Comparative Study on the Pharmacokinetics of Paeoniflorin, White Peony Root Water Extract, and Taohong Siwu Decoction After Oral Administration in Rats.

BACKGROUND AND OBJECTIVE: Taohong Siwu Decoction (TSD) is a classic traditional Chinese medicine (TCM) compound with pharmacological effects such as vasodilation and hypolipidemia. Paeoniflorin (PF) is one of the active ingredients of TSD. The aim of this study was to evaluate the pharmacokinetics of PF in herbal extracts and their purified forms in rats. METHOD: A sensitive and rapid high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS-MS) method for the determination of PF in rat plasma was developed. Rats were divided into three groups, and given PF solution, water extract of white peony root (WPR), or TSD by gavage. At different predetermined timepoints after gavage, blood was collected from the orbital vein. The pharmacokinetic parameters of PF in the plasma of rats in the three groups was determined. RESULTS: The pharmacokinetic studies showed that the time to reach maximum concentration (Tmax) of PF in the purified forms group was relatively high, while the half-lives (T½) of PF in the TSD and WPR groups were longer. Among the three groups, PF in the purified forms group had the maximum area under the concentration-time curve (AUC0-t = 732.997 µg/L·h) and the largest maximum concentration (Cmax = 313.460 µg/L), which showed a significant difference compared with the TSD group (P < 0.05). Compared with the purified group, the clearance (CLz/F = 86.004 L/h/kg) and the apparent volume of distribution (Vz/F = 254.787 L/kg) of PF in the TSD group increased significantly (P < 0.05). CONCLUSIONS: A highly specific, sensitive, and rapid HPLC-MS-MS method was developed and applied for the determination of PF in rat plasma. It was found that TSD and WPR can prolong the action time of paeoniflorin in the body.

Extract of Danggui-Shaoyao-San ameliorates cognition deficits by regulating DHA metabolism in APP/PS1 mice.

ETHNOPHARMACOLOGICAL RELEVANCE: The traditional Chinese medicine formula Danggui-Shaoyao-San (DSS) has been reported to show therapeutic effect on alleviating the symptoms of Alzheimer's disease (AD). AIM OF THE STUDY: The present study aims to investigate the relation between DSS treatment of AD and DHA metabolism and evaluates its neuroprotective effect on cognitive in APP/PS1 mice. MATERIAL AND METHODS: DSS (1.6, 3.2, 6.4 g/kg/day) or Aricept (3 mg/kg/day) was orally administered (i.g.) to APP/PS1 mice, and saline was orally administered to Wild-type (WT) male mice as control group. Then, the Morris water maze (MWM) test, Y-maze spontaneous alternation test, open filed test and fear conditioning test were conducted for evaluation of learning and memory abilities. The DHA content was assessed by HPLC-MS/MS. Physiological indices were determined, including triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), ROS level, activity of superoxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA), PEG2, TXB2 and LTB4. The expressions of COX-1, COX-2, cPLA2, iPLA2, 15-LOX, and were assessed by Western blot. RESULTS: APP/PS1 mice showed serious cognitive impairment in behavioral tests. However, treatment of DSS extract significantly ameliorated the cognitive deficits of APP/PS1 mice. Biochemical measurements showed the increases in TG, TC, LDL-c and the decrease in HDL-c in APP/PS1 mice compared with WT mice, and DSS extract significantly retarded these changes. Low content of DHA, low expression of iPLA2 and 15-LOX were observed both in hippocampus and cortex of APP/PS1 mice, while DSS extract significantly restored these changes. Additionally, the abnormal activity of SOD and ROS level, the decreased levels of MDA and GSH were observed in APP/PS1 mice, while DSS extract prominently lessened these changes. Moreover, DSS extract decreased the level of PEG2, TXB2 and LTB4 and also attenuated the expression of cPLA2, COX-1 and COX-2 in hippocampus as well as cortex of APP/PS1 mice. CONCLUSIONS: Based on these results, we suggest that DSS play a positive effective role in increasing DHA content by up-regulating iPLA2 and 15-LOX, resulting in ameliorating oxidative stress and inflammation and finally ameliorating cognition deficits in APP/PS1 mice.

Effects of Xingnaojing Injection on Adenosinergic Transmission and Orexin Signaling in Lateral Hypothalamus of Ethanol-Induced Coma Rats.

