Nobuyasu Sekiya

Cholesterol-fed rabbit as a unique model of nonalcoholic, nonobese, non-insulin-resistant fatty liver disease with characteristic fibrosis

BackgroundThe number of patients suffering from metabolic syndrome is increasing rapidly. Metabolic syndrome causes severe pathological changes in various organs, including the liver, and its main phenotype is nonalcoholic fatty liver disease (NAFLD). NAFLD has a broad spectrum ranging from simple fatty change to severe steatohepatitis with marked fibrosis. Recently, several experimental animal models for NAFLD have been proposed. However, most were established by rather artificial conditions such as genetic alteration. In the present study, we tried to establish a unique animal model mimicking some of the physiopathological features of NAFLD using high-cholesterol-fed rabbits.MethodsMale rabbits fed with standard rabbit food containing 1% cholesterol for 8 weeks and 12 weeks were compared to controls (six rabbits/group). The weight of food was strictly restricted to 100 g/rabbit per day.ResultsBody weights and fasting plasma insulin levels showed no significant differences among the groups. In contrast, characteristic fine fibrosis was extended from perivenular to pericellular areas, and microvesicular fatty change with ballooning degeneration was observed in perivenular areas in livers of the cholesterol-fed rabbits. Increase of serum cholesterol level, activation of hepatic stellate cells, and exposure to oxidative stress were also recognized.ConclusionsCholesterol-fed rabbits share several physiopathological features of NAFLD. Because this model did not show insulin resistance or obesity, it may be useful for elucidating the mechanism of NAFLD related mainly to hyperlipidemia.

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.

Keishi-bukuryo-gan prevents the progression of atherosclerosis in cholesterol-fed rabbit

In this study, we examined whether in vivo keishi‐bukuryo‐gan (a Kampo formulation) could prevent the progression of atherosclerosis in cholesterol‐fed rabbits, an animal model for hypercholesterolaemia. Sixteen male Japanese white rabbits (2 kg body weight) were divided into two groups. Group A (n = 8) was fed standard rabbit chow containing 1% cholesterol for 8 weeks. Group B (n = 8) was fed standard rabbit chow containing 1% cholesterol and 1% keishi‐bukuryo‐gan for 8 weeks. At the end of the experiment, average plasma concentrations of total‐cholesterol and IDL‐cholesterol were 2055.9 ± 201.8 mg/dL and 408.1 ± 62.6 mg/dL in group A and 1950.5 ± 126.3 mg/dL and 407.6 ± 56.6 mg/dL in group B, respectively. The percentage of the surface area of the total thoracic aorta with visible plaque was significantly reduced by keishi‐bukuryo‐gan administration; group A was 33.2% ± 5.3% and group B was 14.3% ± 2.9%. β ‐very low density lipoprotein (VLDL) and low density lipoprotein (LDL) isolated from cholesterol fed rabbits treated with keishi‐bukuryo‐gan (group B) were shown to be highly resistant to oxidative modification by cupric ion. Sera isolated from rabbits administered keishi‐bukuryo‐gan had reduced lipid peroxide formation compared with those from rabbits without keishi‐bukuryo‐gan. Thus, keishi‐bukuryo‐gan prevents the progression of atherosclerosis in cholesterol‐fed rabbits in vivo by limiting oxidative LDL modification. Copyright

HOCHUEKKITO IMPROVES SYSTEMIC INFLAMMATION AND NUTRITIONAL STATUS IN ELDERLY PATIENTS WITH CHRONIC OBSTRUCTIVE PULMONARY DISEASE

