背景:Hedgehog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。据报道,lncRNA母体表达基因3(lncRNAMeg3)可以通过调节hedgehog信号通路中smoothened(Smo)的表达来调节肝纤维化。然而,lncRNAMeg3通过调节hedgehog信号通路在单侧输尿管梗阻(UUO)导致的肾纤维化中的具体作用尚未见报道.因此,本研究旨在阐述lncRNAMeg3通过Hedgehog通路对UUO诱导的大鼠肾纤维化的影响。
方法:收集慢性肾脏病患者的外周血(CKD,CKD组)和同期健康志愿者(正常组)。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,收集了他们的肾组织和血清。接下来,定量实时聚合酶链反应(qRT-PCR)用于检测患者血清和大鼠肾组织中lncRNAMeg3的表达水平;用于检测血尿素氮(BUN)水平的试剂盒,肌酐(Cr),羟脯氨酸(HYP),各组大鼠24小时尿蛋白(24-up);苏木精和伊红(HE)染色和Masson染色观察大鼠肾脏组织和肾脏纤维化水平;westernblot测量III型胶原(ColIII)的水平,α-平滑肌肌动蛋白(α-SMA),纤连蛋白,E-cadherin,索尼克刺猬(Shh),patched(Ptch)蛋白,平滑(Smo)蛋白和神经胶质瘤相关癌基因同源物1(Gli1)蛋白表达。
结果:LncRNAMeg3在CKD患者和UUO大鼠中高表达(p<0.01)。与UUO+shNC组相比,敲低lncRNAMeg3改善肾损伤,减轻病理性肾脏病变,减少肾脏纤维化和相关蛋白质水平。它抑制了UUO大鼠肾组织中的hedgehog通路(p<0.05和p<0.01)。
结论:LncRNAMeg3可通过激活Hedgehog通路加重UUO诱导的大鼠肾纤维化。
BACKGROUND: 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.
METHODS: 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.
RESULTS: LncRNA Meg3 was highly expressed in CKD patients and UUO rats (p < 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 (p < 0.05 and p < 0.01).
CONCLUSIONS: LncRNA Meg3 can aggravate UUO-induced rat renal fibrosis by activating the hedgehog pathway.