Zibu Piyin Recipe Research Articles

ZiBu PiYin recipe regulates central and peripheral Aβ metabolism and improves diabetes-associated cognitive decline in ZDF rats

Ethnopharmacological relevanceCognitive impairment caused by central neuropathy in type 2 diabetes mellitus (T2DM), namely diabetes-associated cognitive decline (DACD), is one of the common complications in patients with T2DM. Studies have shown that brain β-amyloid (Aβ) deposition is a typical pathological change in patients with DACD, and that there is a close relationship between intestinal microorganisms and cognitive impairment. However, the specific mechanism(s) of alteration in Aβ metabolism in DACD, and of the correlation between Aβ metabolism and intestinal microorganisms remain unknown. Aim of the studyRevealing the mechanism of ZBPYR regulating Aβ metabolism and providing theoretical basis for clinical evaluation and diagnosis of DACD. Materials and methodsWe characterized Aβ metabolism in the central and peripheral tissues of Zucker diabetic fatty (ZDF) rats with DACD, and then explored the preventive and therapeutic effects of ZiBu PiYin Recipe (ZBPYR). Specifically, we assessed these animals for the formation, transport, and clearance of Aβ; the morphological structure of the blood-brain barrier (BBB); and the potential correlation between Aβ metabolism and intestinal microorganisms. ResultsZBPYR provided improvements in the structure of the BBB, attenuation of Aβ deposition in the central and peripheral tissues, and a delay in the development of DACD by improving the expression of Aβ production, transport, and clearance related protein in ZDF rats. In addition, ZBPYR improved the diversity and composition of intestinal microorganisms, decreased the abundance of Coprococcus, a bacterium closely related to Aβ production, and up regulate the abundance of Streptococcus, a bacterium closely related to Aβ clearance. ConclusionThe mechanism of ZBPYR ability to ameliorate DACD may be closely related to changes in the intestinal microbiome.

Structural Alteration of Gut Microbiota During the Amelioration of Chronic Psychological Stress-Aggravated Diabetes-Associated Cognitive Decline by a Traditional Chinese Herbal Formula, ZiBu PiYin Recipe.

Chronic psychological stress (PS) hinders the treatment of diabetes-associated cognitive decline (DACD). However, the impact of chronic PS on the risk of developing DACD remains unclear. There is growing evidence that gut flora interventions are promising targets for treating stress-related diseases. We examined whether chronic PS triggers or exacerbates the onset of DACD in rats and aimed to elucidate whether ZiBuPiYin recipe (ZBPYR) prevents and treats chronic PS-aggravated DACD by dynamically maintaining the components of the gut microbiota. We performed chronic PS (restraint, rotation, and congestion) on ZDF rats to establish a model. Cognitive function was evaluated by behavioral experiments, and activation of the hypothalamic-pituitary-adrenal axis was detected by ELISA. Weekly feces from rats were collected for 16 S RNA sequencing. We found that chronic PS promoted cognitive abnormalities and exacerbated DACD phenotypes. Additionally, chronic PS altered intestinal flora diversity, dynamically elevating the abundance of Alistipes and Coprococcus; enriching Module 1 (Dorea, Blautia, Ruminococcus) and Module 48 (Blautia); and inhibiting Module 20 (Lactobacillus, SMB53), and Module 42 (Akkermansia). ZBPYR significantly alleviated hyperglycemia and cognitive impairment in chronic PS-aggravated DACD rats and dynamically reduced the abundance of Alistipes and Coprococcus; significantly enriched Module 3 (Ruminococcus) and Module 45 (Lactobacillus, Coprococcus, SMB53); and suppressed Module 2 (Lactobacillus), Module 16 (Turicibacter, Trichococcus, Lactobacillus, 02d06, Clostridium), Module 23 (Bifidobacterium), and Module 43 (Clostridium). ZBPYR might prevent and treat chronic PS-aggravated DACD by dynamically regulating Lactobacillus, Alistipes, and Coprococcus.

