Arginase inhibition attenuates arteriogenesis and interferes with M2 macrophage accumulation

Abstract

L-Arginine is the common substrate for nitric oxide synthases (NOS) and arginase. Whereas the contribution of NOS to collateral artery growth (arteriogenesis) has been demonstrated, the functional role of arginase remains to be elucidated and was topic of the present study. Arteriogenesis was induced in mice by ligation of the femoral artery. Laser Doppler perfusion measurements demonstrated a significant reduction in arteriogenesis in mice treated with the arginase inhibitor nor-NOHA (N ω -hydroxy-nor-arginine). Accompanying in vitro results on murine primary arterial endothelial cells and smooth muscle cells revealed that nor-NOHA treatment interfered with cell proliferation and resulted in increased nitrate/ nitrite levels, indicative for increased NO production. Immuno-histological analyses on tissue samples demonstrated that nor-NOHA administration caused a significant reduction in M2 macrophage accumulation around growing collateral arteries. Gene expression studies on isolated growing collaterals evidenced that nor-NOHA treatment abolished the differential expression of Icam1 (intercellular adhesion molecule 1). From our data we conclude that arginase activity is essential for arteriogenesis by promoting perivascular M2 macrophage accumulation as well as arterial cell proliferation.

Transcriptional and Metabolomic Analysis of L-Arginine/Nitric Oxide Pathway in Inflammatory Bowel Disease and Its Association with Local Inflammatory and Angiogenic Response: Preliminary Findings

“…Hypoxia-induced ARG2 overexpression supports cell proliferation in osteosarcoma [61]. Arginase activity has been shown to be necessary for arterial cell proliferation, acting via promoting perivascular M2 macrophage accumulation [62].…”

Contribution of the Potassium Channels KV1.3 and KCa3.1 to Smooth Muscle Cell Proliferation in Growing Collateral Arteries

“…Thereafter, RNA was purified with RNeasy MinElute columns (Qiagen) and reverse transcribed to cDNA using the QuantiTect ® Reverse Transcription Kit (Qiagen) according to the manufacturer´s procedure. The qRT-PCR was performed as previously described [32] using the Power SYBR Green Kit (Life Technologies) and a StepOnePlus cycler (Life Technologies) and the following primers: 18S rRNA forward 5 -GGACAGGATTGACAGATTGATAG-3 , reverse 5 -CTCGTTCGTTATCGGAATTAAC-3 , αSM-actin forward 5 -GAGCATCCGACACTGCTG-3 , reverse 5 -GTACGTCCAGAGGCATAG-3 , fibroblast growth factor receptor-1 (Fgfr1) forward 5 -CTTGCCGTATGTCCAGATCC-3 , reverse 5 -TCCGTAGATGAAGCACCTCC-3 , platelet derived growth factor b (Pdgfrb) forward 5 -AGGACAACCGTACCTTGGGTGACT-3 , reverse 5 -CAGTTCTGACACGTACCGGGTCTC-3 , early growth response 1 (Egr1) forward 5 -CGAACAACCCTATGAGCACCTG-3 , and reverse 5 -CAGAGGAAGACGATGAAGCAGC-3 . To control specific amplification, melt curve analyses and agarose gels were performed.…”

Interactions between Macrophages and Cyst-Lining Epithelial Cells Promote Kidney Cyst Growth in Pkd1-Deficient Mice

“…The doses of nor-NOHA used were 10 and 20 mg/kg, after reference to previous studies. [10][11][12][13] Control PKD mice were treated with normal saline (1.5 ml/g, daily) via i.p. injection.…”

“…The balance of ARG1 and NOS2 activities in arginine metabolism closely relates to cellular functions and behaviors, 12,[20][21][22] and Nor-NOHA was applied to inhibit ARG1 activity in polycystic kidneys ( Figure 5A). In general, short-term nor-NOHA treatment failed to improve survival rates in PKD mice ( Figure 5B).…”