Acute alcohol exposure induces unconscious condition such as coma whose main physical manifestation is the loss of righting reflex (LORR). Xingnaojing Injection (XNJI), which came from Chinese classic formula An Gong Niu Huang Pill, is widely used for consciousness disorders in China, such as coma. Although XNJI efficiently shortened the duration of LORR induced by acute ethanol, it remains unknown how XNJI acts on ethanol-induced coma (EIC). We performed experiments to examine the effects of XNJI on orexin and adenosine (AD) signaling in the lateral hypothalamic area (LHA) in EIC rats. Results showed that XNJI reduced the duration of LORR, which implied that XNJI promotes recovery form coma. Microdialysis data indicated that acute ethanol significantly increased AD release in the LHA but had no effect on orexin A levels. The qPCR results displayed a significant reduction in the Orexin-1 receptors (OX1R) expression with a concomitant increase in the A1 receptor (A1R) and equilibrative nucleoside transporter type 1 (ENT1) expression in EIC rats. In contrast, XNJI reduced the extracellular AD levels but orexin A levels remained unaffected. XNJI also counteracted the downregulation of the OX1R expression and upregulation of A1R and ENT1 expression caused by EIC. As for ADK expression, XNJI but not ethanol, displayed an upregulation in the LHA in EIC rats. Based on these results, we suggest that XNJI promotes arousal by inhibiting adenosine neurotransmission via reducing AD level and the expression of A1R and ENT1.

Modified lingguizhugan decoction incorporated with dietary restriction and exercise ameliorates hyperglycemia, hyperlipidemia and hypertension in a rat model of the metabolic syndrome.

BACKGROUND: Modified Lingguizhugan Decoction (MLD) came from famous Chinese medicine Linggui Zhugan Decoction. The MLD is used for the treatment of metabolic syndrome in the clinical setting. Our study focuses on the comprehensive treatment of MLD incorporated with dietary restriction and exercise in a rat model of the metabolic syndrome (MS). METHODS: Rats were divided into five groups: control group (Cont), high-fat diet group (HFD), high-fat diet incorporated with dietary restriction group (HFD-DR), exercise incorporated with dietary restriction group (HFD-DR-Ex) and MLD incorporated with dietary restriction and exercise group (HFD-DR-Ex-MLD). Treatments were conducted for 1 week after feeding high-fat diet for 12 weeks. The effects of treatments on high fat diet-induced obesity, hyperglycemia, hyperlipidemia, hypertension, hepatic injury and insulin resistance in rats of MS were examined. In addition, the tumor necrosis factor-α (TNF-α), leptin and protein kinase B (PKB) in rats serum and liver were also examined by enzyme-linked immunosorbent assay (ELISA). RESULTS: After a week's intervention by dietary restriction, dietary restriction incorporated with exercise or MLD, compared with HFD rats, the relative weight of liver and fat, levels of triglyceride, total cholesterol, low-density lipoprotein, free fatty acid, aspartate aminotransferase, glutamic-pyruvic transaminase and alkaline phosphatase, insulin, were significantly decreased (p < 0.05 or 0.01). This treatment also inhibited abnormal increases of TNF-α, leptin and PKB in serum and liver. CONCLUSION: MLD incorporated with dietary restriction and exercise treatment exhibit effects in alleviating high-fat diet-induced obesity, hyperglycemia, hyperlipidemia, hypertension, hepatic injury and insulin resistance, which are possibly due to the down-regulation of TNF-α, leptin and PKB.

Alteration of glutamate/GABA balance during acute alcohol intoxication in rats: effect of Xingnaojing injection.

ETHNOPHARMACOLOGICAL RELEVANCE: Xingnaojing Injection (XNJI) is a modern Chinese formula came from famous Chinese medicine An Gong Niu Huang Pill. XNJI has been used for treatment of cerebral diseases and stroke in China, and is approved by the State Food and Drug Administration of China for the treatment of acute alcohol intoxication (AAI). XNJI belongs to the ethnopharmacological family of medicines. In this study, we investigated the mechanisms of the XNJI effect on AAI. AIM OF THE STUDY: To investigate the effects of XNJI on glutamate, gamma-aminobutyric acid (GABA) and related receptor in lateral hypothalamic area (LHA) of AAI rat. MATERIAL AND METHODS: Adult male Sprague-Dawley rats were implanted with microdialysis probes in LHA. Rats were randomly divided into control, model, 1.36mg/kg XNJI, 0.68mg/kg XNJI and 0.34mg/kg XNJI groups. During microdialysis, baseline samples were collected from 1h to 2.5h; thereafter, the rats were given an intraperitoneal injection of 52% ethanol, 5.2g/kg, or saline for control group. Twenty minutes later, three doses of XNJI was given by unilateral injection respectively, while saline for control and model groups, and samples were collected for the next 4h. The extracellular glutamate and GABA levels were measured in the LHA by a high performance liquid chromatography coupled with fluorescence detector (HPLC-FLU). The expression levels of related receptors N-methyl-d-aspartate receptor (NR) subunit NR2A, NR2B and GABAA were analyzed by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Ethanol (5.2g/kg) significantly decreased the extracellular levels of glutamate and increased extracellular GABA in LHA. On the other hand ethanol significantly decreased NR2A and NR2B mRNAs expression, and increase GABAA mRNA expression. XNJI could increase the extracellular level of glutamate and decrease that of GABA; moreover, induced an increase in NR2A and NR2B mRNA expression, and a decrease in GABAA mRNA expression in LHA. CONCLUSIONS: The current changes in glutamate, GABA and mRNA expressions of related receptors in LHA after injection of XNJI suggest that changes in these neurotransmitters and receptors as a potential mechanism of action for AAI.