ZiBuPiYin recipe ameliorates diabetes-associated cognitive decline by improving neuronal mitochondrial function in chronic psychologically stressed zucker diabetic fatty rats

Ethnopharmacological relevanceZibu Piyin Recipe (ZBPYR) is a traditional Chinese medicine compound composed of 12 kinds of Chinese herbal medicines including red ginseng and yam. Long-term basic and clinical applications have proved that ZBPYR can prevent and treat cognitive dysfunction. Previous studies showed that chronic psychological stress can increase the risk of type 2 diabetes mellitus (T2DM), and lead to cognitive decline. Mitochondrial dysfunction plays a key role in chronic psychological stress-induced diabetes mellitus. While the mechanism of mitochondrial dysfunction and insulin resistance in diabetes-associated cognitive decline (DACD) is unclear. Aim of the studyOur previous research found that a ZiBuPiYin recipe (ZBPYR) has significant pharmacological effects against DACD. The present study investigated changes in mitochondrial dysfunction in the brain and the mechanism of insulin resistance and mitochondrial damage to explore the relationship between neuronal mitochondrial dysfunction and insulin resistance in chronic psychologically stressed DACD rats. Materials and methodsZucker diabetic fatty (ZDF) rats with spontaneous T2DM and rats with diabetic cognitive impairment that was induced by chronic psychological stress were used in in vivo experiments. PC12 cells that were damaged by rotenone were used for the in vitro experiment. ResultsThe findings indicated that the number of mitochondria decreased, morphology and membrane potential were damaged, and reactive oxygen species increased in the cortex and hippocampus in psychologically stressed DACD rats. Protein kinase Cβ2 (PKCβ2) activation and insulin resistance were markedly induced by chronic psychological stress, together with decreases in peroxisome proliferator-activated receptor-γ coactivator 1α (PGC-1α) and mitochondrial fusion protein 2 (Mfn2). Furthermore, ZBPYR exerted protective effects both in in vivo and in vitro. ConclusionMitochondrial damage and insulin resistance were observed in the brain in chronic psychologically stressed DACD rats. The ZBPYR significantly improved brain mitochondrial damage and insulin resistance in chronic psychologically stressed DACD rats. These results provide novel insights for the development of ZBPYR as a traditional Chinese medicine for the treatment of chronic psychological stress and DACD.

Liver mitochondria-associated endoplasmic reticulum membrane proteomics for studying the effects of ZiBuPiYin recipe on Zucker diabetic fatty rats after chronic psychological stress.

Type 2 diabetes mellitus (T2DM) is a complex metabolic disease with multiple etiologies, involving both genetic and environmental factors. With changes associated with modern life, increasing attention has been paid to chronic psychological stressors such as work stress. Chronic psychological stress can induce or aggravate diabetes mellitus, and conversely, with the deterioration of T2DM, patients often experience different degrees of depression, anxiety, and other negative emotions. In order to clarify the role of ZiBuPiYin recipe (ZBPYR) in regulating the liver mitochondria-associated endoplasmic reticulum membrane proteome to improve T2DM with chronic psychological stress, differentially expressed proteins (DEPs) were identified among Zucker lean littermates (control group), chronic psychological stress T2DM rats (model group), and ZBPYR administration rats (ZBPYR group) through iTRAQ with LC-MS/MS. Using Mfuzz soft clustering analysis, DEPs were divided into six different clusters. Clusters 1-6 contained 5, 68, 44, 57, 28, and 32 DEPs, respectively. Given that ZBPYR can alleviate T2DM symptoms and affect exploratory behavior during T2DM with chronic psychological stress, we focused on the clusters with opposite expression trends between model:control and ZBPYR:model groups. We screened out the DEPs in clusters 1, 3, and 4, which may be good candidates for the prevention and treatment of T2DM with chronic psychological stress, and further conducted bioinformatics analyses. DEPs were mainly involved in the insulin signaling pathway, oxidative phosphorylation, tricarboxylic acid cycle, amino acid metabolism, lysosome-related processes, and lipid metabolism. This may indicate the pathogenic basis of T2DM with chronic psychological stress and the potential therapeutic mechanism of ZBPYR. In addition, two key proteins, lysosome-associated protein (Lamp2) and tricarboxylic acid cycle-related protein (Suclg1), may represent novel biomarkers for T2DM with chronic psychological stress and drug targets of ZBPYR. Western blot analyses also showed similar expression patterns of these two proteins in liver MAMs of the model and ZBPYR groups.

