CAJ | 학술논문

目的通过动物实验模型观察转录因子GATA-3在肾脏纤维化中的作用,通过对肾脏纤维化信号通路的干预,进一步了解逆转纤维化进程的策略方向.方法将36只雄性SPF级Wistar大鼠随机分为单侧输尿管梗阻组(UUO)和正常对照组,单侧输尿管梗阻组按照处理因素再分为GATA-3 反义寡核苷酸组(AS )、GATA-3 无意义寡核苷酸组(NS )和生理盐水组,每组6只.正常对照组18只大鼠采取同样的分组方法 .分别在试验第1、7、14天处死每小组2只大鼠,HE 和Masson染色观察肾脏纤维化的病理变化并评估纤维化程度,免疫组化方法观察GATA-3、 Smad7等阳性表达情况并统计面积.RT-PCR 方法观察大鼠肾脏组织GATA-3及Smad7 mRNA 表达同时计算灰度值.结果 GATA-3反义寡核苷酸处理的大鼠无论是在梗阻组及正常对照组均观察到肾脏纤维化较其他两个处理组轻微,免疫组化结果显示,在梗阻组及正常对照组组均有GATA-3的表达,通过RT-PCR进一步得以证实,在梗阻组大鼠的肾脏组织中均检测到GATA-3及Smad7 RNA 的表达,反义寡核苷酸处理的大鼠Smad7的表达无论是在梗阻组还是正常对照组中均明显低于其他两个处理因素.结论转录因子GATA-3通过减少BMP-7诱导的Smad7的表达,使得Smad7阻断TGF-β的信号转导作用削弱,加剧了肾脏纤维化的发生发展,进一步证实通过对纤维化信号通路中的转录因子进行干预影响纤维化的进程是可行的.
목적 통과동물실험모형관찰전록인자GATA-3재신장섬유화중적작용,통과대신장섬유화신호통로적간예,진일보료해역전섬유화진정적책략방향.방법 장36지웅성SPF급Wistar대서수궤분위단측수뇨관경조조(UUO)화정상대조조,단측수뇨관경조조안조처리인소재분위GATA-3 반의과핵감산조(AS )、GATA-3 무의의과핵감산조(NS )화생리염수조,매조6지.정상대조조18지대서채취동양적분조방법 .분별재시험제1、7、14천처사매소조2지대서,HE 화Masson염색관찰신장섬유화적병리변화병평고섬유화정도,면역조화방법 관찰GATA-3、 Smad7등양성표체정황병통계면적.RT-PCR 방법 관찰대서신장조직GATA-3급Smad7 mRNA 표체동시계산회도치.결과 GATA-3반의과핵감산처리적대서무론시재경조조급정상대조조균관찰도신장섬유화교기타량개처리조경미,면역조화결과 현시,재경조조급정상대조조조균유GATA-3적표체,통과RT-PCR진일보득이증실,재경조조대서적신장조직중균검측도GATA-3급Smad7 RNA 적표체,반의과핵감산처리적대서Smad7적표체무론시재경조조환시정상대조조중균명현저우기타량개처리인소.결론 전록인자GATA-3통과감소BMP-7유도적Smad7적표체,사득Smad7조단TGF-β적신호전도작용삭약,가극료신장섬유화적발생발전,진일보증실통과대섬유화신호통로중적전록인자진행간예영향섬유화적진정시가행적.
Objectives Through the animal experiment model observe the action of transcription factor GATA-3 in renal fibrosis,and through interfere in the signal iteral of renal fibrosis to get the deeper message of strategy and direction for reversal renal fibrosis.Methods Thirty six male specified-pathogens free Wistar rats were randomly divided into two groups:the Unilateral Ureteral Obstruction group and normal comparison group.the Unilateral Ureteral Obstruction group according to the management agent random subdivision into three group:GATA-3 antisense oligonucleotide group(AS),GATA-3 nonsense oligonucleotide group (NS) and sodium chloride group.the nomal comparison group 18 rats also adopt the same method divide into three groups.And at the time of study 1d、7d、14d executes two rats of each subgroup, Histological changes of renal tubular interstitium were observed by HE and Masson staining. Immunohistochemistry was performed on renal tissue for observe the expression of GATA-3、 Smad7 and so on at each time point. RT-PCR to observe the expression of GATA-3、Smad7 mRNA in renal fibrosis and calculate gray scale.Results Regardless of Ureteral Obstruction group and nomal comparison group ,the rat treat with GATA-3 antisense oligonucleotide observe that renal fibrosis little than other two treatment, Iminunohistochemistry observe that transcription factor GATA-3 expression in Ureteral Obstruction group and nomal group ,RT-PCR further infirm that result .At Ureteral Obstruction group detect the expression of GATA-3 and Smad7 RNA;treat with GATA-3 antisense oligonucleotide observe that Smad7 expression in Ureteral Obstruction group and nomal comparison group even less than the other two treatment.Conclusions Transcription factor GATA-3 through reduce the expression of Smad7 induced by BMP-7 ,and impair the Smad7's function blockade TGF-β signal transduction,to intensify the renal fibrosis generate and development ,further infirm that interfere in the transcription factor of renal fibrosis signal path to effect the fibrosis's proceeding is feasible.