The effects of Dai-kenchu-to (DKT), a Chinese medicine, on spontaneous activity of mouse small intestine were investigated. Experiments were carried out with tension recording and intracellular recording. DKT contracted mouse longitudinal smooth muscles in a dose dependent manner (0.1-10 mg/ml). Low concentration of DKT (0.1 mg/ml) did not contract the longitudinal muscles of mouse small intestine. DKT (0.1 mg/ml) inhibited contraction elicited by transmural nerve stimulation (TNS). DKT (1 mg/ml) evoked relaxation before contraction. The initial relaxation was abolished by Nω-nitro-L-arginine (L-NNA). DKT (10 mg/ml)-induced contraction had two components: a transient rapid contraction and a following slow contraction. Atropine inhibited DKT (1 mg/ml)-induced contraction to about 50% of control. In the presence of atropine, tetrodotoxin (TTX) inhibited the contraction elicited by DKT (1 mg/ml) to about 80%. DKT depolarized the membrane and decreased the amplitude of pacemaker potentials recorded from in situ myenteric interstitial cells of Cajal (ICC-MY) with no alteration to the frequency, duration and maximum rates of rise in the presence of nifedipine and TTX. The same results were obtained in slow waves recorded from circular smooth muscle cells. These results indicate that DKT evoked both contraction and relaxation by releasing acetylcholine, nitric oxide and other excitatory neurotransmitters in mouse small intestine. DKT had no effects on pacemaker mechanisms and electrical coupling between ICC-MY and smooth muscle cells in mouse small intestine. The results also suggest that DKT may contract smooth muscles by depolarizing the membrane directly.
The effects of 5-hydroxytryptamine (5-HT) on electrical responses of the membrane were investigated in circular smooth muscle isolated from the guinea-pig stomach antrum. Small segment of circular muscle tissue produced a periodical generation of slow potentials at frequency of 0.1-2 cycles min-1, during random generation of unitary potentials. Application of 5-HT (10-7-10-5 M) hyperpolarized the membrane and either increased or decreased the frequency of slow potentials, both with associated increase in amplitude of slow potential. These effects of 5-HT were abolished by methysergide. Nω-nitro-L-arginine (L-NA) increased the frequency of spontaneously generated slow potentials and also increased the frequency of slow potentials generated during stimulation with 5-HT, suggesting an involvement of the increased production of nitric oxide (NO) by 5-HT. Atropine did not alter spontaneous and 5-HT-induced electrical responses. The hyperpolarization produced by 5-HT was associated with a decrease in input resistance and time constant of the membrane. The amplitude of the 5-HT-induced hyperpolarization was increased in low [K+]o solution and decreased in high [K+]o solution or in the presence of glybenclamide, suggesting that the hyperpolarization was produced by activation of ATP-sensitive K-channels. The increase in amplitude of slow potentials by 5-HT may be secondary due to hyperpolarization of the membrane. The inhibition by 5-HT of the frequency of slow potentials may be partly due to the increased release of NO, however the mechanism by which dual effects of 5-HT on the frequency of slow potentials remains unsolved.
Mechanisms underlying β-adrenoceptor (β-AR)-mediated vascular relaxation were studied in the isolated rat abdominal aorta. In the endothelium-denuded helical preparations, a non-selective β-AR agonist isoprenaline elicited a concentration-dependent relaxation. In the absence of β-AR antagonists, isoprenaline-induced relaxation was not practically affected by an adenylyl cyclase inhibitor SQ 22,536 (300 μM), but was strongly diminished by high-KCl (80 mM). Isoprenaline-induced relaxation in the presence of SQ 22,536 was significantly diminished by iberiotoxin (IbTx, 0.1 μM), but was not affected by 4-aminopyridine (4-AP, 3 mM). Isoprenaline-induced relaxation was not also affected by SQ 22,536 (300 μM) even in the presence of CGP20712A (a β1-selective antagonist) and ICI-118,551 (a β2-selective antagonist) (0.1 μM for each), but was strongly diminished by high-KCl. By contrast, SQ 22,536-resistant, isoprenaline-induced relaxation in the presence of CGP20712A plus ICI-118,551 was not affected by IbTx (0.1 μM), but was inhibited significantly by 4-AP (3 mM). These results suggest that in rat abdominal aortic smooth muscle: 1) both β1-/β2-AR- and β3-AR-mediated relaxations substantially involve cAMP-independent mechanisms; 2) β1-/β2-AR-mediated, cAMP-independent relaxant mechanisms are partly attributed to the large-conductance, Ca 2+-sensitive K+ (MaxiK, BK) channel whereas β3-AR-mediated relaxant mechanisms are attributed to Kv channel.