Protective effect of keishi-bukuryo-gan and its constituent medicinal plants against nitric oxide donor-induced neuronal death in cultured cerebellar granule cells.

Keishi-bukuryo-gan (Gui-Zhi-Fu-Ling-Wan) (KBG) is a traditional Chinese/Japanese medical (Kampo) formulation that has been administered to patients with "Oketsu" (blood stagnation) syndrome. In the process of neuronal cell death induced by brain ischemia, excessive generation of nitric oxide (NO) free radicals is implicated in the neurotoxicity. In the present study, we examined the protective effects of KBG and its constituent medicinal plants against NO donors, sodium nitroprusside (SNP) and 2,2'-(hydroxynitrosohydrazino)bis-ethanamine (NOC18)-induced neuronal death in cultured rat cerebellar granule cells (CGCs). MTT assay showed cell viability to be significantly increased by the addition of KBG extract (KBGE) (100 microg/ml), Cinnamomi Cortex extract (CCE) (3, 10 and 30 microg/ml), Paeoniae Radix extract (PRE) (100 microg/ml) and Moutan Cortex extract (MCE) (10 and 30 microg/ml) compared with exposure to SNP (30 microM, 24 h) only. Also, cell viability was significantly increased by the addition of KBGE (100 and 300 microg/ml), CCE (30 and 100 microg/ml), PRE (100 and 300 microg/ml) and MCE (30 and 100 microg/ml) compared with exposure to NOC 18 (100 microM, 48 h) only. Persicae Semen extract and Hoelen extract did not protect against NO donor-induced neuronal death. These results suggest that KBG has protective effect against NO-mediated neuronal death in cultured CGCs and that it is derived from Cinnamomi Cortex, Paeoniae Radix and Moutan Cortex.

Effects of Keishi-bukuryo-gan on vascular function and hemorheological factors in spontaneously diabetic (WBN/kob) rats.

Keishi-bukuryo-gan (Gui-zhi-fu-ling-wan) is a formula used for the improvement of blood circulation. Recently it has often also been used for arteriosclerosis. One of the mechanisms involved is thought to be the improvement of endothelial dysfunction, but the details are still unclear. In this study, the effect of Keishi-bukuryo-gan on vascular function and hemorheological factors in spontaneously diabetic (WBN/kob) rats was studied. Rats were given Keishi-bukuryo-gan in chow for 30 weeks. Body weight, blood glucose, endothelium-dependent/-independent relaxation, vasocontraction by free radical-induced and contractive prostanoids, triglyceride, advanced glycation endproduct, lipid peroxides, serum NO2-/NO3- and blood viscosity were measured. The results indicated that Keishi-bukuryo-gan caused a decrease in endothelium-dependent relaxation by acetylcholine to become significantly increased, and vasocontraction induced by free radicals and contractive prostanoids was significantly decreased. Furthermore, serum NO2-/NO3- and blood viscosity were significantly decreased. From these results, it was supposed that Keishi-bukuryo-gan exerted a protective effect on the endothelium. The WBN/kob rat is a useful study model for the complications of human diabetes, and Keishi-bukuryo-gan showed a protective effect against vascular injury in the susceptible rat.

Comparative efficacy of Keishi-bukuryo-gan and pentoxifylline on RBC deformability in patients with "oketsu" syndrome.

Keishi-bukuryo-gan (Gui-Zhi-Fu-Ling-Wan) (KBG) is one of the prescriptions in Japanese traditional medicine for improving the "oketsu" syndrome, so-called blood stasis syndrome. "Oketsu" syndrome is an important pathological conception in Japanese traditional medicine and often accompanies cerebro-vascular disorders. Previously, we were able to reveal a deterioration of RBC (Red blood cell) deformability and viscoelasticity in patients with "oketsu" syndrome. The purpose of the present study was to evaluate whether KBG has an effect on RBC deformability in comparison with pentoxifylline (PXF). The subjects were 30 male patients with multiple lacunar infarctions. Eighteen patients (44-79 yrs, mean +/- SD, 66.1 +/- 10.7 yrs) were treated with 12 g of KBG daily for 4 weeks (KBG group). Twelve patients (59-78 yrs, 70.7 +/- 6.4 yrs) were treated with 300 mg of PXF daily for 4 weeks (PXF group). Based on the "oketsu" score, the patients of each group were divided into two subgroups, a non-"oketsu" group ("oketsu" score 20 points or less) and an "oketsu" group ("oketsu" score 21 points or higher). KBG had significant effects on RBC deformability as evaluated by filtration method. KBG also significantly increased intracellular ATP content, as did PXF. Moreover, KBG was more effective for patients with a more severe "oketsu" state. However, PXF was effective only in patients with "oketsu" syndrome, who might have deteriorated RBC deformability. In conclusion, the effect of KBG on RBC deformability was by no means inferior to PXF.

Effects of Choto-san on microcirculation, serum nitric oxide and lipid peroxides in patients with asymptomatic cerebral infarction.

The effects of Choto-san on microcirculation, serum nitric oxide and lipid peroxides were investigated. Fifteen patients with asymptomatic cerebral infarction were investigated before and after a four-week administration of Choto-san. The variables of microcirculation of the bulbar conjunctiva, which were the internal diameter of vessels, flow velocity and flow volume rate, were increased. Serum NO2(-)/NO3(-) tended to increase and lipid peroxides were decreased. Total serum cholesterol was also decreased. These results suggest that Choto-san may improve microcirculation affected by endothelial function and fat metabolism.

Effects of Shimotsu-to on the microcirculation of the bulbar conjunctiva and hemorheological parameters in normal subjects.

The effects of 'Shimotsu-to' (Si-Wu-Tang in Chinese), one of the most important prescriptions for 'ketsukyo' and 'oketsu' syndrome in traditional Chinese medicine, on the microcirculation of bulbar conjunctiva and the hemorheological parameters were examined in ten healthy volunteers. After one hour of oral administration of Shimotsu-to extract, the blood flow rate and the blood flow volume significantly increased and the DEA (maximum diameter of the column of intravascular erythrocyte aggregation) decreased. The whole blood viscosity declined at middle and high shear rates, but both the plasma viscosity and the erythrocyte deformability were not effected. These results suggest that Shimotsu-to has a salutary effect on the microcirculation through a decrease in the whole blood viscosity.