Mechanism of Key Ingredient of Astragalus membranaceus on Lung Adenocarcinoma via PI3K/AKT Signaling Clarified by Utilizing Network Pharmacology Approach and Experimental Validation.

OBJECTIVE: To investigate the mechanism of the effect of Astragalus membranaceus (A. membranaceus) on lung adenocarcinoma at the molecular level to elucidate the specific targets according to the network pharmacology approach. METHODS: The active components of A. membranaceus and their potential targets were collected from the Traditional Chinese Medicine Systems Pharmacology Database. Lung adenocarcinoma-associated genes were acquired based on GeneCards, Online Mendelian Inheritance in Man (OMIM), PharmGKB, and Therapeutic Targets databases. The PI3K/AKT signaling pathway-related genes were obtained using Reactome portal. Networks of "ingredient-target" and "ingredient-target-pathway-disease" were constructed using the Cytoscape3.6.0 software. The relationships among targets were analyzed according protein-protein interaction (PPI) network. Finally, molecular docking was applied to construct the binding conformation between active ingredients and core targets. Cell counting kit 8 (CCK8) and Western blot assays were performed to determine the mechanism of the key ingredient of A. membranaceus. RESULTS: A total of 20 active components and their 329 targets, and 7,501 lung adenocarcinoma-related genes and 130 PI3K/AKT signaling pathway-related genes were obtained. According to Venn diagram and PPI network analysis, 2 mainly active ingredients, including kaempferol and quercetin, and 6 core targets, including TP53, MAPK1, EGF, AKT1, ERBB2, and EGFR, were identified. The two important active ingredients of A. membranaceus, kaempferol and quercetin, exert the therapeutic effect in lung adenocarcinoma partly by acting on the 6 core targets (TP53, MAPK1, EGF, AKT1, ERBB2, and EGFR) of PI3K/AKT signaling pathway. Expressions of potential targets in lung adenocarcinoma and normal samples were analyzed by using UALCAN portal and found that ERBB2 was overexpressed in lung adenocarcinoma tissues and upregulation of it correlated with clinicopathological characteristics. Finally, quercetin repressed viabilities of lung adenocarcinoma cells by targeting ERBB2 on PI3K/AKT signaling confirmed by CCK8 and Western blot. CONCLUSION: Our finding unraveled that an active ingredient of A. membranaceus, quercetin, significantly inhibited the lung adenocarcinoma cells proliferation by repressing ERBB2 level and inactivating the PI3K/AKT signaling pathway.

[Effect of Tongli Shuji acupuncture on the expression of Jagged2 and Notch2 in a rat model of permanent ischemia of the right middle cerebral artery].

OBJECTIVE: To investigate the effect of Tongli Shuji acupuncture on the behavioristics, the percentage of cerebral infarct volume, and the expression of Jagged2 and Notch2 proteins in the hippocampus of the ischemic hemisphere in rats with permanent ischemia of the right middle cerebral artery, as well as the mechanism of Tongli Shuji acupuncture in improving ischemic cerebrovascular disease. METHODS: Sprague-Dawley rats were randomly divided into sham-operation group, model control group, medication group, routine acupuncture group, and Tongli Shuji group, with 10 rats in each group. The suture method was used to establish a model of permanent ischemia of the right middle cerebral artery. The rats in the medication group were given Citicoline (0.4 mg/kg) by gavage, those in the routine acupuncture group were given acupuncture at"Baihui"(GV20) and"Dazhui" (GV14) 15 minutes per day, and those in the Tongli Shuji group were given acupuncture at "Zhengying "(GB17), "Tianjing"(SJ10), and"Huantiao"(GB30) at the left side. The neurological deficit score was determined on days 1, 7, 14, 21 and 28 after surgery, the percentage of cerebral infarct volume was measured after treatment, and Western blot was used to measure the protein expression of Jagged2 and Notch2 in the right hippocampal tissue. RESULTS: Compared with the sham-operation group, the model control group had significant increases in the neurological deficit score on day 1 after surgery (P<0.001). Compared with the model control, medication and routine acupuncture groups, the Tongli Shuji group had significant reductions in the neurological deficit score on day 21 after surgery (P<0.01). Compared with the sham-operation group, the model control group had a significant increase in the percentage of cerebral infarct volume(P<0.05)．The Tongli Shuji group had a significant reduction in the percentage of cerebral infarct volume compared with the medication groups (P<0.05). Compared with the model control group, the routine acupuncture group had a significant increase in the expression of hippocapmal Jagged2(P<0.01)．Compared with the model control, medication and routine acupuncture groups, the Tongli Shuji group had significant increases in the expression of hippocampal Jagged2 and Notch2 (P<0.01). CONCLUSION: Acupuncture can improve the behavioral manifestations of neurological deficit in rats with permanent ischemia of the right middle cerebral artery. Tongli Shuji acupuncture can significantly up-regulate the protein expression of Jagged2 and Notch2 in the hippocampal tissue of the ischemic region, possibly by activating the Notch signaling pathway to exert a neuroprotective effect.

