Uncaria rhynchophylla ameliorates unpredictable chronic mild stress-induced depression in mice via activating 5-HT1A receptor: Insights from transcriptomics.

BACKGROUND: Depression is a pervasive or persistent mental disorder that causes mood, cognitive and memory deficits. Uncaria rhynchophylla has been widely used to treat central nervous system diseases for a long history, although its efficacy and potential mechanism are still uncertain. PURPOSE: The present study aimed to investigate anti-depression effect and potential mechanism of U. rhynchophylla extract (URE). STUDY DESIGN AND METHODS: A mouse depression model was established using unpredictable chronic mild stress (UCMS). Effects of URE on depression-like behaviours, neurotransmitters, and neuroendocrine hormones were investigated in UCMS-induced mice. The potential target of URE was analyzed by transcriptomics and bioinformatics methods and validated by RT-PCR and Western blot. The agonistic effect on 5-HT1A receptor was assayed by dual-luciferase reporter system. RESULTS: URE ameliorated depression-like behaviours, and modulated levels of neurotransmitters and neuroendocrine hormones, including 5-hydroxytryptamine (5-HT), 5-hydroxyindole acetic acid (5-HIAA), dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), corticosterone (CORT), corticotropin-releasing hormone (CRH), and adrenocorticotropic hormone (ACTH), in UCMS-induced mice. Transcriptomics and bioinformatics results indicated that URE could regulate glutamatergic, cholinergic, serotonergic, and GABAergic systems, especially neuroactive ligand-receptor and cAMP signaling pathways, revealing that Htr1a encoding 5-HT1A receptor was a potential target of URE. The expression levels of downstream proteins of 5-HT1A signaling pathway 5-HT1A, CREB, BDNF, and PKA were increased in UCMS-induced mice after URE administration, and URE also displayed an agonistic effect against 5-HT1A receptor with an EC50 value of 17.42 µg/ml. CONCLUSION: U. rhynchophylla ameliorated depression-like behaviours in UCMS-induced mice through activating 5-HT1A receptor.

Shenqi Xingnao Granules ameliorates cognitive impairments and Alzheimer's disease-like pathologies in APP/PS1 mouse model.

Objective: Alzheimer's disease (AD) is along with cognitive decline due to amyloid-ß (Aß) plaques, tau hyperphosphorylation, and neuron loss. Shenqi Xingnao Granules (SQXN), a traditional Chinese medicine, significantly ameliorated the cognitive function and daily living abilities of patients with AD. However, till date, no study has investigated the mechanism of action of SQXN on AD. The present study aimed to verify the effects of SQXN treatment on cognitive impairments and AD-like pathologies in APP/PS1 mice. Methods: Four-month-old APP/PS1 transgenic (Tg) mice were randomly divided into a model group and SQXN-treated (3.5, 7, 14 g/kg per day) groups. Learning-memory abilities were determined by Morris water maze and object recognition test. All mice were sacrificed and the brain samples were collected after 75 d. The soluble Aß contents were detected by Elisa kit; The levels of expression of NeuN, APP, phosphorylated tau and related protein were measured by Western blotting; The inflammation factors were detected by the proinflammatory panel kit. Results: Four-month-old APP/PS1 mice were administered SQXN by oral gavage for 2.5 months. Using the Morris water maze tests and Novel object recognition, we found that SQXN restored behavioral deficits in the experimental group of Tg mice when compared with the controls. SQXN also inhibited neuronal loss (NeuN marker). SQXN treatment decreased soluble Aß42 through inhibiting the expression of sAPPß and BACE-1 without regulating full-length amyloid precursor protein (FL APP). Insulin degrading enzyme (IDE), the Aß degrading enzyme, were increased by SQXN. In addition, SQXN reduced hyperphosphorylated tau protein levels and prevented excessive activation of p-GSK-3ß in the brain of APP/PS1 mice. Compared with APP/PS1 transgenic negative mice, IFN-γ, IL-1ß, IL-2, IL-4, IL-5, IL-6, IL-12p70, KC/GRO and TNF-α were not obviously changed in the brain of 6.5-month-old APP/PS1 transgenic (Tg) mice. However, SQXN could inhibited the expression of IL-2. Conclusion: These results demonstrate that SQXN ameliorates the cognitive impairments in APP/PS1 mice. The possible mechanisms involve its inhibition of neuronal loss, soluble Aß deposition, tau hyperphosphorylation and inflammation.