ZiBuPiYin Recipe Prevented and Treated Cognitive Decline in ZDF Rats With Diabetes-Associated Cognitive Decline via Microbiota-Gut-Brain Axis Dialogue.

Gut microbiota is becoming one of the key determinants in human health and disease. Shifts in gut microbiota composition affect cognitive function and provide new insights for the prevention and treatment of neurological diseases. Diabetes-associated cognitive decline (DACD) is one of the central nervous system complications of type 2 diabetes mellitus (T2DM). ZiBuPiYin recipe (ZBPYR), a traditional Chinese medicine (TCM) formula, has long been used for the treatment of T2DM and prevention of DACD. However, the contribution of ZBPYR treatment to the interaction between the gut microbiota and metabolism for preventing and treating DACD remains to be clarified. Here, we investigate whether the gut microbiota plays a key role in ZBPYR-mediated prevention of DACD and treatment of T2DM via incorporating microbiomics and metabolomics, and investigate the links between the microbiota–gut–brain axis interaction and the efficacy of ZBPYR in ZDF rats. In the current study, we found that ZBPYR treatment produced lasting changes in gut microbiota community and metabolites and remotely affected hippocampus metabolic changes, thereby improving memory deficits and reversing β-amyloid deposition and insulin resistance in the brain of ZDF rats from T2DM to DACD. This may be related to a series of metabolic changes affected by gut microbiota, including alanine, aspartic acid, and glutamic acid metabolism; branched-chain amino acid metabolism; short-chain fatty acid metabolism; and linoleic acid/unsaturated fatty acid metabolism. In summary, this study demonstrates that prevention and treatment of DACD by ZBPYR partly depends on the gut microbiota, and the regulatory effects of bacteria-derived metabolites and microbiota–gut–brain axis are important protective mechanisms of ZBPYR.

Neuroprotective Effects of ZiBuPiYin Recipe on db/db Mice via PI3K-Akt Signaling Pathway by Activating Grb2.

Background Diabetes-associated cognitive decline (DACD) is one of the nervous system dysfunctions induced by diabetes mellitus with cognitive impairment as the major symptom. In a previous preliminary proteomic study, we found that endoplasmic reticulum processing and PI3K-Akt signaling pathway might be impaired in DACD pathogenesis. In addition, growth factor receptor-bound protein 2 might be a crucial protein as a molecular target of the neuroprotective effects of ZiBuPiYin recipe (ZBPYR). Methods In this study, 6-8 weeks aged db/db mice were treated with excipients or ZBPYR for 6 weeks. Body weight and RBG were recorded weekly. Oral glucose tolerance and insulin tolerance tests were used to assess insulin sensitivity. Morris water maze (MWM) tests were used to assess memory function. The expression of Grb2, Gab2, Akt, and GSK3β in mouse hippocampus and cerebral cortex were analyzed by Western blotting. Results ZBPYR not only significantly reduced RGB and improved glucose tolerance and insulin resistance, but also improved spatial cognition in DACD mice. The expression of Grb2 and Gab2 in hippocampus and cerebral cortex of db/db mice was upregulated after treated with ZBPYR, and then affected the PI3K/Akt signaling pathway, and inhibited GSK3β overactivity. Conclusions This study showed that ZBPYR could enhance the memory and learning ability of db/db mice. Such neuroprotective effect might be related to the activation of Grb2-PI3K/Akt signaling which might provide a novel therapeutic target for the clinical treatment of DACD.

