Intestinal Absorption of Berberine and 8-Hydroxy Dihydroberberine and Their Effects on Sugar Absorption in Rat Small Intestine

The intestinal absorption ofberberine （Ber） and its structural modified compound 8-hydroxy dihydroberberine （Hdber） was compared, and their effects on the intestinal absorption of sugar by per- fusion experiment were investigated in order to reveal the mechanism of low dose and high activity of Hdber in the treatment of hyperglycemia. The absorption of Hdber and Ber in rat small intestine was measured by in situ perfusion. High performance liquid chromatography （HPLC） was used to determine the concentrations of Hdber and Ber. In situ perfusion method was also used to study the effects of Hdber and Ber on sugar intestinal absorption. Glucose oxidase method and UV spectrophotometry were applied to examine the concentrations of glucose and sucrose in the perfusion fluid. The results showed that the absorption rate of Ber in the small intestine was lower than I0%, but that of Hdber was larger than 70%. Both Hdber and Ber inhibited the absorption of glucose and sucrose at the doses of 10 and 20 ~tg/mL. However, Hdber presented stronger activity than Bet （P〈0.01）. It is suggested that Hdber is ab- sorbed easily in rat small intestine and that its inhibitory effect on the absorption of sugar is better than Ber.

Induction of pancreatic duct cells of neonatal rats into insulin-producing cells with fetal bovine serum: A natural protocol and its use for patch clamp experiments

AIM: To induce the pancreatic duct cells into endocrine cells with a new natural protocol for electrophysiological study.METHODS: The pancreatic duct cells of neonatal rats were isolated, cultured and induced into endocrine oells with 15% fetal bovine serum for a period of 20 d. During this period, insulin secretion, MTT value, and morphological change of neonatal and adult pancreatic islet cells were comparatively investigated. Pancreatic β-cells were identified by morphological and electrophysiological characteristics, while ATP sensitive potassium channels(KATP), voltage-dependent potassium channels (KV), and voltage-dependent calcium channels (KCA) in β-cells were identified by patch clamp technique.RESULTS: After incubation with fetal bovine serum, the neonatal duct cells budded out, changed from duct-like cells into islet clusters. In the first 4 d, MTT value and insulin secretion increased slowly (MTT value from 0.024±0.003 to0.028±0.003, insulin secretion from 2.6±0.6to 3.1±0.8 mIU/L). Then MTT value and insulin secretion increased quickly from d 5 to d 10 (MTT value from 0.028±0.003 to 0.052±0.008, insulin secretion from 3.1±0.8to 18.3±2.6 mIU/L), then reached high plateau (MTT value ＞0.052±0.008, insulin secretion ＞18.3±2.6 mIU/L).In contrast, for the isolated adult pancreatic islet cells,both insulin release and MTT value were stable in the first 4 d (MTT value from 0.029±0.01 to 0.031±0.011,insulin secretion from 13.9±3.1 to 14.3±3.3 mIU/L), but afterwards they reduced gradually (MTT value ＜0.031±0.011, insulin secretion ＜8.2±1.5 mIU/L), and the pancrearic islet cells became dispersed, broken or atrophied correspondingly. The differentiated neonatal cells were identified as pancreatic islet cells by dithizone staining method, and pancreatic β-cells were further identified by both morphological features and electrophysiological characteristics, i.e. the existence of recording currents from KATP KV, and KCA.CONCLUSION: Islet cells differentiated from neonatal pancreatic duct cells with the new natural protocol are more advantageous in performing patch clamp study over the isolated adult pancreatic islet cells.

Clinical Study of Electro-acupuncture Treatment with Different Intensities for Functional Constipation Patients