[Progress of Experimental Researches on Mechanisms of Acupuncture Underlying Improvement of Ischemic Cerebral Vascular Disease by Regulating Different Intracellular Signaling Pathways].

China is in the "high death zone" of the world cerebrovascular disease (CVD) map. The prevention and treatment of ischemic CVD (ICVD) have become the top priority in clinical practice. It has been demonstrated that acupuncture therapy has a positive role in bettering clinical symptoms of ICVD patients. Findings of recent experimental studies displayed that electroacupuncture (EA) intervention is effective in reducing the cerebral infarcted volume and cellular injury, promoting proliferation, migration and differentiation of nerve stem cells, facilitating the regeneration of nerve tissue, bettering dysneuria and limb locomotor ability, etc. in ICVD rats, which are closely associated with its effects in regulating different intracellular signaling pathways. In the present study, we review the progress of recent experimental studies on the underlying mechanisms of EA in improving ICVD from six major signaling pathways including Notch, mitogen-activated protein kinases (MAPK)/ extracellular signal regulated protein kinases (ERK), phosphatidy linositol-3-kinase (PI 3 K)/ protein kinase B (Akt), Janus kinase-signal transducer and activator of transcription (JAK-STAT), erythropoietin-producing hepatocyte receptor (Eph)/Ephrin, nuclear factor of kappa B (NF-κB) and their compositions, which may provide new therapeutic targets for acupuncture treatment of ICVD. More attention should be paid to the comparison of the efficacy of acupuncture of different acupoint combinations (traditional acupoint formulas) in relieving ICVD.

[Rasch analysis on stroke-specific quality of life (SS-QOL) scale of acupuncture intervention on stroke].

OBJECTIVE: To optimize the evaluation level of active motor threshold and the functional domain of upper limbs of stroke-specific quality of life (SS-QOL) scale with Rasch analysis. METHODS: Sixty patients with acute ischemic stroke that were in accord with research criterid were randomly divided into a test group (30 cases) and a control group (30 cases). Acupuncture treatment and routine western medicine were applied on the test group, and single treatment of routine western medicine was applied on the control group. Selected acupoints were MS 5, Fengchi (GB 20), Hegu (LI 4), etc. Active motor threshold and the functional domain of upper limbs of SS-QOL were self-tested by patients after one treating course. The characteristics of the above mentioned items were tested with Rasch model. RESULTS: The statistical result on fitness of active motor threshold and the functional domain of upper limbs of SS-QOL showed that every reference of samples and items wosin accord with the Rasch model and has well inner reliability and validity. The Infit and Outfit MnSq values of active motor threshold and the functional domain of upper limbs of SS-QOL are basically between 0.5 and 1.5. CONCLUSION: The application of Rasch analysis on the assessment of patient reported outcome (PRO) has optimized the PRO scale (the activity and upper limb function domain of SS-QOL scale) and enhanced evaluation level of active motor threshold and the functional domain of upper limbs of SS-QOL scale.

[Studies on dynamic changes in traditional Chinese medicine syndrome patterns for stroke using data-driven and model-driven approaches: a review].

Many clinical studies showed that the traditional Chinese medicine (TCM) syndromes in stroke have been dynamically changing since the onset of the disease. The changing of TCM syndromes can be attributed to multiple correlative factors such as age, sex, area distribution, underlying diseases, and constitutional factor. Data-driven methods involving multivariate statistical methods and descriptive approach have been used to analyze the regularity of dynamically changed TCM syndromes of stroke. However, expressing non-linear relationship between symptom or correlative factors and syndrome patterns by data-driven models is challenging. Model-driven methods involving artificial neural networks and Bayesian networks are new methods for studying the changes in TCM syndromes in patients with stroke. In this review, the authors summarized the studies of dynamically changed patterns of stroke syndromes based on data-driven methods and some clinical trials on TCM syndromes based on model-driven methods. Further studies are needed to improve the understanding of the dynamically changing regularity of TCM syndromes for stroke by using model-driven methods so as to develop appropriate and timely TCM treatments.