The ZiBuPiYin recipe regulates proteomic alterations in brain mitochondria-associated ER membranes caused by chronic psychological stress exposure: Implications for cognitive decline in Zucker diabetic fatty rats.

Chronic psychological stress (PS) cumulatively affects memory performance through the deleterious effects on hypothalamic-pituitary-adrenal axis regulation. Several functions damaged in cognitive impairment-related diseases are regulated by mitochondria-associated ER membranes (MAMs). To elucidate the role of ZiBuPiYin recipe (ZBPYR) in regulating the MAM proteome to improve PS-induced diabetes-associated cognitive decline (PSD), differentially expressed MAM proteins were identified among Zucker diabetic fatty rats, PSD rats, and PS combined with ZBPYR administration rats via iTRAQ with LC-MS/MS. Proteomic analysis revealed that the expressions of 85 and 33 proteins were altered by PS and ZBPYR treatment, respectively. Among these, 21 proteins were differentially expressed under both PS and ZBPYR treatments, whose functional categories included energy metabolism, lipid and protein metabolism, and synaptic dysfunction. Furthermore, calcium signaling and autophagy-related proteins may play roles in the pathogenesis of PSD and the mechanism of ZBPYR, respectively. Notably, KEGG pathway analysis suggested that ‘Alzheimer's disease’ and ‘oxidative phosphorylation’ pathways may be impaired in PSD pathogenesis, while ZBPYR could play a neuroprotective role through regulating the above pathways. Overall, exposure to chronic PS contributes to the evolution of diabetes-associated cognitive decline and ZBPYR might prevent and treat PSD by regulating the MAM proteome.

Effect of the ZiBuPiYin Recipe on Diabetes-Associated Cognitive Decline in Zucker Diabetic Fatty Rats After Chronic Psychological Stress.

BackgroundCognitive impairment is a complication of type 2 diabetes mellitus (T2DM) that affects the central nervous system (CNS). Studies have shown that chronic psychological stress may promote the development of T2DM into diabetes-associated cognitive decline (DACD). Previously, cognitive impairment in T2DM was correlated predominantly with insulin resistance in the medial prefrontal cortex (mPFC).AimsWe examined the effect of the ZiBuPiYin recipe (ZBPYR) on Zucker diabetic fatty (ZDF) rats and explored the impact of chronic stress on altered β-amyloid (Aβ) metabolism through insulin receptor substrate (IRS) 1/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway after the induction of chronic psychological stress.Main MethodsAfter chronic psychological stress and drug treatment, cognitive function was observed via behavioral experiments. The activation of the hypothalamus-pituitary-adrenal (HPA) axis and levels of Aβ were detected by enzyme-linked immunosorbent assay, and the expression of related proteins was evaluated by Western blotting.Key FindingsZBPYR treatment significantly decreased anxious-like behaviors and plasma corticosterone (CORT) levels, and ameliorated learning and memory impairments of ZDF rats after chronic psychological stress. ZBPYR also reduced the deposition of Aβ in the mPFC, improved brain insulin resistance, and modulated the mTOR-autophagy pathway.SignificanceZBPYR may be a potential therapeutic application for the treatment of DACD induced by chronic psychological stress.

Pharmacokinetic Assessments of Liquiritin, Protocatechuic Aldehyde and Rosmarinic Acid in Rat Plasma by UPLC-MS-MS After Administration of ZibuPiyin Recipe.