Synergism and rules from combination of Baicalin, Jasminoidin and Desoxycholic acid in refined Qing Kai Ling for treat ischemic stroke mice model.

Refined Qing-Kai-Ling (QKL), a modified Chinese medicine, consists of three main ingredients (Baicalin, Jasminoidin and Desoxycholic acid), plays a synergistic effect on the treatment of the acute stage of ischemic stroke. However, the rules of the combination and synergism are still unknown. Based on the ischemic stroke mice model, all different kinds of combination of Baicalin, Jasminoidin, and Desoxycholic acid were investigated by the methods of neurological examination, microarray, and genomics analysis. As a result, it confirmed that the combination of three drugs offered a better therapeutical effect on ischemic stroke than monotherapy of each drug. Additionally, we used Ingenuity pathway Analysis (IPA) and principal component analysis (PCA) to extract the dominant information of expression changes in 373 ischemia-related genes. The results suggested that 5 principal components (PC1-5) could account for more than 95% energy in the gene data. Moreover, 3 clusters (PC1, PC2+PC5, and PC3+PC4) were addressed with cluster analysis. Furthermore, we matched PCs on the drug-target networks, the findings demonstrated that Baicalin related with PC1 that played the leading role in the combination; Jasminoidin related with PC2+PC5 that played a compensatory role; while Desoxycholic acid had the least performance alone which could relate with PC3+PC4 that played a compatible role. These manifestations were accorded with the principle of herbal formulae of Traditional Chinese Medicine (TCM), emperor-minister-adjuvant-courier. In conclusion, we firstly provided scientific evidence to the classic theory of TCM formulae, an initiating holistic viewpoint of combination therapy of TCM. This study also illustrated that PCA might be an applicable method to analyze the complicated data of drug combination.

[Using markov model to cost-effectiveness analysis of traditional Chinese medicine interference].

Mild cognitive impairment (MCI) is the transition state between normal aging and AD. There are better detection and treatment methods to screen a population of patients with MCI. Through intervention, the probability of MCI conversion to AD can be significantly reduced. This paper first introduces the concept of cost-effectiveness analysis, reconsiders the concept of cost for the particularity of MCI, and uses QALY to evaluate the health effects of the quality of life. Then measure the health quality of life of elderly MCI population, and use Markov model to study the cost of intervention with traditional Chinese medicine-effectiveness analysis. Finally, according to the QALY measure and CEA results, we draw the conclusion that it's helpful to get early intervention in MCI.

Comparison of the network structural characteristics of calcium signaling pathway in cerebral ischemia after intervention by different components of Chinese medicine.

OBJECTIVE: To explore the network control mechanism of the calcium signaling pathway in cerebral ischemic injury after intervention by the main components of Qingkailing (see text), i.e. Baicalin, Jasminoidin and their combination. METHODS: Thirty mice were randomly divided into 5 groups, a baicalin group, a Jasminoidin group, a baicalin plus Jasminoidin group, a nimodipine group, and a model group (n = 6). The global cerebral ischemia-reperfusion mouse model was established. The mice were administrated respectively by injection of baicalin, Jasminoidin, mixture of baicalin and Jasminoidin, and nimodipine into the caudal vein, with the model group given no any drug. Three hours after operation, the brain was removed and sectioned. After calculation of cerebral ischemic area by 2,3,5-triphenyltetrazolium staining, the percentage of infarct volume was calculated. The total RNA of the mouse brain tissue was extracted to obtain the whole genome expression profile, and the differentially expressed genes related to the calcium signaling pathway was analyzed with Bayesian network structures. RESULTS: Compared with the model group, the ischemic area was significantly reduced in the baicalin group, the Jasminoidin group, the Baicalin plus Jasminoidin group (all P < 0.05). The ischemic area in the baicalin plus Jasminoidin group was smaller than the other three groups (all P < 0.01). In the gene regulatory network structures of calcium signaling pathway, the average length and equitability were the highest in the baicalin plus Jasminoidin group, followed by the nimodipine group. CONCLUSION: Compared with a single component, combination of Baicalin and Jasminoidin can more obviously intervene in the overall expression of calcium signaling pathway, and the mechanism is related with the aggregation characteristic of the gene expression network.

