Electroacupuncture alleviates ventilator-induced lung injury in mice by inhibiting the TLR4/NF-κB signaling pathway.

OBJECTIVE: This study was aimed to explore the protective effect of electroacupuncture (EA) pretreatment at Zusanli point (ST36) on ventilation-induced lung injury (VILI) and its potential anti-inflammatory mechanism. METHODS: High tidal volume ventilation was used to induce the VILI in mice, and EA pretreatment at ST36 was given for 7 consecutive days. The wet/dry ratio and pathological injury score of lung tissue, and total protein content of pulmonary alveolar lavage fluid (BALF) were detected after 4 h of mechanical ventilation (MV). Meanwhile, the expressions of TLR4 and NF- κB in lung tissue were evaluated by Western Blot, and the inflammatory factors in lung tissue were detected by ELISA. RESULTS: After four hours of mechanical ventilation, mice with ventilator-induced lung injury showed significant increases in lung wet/dry ratio, tissue damage scores, and protein content in bronchoalveolar lavage fluid. Pro-inflammatory cytokines (IL-6, IL-1ß, TNF-α) and TLR4/NF-κB expression levels in the lung were also markedly elevated (P < 0.05). Conversely, ST36 acupuncture point pre-treatment significantly reduced these parameters (P < 0.05). CONCLUSION: EA pretreatment at ST36 could alleviate the inflammatory response for VILI via inhibiting TLR4/NF- κB pathway.

Efficient Approach for the Extraction and Identification of Red Pigment from Zanthoxylum bungeanum Maxim and Its Antioxidant Activity.

Red pigment (RP) was extracted from the peels of Zanthoxylum bungeanum Maxim (PZB) by ultrasonic-assisted extraction (UAE) in this work. Box⁻Behnken design⁻response surface methodology (BBD-RSM) was employed to research the efficiency of the RP extraction. Based on the optimization of RSM, results showed that the optimal extraction conditions were as follows: liquid⁻solid ratio of 31 mL/g, extraction time of 41 min, and extraction temperature of 27 °C, and under these conditions, the actual absorbance value was 0.615 ± 0.13%, highly agreeing with the predicted value by the model. Furthermore, ultra-performance liquid chromatography⁻mass spectrometry (UPLC-MS) was used to separate and analyze RP. The compounds of RP were mainly flavonoids, and there were five compounds detected for the first time in PZB. In addition, RP showed significant antioxidant activities in vitro, which could be developed for anti-aging candidate drugs and functional foods. In conclusion, ultrasound-assisted extraction with BBD-RSM and chromatographic separation technology with UPLC-MS are efficient strategies for the isolation and identification of RP from PZB.

[Effect of Acupuncture on Histology of Thymus and Expressions of ANP and NPR-A in Anxiety Rats].

OBJECTIVE: To explore possible mechanism of electroacupuncture (EA) for regulating immune function in anxiety disorder (AD) rats by observing the effect of acupuncture on the histology of thymus and expressions of atrial natriuretic peptide (ANP) and natriuretic peptide receptor type A (NPR- A) in thymus. METHODS: Totally 34 SD healthy rats were randomly divided into the blank control group (n = 10), the model group (n = 12), the EA group (n = 12). Anxiety model was established in rats of the model group and the EA group by using chronic unpredictable stress (CUS) stimulation. EA (15/25 Hz) at Neiguan (PC6) and Shenmen (HT7) was performed in the EA group, with 15-min needle retaining, once every other day, 15 days in total. Needle was fixed at same acupoints for 15 min without electric stimulus in the other two groups. Anxiety-like behavior was measured by elevated plus-maze (EPM) test. Pathological changes of thymus tissue were observed by optical microscope. Expressions of ANP and NPR-A in thymus were measured by immunohistochemical assay. RESULTS: The thymus tissue in the model group was severely atrophied, with unclear structure of thymic lobules, unclear margin of thymic medulla, loosely arranged lymphocytes ,and obviously enlarged volume of thymic corpuscle. The thymus tissue in the EA group was mildly atrophied, with existent structure of thymic lobules, clear margin of thymic medulla, densely arranged lymphocytes in cortical region, and widened medullary area. Com- pared with the blank control group, the percentage of open-arms entries (OE%) in the total QE times ob- viously decreased in the model group (P < 0.05), ANP expression obviously increased (P < 0.05), and NPR-A expression obviously decreased (P < 0.01). Compared with the model group, OE% was obviously elevated (P < 0.05), ANP expression obviously decreased (P < 0.05), and NPR-A expression obviously increased (P < 0.01) in the EA group. CONCLUSION: EA not only could reduce anxiety of rats, but also could improve chronic stress induced thymus injury through intervening synthesis and secretion of ANP, as well as the expression of NPR-A (a specific receptor of ANP).