The three analytes of the Traditional Chinese Medicine ZibuPiyin Recipe (ZBPYR), namely, liquiritin, protocatechuic aldehyde and rosmarinic acid, may synergistically play an important role in regulating memory and learning. However, the pharmacokinetic behaviors of these compounds after their co-administration remain unclear. To this end, a selective and sensitive ultra-performance liquid chromatography-tandem mass spectrometry method was developed and validated in rat plasma for the study of these three major bioactive ingredients in ZBPYR. The analytes in the plasma samples were separated on a Shiseido Capcell core C18 column using bendrofluazide as an internal standard, with a gradient mobile phase system of acetonitrile-water containing 0.1% formic acid. Electrospray ionization in the negative-ion mode and multiple reaction monitoring were used to identify and quantify the three analytes. All of the calibration curves showed good linearity (r > 0.992) over the concentration range, with a lower limit of quantification of 5 ng/mL. The precision of the analytical method was evaluated by intra- and inter-day assays, and the percentage of relative standard deviation (SD) was within 15%. Satisfactory extraction efficiency (between 83.4 and 99.4%) and matrix effects (76.4-107.4) were obtained by liquid-liquid extraction. The pharmacokinetic results showed that the three bioactive ingredients were rapidly absorbed and had a short terminal half-life in rats after oral administration of ZibuPiyin recipe. This UPLC-MS-MS study method used in this study may be useful for assessing the pharmacokinetic characteristics of various compounds, which would be helpful in determining their clinical potential.

The Alteration of ZiBuPiYin Recipe on Proteomic Profiling of Forebrain Postsynaptic Density of db/db Mice with Diabetes-Associated Cognitive Decline.

Diabetes-associated cognitive decline (DACD) is a brain injury induced by diabetes mellitus, with cognitive impairment as the major symptom. Growing evidence has revealed that DACD is correlated with disruptions in synapses involved in cognition. Within synapses, more specifically in areas of postsynaptic density (PSD), there is a high concentration of proteins that receive and transduce synaptic information. In the present study, to identify the differentially expressed PSD proteins among DACD mice, ZiBuPiYin recipe (ZBPYR)-treated DACD mice and control mice, we applied isobaric tags for relative and absolute quantitation (iTRAQ) with LC-MS/MS technology, by which three biological replicates and three technical replicates were examined. A total of 24 and 23 differentially expressed proteins were observed in control versus DACD mice and in DACD versus ZBPYR-treated DACD mice, respectively. Notably, we found 'Protein processing in endoplasmic reticulum' and 'PI3K-Akt signaling pathway' might be impaired in DACD pathogenesis, while Growth factor receptor-bound protein 2 might be a crucial protein as a molecular target of the neuroprotective effects of ZBPYR. To our knowledge, this is the first study to provide a reference proteome map for DACD and ZBPYR-treated DACD mouse forebrain PSD to aid understanding the underlying mechanisms of DACD and ZBPYR.

ZiBu PiYin recipe prevents diabetes-associated cognitive decline in rats: possible involvement of ameliorating mitochondrial dysfunction, insulin resistance pathway and histopathological changes.

BackgroundDisturbance in energy metabolism, as a key factor in diabetes-associated cognitive decline (DACD), has become a promising therapeutic target of Chinese medicine ZiBu PiYin Recipe (ZBPYR). However, it is still not clear how ZBPYR affects the mitochondrial function in DACD rats’ brains, which is considered as the crucial cell organelle to supply energy for the brain.MethodsType 2 diabetes mellitus (T2DM) rat models were established by using high fat diet and streptozotocin (STZ) (30 mg/kg, ip). The evaluation of insulin sensitivity was performed by oral glucose tolerance and insulin tolerance test. After 7 weeks, the T2DM rats were treated with vehicle or ZBPYR for 11 weeks and morris water maze (MWM) test were used to evaluate memory function. The ultra structural changes of prefrontal cortex (PFC) and hippocampus were examined by transmission electron microscopy (TEM). The mitochondrial membrane potential (ΔΨm) and reactive oxygen species (ROS) were measured with JC-1 and DCFDA assay. The levels of insulin proteins were quantified by Western Blot analysis and the markers of histopathological changes were detected by immunohistochemistry.ResultsZBPYR could alleviate learning and memory impairment of DACD rats. TEM showed that ZBPYR prevented mitochondrial ultra-structural alterations and number changes in the PFC and hippocampus of the DACD rats. In addition, ZBPYR significantly increased ΔΨm and lowered the levels of ROS. Further investigation indicated that ZBPYR suppressed the release of cytochrome c from mitochondria, strengthened insulin signaling and inhibited GSK3β over-expression. These positive effects were associated with reduced Aβ1-42 deposition and restored expression levels of microtubule-associated protein MAP2.ConclusionZBPYR showed excellent protective effect against DACD via ameliorating mitochondrial dysfunction, insulin resistance and histopathological changes.