To the Editor: Chronic obstructive pulmonary disease (COPD) has significant systemic effects, such as weight loss and nutritional abnormalities. Although not much is known regarding the underlying mechanisms of systemic effects, any new therapeutic strategy would be required to improve this chronic inflammatory process. The traditional herbal medicine Hochuekkito (Bu-Zhong-Yi-Qi-Tang) could improve the quality of life and immunological status of elderly patients and be useful in protecting infection. The effects of Hochuekkito on systemic inflammation and nutritional status in elderly patients with COPD were investigated. It has been reported that Hochuekkito inhibits rhinovirus infection, one of the major causes of common colds, and modulates airway inflammation by reducing cytokine production. In the present study, the effects of Hochuekkito on systemic inflammation and nutritional status was investigated in 71 elderly patients with moderate to severe COPD. COPD was diagnosed based on past history, physical examination, and spirometric data. Patients with COPD frequently experience common colds and exacerbations in winter season in Japan. This study was started in September to December 2004 and 2005, so that it would cover the winter for 6 months of the study period, and subjects would have more chances to acquire common colds and experience exacerbations. Health-related quality of life was evaluated according to the Symptoms score of the St. George’s Respiratory Questionnaire (SGRQ). This report is based on the results of a prospective, multicenter (n 5 20), parallel-group, randomized study. The results from Chiba University have been reported previously. The study group (n 5 34) was given 7.5 g of Hochuekkito extract daily in addition to inhaled bronchodilators, inhaled glucocorticosteroids, or both, whereas the control group (n 5 37) continued the previous prescription of inhaled bronchodilators, inhaled glucocorticosteroids, or both. The daily dose of Hochuekkito extract was divided into three doses of 2.5 g each taken orally 30 minutes before each meal for 6 months. There were no significant differences between the two groups regarding age, sex, body mass index, pulmonary function, previous prescriptions, body weight, or Symptoms score of the SGRQ at the start of the study. Body weight increased from baseline (50.9 2.0 kg) to the end of the trial (52.2 1.9 kg) in the study group (Po.05), and remained unchanged in the control group. The Symptoms score decreased with treatment with Hochuekkito in the study group (from 59.8 4.1 to 45.6 3.3; Po.05), but did not in the control group, indicating that Hochuekkito improved patient quality of life. The mean number of common colds for 6 months was significantly lower in the study group (2.04 0.46) than in the control group (3.42 0.49) (Po.05). The number of exacerbations over 6 months was also significantly lower in the study group (0.12 0.06) than in the control group (0.52 0.13) (Po.05). Serum inflammatory (C-reactive protein (CRP), tumor necrosis factor alpha (TNF-a), and interleukin (IL)-6) and nutritional (prealbumin) markers were examined before and 6 months after the treatments. CRP, TNF-a, and IL-6 were not significantly different between the groups in the baseline period. Blood levels of CRP, TNF-a, and IL-6 correlated negatively with percentage predicted forced expiratory volume in 1 minute in the baseline period. CRP, TNF-a, and IL-6 decreased after Hochuekkito treatment but remained unchanged in the control group. Serum prealbumin correlated positively with body mass index and was not significantly different between groups in the baseline period. In the study group, serum prealbumin level increased and was unchanged in the control group (Figure 1). Any apparent adverse effect due to Hochuekkito therapy was not observed during the study period. These data indicate that Hochuekkito treatment improves systemic inflammation and nutritional status and decreases the chances of the common cold and

Evidence-based efficacy of Kampo formulas in a model of non alcoholic fatty liver.

Data on the efficacy of herbal compounds are often burdened by the lack of appropriate controls or a limited statistical power. Treatments to prevent the progression of non alcoholic fatty liver disease (NAFLD) to steatohepatitis (NASH) remain unsatisfactory. A total of 56 rabbits were arrayed into 7 groups fed with standard rabbit chow (SRC), SRC with 1% cholesterol, or each of the five experimental treatments (Kampo formulas 1% keishibukuryogan [KBG], 1% orengedokuto [OGT], and 1% shosaikoto [SST]; vitamin E [VE]; or pioglitazone [PG]) in a 1% cholesterol SRC. We analyzed changes after 12 weeks in plasma and liver lipid profiles, glucose metabolism, adipocytokines, oxidative stress, and liver fibrosis. Data demonstrated that all five treatments were associated with significant amelioration of lipid profiles, oxidative stress, and liver fibrosis compared to no supplementation. KBG was superior to VE and PG in the reduction of liver total cholesterol (P < 0.01) and lipid peroxidase levels (P < 0.05), urinary 8-hydroxy-2′-deoxyguanosine (P < 0.05), hepatic α-smooth muscle actin positive areas (P < 0.01) and activated stellate cells (P < 0.01). In conclusion, there was a statistically significant benefit of Kampo formulas (KBG in particular) on a dietary model of NAFLD/NASH. Future studies need to be directed at the mechanisms in the treatment of NASH.

Effect of Uncariae ramulus et Uncus on endothelium in spontaneously hypertensive rats.

The protective effect of the extract of Uncariae ramulus et Uncus (URE) against endothelium disorder due to hypertension was investigated. We administered low (150 mg/kg/day) and high (450 mg/kg/day) doses of URE orally to spontaneously hypertensive rats for 8 weeks. Endothelium dependent vasodilatation by acetylcholine increased significantly in the high URE group compared with the control group. Endothelium dependent vasocontraction by xanthine oxidase decreased significantly in the high URE group compared with the control group. Serum NO2-/NO3- were tended to increase in the high URE group. It is suggested that URE may have a protective effect for the endothelium against the influence of hypertension.