Functional constipation （FC） is a common functional bowel disorder disease that affects life quality of a large number of people. This study aimed to explore the impact of different intensities of electro-acupuncture （EA） treatment for FC patients. Totally, 111 patients with FC meeting the Rome III criteria were randomly assigned to different intensities of EA groups （low and high intensity of EA groups） and medicine-controlled （MC） group. In EA groups, patients were treated with EA at quchi （Llll） and shangjuxu （ST37） bilaterally for 4 weeks, 5 times/week in the first 2 weeks, and 3 times/week in the last 2 weeks. In MC group, 5 mg mosapride citrate was administered orally 3 times/day for 4 weeks. Spontaneous bowel movement frequency each day was recorded using a consti- pation diary. Self-rating anxiety scale （SAS） and self-rating depression scale （SDS） were used to assess the patients＇ psychological state. Cortisol （CORT）, substance P （SP）, and vasoactive intestinal polypep- tide （VIP） were evaluated at baseline and at the end of 4 weeks after treatment. As compared with the baseline, there was statistically significant increase in stool frequency every week （P〈0.01）, but there was no statistically significant difference among the three groups. As compared with the baseline, after 4 weeks of EA therapy, the scores of SDS and serum levels of CORT were decreased significantly in low intensity of EA group （P〈0.01）, and the serum levels of SP and VIP were increased significantly （P〈0.05）; the scores of SAS and SDS and serum levels of CORT were decreased significantly in high intensity of EA group （P〈0.05）, and the serum levels of SP and VIP were increased significantly （P〈0.05）; the serum levels of CORT and VIP were increased significantly in MC group （P〈0.05）. As compared with MC group, after 4 weeks of treatment, the serum levels of SP were signifcicantly in- creased in low intensity of EA group （P〈0.01）. Low and high intensities of EA could increase the stool frequency, improve the FC patient＇s anxiety and depression, reduce the serum levels of CORT, and in- crease the serum levels of SP and VIP effectively. It is concluded that both low and high intensities of EA are effective for FC patients, but there is no significant difference between the low and high intensi- ties of EA.

Effect of Ethanol Extract of Rhodiola Rosea on the Early Nephropathy in Type 2 Diabetic Rats

Summary： This study aimed to investigate the therapeutical effects of Rhodiola rosea extract on rats with type 2 diabetic nephropathy （DN）. The rat type 2 DN model was established by high fat and high calorie feeding and intravenous injection of streptozocin （STZ）. Wistar rats were randomly divided into normal group, control group, low dose Rhodiola rosea group, high dose Rhodiola rosea group and Cap- topril group. Oral glucose tolerance test （OGTT） was performed to determine the impairment of glucose tolerance in the established animal model. A series of parameters including fasting blood glucose （FBG）, total cholesterol （TC）, triglyceride （TG）, creatinine clearance rate （Ccr）, 24-h urinary albumin （UA）, the ratio of kidney mass/body weight （renal index） and glomerular area were examined after 8 weeks. Moreover, the expression of transforming growth factor （TGF）-β1 in renal tissues was detected by using immunohistochemisty. At the end of the eighth week, FBG, TC, TG, Ccr, 24-h urinary albumin, the ratio of kidney mass/body weight and glomerular area were significantly reduced in Rhodiola rosea extract treatment groups as compared with those in control group. TGF-β1 expression in renal tissues of Rhodiola rosea extract treatment groups was also significantly decreased as compared with that of con- trol group. These results indicate that Rhodiola rosea extract may have a protective effect on early nephropathy in diabetic rats, which might be related to the decrease of the renal expression of TGF-β1.

Chinese Herbal Medicine in the Treatment of Nonalcoholic Fatty Liver Disease

Chinese herbal medicine has developed new therapies for nonalcoholic fatty liver disease （NAFLD） based on its unique theory system and substantial herb remedies. In this review, 21 traditional Chinese herbs were introduced for their potential benefit in the treatment of NAFLD. Majority of them are evaluated by experimental studies and few by multicenter clinical trials. Herbal monomers as berberine and resveratrol, extracts from Polygonum hypoleucum Ohwi, and Artemisia sacrorum Ledeb., and formulae including Yinchenhao Decoction （~.~.~, YCHD）, Qushi Huayu Decoction （~＇~. ~.~, QSHYD）, and Danning Tablet （）~？）~） were discussed in detail on their therapeutic potentials. Most of these herbal medicines were proved to improve biochemical and histological changes of NAFLD both in vitro and in vivo. Also, their therapeutic activities were associated with inhibiting lipid accumulation through adenosine monophosphate-activated protein kinase activation or upregulating low-density lipoprotein receptor （LDLR） expression, alleviating lipid peroxidation, and reducing the production of inflammatory cytokines. Although the efficacy and safety of these herbal medicines needed to be evaluated in multicenter large-scale clinical trials, Chinese medicine is promising and effective for preventing and treating NAFLD disease.