Comparison of microwave‐assisted and heat reflux extraction techniques for the extraction of ten major compounds from Zibu Piyin Recipe using ultra high performance liquid chromatography with tandem mass spectrometry

Microwave-assisted extraction and efficient ultra-high performance liquid chromatography with triple quadrupole mass spectrometry were previously used to quickly extract and simultaneously quantify ginsenoside Rf, Ro, and Rd, 20(S)-ginsenoside-Rg2 , 20(R)-ginsenoside-Rg2 , tanshinone IIA, cryptotanshinone, dihydrotanshinone I, lithospermic acid, and osthole from Zibu Piyin Recipe. We here showed that heat reflux extraction provides higher extraction efficiency of these target compounds but is more time consuming. Chromatographic separation was achieved on an Agilent ZORBAX RRHD Eclipse Plus C18 column with a gradient mobile phase consisting of water/0.5% formic acid and acetonitrile at a flow rate of 0.2 mL/min, and detection was performed by positive and negative ion multiple-reaction monitoring mode. All analytes showed good linearity (r, 0.9989-0.9999) within the test range, with a limit of detection of 0.002-0.180 μg/mL. The overall intra- and interday variations of the ten compounds were ≤2.9%, and the accuracy was evaluated using a recovery test at three concentrations and was in the range 97.61-103.18% (RSD ≤ 4.25%). The analytical results showed remarkable differences in the concentrations of the ten compounds extracted from Zibu Piyin Recipe by microwave-assisted extraction and heat reflux extraction. These findings provide important information for determining the quality of Zibu Piyin Recipe.

Correction: Simultaneous determination of eight ginsenosides in Zibu Piyin Recipe using ultra-performance liquid chromatography coupled with triple-quadrupole mass spectrometry (UPLC-MS/MS)

Correction for ‘Simultaneous determination of eight ginsenosides in Zibu Piyin Recipe using ultra-performance liquid chromatography coupled with triple-quadrupole mass spectrometry (UPLC-MS/MS)’ by Hui-Ying Xu et al., Anal. Methods, 2015, 7, 7527–7533.

Ultra high performance liquid chromatography with mass spectrometry for the rapid analysis and global characterization of multiple constituents from Zibu Piyin Recipe.

Zibu Piyin Recipe is a traditional Chinese medicine that has been used for the clinical treatment of memory loss in China. However, the chemical components have not been thoroughly studied so far. To quickly identify the chemical components and understand the chemical profiles related to cognition improvement activity, ultra high performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry has been applied. The analysis was performed on an Agilent 1290 ultra-high performance liquid chromatography system with a Poroshell 120 EC-C18 column (3.0 × 150 mm, 2.7 μm) using gradient elution. Using the optimized method, 155 chemical components in Zibu Piyin Recipe were tentatively identified. Among them, 112 components in negative ion mode, 73 constituents in positive ion mode, and 30 compounds in both modes could be detected. The major components were identified from Red Ginseng, Dan shen Root, White Paeony Root, liquorice root, Radix Polygalae, Tangerine Red Epicarp, Grassleaf Sweetflag Rhizome, Indian Buead, and lotus seed. These results suggest that the established rapid and robust method will be useful for identifying multiple constituents of traditional Chinese medicine prescriptions. This method could provide helpful chemical information for further in vivo pharmacological mechanism research of Zibu Piyin Recipe and new drug development for cognition improvement.