Bear Bile Powder Improves Ulcerative Colitis by Protecting the Intestinal Mechanical Barrier and Regulating Intestinal Flora.

BACKGROUND: Bear Bile Powder (BBP) is a traditional Chinese medicine. It has been widely used in clinical practices and has shown a good anti-inflammatory effect. However, its effectiveness in treating Ulcerative Colitis (UC) has not yet been studied. OBJECTIVE: To explore the therapeutic effect of BBP on ulcerative colitis and its potential mechanism by combining acute ulcerative colitis mouse models and comprehensively observing various physiological and biochemical indexes of mice. METHODS: The acute ulcerative colitis model was induced by drinking water containing dextran sulfate sodium salt (DSS) for 7 days. Studies were divided into Control, DSS, DSS+ Sulfasalazine (SASP, 450 mg/kg), and DSS + bear bile powder group (BBP, 320 mg/kg). The Disease Activity Index (DAI) and colonic tissue damage of mice were evaluated. Tissue immunofluorescence and western blot were used to determine related tight Junction Proteins (TJs), and 16S V34 amplicon was used to analyze intestinal microorganisms. The therapeutic effect of BBP on ulcerative colitis model mice was studied comprehensively. RESULTS: After treatment, BBP can significantly improve the physiological condition of acute UC mice and reduce DAI fraction. Compared with the DSS group, the BBP group significantly increased the colon length and significantly decreased the injury fraction of acute UC mice. Regarding the intestinal mechanical barrier, BBP significantly increased the expression of ZO-1, Occludin, and Claudin 1 protein in colon tissue. In terms of microbial community, the intestinal microbial diversity of mice decreased after the administration of BBP, but there was no significant difference in structural composition between the BBP group and the Control group. By comparing the four groups of species with significant differences, it was found that the BBP group significantly reduced the abundance of specific harmful microorganisms at the order, family, genus, and species levels. CONCLUSION: Oral administration of a certain dose of BBP can significantly improve the symptoms of ulcerative colitis in mice. Part of the reason may be that it increases the expression of tight junction proteins, regulates specific flora in the intestine of mice, and maintains intestinal barrier homeostasis. In the future, the clinical application value of BBP will be explored, and BBP will be developed as a drug with the potential to treat UC and alleviate the pain of UC patients.

Peptide mediated therapy in fibrosis: Mechanisms, advances and prospects.

Fibrosis, a disease characterized by an excess accumulation of extracellular matrix components, could lead to organ failure and death, and is to blame for up to 45 % of all fatalities in developed nations. These disorders all share the common trait of an unchecked and increasing accumulation of fibrotic tissue in the affected organs, which leads to their malfunction and eventual failure, even if their underlying causes are highly diverse and, in some cases, remain unclear. Numerous studies have identified activated myofibroblasts as the common cellular elements ultimately responsible for the replacement of normal tissues with nonfunctional fibrotic tissue. The transforming growth factor-ß pathway, for instance, plays a significant role in practically all kinds of fibrosis. However, there is no specific drug for the treatment of fibrosis, several medications with anti-hepatic fibrosis properties are still in the research and development stages. Peptide, which refers to a substance consisting of 2-50 amino acids, is characterized by structural diversity, low toxicity, biological activities, easy absorption, specific targeting, few side effects, and has been proven to be effective in anti-fibrosis. Here, we summarized various anti-fibrosis peptides in fibrosis including the liver, lungs, kidneys, and other organs. This review will provide a new insight into peptide mediated anti-fibrosis and is helpful to creation of antifibrotic medications.

Effects of Musk Volatile Compounds on Attenuated Nerve Injury and Improving Post-cerebral Ischemic Exercise Functions.