[Effects of Astragalus injection in reversing left ventricular hypertrophy induced by renal hypertension in rats].

OBJECTIVE: To explore whether the Astragalus injection (AI) has effect for reversing left ventricular hypertrophy and myocardial fibrosis induced by renal vascular hypertension in rats. METHODS: Thirty male SD rats were randomized equally into three groups: the AI group, the control group and the sham-operated group. All rats, except those in the sham-operated group, were established into the hypertension models by two kidney one clip (2K1C) operation. Blood pressure was measured before operation and every 4 weeks after operation. AI intervention was given to rats in the AI group starting from the 4th week of experiment at dose of 8 g/kg by peritoneal injecting once a day for 8 weeks, while for rats in the other 2 groups, equal volume of normal saline was given instead. All rats were sacrificed 12 weeks after operation by cervical breaking. And indexes including left ventricular mass index (LVMI), left ventricular wall thickness (LVWT), inter-ventricular septal thickness (IVST), left ventricular diameter (LVD), cardiomyocytes diameter (CCD), collagen volume fraction (CVF), and peri-vascular volume collagen area (PVCA) in rats were measured. RESULTS: Blood pressure was not different in the three groups before operation (P>0.05), whereas it rose in the control group and the AI group 4, 8 and 12 weeks after operation correspondingly, showing no difference between the two groups, but significantly higher than that in the sham-operated group (P<0.05). The related indexes in the sham-operated group, control group and AI group were: LVMI, 2.71 +/- 0.24, 3.42 +/- 0.26, 3.13 +/- 0.23, respectively; LVWT (mm), 2.25 +/- 0.42, 4.26 +/- 0.48, 3.28 +/- 0.36; IVST (mm), 2.13 +/- 0.38, 3.98 +/- 0.32, 3.02 +/- 0.28; and LVD (mm), 3.76 +/- 0.29, 2.18 +/- 0.27, 2.82 +/- 0.20 respectively. Comparisons showed that LVMI, LVWT and IVST were significantly higher, but LVD was significantly lower in the control group than those in the sham-operated group (P<0.05); LVMI, LVWT and IVST were significantly lower but LVD was significantly higher in the AI group than those in the control group (P<0.05). CCD, CVF and PVCA in the three groups (in the fore-mentioned order) were: CCD (microm), 14.54 +/- 2.25, 19.56 +/- 2.53, 16.58 +/- 2.46; CVF(%), 3.83 +/- 1.40, 11.21 +/- 2.96, 7.83 +/- 1.67; PVCA (%), 15.71 +/- 3.85, 30.58 +/- 6.25, 21.76 +/- 4.36, respectively. These indexes showed that CCD, CVF, PVCA in the control group were significantly higher than those in the sham-operated group (P<0.05); and those were significantly lower in the AI group than in the control group (P<0.05). CONCLUSION: AI intervention can reverse the left ventricular hypertrophy and myocardial fibrosis induced by renal vascular hypertension in rats.

[The effect of borneol on level of HA and 5-HT in rat's hypothalamus].

OBJECTIVE: To observe the effect of borneol on level of HA and 5-HT in rat's hypothalamus. METHODS: The levels of HA and 5-HT in rat's hypothalamus were determined before and after p.o. with different amount of borneol by high performance liquid chromatography with electrochemical detection (HPLC-ECD), and the borneol's effect on the levels of HA and 5-HT was also studied. RESULTS: The level of HA in rat's hypothalamus after different dose of borneol were higher than before administration. The level of HA in 20 min group after middle dose increased significantly comparing with before administration (P < 0.01), the others of group after middle dose, the 45 min group after high dose, the 20 and 45 min after low dose were also increased significantly than before administration (P < 0.05). After different dose of borneol, the level of 5-HT in rat's hypothalamus changed as follows: the level of 5-HT after high dose were higher than before administraton (P <0.05 or P < 0.01 ); the level of 5-HT after 5, 20 and 45 min of middle and low dose incrased significantly (P <0.05 or P < 0.01). CONCLUSION: Borneol could increase the levels of HA and 5-HT in rat's hypothalamus.