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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54只雄性SPF级Wistar大鼠随机分为单侧输尿管梗阻组(unilateral ureteral obstruction group,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(antisense oligonu-cleotide group,AS),GATA-3无意义寡核苷酸组(nonsense oligonucleotide group,NS)和生理盐水组,每组9只。正常对照组27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3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的肾脏纤维化现象均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进一步证实,在梗阻组大鼠的肾脏组织中均有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随梗阻时间的延长显著下降。结论:转录因子GATA-3通过减少BMP-7诱导的Smad7的表达,使Smad7阻断转化生长因子(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。
OBJECTIVE: To observe the role of GATA-3 in renal fibrosis through animal experimental model and to further understand the strategic direction of reversing fibrosis by intervention of renal fibrosis signaling pathway. Methods: Fifty-four male SPF Wistar rats were randomly divided into unilateral ureteral obstruction group (UUO) and normal control group. Unilateral ureteral obstruction group was divided into GATA-3 antisense oligonucleotide Antisense oligonu-cleotide group (AS), GATA-3 nonsense oligonucleotide group (NS) and normal saline group. The normal control group of 27 rats to take the same grouping method. On the 1st, 7th and 14th day of the experiment, 3 rats in each group were sacrificed. The pathological changes of renal fibrosis were observed by HE and Masson staining and the degree of fibrosis was evaluated. The expression of GATA-3 and Smad7 were observed by immunohistochemistry Statistical area. RT-PCR method was used to observe the expression of GATA-3 and Smad7 mRNA in rat kidney while calculating the gray value. Results: GATA-3 antisense oligodeoxynucleotide-treated rats both in the obstruction group and the normal control group of renal fibrosis were slightly less than the other two treatment groups, immunohistochemistry results showed that in the obstruction group and the normal control The expression of GATA-3 and GATA-3 in both groups were further confirmed by RT-PCR. The expression of GATA-3 and Smad7 mRNA in the kidney of obstructive rats and the expression of Smad7 in antisense oligonucleotide-treated rats No matter in the obstruction group or the normal control group, the obstruction time was significantly decreased. CONCLUSION: GATA-3 attenuates BMP-7-induced Smad7 expression and attenuates the signal transduction of TGF-β by Smad7, which aggravates the occurrence and development of renal fibrosis. It is feasible to interfere with the process of fibrosis through the intervention of transcription factors in the fibrosis signaling pathway.