BACKGROUND: Reperfusion Injury Acute ischemic stroke is increasing in people recently and Musk, as a commonly used Traditional Chinese Medicine (TCM), has been suggested as a potential agent against acute ischemic stroke, but the efficacies and underlying mechanisms of it remain unknown. OBJECTIVE: This study was aimed to test the hypotheses that volatile compounds of musk could attenuate nerve injury and identify the bioactive compounds and potential mechanisms of Musk. METHODS: Transient middle cerebral artery occlusion (MCAO) model in vivo in Sprague-Dawley rats (SD rats) was used to test this hypothesis. Collecting ingredients of Musk and their related targets were discerned from the Gas chromatography-olfactory mass spectrometry (GC-O-MS) experiment. Then the potential mechanisms and targets of the compounds were searched by network pharmacology techniques. Finally, the pathway was verified by Western Bolt (WB). RESULTS: First, Musk treatment significantly up-regulated the relative levels of AKT1, PI3KA, and VEGFA in the hippocampus, and improved the sport functions in the post-MCAO ischemic rats in vivo. Next, twenty potential flavor active compounds were recognized by GC-O-MS. A total of 89 key targets including HIF-1, PIK3CA, TNF signaling pathway, and VEGF were identified. AKT1, HIF1A, PIK3CA, and VEGFA were viewed as the most important genes, which were validated by molecular docking simulation. CONCLUSION: The Volatile compounds of musk can attenuate nerve injury and improving post-cerebral ischemic exercise functions by HIF1A pathways, and the combined data provide novel insight for Musk volatile compounds developed as new drug for improving reperfusion injury in acute ischemic stroke.

Application of response surface methodology (RSM) for optimization of the supercritical CO2 extract of oil from Zanthoxylum bungeanum pericarp: Yield, composition and gastric protective effect.

Supercritical carbon-dioxide (SC-CO2) is a promising two-phase technology for flavor components (volatile oil and alkylamides) extract from Zanthoxylum bungeanum pericarp. However, the gastric protective effect of SC-CO2 extract from Z. bungeanum (SZB) have not been systematically investigated. In this study, response surface methodology (RSM) was employed to optimize the yield of SZB, and the average yield of 11.07 % were obtained under optimal parameters (30 MPa, 43 °C and time 75 min). Here, limonene, linalool and hydroxy-α-sanshool were identified as the main compounds of SZB by GC-MS and UPLC-Q-Extractive Orbitrap/MS analysis. When the gastric protective effect of SZB (5, 10 and 20 mg/kg, p.o.) were evaluated, significant increase in body weight and organ indexes of rat, and decreased gastric lesion were observed. Furthermore, nineteen serum metabolites were regarded as the potential biomarkers for the gastric protective effect of SZB. Collectively, this study provides a comprehensive perspective into the chemical composition analysis and gastric protective effect of Z. bungeanum SC-CO2 extract.

Gut Microbiota Regulate Astrocytic Functions in the Brain: Possible Therapeutic Consequences.

Astrocytes are essential for maintaining the homeostasis of the central nervous system (CNS). Astrocytic dysfunction has been implicated in the progression of several neurodegenerative and psychiatric diseases; however, a multitude of factors and signals influencing astrocytic activity have not been entirely elucidated. Astrocytes respond to local signals from the brain, but are also indirectly modulated by gut microbiota. Previous studies revealed that most of the CNS diseases triggered by astrocytic dysfunction are closely associated with the dysbiosis of gut microbiome. Emerging data from preclinical and clinical studies suggest that the maturation and functioning of astrocytes rely on gut microbiota, which plays a pivotal role in the decrease of astrocytic activation and may alleviate symptoms of brain diseases. Herein, we discuss the most recent advances concerning the complex connections between astrocytes and gut microbiota, which are involved in the immune, neurotransmission and neuroendocrine pathways. Deciphering these pathways will facilitate a better understanding of how perturbed gut microbiota contributes to the dysfunction of astrocytes and open therapeutic opportunities for the treatment of brain diseases.

Identification of Active Compounds and Mechanism of Huangtu Decoction for the Treatment of Ulcerative Colitis by Network Pharmacology Combined with Experimental Verification.

INTRODUCTION: Huangtu decoction (HTD) has been widely used in the treatment of gastrointestinal bleeding, ulcerative colitis (UC) and gastrointestinal tumors in China, but its active compounds and mechanism are still not clear yet. The present research aimed to identify the active compounds and mechanism of HTD for the treatment of UC. METHODS: Firstly, the chemical compounds of HTD were qualitatively identified based on Q Exactive Orbitrap LC-MS/MS, and their potential targets were predicted through SwissTargetPrediction. Secondly, the differential expressed genes (DEGs) in colon tissues of UC patients and normal controls were retrieved from the GEO database. Thirdly, the overlapping targets of DEGs and the predicted targets were obtained and subjected to GO and KEGG analysis. Finally, the key targets in the most significantly enriched pathway were verified by in vivo experiment, and the protein and mRNA expressions of matrix metalloproteinase-1 (MMP1), MMP3, MMP7, MMP9 and MMP12 were determined by immunohistochemistry (IHC), Western blotting (WB) and quantitative real-time-PCR (qRT-PCR). RESULTS: A total of 47 compounds were identified and 29 overlapping targets were obtained from HTD extract. The most significantly enriched pathway of overlapping targets involved was MMP. HTD improved the pathological damage in colon tissues of DSS-induced UC model and significantly decreased the serum levels of IL-1ß and IL-6. The protein and mRNA expressions of MMP1, MMP3 and MMP9 in colon tissues were significantly decreased after HTD treatment. CONCLUSION: HTD treatment can alleviate the colonic inflammation via inhibiting MMPs including MMP1, MMP3 and MMP9.