Effect of Huanglian Jiedu Decoction (黄连解毒汤) on Glucose Transporter 4 Expression in Adipose and Skeletal Muscle Tissues of Insulin Resistant Rats

Objective： To investigate the effects of Huanglian Jiedu Decoction （黄连解毒汤, HLJDD） on glucose transporter 4 （GLUT4） protein expressions in insulin-resistant murine target tissues. Methods： The experimental male Wistar rats were established into insulin resistant models by injecting streptozotocin （STZ 30 mg/kg） via caudal vein and feeding them with high fat high caloric diet, and randomly divided into the model group, the aspirin group and the HLJDD group. Besides, a normal group was set up for control. Changes of body weight （BW）, levels of serum fasting blood glucose （FBG）, serum fasting insulin （FINS） and oral glucose tolerance test （OGTT） were routinely determined. The expression of GLUT4 protein in adipose and skeletal muscle tissues before and after insulin stimulation was determined with Western blot. Results： In the HLJDD group after treatment, BW and FBG got decreased, OGTT improved, and the expression and translocation of GLUT4 protein elevated obviously, either before or after insulin stimulation, as compared with those in the model group, showing significant differences respectively. Conclusion： The mechanism of improving insulin resistance by HLJDD is probably associated with its effect in elevating GLUT4 protein expression and translocation in adipose and skeletal muscle tissues of insulin resistant rats.

Effects of Acupuncture on Progesterone and Prolactin in Rats of Embryo Implantation Dysfunction

Objective： To investigate the effect of acupuncture on progesterone （P4） and prolactin （PRL） in rats of embryo implantation dysfunction （EID）. Methods： On the first day of pregnancy, 72 female Wistar rats were randomly allocated into the normal group, the EID model group, the acupuncture group and the P4 group （18 in each group）. The normal group was injected sesame oil, while the other three groups were given mifepristone to establish the EID model. The acupuncture group and the P4 group were given treatment of acupuncture and P4 injection, respectively. The serum of P4 and PRL were detected by radioimmunoassay, and the mRNA and protein expressions of P4 receptor （PR） and PRL receptor （PRLR） were detected using real-time polymerase chain reaction and immunohistochemical method, respectively. Results： Compared with the normal group, the serum levels of P4 and PRL as well as the mRNA and protein expression levels of PR and PRLR in the EID model group were significantly lowered （P〈0.01 or P〈0.05）. The above indices in the acupuncture group and the P4 group were significantly elevated compared with the EID model group （P〈0.01 or P〈0.05）. Conclusion： Acupuncture can promote embryo implantation effectively, which might be related to the effects of acupuncture on upregulating the P4 and PRL levels in serum and the PR and PRLR expression levels in rats.

Synergistic Cytotoxicity of Ampelopsin Sodium and Carboplatin in Human Non-Small Cell Lung Cancer Cell Line SPC-A1 by G1 Cell Cycle Arrested

Objective： To evaluate the cytotoxic effects of ampelopsin sodium（Amp-Na） and carboplatin（CBP） used alone or in combination on human non-small cell lung cancer（NSCLC） cells SPC-A1 in vitro and its related mechanism. Methods： Cytotoxic effects were assessed by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） assays. The synergistic effects of the drugs were calculated with coefficient of drug interaction（CDI）. Cell cycle was determined by flow cytometry（FCM）. The levels of p53, p21, cyclin E, cyclin D1, and phosphorylated cyclin-dependent kinase-2（p-CDK2） were evaluated by Western blot. Results： Amp-Na（6.25–200 μg/m L） and CBP（3.13–100 μg/m L） alone exhibited prominent cytotoxic activity in a concentration-dependent manner on SPC-A1 cells with 50% inhibitive concentration values of 57.07±14.46 and 34.97±6.30 μg/m L, respectively. Drug combinations were associated with significantly higher cytotoxic effects than each drug alone（P〈0.05 or 0.01）. The CDI analysis confirmed the synergy of Amp-Na and CBP on inhibiting cancer cell viability across a wide concentration range（CDI 〈1）. FCM and Western blot showed that synergistic cytotoxic effects of Amp-Na and CBP were related to G1 arrested which mainly mediated by p21 through the inhibition of CDK2 activity independent of the p53 tumor suppressor pathway. Conclusions： AmpNa exhibits anticancer activities and enhances the antitumor activities of CBP through up-regulation of p21 and inhibition of CDK2 activity in human NSCLC cells SPC-A1. These results suggest that Amp-Na may be applied to enhance the anticancer action of CBP.