Clinical Evaluation of the Effect of Daio (Rhei Rhizoma) on the Progression of Diabetic Nephropathy with Overt Proteinuria

We studied the effect of traditional herbal medicines containing daio (Rhei Rhizoma) on the long-term progression of diabetic nephropathy with overt proteinuria in eight patients [mean age 60 (45-73) years; duration of diabetes 18 (7-36) years]. At the beginning of the study, mean HbA1c was 8.2% and mean serum creatinine was 1.0 +/- 0.3 mg/dl. Everypatient had diabetic neuropathy and retinopathy. Three of the patients had hypertension and four had ischemic heart disease. After 107 +/- 25 months, the mean serum creatinine level had significantly increased to 4.8 +/- 2.6 mg/dl. The mean serum creatinine levels of five patients not advancing to dialysis treatment increased from 1.2 +/- 0.3 to 3.2 +/- 1.0 mg/dl, and the three patients requiring dialysis increased from 0.8 +/- 0.1 to 7.5 +/- 2.1 mg/dl. In the control group, treated without traditional herbal medicines, the mean serum creatinine level had significantly increased from 1.0 +/- 0.3 to 9.5 +/- 1.9 mg/dl after 71 +/- 12 months. All of the control group required dialysis treatment. Diabetic nephropathy with overt proteinuria is reported to develop into renal failure after 6-7 years. In this retrospective study, traditional herbal medicines with Daio were considered to be effective in prolonging the pre-dialysis period of diabetic nephropathy.

Measurement of plasma procyanidin B-2 and procyanidin B-3 levels after oral administration in rat.

Using a high-performance liquid chromatographic method and mass spectrometry analysis, we successfully measured the absorption of orally administered procyanidin B-2 and procyanidin B-3 isolated from Cinnamonomi cortex (the bark of Cinnamomum cassia Blume) in the rat plasma. This method used a TSK- GEL ODS-80TS column, two solvents (A: 0.01% acetic acid; B: methanol with 0.01% acetic acid) in a linear gradient at a flow-rate of 1.0 ml/min, and fluorescence detection at excitation and emission wavelengths of 220 and 327 nm.

Effect of Seihai-to, a Kampo medicine, in relapsing aspiration pneumonia an open-label pilot study

Two published case reports described palliation of disease after Seihai-to therapy for refractory aspiration pneumonia caused by recurrent laryngeal nerve paralysis and cerebrovascular disease. We undertook an open-label trial in patients with relapsing aspiration pneumonia. Fifteen patients with relapsing aspiration pneumonia were randomly divided into conventional therapy group (n = 8) or Seihai-to group (n = 7). In Seihai-to group, patients were treated with Seihai-to in addition to conventional therapy (Western medicines). Frequency of feverish days and antibiotics-use, CRP value and chest CT or X-ray findings were compared between the two groups during the study period of 16 weeks. In the Seihai-to group, the latency of swallowing reflex was measured in 6 patients before and after administration of Seihai-to. The mean values of fever index, CRP value and antibiotics-use in the Seihai-to group were decreased significantly, compared with those of the conventional therapy group. However, the latency of the swallowing reflex after 4 weeks of treatment was not significantly changed (p = 0.249), compared with the latency before administration of Seihai-to. No adverse reaction was observed in either group. Seihai-to was effective in reducing relapse of aspiration pneumonia in this small group. Seihai-to might not improve the swallowing reflex, but might instead improve a defense mechanism or excessive inflammation caused by pneumonia in the lower airway. Further evaluation of Seihai-to therapy for patients with aspiration pneumonia in a larger population is warranted.

Inhibitory effects of Choto-san (Diao-teng-san), and hooks and stems of Uncaria sinensis on free radical-induced lysis of rat red blood cells.

The present study is designed to test our hypothesis that the ingestion of Uncaria sinensis (US), the main medicinal plant of Choto-san (Diao-teng-san, CS), would protect red blood cell (RBC) membrane from free radical-induced oxidation if polyphenolics in US could be absorbed and circulated in blood. When incubated with RBC suspension, Choto-san extract (CSE) and Uncaria sinensis extract (USE) exhibited strong protection for RBC membrane against hemolysis induced by 2,2-azo-bis (2-amidinopropane) dihydrochloride (AAPH), an azo free-radical initiator. The inhibitory effect was dose-dependent at concentrations of 50 to 1000 microg/mL. Ingestion of 200 mg of USE was associated with a significant decrease in susceptibility of RBC to hemolysis in rats. Furthermore, caffeic acid, an antioxidative hydroxycinnamic acid, was identified in rat plasma after administration of URE.