Department of Nephrology, the Second People's Hospital of Three Gorges University (Yichang Second People's Hospital), 443000 Yichang, Hubei, China.
Institute of Nephrology of Integrated Chinese and Western Medicine of Three Gorges University, 443000 Yichang, Hubei, China.
Discov Med. 2024 Mar;36(182):604-612. doi: 10.24976/Discov.Med.202436182.57.
The hedgehog signaling pathway exerts vital functions in regulating epithelial-to-mesenchymal transition (EMT) in renal interstitial fibrosis (RIF). It was reported that lncRNA-maternally expressed gene 3 (lncRNA Meg3) can regulate hepatic fibrosis by regulating the expression of smoothened (Smo) in the hedgehog signaling pathway. However, the specific role of lncRNA Meg3 in renal fibrosis resulting from unilateral ureteral obstruction (UUO) by regulating the hedgehog signaling pathway has not been reported. Hence, this research aimed to expound the effects of lncRNA Meg3 on renal fibrosis induced by UUO in rats via the hedgehog pathway.
Peripheral blood was collected from patients with chronic kidney disease (CKD, CKD group) and healthy volunteers (Normal group) at the same period. In addition, 6-week-old male Sprague-Dawley (SD) rats were divided to Sham, UUO, UUO+shRNA Negative control (shNC), and UUO+sh-Meg3 groups, and their kidney tissues and serum were gathered. Next, quantitative real-time polymerase chain reaction (qRT-PCR) was employed for detecting the lncRNA Meg3 expression level in the serum of patients and renal tissue of rats; kits for testing levels of blood urea nitrogen (BUN), creatinine (Cr), hydroxyproline (HYP), and 24-hour urine protein (24-up) in rats of each group; hematoxylin and eosin (HE) staining and Masson staining for observing kidney tissue and renal fibrosis level in rats; western blot for measuring levels of collagen type III (Col III), α-Smooth muscle actin (α-SMA), fibronectin, E-cadherin, sonic hedgehog (Shh), patched (Ptch) protein, smoothened (Smo) protein and glioma-associated oncogene homolog 1 (Gli1) protein expression.
LncRNA Meg3 was highly expressed in CKD patients and UUO rats ( < 0.01). In contrast to the UUO+shNC group, knocking down lncRNA Meg3 improved renal injury, relieved pathological renal lesions, and reduced kidney fibrosis and related protein levels. It inhibited the hedgehog pathway in kidney tissues of UUO rats ( < 0.05 and < 0.01).
LncRNA Meg3 can aggravate UUO-induced rat renal fibrosis by activating the hedgehog pathway.
刺猬信号通路在调节肾间质纤维化（RIF）中的上皮间质转化（EMT）中发挥重要作用。有报道称，长链非编码 RNA 母源表达基因 3（lncRNA Meg3）可通过调节刺猬信号通路中的 smoothened（Smo）表达来调节肝纤维化。然而，lncRNA Meg3 通过调节刺猬信号通路在单侧输尿管梗阻（UUO）引起的肾纤维化中的具体作用尚未报道。因此，本研究旨在通过刺猬途径阐述 lncRNA Meg3 对 UUO 诱导的大鼠肾纤维化的影响。
同期采集慢性肾脏病（CKD，CKD 组）患者和健康志愿者（正常组）外周血。此外，将 6 周龄雄性 Sprague-Dawley（SD）大鼠分为 Sham、UUO、UUO+shRNA 阴性对照（shNC）和 UUO+sh-Meg3 组，采集其肾组织和血清。然后，采用实时定量聚合酶链反应（qRT-PCR）检测患者血清和大鼠肾组织中 lncRNA Meg3 的表达水平；试剂盒检测各组大鼠血尿素氮（BUN）、肌酐（Cr）、羟脯氨酸（HYP）和 24 小时尿蛋白（24-up）水平；苏木精和伊红（HE）染色和 Masson 染色观察大鼠肾组织和肾纤维化程度；Western blot 检测各组大鼠胶原 III（Col III）、α-平滑肌肌动蛋白（α-SMA）、纤维连接蛋白、E-钙黏蛋白、刺猬（Shh）、 patched（Ptch）蛋白、 smoothened（Smo）蛋白和神经胶质瘤相关癌基因同源物 1（Gli1）蛋白表达水平。
lncRNA Meg3 在 CKD 患者和 UUO 大鼠中高表达（<0.01）。与 UUO+shNC 组相比，敲低 lncRNA Meg3 可改善肾损伤，减轻病理肾脏病变，并降低肾脏纤维化和相关蛋白水平。它抑制了 UUO 大鼠肾组织中的刺猬通路（<0.05 和 <0.01）。
lncRNA Meg3 可通过激活刺猬通路加重 UUO 诱导的大鼠肾纤维化。