A comparative study on the individual and combined effects of baicalin and jasminoidin on focal cerebral ischemia-reperfusion injury.

To compare the individual effects of baicalin and jasminoidin with the combined effect of them on cerebral ischemia-reperfusion injury, and test whether the combined administration of baicalin and jasminoidin can improve the therapeutic effect. Male Sprague-Dawley rats underwent focal cerebral ischemia for 1.5 h and reperfusion for 24 h. Just before reperfusion, tested drugs (baicalin, jasminoidin, a drug combination consisting of baicalin and jasminoidin, or nimodipine) were intravenously treated. Diffusion weighted imaging (DWI) of magnetic resonance imaging (MRI), behavior examination, 2,3,5-triphenyltetrazolium chloride (TTC) staining, histological examination, and real-time PCR for BDNF and caspase-3 were performed. All of the drug treatments could significantly ameliorate the results of TTC and histological examination, and the baicalin/jasminoidin combination did so most prominently. This combination could also significantly ameliorate DWI of MRI and behavior examination results, and promote the expression of BDNF and inhibit the expression of caspase-3. On the whole, both baicalin and jasminoidin have a preventive effect against ischemic stroke, although their effects are not very strong. However, the combination of baicalin and jasminoidin can significantly improve their effectiveness. This may be related to its better regulation on the BDNF and caspase-3.

[Pharmacological evaluation of baicalin and jasminoidin and their combination on focal cerebral ischemia-reperfusion injury].

OBJECTIVE: To elucidate the therapeutic effect and the influence on PI3K-Akt-PKB-BAD-CREB-PCREB pathway in focal cerebral ischemia rat responses before and after treatment with baicalin and jasminoidin given alone or in combination. METHOD: Rat model of ischemia reperfusion was established with thread. Generally accepted methods were used, including TTC staining, behavior test, as well as micro and ultra microscopy which can dynamically and accurately monitor pathological and physiological changes after cerebral ischemia on earlier period, to evaluate the brain injury induced by ischemia and the attenuations by the drugs. The difference of PI3K-Akt-PKB-BAD-CREB-PCREB expression was detected by western-blot technology. RESULT AND CONCLUSION: The combination of baicalin and jasminoidin composition can be potential neuroprotective agent. TTC staining technology combined with behavior grade and ultrmicro-structure observation on brain tissue is effective method to evaluate protective agent, which is related to signal transduction PI3K-Akt-PKB-BAD-CREB-PCREB pathway. The results provide benofical basis for revealing the complex of therapeutic mechanism of traditional Chinese medicine Qingkai Ling (QKL).

[Application of magnetic resonance DW imaging technique in studying treatment of cerebral ischemia in rats by single or combined use of jasminoidin and cholalic acid].

OBJECTIVE: To estimate the therapeutic effect of single or combined use of jasminoidin and cholalic acid on focal cerebral ischemia rat with magnetic resonance-diffusion-weighted imaging (MR-DWI) technique, ultra-microscopy, and neuro-behavior scoring. METHODS: The model of cerebral ischemia-reperfusion injury was induced by string method. Three hours after reperfusion, MR-DWI was applied with ultra-microscopy and neuro-behavior test to give evaluation on cerebral ischemic rats, and pathologic, ultramicroscopic observation of tissue were taken as adjuvant measures to comprehensively evaluate the pharmacological effect on ischemia-reperfusion rats and delimit the efficacy of the two different components and their combination. RESULTS: Compared with the model group, ADC and DCavg values of the foci in all the treated groups had the incrensing trend. There was significant difference arund the foci in the group of combined use of jasminoidin and cholalic acid (P < 0.05). CONCLUSION: Combined use of jasminoidin and cholalic acid had protective effects on nerve and brain. MR-DWI technique accompanied with ultramicroscopic observation of tissues and neuro-behavior test is an effective method for evaluating the effect of neuro-protective agent.