The Volatile Oil of Zanthoxylum bungeanum Pericarp Improved the Hypothalamic-Pituitary-Adrenal Axis and Gut Microbiota to Attenuate Chronic Unpredictable Stress-Induced Anxiety Behavior in Rats.

BACKGROUND: Anxiety disorders (ADs) are the most prevalent mental disorders worldwide. Stress-induced activation of the hypothalamic-pituitary-adrenal (HPA) axis and dysbiosis of gut microbiota seem to contribute to the onset of ADs. This study was designed to investigate the ameliorative effect of volatile oil of Zanthoxylum bungeanum (VOZB) on chronic unpredictable stress (CUS) induced anxiety behavior, as well as the altered HPA axis and gut microbiota. METHODS: Experimental rats were exposed to the CUS for 14 consecutive days. Meanwhile, VOZB was administered at doses of 50, 100 and 200 mg/kg/day for 14 days. The anxiety behavior was evaluated by elevated plus-maze (EPM) and open field (OF). The protein expressions and mRNA levels of corticotropin-releasing hormone (CRH) and glucocorticoid receptor (GR) in hypothalamus was determined, as well the hormone levels of HPA axis in serum. Furthermore, gut microbiota was detected by16S rRNA gene sequencing. The chemical constituents of VOZB were identified by GC-MS analysis. RESULTS: VOZB treatment (100 and 200 mg/kg/day) increased the ratio of open-arm entries and time in EPM test, as well as the central zone entries and time in OF test. Moreover, VOZB treatment reduced the protein expressions and mRNA levels of CRH, but elevated those of GR in hypothalamus. Similarly, the hormone levels of the HPA axis in serum were decreased by VOZB treatment. Besides, VOZB treatment restored the CUS-induced dysbiosis of gut microbiota, raising the Sobs and Chao indexes, inhibiting Lachnospiraceae, but facilitating Bacteroidales_S24-7_group, Lactobacillaceae, and Prevotellaceae. Additionally, Sobs and Chao indexes were negatively correlated to the serum corticosterone and CRH levels. CONCLUSION: VOZB showed an ameliorative effect on CUS-induced anxiety behavior, potentially via inhibiting activation of the HPA axis and restoring the dysbiosis of gut microbiota, thus improving the stress-induced abnormality of the microbiota-gut-brain axis.

Improvement of intestinal flora: accompany with the antihypertensive effect of electroacupuncture on stage 1 hypertension.

BACKGROUND: Increasing evidence have indicated the relationship between intestinal dysbiosis and hypertension. We aimed to evaluate the effect of the electroacupuncture (EA) on intestinal microbiota in patients with stage 1 hypertension. METHODS: 93 hypertensive patients and 15 healthy subjects were enrolled in this study. Applying a highly accurate oscillometric device to evaluate the antihypertensive effect of EA. 16S rRNA sequencing was used to profile stool microbial communities from Healthy group, Before treatment (BT) group and After treatment (AT) group, and various multivariate analysis approaches were used to assess diversity, composition and abundance of intestinal microbiota. RESULTS: In this study, EA significantly decreased the blood pressure (BP) of hypertensive patients. Higher abundance of Firmicutes and lower Bacteroidetes abundance were observed in the BT group compared to the Healthy group. And EA treatment significantly decreased the Firmicutes/Bacteroidetes ratio compared to the BT group. Moreover, at the genus level, there was an increased abundance of Escherichia-Shigella in patients with hypertension, while Blautia were decreased, and EA reversed these changes. CONCLUSIONS: Our study indicates that EA can effectively lower BP and improve the structure of intestinal microbiota which are correlate with the alteration of blood pressure by electroacupuncture. TRIAL REGISTRATION: Clinicaltrial.gov, NCT01701726. Registered 5 October 2012, https://clinicaltrials.gov/ct2/show/study/NCT01701726.

