Hange-shashin-to Raises Levels of Somatostatin, Motilin, and Gastrin in the Plasma of Healthy Subjects.

Abstract

Somatostatin is a gastrointestinal motility regulator and functions as an inhibitor of hormone release in digestive organs. This peptide participates in the regulation of gastrointestinal motility with motilin. [1][2][3][4][5][6] Somatostatin as a gut regulatory peptide stimulates migrating motor complex (MMC)-like activity and its analogue was the first to be used therapeutically among the gastrointestinal peptides.7) Motilin, a peptide connected with somatostatin, has a powerful fundic pouch motor-stimulating activity and is one of the most important factors controlling the regular occurrence of phase 3 contractions of the MMC. 8,9) Another gut regulatory peptide, gastrin, stimulates acid secretion, and its release is mediated by various pathways such as direct stimulation of G cells, mediation by cholinergic nerves, stimulation of gastrin-releasing peptide, stomach pH, etc. 10)Vasoactive intestinal peptide (VIP) is widely distributed in the central and peripheral nervous system. This peptide has a vasodilating effect in many vascular beds, including the peripheral systemic and splanchnic beds and the cerebral arteries, and is an important neurotransmitter for the enteric nervous system. 11,12) Hange-shashin-to (EK-14), a Kampo herbal medicine, is prepared from seven crude herbs: Pinelliae Tuber, Scutellariae Radix, Zingiberis Rhizoma, Ginseng Radix, Glycyrrhizae Radix, Zizyphi Fructus, and Coptidis Rhizoma. Hangeshashin-to is frequently used for acute or chronic gastrointestinal catarrh, fermentative diarrhea, and acute gastroenteritis. This medicine has been clinically evaluated as more effective in the treatment of chronic hypofunction of the gastrointestinal system than is Rikkunshi-to in the treatment of hyperfunctioning conditions. Recently, Hange-shashin-to has been reported to be effective against diarrhea as a side effect of carcinostatic drugs. 13)Based on empirical effects of Kampo, some gut-targeting medicines have been determined to act at the gut regulatory hormone level. Ninjin-to, Rikkunshi-to, and Dai-kenchu-to affect gastrointestinal motility and based on empirical evidence their effects are assumed to be due to changes in the levels of somatostatin-, motilin-, gastrin-, or VIP-immunoreactive substances (IS) in plasma.14-18) Furthermore, abnormalities of gastrointestinal function is presumed to result from obstruction of the autonomic nervous system and changes in hormone levels. Thus we examined the plasma levels of gut regulatory peptides (somatostatin, motilin, gastrin, and VIP).Radioimmunoassays (RIA) to detect motilin have been developed by several groups using 125 I-motilin. [19][20][21] However, in terms of safety, sensitivity, and ease of handling, RIA methods are still less than satisfactory. We developed a sensitive and specific double-antibody enzyme immunoassay (EIA) for detecting motilin, using motilin-linked b-D-galactosidase as a marker antigen, a secondary antibody-coated immunoplate, and 4-methylumbelliferyl-b-D-galactopyranoside as a fluorogenic substrate.The purpose of this...

Comparison of the Effects of Pantethine and Fursultiamine on Plasma Gastrointestinal Peptide Levels in Healthy Volunteers

“…Synthetic human CGRP and its fragment , VIP, motilin and SP were purchased from Peptide Institute, Inc. (Osaka, Japan). VIP fragment (11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28) was supplied by Professor H. Yajima (Kyoto University, Kyoto, Japan). Antiserum to CGRP (14160) was purchased from Peptide Institute, Inc. (Osaka, Japan), VIP (T-4116) from Peninsula Laboratories (California, U.S.A.), and motilin (Y121) and SP (Y150) from Yanaihara Institute (Shizuoka, Japan).…”

“…The recovery of plasma CGRP-, VIP-, motilin-and SP-IS was greater than 90% using this extraction procedure. [15][16][17][18] Enzyme Immunoassays for CGRP-, VIP-, Motilin-and SP-IS Plasma peptide levels were measured using a highly sensitive enzyme immunoassay for CGRP, 15) VIP, 16) motilin 17) and SP 18) as described previously. The assay was performed by a delayed addition method.…”

“…An anti-rabbit IgG-(55641, ICN Pharmaceuticals, Inc., Ohio, U.S.A.) coated immunoplate (Nunc-Immuno Module Maxisorp F8, InterMed, Denmark) was used to separate bound and free antigens. Human CGRP fragment (8-37), VIP fragment (11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28), motilin and SP were conjugated with b-galactosidase by N-(emaleimido-caproyloxy)-succimide according to the methods of Kitagawa et al 19) The enzyme immunoassays for CGRP-, VIP-, motilin-and SP-IS were specific and highly sensitive, with detection limits of 4.0, 8.0, 0.25 and 0.25 pg/ml, respectively.…”

Effects of Pirenzepine on Dai-kenchu-to-Induced Elevation of the Plasma Neuropeptide Levels in Humans

“…For the EIA system, plasma samples were concentrated five-fold with SepPak C 18 cartridges. The recovery of plasma gastrin-, 19) somatostatin-, 20) motilin-, 21) VIP-, 22) CGRP-, 23) and substance P-IS 24) was Ͼ90% with this extraction procedure (data not shown).…”

“…The fluorescent product 4-methylumbelliferon was measured with an MTP-100F microplate reader (Corona Electric, Ibaraki, Japan). Fragment gastrin I (2-17), human somatostatin, porcine motilin, fragment VIP (11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28), human CGRP (8-37), and substance P were conjugated with b-D-galactosidase (Boehringer Mannheim, Mannheim, Germany) with N-(e-maleimidocaproyloxy)-succimide according to the method Kitagawa et al 25) The EIAs for gastrin, somatostatin, motilin, VIP, CGRP, and substance P were specific and highly sensitive to detection limits of 0.04, 0.10, 0.80, 1.00, 0.08, and 0.40 fmol/well, respectively.…”