Sun Lei, Shen Yun-Lin, Liu Hua-Jie, Hu Yu-Jie, Kang Yu-Lin, Huang Wen-Yan
a Department of Nephrology and Rheumatology , Shanghai Children's Hospital, Children's Hospital of Shanghai Jiaotong University , Shanghai , P.R. China.
Ren Fail. 2016;38(2):276-81. doi: 10.3109/0886022X.2015.1120118. Epub 2015 Dec 10.
To investigate the expression of response gene to complement 32 (RGC32) in rat with acute kidney injury (AKI) and to explore the role of RGC32 in renal injury and repair induced by ischemia reperfusion. Rats were randomly divided into two groups, including sham operation group (n = 48) and acute ischemia reperfusion injury (IRI) group (n = 48). Rats were sacrificed following reperfusion 2 h, 6 h, 24 h, 48 h, 72 h, 1 week (w), 2 w, and 4 w. The distribution and expression of RGC32 in renal tissue were observed by means of immunohistochemistry. The mean density of the images detected by Image-Pro Plus 6 was designated as the representative RGC32 expression levels. Meanwhile, RGC32 mRNA expression was measured by qPCR. RGC32 mainly expressed in cytoplasm of proximal tubular epithelial cells. However, RGC32 did not express in renal interstitium and vessels. The expression levels of RGC32 measured by immunohistochemistry at different reperfusion time were 0.0168 ± 0.0029, 0.0156 ± 0.0021, 0.0065 ± 0.0013, 0.0075 ± 0.0013, 0.0096 ± 0.0014, 0.0132 ± 0.0016, 0.0169 ± 0.0014, 0.0179 ± 0.0022, respectively. Compared with the sham group, the level of RGC32 expression in IRI group was significant lower at 24 h, 48 h, 72 h after IRI (p < 0.05). The expression levels of RGC32 mRNA at different reperfusion time measured by qPCR were corroborated the immunohistochemistry finding. The in vitro experiments show the expression of α-SMA and extracellular matrix expression increased signification when the RGC32 was silenced. Our data showed that the RGC32 expression in AKI rat decreased significantly reduces with different reperfusion time and performs a time-dependent manner. RGC32 may play an important role in the pathogenesis of AKI following IRI and repair in rat.
研究急性肾损伤（AKI）大鼠中补体反应基因32（RGC32）的表达，并探讨RGC32在缺血再灌注诱导的肾损伤及修复中的作用。将大鼠随机分为两组，包括假手术组（n = 48）和急性缺血再灌注损伤（IRI）组（n = 48）。在再灌注2小时、6小时、24小时、48小时、72小时、1周（w）、2周和4周后处死大鼠。采用免疫组织化学方法观察RGC32在肾组织中的分布及表达。将Image-Pro Plus 6检测图像的平均密度作为RGC32的代表性表达水平。同时，采用qPCR检测RGC32 mRNA表达。RGC32主要表达于近端肾小管上皮细胞的细胞质中。然而，RGC32在肾间质和血管中不表达。不同再灌注时间免疫组织化学检测的RGC32表达水平分别为0.0168±0.0029、0.0156±0.0021、0.0065±0.0013、0.0075±0.0013、0.0096±0.0014、0.0132±0.0016、0.0169±0.0014、0.0179±0.0022。与假手术组相比，IRI组在IRI后24小时、48小时、72小时的RGC32表达水平显著降低（p < 0.05）。qPCR检测不同再灌注时间的RGC32 mRNA表达水平证实了免疫组织化学结果。体外实验显示，当RGC32沉默时，α-SMA的表达和细胞外基质表达显著增加。我们的数据表明，AKI大鼠中RGC32的表达随不同再灌注时间显著降低，并呈时间依赖性。RGC32可能在大鼠IRI后的AKI发病机制及修复中起重要作用。