Research progress regarding potential effects of traditional Chinese medicine on postoperative intestinal obstruction.

OBJECTIVES: Postoperative intestinal obstruction is a common postoperative complication with typical symptoms of abdominal pain, vomiting, abdominal distension and constipation. The principal aim of this paper is to provide a full-scale review on the categories and characteristics of postoperative intestinal obstruction, pathophysiology, effects and detailed mechanisms of compounds and monomers from traditional Chinese medicine for treating postoperative intestinal obstruction. Moreover, the possible development and perspectives for future research are also analyzed. METHODS: Literature regarding postoperative intestinal obstruction as well as the anti-pio effect of aqueous extracts and monomers from traditional Chinese medicine in the last 20 years was summarized. KEY FINDINGS: To date, approximately 30 compounds and 25 monomers isolated from traditional Chinese medicine including terpenes, alkaloids, polysaccharides, flavonoids, phenylpropanoids and quinones, have exerted significant antipio effect. This paper reviews the effective doses, models, detailed mechanisms, and composition of these traditional Chinese medicine compounds, as well as the structure of these monomers. Moreover, challenges existed in the current investigation and further perspectives were discussed as well, hoping to provide a reference for future clinical treatment of postoperative intestinal obstruction and the development of new drugs. CONCLUSIONS: Above all, the convincing evidence from modern pharmacology studies powerfully supported the great potential of traditional Chinese medicine in the management of postoperative intestinal obstruction. Regrettably, less attention was currently paid on the mechanisms of traditional Chinese medicine compounds and monomers with antipio effect. Consequently, future study should focus on monomer-mechanism and structure-function relationship.

Hydroxy-α-sanshool isolated from Zanthoxylum bungeanum attenuates learning and memory impairments in scopolamine-treated mice.

Learning and memory impairments are common symptoms of dementia in neurodegenerative disorders. Occasionally, we found that Zanthoxylum bungeanum pericarps (ZBP) significantly activated the spontaneous activity of the hippocampus (HIPP) and paraHIPP (P < 0.001, uncorrected), implying the potential ability of ZBP to improve cognitive impairments. Thus, this study aimed to investigate the improving effect of hydroxy-α-sanshool (HAS), a characteristic ingredient of ZBP, against scopolamine (1 mg kg-1, i.p.)-induced learning and memory deficits. HAS (5 mg kg-1, p.o.) markedly reversed scopolamine-induced cognitive impairments, as indicated by its performance in the passive avoidance test and Morris water maze test (P < 0.01). Furthermore, HAS (2.5 and 5.0 mg kg-1, p.o.) also dose-dependently prevented changes in hippocampal neuronal morphology and apoptosis, inhibited acetylcholinesterase (AChE) activity, increased the acetylcholine (ACh) content, and increased the protein and mRNA expression of brain-derived neurotrophic factor (BDNF) and phospho-cAMP response element-binding (p-CREB) compared with those in the model group (P < 0.05 & P < 0.01). These findings demonstrated that HAS attenuated scopolamine-induced cognitive impairments mainly by enhancing the activity of the cholinergic system and increasing the CREB/BDNF signalling pathway.

Antiobesity, Regulation of Lipid Metabolism, and Attenuation of Liver Oxidative Stress Effects of Hydroxy-α-sanshool Isolated from Zanthoxylum bungeanum on High-Fat Diet-Induced Hyperlipidemic Rats.