Fingerprint analysis and multi-component determination of Zibu Piyin recipe by HPLC with DAD and Q-TOF/MS method

Zibu Piyin recipe (ZBPYR), a traditional Chinese medicine formula, is used for curing dementia caused by diabetes. For quality control of ZBPYR, fingerprint analysis and qualitative analysis using high-performance liquid chromatography (HPLC) with a diode-array detector, and confirmation using HPLC coupled with electrospray ionisation quadrupole time-of-flight tandem mass spectrometry (HPLC-Q-TOF-MS) were undertaken. HPLC fingerprint consisting of 34 common peaks was developed among 10 batches of ZBPYR, in which 7 common peaks were identified in comparison with the authentic standards and detected simultaneously. Furthermore, these seven compounds were verified by HPLC-Q-TOF-MS methods. The method can be applied to the quality control of ZBPYR.

Simultaneous determination of eight ginsenosides in Zibu Piyin Recipe using ultra-performance liquid chromatography coupled with triple-quadrupole mass spectrometry (UPLC-MS/MS)

A simple, sensitive, selective, and reliable UPLC-MS/MS method was developed and validated for the simultaneous detection and quantification of eight compounds found in Zibu Piyin Recipe (ZBPYR).

ZiBuPiYin Recipe Protects db/db Mice from Diabetes-Associated Cognitive Decline through Improving Multiple Pathological Changes

Multiple organ systems, including the brain, which undergoes changes that may increase the risk of cognitive decline, are adversely affected by diabetes mellitus (DM). Here, we demonstrate that type 2 diabetes mellitus (T2DM) db/db mice exhibited hippocampus-dependent memory impairment, which might associate with a reduction in dendritic spine density in the pyramidal neurons of brain, Aβ1-42 deposition in the prefrontal cortex (PFC) and hippocampus, and a decreased expression of neurostructural proteins including microtubule-associated protein (MAP2), a marker of dendrites, and postsynaptic density 95 (PSD95), a marker of excitatory synapses. To investigate the effects of the ZiBuPiYin recipe (ZBPYR), a traditional Chinese medicine recipe, on diabetes-related cognitive decline (DACD), db/db mice received daily administration of ZBPYR over an experimental period of 6 weeks. We then confirmed that ZBPYR rescued learning and memory performance impairments, reversed dendritic spine loss, reduced Aβ1-42 deposition and restored the expression levels of MAP2 and PSD95. The present study also revealed that ZBPYR strengthened brain leptin and insulin signaling and inhibited GSK3β overactivity, which may be the potential mechanism or underlying targets of ZBPYR. These findings conclude that ZBPYR prevents DACD, most likely by improving dendritic spine density and attenuating brain leptin and insulin signaling pathway injury. Our findings provide further evidence for the effects of ZBPYR on DACD.

The effects of Zibu Piyin Recipe components on scopolamine-induced learning and memory impairment in the mouse

Ethnopharmacological relevanceThe Zibu Piyin Recipe (ZBPYR) is derived from Zicheng Decoction, a traditional Chinese medicine formula recorded in the book of Bujuji, written by Wu Cheng in the Qing dynasty and used for clinical treatment of amnesia. Our aim was to study the effects of Zibu Piyin Recipe (ZBPYR) fractions on scopolamine-induced learning and memory impairment in the mouse. Materials and methodsCrude extracts were prepared using various solvents, and individual fractions produced following D101 macroporous resin column chromatography. The passive avoidance task, step down test and Morris water maze test were then performed in mice for the evaluation of learning and memory alterations. The effective fractions were then analyzed using GC–MS and polysaccharide measurement methods, respectively. ResultsThe treatment group latency for the alcohol precipitation from water part (EP) and 95% ethanol part (95%E) following D101 macroporous resin column chromatography was significantly prolonged when compared to that of the scopolamine treated groups for both the passive avoidance task and step down test. In the Morris water maze tests, treatment with EP and 95%E resulted in a significantly shorter escape latency time (from the fourth day and the second day) and swimming distance (on the third day and from the third day) in scopolamine-induced mice. In the memory retention test, treatment with EP and 95%E dramatically shortened the latency to cross platform location and increased the numbers of platform location crosses in the scopolamine-induced mice. The polysaccharide content in EP was determined to be 69.79%. The 95%E was found to mainly contain asarone, α-cadinol, isocalamendiol, 2,4,7,14-tetramethyl-4-vinyl-tricyclo[5.4.3.0(1,8)]tetradecan-6-ol, 3-isopropyl-6,7-dimethyltricyclo[4.4.0.0(2,8)]decane-9,10-diol, 2-methyl-9-(prop-1- -en-3-ol-2-yl)-bicyclo[4.4.0]dec-2-ene-4-ol, diepicedrene-1-oxide, 7-methoxy-6-(3- -methyl-2-oxobutyl)-2H-1-benzopyran-2-one and diisooctyl phthalate when assessed using GC–MS analysis. ConclusionThese findings suggest that the polysaccharide and volatile oil present in ZBPYR exhibit ameliorating effects on scopolamine-induced memory dysfunction.