Jiaotai Pill(交泰丸) Enhances Insulin Signaling through Phosphatidylinositol 3-Kinase Pathway in Skeletal Muscle of Diabetic Rats

objective： To investigate the effect of Jiaotai Pill （交泰丸, JTP） at different constitutional proportions on insulin signaling through phosphatidylinositol 3-kinase （PI3K） pathway in the skeletal muscle of diabetic rats. Methods： The rat model of type 2 diabetes mellitus （T2DM） was established by intravenous injection of a small dose of streptozotoein plus high fat diet feeding. JTP at the same dosage of cinnamon and the increasing dosage of Coptis chinensis was administered to diabetic rats for nine weeks respectively. Plasma glucose and insulin levels were assayed. The expressions of proteins were determined by Western blot method. Results： All the three formulations of JTP decreased plasma glucose and fasting insulin levels as well as increased the protein expressions of insulin receptor β （InsR β） subunit, insulin receptor substrate-1 （IRS-1）, PI3K p85 subunit and glucose transporter 4 （GLUT4） in skeletal muscle. Meanwhile, JTP increased the tyrosine phosphorylation of InsRβ subunit and IRS-1, and reduced the serine phosphorylation of IRS-1 in skeletal muscle. Interestingly, the effect of JTP on improving insulin sensitivity was not dose-dependent. In contrast, JTP containing the least amount of Coptis chinensis exhibited the best effect. Conclusion： JTP at different constitutional proportions attenuates the development of diabetes in a rat model of T2DM. The mechanism might be associated with enhancing insulin signaling through PI3K pathway in the skeletal muscle.

Coptidis Rhizoma-Contained Traditional Formulae for Insomnia:A Potential to Prevent Diabetes?

It has been well known that insomnia and sleep deprivation can lead to many metabolic diseases such as type 2 diabete mellitus （T2DM）o Chinese medicine （CM） has been used to improve insomnia and showed many good therapeutic effects. Some Coptidis Rhizoma-contained traditional formulae have been found to exert obvious effects on insomnia, such as Jiaotai Pill （交泰丸）, Huanglian Ejiao Decoction （黄连阿胶汤）, Huanglian Wendan Decoction （黄连温胆汤）, Niuhuang Qingxin Pill （牛黄清心丸）, and Zhusha Anshen Pill （朱砂安神丸）. Coptidis Rhizoma, a traditional herbal medicine which can purge Xin （Heart） fire, is the most important component of these formulas. Studies have found that these formulae could improve T2DM symptoms. We hypothesized that treating insomnia theoretically may help to prevent diabetes and organized many experimental studies in this review and concluded that Coptidis Rhizoma-contained traditional formulae for insomnia might be a potential to prevent diabetes.

Type 2 diabetic patients with non-alcoholic fatty liver disease exhibit significant haemorheological abnormalities

Haemorheological abnormalities have been described in diabetes mellitus,as well as in non-alcoholic fatty liver disease(NAFLD).However,the relationship between the changes in liver fat content and haemorheology is unknown.The current study aims to show the correlation between haemorheological parameters and intrahepatic lipid content(IHLC)in patients with type 2 diabetes.The serum biochemical markers,such as fasting plasma glucose(FPG),haemoglobin A1c(HbA1c),liver enzymes,lipid profiles,and haemorheological properties,were examined.IHLC was quantified using proton magnetic resonance spectroscopy(1 H-MRS)scanning of the liver.A significant correlation was observed between IHLC and whole blood viscosity at high,middle,and low shear rates.IHLC also positively correlated with haematocrit,the reduced whole blood viscosity at low and middle shear rates,and the erythrocyte aggregation index.Diabetic patients with NAFLD exhibited significant haemorheological abnormalities compared with patients without NAFLD.In summary,haemorheological disorders are linked to non-alcoholic fatty liver in type 2 diabetes.