Zanthoxylum bungeanum is a traditional Chinese medicine (TCM) used to relieve pain, dispel dampness, stop diarrhea, and prevent itching. The aim of this study was to investigate the antiobesity and hypolipidemic effects of hydroxy-α-sanshool (HAS) isolated from Z. bungeanum on hyperlipidemic rats. Wistar rats (n = 48) were randomly divided into six groups: (1) normal diet rats (ND), (2) high-fat diet- (HFD-) treated rats, (3) HFD+fenofibrate-treated rats (HFD+FNB), (4) HFD+low dose of HAS-treated rats (HFD+LD, 9 mg/kg), (5) HFD+middle dose of HAS-treated rats (HFD+MD, 18 mg/kg), and (6) HFD+high dose of HAS-treated rats (HFD+HD, 36 mg/kg). The body weight and food intake of the rats were recorded during the treatment period. After 4 weeks of HAS treatment, abdominal adipose tissues were observed and total cholesterol (T-CHO), triglycerides (TG), high-density lipoprotein (HDL) cholesterol (HDL-C), and low-density lipoprotein (LDL) cholesterol (LDL-C) of serum and liver tissues were determined. Furthermore, histochemical examinations using oil red O and hematoxylin-eosin staining (H&E) were carried out and levels of malondialdehyde (MDA) and glutathione (GSH) and activities of superoxide dismutase (SOD) in the liver were determined. After HFD feeding, the body weight gain and food efficiency ratio of HFD rats were significantly enhanced (p < 0.05vs. ND rats) and HAS treatment (18 and 36 mg/kg) significantly decreased the body weight gain and food efficiency ratio (p < 0.05vs. HFD rats). In addition, HAS treatment could decrease the abdominal adipose tissues and liver adipocytes. Furthermore, HAS treatment significantly decreased the T-CHO, TG, and LDL-C, whereas it increased HDL-C (p < 0.05vs. HFD rats) in serum and the liver. HAS treatment increased the GSH level and SOD activity in the liver (p < 0.05vs. HFD rats), whereas it decreased the levels of MDA (p < 0.05vs. HFD rats). mRNA analyses suggested that HAS treatment increases the expression of Pparg (proliferator-activated receptor γ) and Apoe (peroxisome apolipoprotein E). Immunohistochemistry and Western blotting indicated that HAS stimulation increased the levels of PPARγ and APOE in the liver, as a stress response of the body defense system. These results revealed that HAS exerts antiobesity and hypolipidemic activities in HFD rats by reducing liver oxidative stress and thus could be considered as a potential candidate drug to cure or prevent obesity and hyperlipidemia.

Gut Microbiota: A New Strategy to Study the Mechanism of Electroacupuncture and Moxibustion in Treating Ulcerative Colitis.

Previous studies have confirmed that acupuncture and moxibustion is an effective way for treating ulcerative colitis (UC). However, the exact mechanism is unclear yet. In this study, DSS-induced UC mice were treated by electroacupuncture and moxibustion, and the genome of intestinal flora was subsequently detected by high-throughput sequencing in order to explore the detailed mechanism in terms of intestinal flora. The results indicated that the alpha diversity indices and beta diversity of intestinal flora were improved by electroacupuncture and moxibustion treatments, especially by the moxibustion treatment. These treatments inhibited Streptococcus, Odoribacter, and Allobaculum whereas it facilitated Lactobacillus on genus level. Further correlation analysis showed that the alpha diversity indices were positively correlated with the percentage of Treg cells in CD4+ cells but negatively correlated with the percentage of Th17 in CD4+ cells. These data indicated that both electroacupuncture and moxibustion can promote the intestinal flora diversity, providing a new view to understand the relationship between host and microbiome when using some external therapies.

Electroacupuncture and Moxibustion Improved Anxiety Behavior in DSS-Induced Colitis Mice.

BACKGROUND AND AIMS: Psychological disorders are prevalent in patients with inflammatory bowel disease, but the underlying mechanisms remain unknown. The aim of this study was to study whether electroacupuncture (EA) and moxibustion (MB) can improve anxiety behavior in DSS-induced colitis mice and to investigate whether this effect is related to hypothalamic-pituitary-adrenocortical (HPA) axis. METHODS: The colitis model was established by drinking 2.5% dextran sodium sulfate (DSS). DSS-induced colitis mice were treated by EA or MB. Disease activity index (DAI) was scored; intestinal morphological and pathological structure was observed; anxiety behavior was tested by the elevated plus maze and open field. The concentration of corticotropin-releasing hormone (CRH) and cortisol (CORT) in serum was measured by enzyme-linked immunosorbent assay (ELISA). The protein expression of CRH in the colon and hypothalamus was detected by Western blot (WB). RESULTS: Both EA and MB treatments can improvethe morphology of their distal colonic mucosal epithelia, as well as the disease activity index. Meanwhile, anxiety behavior in colitis mice was improved slightly after EA and MB treatment. In addition, the levels of CRH and CORT in the serum were slightly improved after EA and MB treatment. These effects are further supported by WB results. The expression of CRH in the colon and hypothalamus was increased significantly after treatment, compared with the model group. CONCLUSION: EA and MB were able to regulate the concentration of CRH in serum and protein expression in the peripheral and central at different levels and promote the recovery of the HPA axis that may be the basis for EA and MB to improve colonic pathology and alleviate anxiety behavior in DSS-induced colitis.

Guizhi-Shaoyao-Zhimu decoction possesses anti-arthritic effects on type II collagen-induced arthritis in rats via suppression of inflammatory reactions, inhibition of invasion & migration and induction of apoptosis in synovial fibroblasts.

