Decreased Level of Neuropeptide Y Is Associated With Gastroparesis in Male Diabetic Rats

Rebecca Smiley, Richard McCallum, Mohammed Showkat Ali

Abstract


Background: Substance P (SP) and neuropeptide Y (NPY), excitatory and inhibitory neuropeptides, respectively, may impact gastric motility in patients with diabetic mellitus (DM). We investigated these neuropeptide levels, NPY receptors, total nitric oxide synthase (NOS) levels, and neuronal NOS alpha (nNOSalpha) activation status and levels in streptozotocin-induced type I diabetes in male rats.

Methods: Rats were grouped based on serum glucose and gastric emptying time: normal untreated control (CM), diabetic (DM) and diabetic gastroparesis (DM + GP). Neuropeptide serum levels were determined by enzyme-linked immunosorbent assay (ELISA). Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting measured NPY receptors, Y1 and Y2, and nNOSalpha expression. Low-temperature SDS-PAGE followed by western blotting was used to measure the dimerization of nNOSalpha. An NOS colorimetric assay kit was used to measure total NOS activity.

Results: SP levels were significantly decreased (P < 0.05) in DM and DM + GP compared to CM. NPY levels were significantly decreased (P < 0.05) in DM compared to CM, and DM + GP had a more significantly decreased NPY when compared to both DM and CM. Protein levels of neuropeptide receptor Y1 (NPY-Y1) in the smooth muscle of pylorus were significantly increased in DM, but not in DM + GP when compared to CM. Neuropeptide receptor Y2 (NPY-Y2) was not detected. Changes in nNOSalpha activity and their protein levels, as well as total NOS activity, among the groups were insignificant.

Conclusions: Increased expression of pylorus NPY-1R and decreased serum NPY are present in diabetes. A more pronounced decreased serum NPY with no NPY-1R upregulation in pyloric smooth muscle is associated with gastroparesis. NPY levels show no relationship with nNOSalpha levels, their activation status, or total NOS activity in pyloric smooth muscle. These data suggest a pathophysiological role of severely depleted NPY and absence of NPY-Y1 upregulation for gastroparesis phenotype.




Gastroenterol Res. 2020;13(6):246-252
doi: https://doi.org/10.14740/gr1322

Keywords


Neuropeptide; Substance P; Diabetic gastroparesis

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