小管中TLR4的表达水平与血清TLR4的表达水平进行相关性分析。结果1.与正常肾组织TLR4水平(0.93±0.26)%]比较,TLR4在各型INS患儿肾小球及肾间质的表达系膜增生性肾小球肾炎(MsPNG)型:(0.93±0.21)%、局灶节段性肾小球硬化(FSGS)型:(1.02±0.25)%、膜性肾病(MN)型(1.03±0.09)%、微小病变(MCD)型(1.02±0.27)%]差异无统计学意义(F=0.741,P=0.562);而肾小管TLR4的表达水平明显增高MCD型:(82.94±4.62)%、MN型:(63.54±1.98)%、MsPGN型:(42.32±2.97)%、FSGS型:(22.60±2.07)%],差异有统计学意义(F=1929.842,P<0.01),其中MCD型INS患儿肾小管TLR4表达水平较MN型、MsPGN型、FSGS型明显增高,差异均有统计学意义(均P<0.01);肾小管TLR4在临床分型为激素敏感型肾病综合征(SSNS型)肾脏组织中表达最高,在激素耐药型肾病综合征(SRNS)型表达最低,差异有统计学意义(F=220.951,P<0.01)。2.活动期INS患儿血清TLR4表达MsPNG型:(143.36±12.99)ng/L、FSGS型(75.94±7.29)ng/L、MN型(210.22±14.66)ng/L、MCD型(283.93±21.58)ng/L]显著高于缓解期患儿MsPNG型:(29.51±4.93)ng/L、FSGS型(15.66±3.78)ng/L、MN型(45.40±5.73)ng/L、MCD型(62.29±7.90)ng/L]及对照组2儿童(0.69±0.33)ng/L],差异均有统计学意义(均P<0.01);且缓解期INS患儿血清TLR4的表达水平高于对照组2儿童,差异有统计学意义(F=286.287,P<0.01)。活动期及缓解期INS患儿血清TLR4的表达水平均以MCD型最高,其次为MN型,而FSGS型最低;血清TLR4在临床分型为SSNS型表达最高,SRNS型表达最低,差异有统计学意义(F=147.438,P<0.01)。3.INS患儿肾小管中TLR4的表达水平(62.82±20.94)%]与其活动期血清中TLR4的表达水平(213.26±73.33)ng/L]呈正相关(r=0.852,P<0.05);INS患儿肾小管及活动期血清TLR4的表达水平均与24 h尿蛋白水平(123.05±33.55)mg/kg]均呈正相关(r=0.401、0.427,均P<0.05),与血清清蛋白水平(19.54±3.55)g/L]均呈负相关(r=-0.602、-0.617,均P<0.05)。结论TLR4在INS患儿肾小管及血清中表达增高,且表达水平可能与不同肾脏病理类型及临床分型有关,与疾病活动具有相关性。

Expression and significance of Toll-like receptor 4 in renal tissue and peripheral blood of children with idiopathic nephrotic syndrome

Objective To investigate the expression and significance of Toll-like receptor 4(TLR4)in renal tissue and peripheral blood of children with idiopathic nephrotic syndrome(INS).Methods The renal biopsy tissues of 78 children with INS diagnosed in the First Affiliated Hospital of Xinxiang Medical University from October 2015 to June 2018 and normal renal tissues of 21 children(control group 1)were collected,and the expressions of TLR4 in the renal tissue was detected by using immunohistochemical method.The expression of TLR4 in different renal pathological types and clinical types of INS was compared,and the correlation of TLR4 with 24-hour urinary protein and serum albumin was analyzed.The expression levels of TLR4 in peripheral blood of children with INS before and after treatment(active stage and remission stage)and 23 healthy children(control group 2)were detected by enzyme linked immunosorbent assay(ELISA).The serum expression levels of TLR4 in different renal pathological types and clinical types of INS were compared,and the correlation of TLR4 with 24-hour urinary protein and serum albumin was analyzed;The correlation between TLR4 expression in renal tubules and in the serum of children with INS was also analyzed.Results(1)Compared with the expression of TLR4 in normal renal tissues(0.93±0.26)%],the expression of TLR4 in glomeruli and interstitium of all pathological types of INSmesangial proliferative glomerulonephritis(MsPGN):(0.93±0.21)%,focal segmental glomerulosclerosis(FSGS):(1.02±0.25)%,membranous glomerulonephritis(MN):(1.03±0.09)%,minimal change disease(MCD):(1.02±0.27)%]was not significantly different(F=0.741,P=0.562),but the expression of TLR4 in renal tubulesMCD:(82.94±4.62)%,MN:(63.54±1.98)%,MsPGN(42.32±2.97)%,FSGS:(22.60±2.07)%]was significantly increased(F=1929.842,P<0.01),Especially,the expression of TLR4 in renal tubules of MCD type INS was significantly higher than that of MN,MsPG N and FSGSMCD:(82.94±4.62)%,MN:(63.54±1.98)%,MsPGN:(42.32±2.97)%,FSGS:(22.60±2.07)%],and the differences were statistically significant(all P<0.01).TLR4 expression in renal tubules was the highest in steroid-sensitive nephrotic syndrome(SSNS)type and the lowest in INS patients with steroid-resistant nephrotic syndrome(SRNS)type,and the differences were statistically significant(F=220.951,P<0.01).(2)The expression of serum TLR4 in INS children at the active stageMsPNG:(143.36±12.99)ng/L,FSGS(75.94±7.29)ng/L,MN(210.22±14.66)ng/L,MCD(283.93±21.58)ng/L]was significantly higher than that in INS children at remission stageMsPNG:(29.51±4.93)ng/L,FSGS(15.66±3.78)ng/L,MN(45.40±5.73)ng/L,MCD(62.29±7.90)ng/L]and control group 2(0.69±0.33)ng/L],and the differences were statistically significant(all P<0.01);the expression of serum TLR4 in INS children at remission stage was significantly higher than that in the control group 2(F=286.287,P<0.01).TLR4 had the highest expression level in serum of MCD type INS children at active and remission stages,followed by MN and FSGS successively.The expression of serum TLR4 was highest in SSNS and lowest in SRNS,and the differences were statistically significant(F=147.438,P<0.01).(3)The expression of TLR4 in renal tubules of children with INS(62.82±20.94)%]was positively correlated with the expression of TLR4 in serum(213.26±73.33)ng/L](r=0.852,P<0.05).The expression levels of TLR4 in renal tubules and serum of INS patients at active stage were positively correlated with 24-hour urinary protein level(123.05±33.55)mg/kg](r=0.401,0.427,all P<0.05),and negatively correlated with serum albumin level(19.54±3.55)g/L](r=-0.602,-0.617,all P<0.05).Conclusions The expression of TLR4 in renal tubules and serum of children with INS increases,and may be related to different renal pathological types and clinical types of children with INS,as well as disease activity.