BACKGROUND: Rheumatoid arthritis (RA) is a known intractable chronic inflammatory disease of synovial joints characterized by hyperplasia and consecutive inflammation with a high prevalence.Guizhi-Shaoyao-Zhimu (GSZD) is the first choice for clinical treatment of RA in Chinese traditional medicine. This study is aimed to explore the possible pharmacological mechanisms of anti-arthritic effect of GSZD. METHODS: Type II collagen-induced arthritis (CIA) rat model was used to study the anti-arthritic activity of GSZDin vivo, and toe swelling & arthritis score, serum levels of cytokines, and pathological examinations were carried out. In vitro, TNF-α induced MH7A cells were used to study the possible mechanisms of GSZD. The anti-proliferative effects of GSZD were determined by MMT assay, and pro-apoptotic activity of GSZD in MH7A cells was determined by flow cytometry analysis & DAPI staining. Furthermore, the adhesive and invasive abilities of MH7A cells were determined using cell adhesion and transwell assays. MMPs levels were determined by ELISA assays, and mRNA expressions of Caspase-3, -9, Bax, SOCS1, Bcl-2, JAK2, STAT-3 and -5 were determined using qRT-PCR analysis. Besides, the major chemical components in GSZD were analyzed by HPLC-QqQ-MS analysis. RESULTS: Our results showed GSZD reduced the toe swelling & arthritis score, and serum levels of TNF-α, IL-1ß, IL-6 & IL-17a in CIA rats; pathological examination results indicated GSZD improved ankle joint injury in CIA rats.In vitro, GSZD showed significant anti-proliferative and pro-apoptotic effects on TNF-α stimulated MH7A cells. After GSZD treatment, the adhesive and invasive abilities of MH7A cells were reduced, and secretions of MMPs, IL-6 and IL-8 were also reduced. GSZD decreased the releases of TNF-α and IL-1ß in LPS stimulated RAW 264.7 cells. Further studies showed GSZD up-regulated mRNA expressions of Caspase-3, -9, Bax, and SOCS1, whereas down-regulated mRNA expressions of Bcl-2, JAK2, STAT3 and STAT5. Besides, 13 major chemical components were identified in GSZD extracts through HPLC-QqQ-MS analysis. CONCLUSION: Our results suggested GSZD possesses an anti-rheumatic effect on CIA rats, and the possible mechanism is related to inhibiting inflammatory response, inhibiting invasion and migration of synovial fibroblasts, and inducing apoptosis in synovial fibroblasts.

In silico drug design of inhibitor of nuclear factor kappa B kinase subunit beta inhibitors from 2-acylamino-3-aminothienopyridines based on quantitative structure-activity relationships and molecular docking.

Inhibitor of nuclear factor kappa B kinase subunit beta (IKK-ß), a specific regulator of nuclear factor-κB (NF-κB), is considered a valid target to design novel candidate drugs to treat rheumatoid arthritis and various cancers. In the present study, quantitative structure-activity relationships (QSAR) and molecular docking techniques were used to screen for new IKK-ß inhibitors from a series of 2-acylamino-3-aminothienopyridine analogs. During the two-dimensional QSAR phase, the statistical model partial least square was selected from among two alternatives (r2 = 0.868, q2 (cross-validation) = 0.630). Descriptors with positive or negative contributions were derived from the created model. To build of three-dimensional QSAR models, we used three different fingerprints as analysis precepts for molecular clustering and the subsequent division of training sets and test sets. The best model, which used fingerprint model definition language public keys, was selected for further prediction of the compounds' activities. Favorable physicochemical, structural, electrostatic, and steric properties were derived from the created QSAR models and then used for drug design with an in-house library. Amongst the designed compounds, compounds B01 and B02 showed good predicted activities. Furthermore, after a selecting the protein structure and docking method, docking studies were carried out to reveal the detailed interactions between the ligands and the target protein. Binding affinity was measured and sorted using the value of "-CDOCKER_ENERGY". The high -CDOCKER_ENERGY values of compounds B01 (41.6134 kcal/mol) and B02 (40.1366 kcal/mol) indicated their prominent docking affinities.