Effects of Zibu Piyin Recipe on Endoplasmic Reticulum Stress in the Brain Tissue of Spleen-Yin Deficiency Alzheimer's Disease Rats

The effects of Chinese herbs for Zibu Piyin Recipe (ZBPYR) on spleen-Yin deficiency Alzheimer's Disease (AD) rats were investigated. A new rat model was established based on both AD and the spleen-Yin deficiency syndrome. Then, expressions of glucose- regulated protein78 (GRP78), CCAAT/EBP homologous protein (CHOP) in the hippocampus (CA1, CA3, and DAG regions) and cerebral cortex that were correlated to endoplasmic reticulum stress were determined. The results showed that in contrast to the Spleen-Yin Deficiency AD groups, the expression of the GRP78 protein was obviously upregulated (P < 0.05) in the ZBPYR group; whereas the expression of the CHOP protein was obviously downregulated (P < 0.05). It is concluded that Chinese herbs for ZBPYR improve the ability of learning and memory by reinforcing the survival pathway of ERS, and inhibiting the apoptosis pathway of ERS.

Empirical Study of Proteome of Hippocampus from A Rat Model with Alzheimer's Disease with Spleen Deficiency Syndrome Effected by Zibu Piyin Recipe

Abstract The article is aimed at investigating Alzheimer's disease (AD) with spleen yin deficiency syndrome and identifying the effective therapeutic targets of Zibu Piyin Recipe (ZBPYR) at whole cell protein levels. The rat model of spleen yin deficiency syndrome was set up through combination with improper diet, overstrain, and consumption of body fluid. Then, the polymerized β-amyloid 1–40 was injected into both hippocampi of the rats. The treated group was administered with ZBPYR. Taking the control group as a standard, the changes of proteins in the hippocampus were compared between the model group of AD with spleen yin deficiency syndrome and ZBPYR-treated group by two-dimensional gel electrophoresis. The altered protein spots were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Compared with the control group, 9 protein spots of the hippocampus in the model group of AD with spleen yin deficiency syndrome and 2 spots in ZBPYR-treated group were found to be changed totally. The differential protein spots were identified by MALDI-TOF-MS. The results showed that in the model group of AD with spleen yin deficiency syndrome, the level of annexin III was higher, but the levels of tubulin beta chain 15, dihydropyrimidinase-like 2, and guanine nucleotide-binding protein beta-1 subunit were lower than that of the control group. These differential proteins did not show significant changes between the ZBPYR-treated group and the control group. It is concluded that changes of several proteins in the hippocampus may involve in the pathogenesis of AD with spleen yin deficiency syndrome together. Annexin III, tubulin beta chain 15, dihydropyrimidinase-like 2, and guanine nucleotide-binding protein beta-1 subunit are possibly related to pathological changes of the hippocampus in AD with spleen yin deficiency syndrome. ZBPYR may produce its effects by regulating these proteins in the hippocampus.