Gene expression profile induced by oral administration of baicalin and gardenin after focal brain ischemia in rats.

AIM: To investigate differential gene expression and the pharmacological mechanism of baicalin and gardenin in focal cerebral ischemia in rats with high-density cDNA microarray. METHODS: Rat left middle cerebral arteries were occluded and treated with either baicalin or gardenin. The pharmacological effects were investigated using the difference in infarction areas before and after treatment, which were determined by 2,3,5-triphenyltetrazolium chloride (TTC) staining. Gene expression was demonstrated using a "Biostar40S" gene microarray. Semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to verify the result of the selected genes. RESULTS: Both baicalin and gardenin reduced the infarction areas in focal cerebral ischemia rats (P<0.05). The differential genes were 211, 177, and 70 (upregulated or downregulated) in the model group, baicalin, and gardenin treatment groups compared with the sham-operated group, respectively. Gene expression of RpL19 and Csnk2 underwent an approximately 1.9 and 2.1-fold increase, respectively, verified by semiquantitative RT-PCR, which was the same trend as the cDNA microarray. CONCLUSION: Differential gene expression with respect to the pharmacological effects of baicalin and gardenin on focal cerebral ischemia by cDNA microarray revealed a number of clues with respect to the therapeutic mechanisms of Chinese traditional medicine. In addition, the present study provided theoretical and experimental evidence that will aid future studies examining cerebral ischemia.

[Effect of gardenin on gene expression profile in brain of rats with focal cerebral ischemia].

OBJECTIVE: To explore the pharmacologic mechanism of gardenin in treating cerebral ischemia, by studying its effect on gene expression profile in brain of rats with focal cerebral ischemia (FCI). METHODS: Total RNAs were isolated from rats with FCI and those treated with gardenin. The mRNAs were reversely transcribed to cDNA with incorporation of fluorescent Cy5- or Cy3-dUTP to prepare hybridization probes. The PCR products of 4096 genes were spotted on the chip after a serial treatment. The mixed probes were hybridized to the cDNA microarray. Axon Genepix 4000B and GenePixPro 3.0 software were used to scan and analyze the fluorescent signals. RESULTS: In the group treated with gardenin, there were 70 genes had expression profiles different to that in the model group in the focal cerebral ischemic brain tissue, in which 68 were up-regulated and 2 down-regulated. CONCLUSION: Gardenin has regulatory effect on the gene expression in rats with focal cerebral ischemia, which elucidates part of the pharmacologic mechanism of Qingkailing in molecular level.

[The effect of Baicalin on gene expression profile in rat brain of focal cerebral ischemia].

OBJECTIVE: To explore the difference of genes expression profiles between focal cerebral ischemia tissue and that treated with Baicalin using cDNA microarray. METHOD: The total RNAs were isolated from rat brains of sham-operation, vehicle (focal cerebral ischemia of rat brain) and baicalin-treated groups. mRNAs were reversely transcribed to cDNA with incorporation of fluorescent dUTP (Cy5 or Cy3 dUTP) to prepare hybridization probes. The PCR products of 4096 genes were spotted on the chip after a serial of treatment. The mixed probes were hybridized to the cDNA microarray. Axon Genepix 4000B and GenePixPro 3.0 software were used to scan and analyze the fluorescent signals. RESULT: The expressions of 199 and 12 genes were found up-regulated and down-regulated, respectively, in the vehicle group compared with the sham-operation one. But the numbers of genes whose expressions were up-regulated and down-regulated were 89 and 88, respectively, when comparing the gene expression in the Baicalin-treated rat brain with that in the vehicle group. Moreover, one down-regulated and three up-regulated genes in the vehicle group were up-regulated and down-regulated in the Baicalin-treated group, respectively. Expressions of three up-regulated genes in the vehicle group were further reinforced in the Baicalin-treatment group. CONCLUSION: Multiple pathways and nodes may be involved in the pharmacological effect of Baicalin on brain ischemia.