[Electroacupuncture Intervention Regulates Circadian Rhythms by Down-regulating Per Gene Expression in Hypothalamic Suprachiasmatic Nucleus of Hepatocellular Carcinoma Mice].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on the rhythm of running-wheel activity of hepatocellular carcinoma (HCC) mice and the expression of Per 1 and Per 2 (circadian rhythm genes) in the hypothalamic suprachiasmatic nucleus (SCN), so as to investigate its mechanism underlying regulating circadian rhythm. METHODS: A total of 108 male C 57 BL / 6 J mice were randomly divided into control, HCC model and EA groups which were further assigned to six zeitbeger (environmental light-dark cycle) time (ZT) point (ZT 0, ZT 4, ZT 8, ZT 12, ZT 16 and ZT 20) subgroups. The HCC model was established by injection of H 22 cancer cell (abdominal 3rd generation, 10 µL) suspension into the larger live lobe. Mice of the control group received saline injection of the liver lobe. EA (2 Hz/15 Hz, 0.2 mA) was applied to bilateral "Ganshu" (BL 18) and "Zhiyang" (GV 9) for 15 min, once daily for 10 days. Mice of the control and model groups received the same binding-fixing to those of the EA group. Circadian running-wheel activity of 12 h∶12 h light darkness (LD) cycle (activity onset and acrophase of actogram, amplitude or peak of periodogram) was recorded by using ClockLab (ACT-500) software and analyzed by MATLAB (R 2007 b) before and after EA treatment. The pathological changes of liver cells were observed under light microscope after sectioning and H．E. staining. The expression levels of Per 1 mRNA and Per 2 mRNA in the liver tissues were determined by fluorogenic quantitative real time-PCR. RESULTS: (1) Following modeling, the amplitude of periodogram of running-wheel activity was significantly lowered at ZT 0, ZT 4, ZT 8, ZT 12, ZT 16, and ZT 20 relevant to the control group (P<0.05). After EA intervention, the amplitude of periodogram at ZT 8 (15：00) was considerably increased relevant to the model group (P<0.05), and the acrophase at ZT 8 was remarkably advanced (P<0.05). No significant changes were found in the onset time and periods of periodogram at the 6 time-points after modeling and EA intervention (P>0.05). (2) The expression levels of Per 1 mRNA and Per 2 mRNA in the SCN were significantly up-regulated at the 6 time-points in the model group relevant to the control group (P<0.05), and obviously down-regulated at ZT 8 after EA intervention relevant to the model group (P<0.05)．. CONCLUSION: EA can benignly regulate the rhythm of running-wheel activity of HCC mice, which may be closely related to its effect in down-regulating the expression of circadian rhythm genes Per 1 and Per 2 in the SCN.

[Effects of acupuncture on ANP and CNP in adrenal gland and CORT in plasma in rats with chronic emotional stress anxiety].

OBJECTIVE: To analyze the effects of acupuncture on the level of atrial natriuretic peptide (ANP), C-type natriuretic peptide (CNP) in adrenal gland and the content of corticosterone (CORT) in plasma in rats withchronic emotional stress anxiety, and to explore the partial action mechanism of acupuncture on anxiety disorder. METHODS: Thirty-two healthy Sprague-Dawley (SD) rats, after 7 days of feeding and domestication, were randomly divided into a blank group (10 rats), a model group (11 rats) and an acupuncture group (11 rats). The rats inthe model group and acupuncture group were treated with unpredictable chronic emotional stress (CES) method toestablish the model of anxiety. Rats in the acupuncture group were treated with acupuncture at "Neiguan" (PC 6)and "Shenmen" (HT 7), once every other day, 30 minutes each time. The model establishment and treatment lasted 15 days. Rats in the blank group were treated with identical immobilization but no treatment was given. Theelevated plus maze was used to test the behavioral changes of rats with anxiety; the level of CORT in plasma wasdetected by ELISA, and the expression level of CNP and ANP in adrenal cortex and medulla was detected by immunohistochemical method. RESULTS: (1) The percentage of open-arms time in total time (OT%) in elevated plus maze in the model group was significantly lower than that in the blank group (P<0. 05); the OT% in the acupuncture group was significantly higher than that in the model group (P<0.01). (2) The content of CORT in plasma in the model group was higher than that in the blank group (P<0. 05), while that in the acupuncture group was significantly lower than that in the model group (P<0. 05). (3) The expression of ANP in adrenal medulla and cortex in the model group was lower than that in the blank group (P<0. 01), while the expression of CNP in adrenal medulla and cortex in the model group was higher than that in the blank group (P<0. 01). CONCLUSION: The effects of acupuncture against anxiety are likely to be related to the regulation on the expression of ANP and CNP in adrenal medulla, affecting the release of CORT and inhibition on the activity !f hypothalamic-pituitary-adrenal axis (HPA axis).