Overexpression of lncRNA ANRIL up-regulates VEGF expression and promotes angiogenesis of diabetes mellitus combined with cerebral infarction by activating NF-κB signaling pathway in a rat model

Abstract

ObjectiveThis study aimed to explore the effects of lncRNA ANRIL on vascular endothelial growth factor (VEGF) and angiogenesis in diabetes mellitus (DM) combined with cerebral infarction (CI) through NF-κB signaling pathway.MethodsAdult male Wistar rats were randomly divided into control group and DM + CI group, and the DM + CI group were subdivided into Vector, shANRIL, PDTC, pcDNA-ANRIL, and pcDNA-ANRIL + PDTC groups. VEGF and FMS-like tyrosine kinase (FLT-1) expressions were measured by immunohistochemistry and endothelium dependent microvessel density (MVD) was detected by differentiation 31 (CD31) and para-amiuosalicylic acid (PAS) double staining. The qRT-PCR was applied to measure mRNA expressions of VEGF, FLT-1, Kinase insert domain protein receptor (FLK-1) and NF-κB, and Western blotting was conducted to detected expressions of VEGF, NF-κB and p-I?B/I?B.ResultsCompared with the control group, protein expressions of VEGF, NF-κB, p-I?B/I?B, expression of ANRIL, and mRNA expressions of VEGF, FLT-1 and NF-κB were increased in the DM + CI group. Compared with the Vector group, protein expressions of VEGF, NF-κB, p-I?B/I?B, expression of ANRIL, mRNA expressions of VEGF, FLT-1 and NF-κB, and endothelium dependent MVD were increased in the pcDNA-ANRIL group, while decreased in the shANRIL group and PDTC group. Compared with the pcDNA-ANRIL group, protein expressions of VEGF, NF-κB, p-I?B/I?B, expression of ANRIL, mRNA expressions of VEGF, FLT-1 and NF-κB, and endothelium dependent MVD were decreased in the pcDNA-ANRIL + PDTC group.ConclusionOverexpressed lncRNA ANRIL upregulates VEGF and promotes angiogenesis by activating NF-κB signaling pathway in DM + CI rats.

Plasma level of miR‐99b may serve as potential diagnostic and short‐term prognostic markers in patients with acute cerebral infarction

“…It has been observed that in cerebral infarction mouse models, miR‐145 can regulate the proliferation and migration of endothelial progenitor cells; moreover, miR‐210 was able to activate the NOTCH signaling and improve the angiogenesis after acute cerebral ischemia . VEGF is known as an angiogenic factor, and in the present study, we observed that the plasma levels of VEGF were significantly up‐regulated in patients with ACI, which was consistent with previous findings . More important, we first reported that the plasma level of miR‐99b was negatively correlated with the level of VEGF in patients with ACI, suggesting that down‐regulation of miR‐99b may be involved in the process angiogenesis in brain tissue after cerebral infarction.…”

ANRIL and atherosclerosis

“…VEGF is an important angiogenic factor reported to induce the migration and proliferation of endothelial cells and enhance vascular permeability, thus accelerating endothelial injury . Zhang et al found that ANRIL regulated the expression of VEGF by activating the NF‐κB signalling pathway in a rat model of diabetes mellitus complicated by cerebral apoplexy . McArthur et al demonstrated that ANRIL regulated the expression of VEGF through recruiting PRC2 or p300 and regulating the expression of miR‐200b in a rat model of diabetes mellitus .…”

The correlation of long non‐coding RNA intersectin 1‐2 with disease risk, disease severity, inflammation, and prognosis of acute ischemic stroke

“…For instance, increased lncRNA H19 expression is associated with impaired neurological function and increased TNF‐α level in AIS animal models . Antisense non‐coding RNA in the cyclin‐dependent kinase inhibitor 4 locus (ANRIL), an antisense lncRNA co‐clustered with p15/CDKN2B‐p16/CDKN2A‐p14/ARF, is overexpressed in cerebral infarction rat models and plays a pro‐inflammatory role by activating NF‐κB pathway . Likewise, lncRNA Gm4419 could activate NF‐κB pathway and contributes to cell damage in oxygen‐glucose–deprived cerebral microglial cells .…”

“…23 Antisense non-coding RNA in the cyclin-dependent kinase inhibitor 4 locus (ANRIL), an antisense lncRNA co-clustered with p15/CDKN2B-p16/CDKN2A-p14/ ARF, is overexpressed in cerebral infarction rat models and plays a pro-inflammatory role by activating NF-κB pathway. 24 Likewise, ln-cRNA Gm4419 could activate NF-κB pathway and contributes to cell damage in oxygen-glucose-deprived cerebral microglial cells. 25 Another in vitro and in vivo study discloses that lncRNA SNHG14 elevates the expression of pro-inflammatory factors (such as TNF-α and nitric oxide), thereby aggravating neuron damage by regulating miR-145-5p/PLA2G4A.…”