人巨细胞病毒肝炎综合征婴儿外周血宿主microRNA表达的变化

Expression profile of host microRNA in the peripheral blood in infantile hepatitis syndrome caused by human cytomegalovirus

目的观察人巨细胞病毒（HCMV）所致肝炎综合征婴儿治疗前后外周血microRNA（miRNA）水平变化，探讨宿主miRNA与更昔洛韦（GCV）疗效的关系及临床意义。方法收集HCMV感染所致肝炎综合征婴儿44例为病例组，分为GCV不敏感组（20例）和GCV敏感组（24例）；15例同期体检健康婴儿为健康对照组。分别在GCV治疗前2d和停药后（或用药4周末），采集患儿外周静脉血1mL，提取miRNA，采用SYBRGreenI实时荧光定量PCR方法检测hsa—miR一92a、hsa—miR一96、hsa—miR-433和hsa—miR一183表达水平，结果用相对表达量表示。结果病例组hsa—miR-92a、hsa—miR一96、bsa—miR-433和hsa—miR一183表达水平均高于健康对照组，差异均有统计学意义（P均〈0．05）；治疗前GCV不敏感组hsa—miR-96表达水平（1．24±0．62）低于GCV敏感组患儿（5．43±1，11），差异有统计学意义（t：一2．41，P=0．02）；治疗后GCV不敏感组hsa—miR-96（0．70±0．06）和hsa—miR一183（2．97±0．14）均明显低于敏感组（6．47±1．01、9．84±0．95），差异均有统计学意义（t’=一5．46，P〈0．01；t=一2．52，P=0．019）；GCV不敏感组hsa—miR一183治疗后（2．974-0．14）较治疗前（9．79±1．31）明显下降，hsa—miR-433治疗后（6．20±0．92）较治疗前（10．90±0．77）表达水平下降，差异均有统计学意义（t=一3．54，P=0．002；t：2．45，P=0．027）；GCV敏感组hsa—miR-183表达水平治疗后（9．84±0．95）较治疗前（9．27±0．96）升高，hsa—miR-433治疗后（6．57±0．71）较治疗前（9．88±1．32）下降，差异均有统计学意义（t=-2．54，P=0．018；t=3．70，P〈0．001）。结论GCV敏感患儿用药前后hsa—miR-96表达均高于不敏感患儿，用药后敏感患儿hsa-miR一-83表达也高于不敏感患儿，miRNA对临床治疗婴儿肝炎综合征具有指导意义。

Objective To measure the expressions of mieroRNAs （miRNA） in the peripheral blood in infan- tile hepatitis syndrome（IHS） caused by human cytomegalovirus （HCMV） and analyze the relationship between miR- NAs and the Gancielovir （GCV） effectiveness. Methods Forty-four eases of IHS caused by HCMV were recruited from Xinhua Hospital Affiiiated to Shanghai Jiaotong University School of Medicine as patients group. Patients were fur- ther grouped into GCV nonsensitive group（ n = 20） and GCV sensitive group（ n = 24） ;15 healthy infants were taken as healthy control group. One mL peripheral blood were collected at 2 days before the GCV treatment and termination of GCV treatment （ or 4 weeks after treatment） respectively, miRNAs were extracted by using special kit, which followed by SYBR Green I real-time fluorescence quantitative polymerase chain reaction for testing the relative quantitative （RQ） of hsa-miR-92a,hsa-miR-96, hsa-miR-433 and hsa-miR-183. Results The RQ of hsa-miR-92a, hsa-miR-96, hsa-miR-433 and hsa-miR-183 in patients group were significantly higher than those of healthy control group （ all P 〈 0.05 ） ; Before GCV therapy, the RQ of hsa-miR-96 in GCV nonsensitive group （ 1.24±0.62） was lower than that of GCV sensitive group （5.43±1.11 ） （ t = - 2.41, P = 0.02） ; After GCV treatment, the expression of hsa-miR-96 in GCV nonsensitive group （0.70± 0.06） was lower than that of GCV sensitive group （6.47±1.01 ） （ t = - 5.46,P 〈 0.01） ,while the expression of hsa-miR-183 showed the same tendency （2.97 ±0.14 vs 9.84 ±0.95） （t = -2.52, P =0. 019） ; Furthermore, the expression of hsa-miR-183 and hsa-miR-433 were observed to be down-regulated from 9.79 ± 1. 31 and 10.90±0.77 to 2.97±0.14 and 6.20±0.92,respectively after therapy in GCV nonsensitive group （ t = - 3.54 ,P = 0. 002 ; t = 2.45, P = 0. 027 ）. However, the level of hsa-miR-183 after therapy （ 9.84±0.95 ） was higher than that before treatment （9.27±0.96） in GCV sensitive group（t = -2. 54,P = 0. 018）,while the level of hsa-miR-433 （6.57±0.71 ） after therapy was lower than that before treatment （9.88 ±1.32 ） （ t = 3.70,P 〈 0.001 ）. Conclusions The expression levels of hsa-miR-96 in GCV sensitive group were higher than those in GCV nonsensitive group before or after GCV treatment; the level of hsa-miR-183 in GCV sensitive group was also higher than that in GCV nonsentitive group. The miRNAs expression levels have significance for clinical practice.