ABSTRACT
BACKGROUND: Berberine has been established as a potential drug for inflammation and metabolic disorder. Here, we aimed to explore the effects and the underlying mechanisms of berberine on obesity-induced chronic inflammation. METHODS: Mice were fed with high-fat diet to induce obesity. Inflammation in adipocytes were induced with treatment of free fatty acids. The expression of IL-4, CD206, ARG1 and other markers were used to identify M1 and M2 polarization. The expression of GPR78 and CHOP were used to evaluate endoplasmic reticulum stress. H&E staining was used to reveal the adipose tissue macrophage and adipocytes enlargement. RESULTS: Berberine treatment attenuated endoplasmic reticulum stress and inflammation in obese mice and free fatty acids-treated adipocytes. Overexpression of lncRNA Gomafu partially blocked the protective effects of berberine in free fatty acids-treated adipocytes by increasing endoplasmic reticulum stress. Moreover, Gomafu overexpression partly reversed berberine-induced enhancement of M2 polarization in macrophages. Finally, Gomafu overexpression induced ER stress and inflammation in mice, which were improved by berberine administration. CONCLUSIONS: Berberine improves obesity-induced chronic inflammation by alleviating endoplasmic reticulum stress and consequently promoting macrophage M2 polarization. And these protective effects were mediated at least partly by the suppression of lncRNA Gomafu.
Subject(s)
Anti-Inflammatory Agents/therapeutic use , Berberine/therapeutic use , Inflammation/drug therapy , Macrophages/immunology , Obesity/drug therapy , RNA, Long Noncoding/genetics , Th2 Cells/immunology , Animals , Cell Differentiation , Chronic Disease , Cytokines/metabolism , Disease Models, Animal , Down-Regulation , Endoplasmic Reticulum Chaperone BiP , Endoplasmic Reticulum Stress/drug effects , Humans , Mice , Mice, Inbred C57BL , RAW 264.7 CellsABSTRACT
BACKGROUND: Colorectal Adenomatous Polyp (CAP) was one precursor of colorectal cancer (CRC) and having a high chance of developing into CRC. There was a lack of conclusive chemoprevention evidences to prevention new CAP occurrence in post-polypectomy. Xiaoai Jiedu Decoction, Chinese National Medical Professor (Zhou Zhongying)'s experience formula, has been used to treat new CAP occurrence in post-polypectomy from the 20th century in China. However, clinical research of Xiaoai Jiedu Decoction in the treatment of CAP recurrence was lack. We design this study to evaluate the efficacy and safety of Xiaoai Jiedu Decoction in the treatment of new CAP occurrence in post-polypectomy on colonoscopy. METHODS/DESIGN: A randomized, controlled, blind and multicenter trial to evaluate the efficacy and safety of Xiaoai Jiedu Decoction is proposed. CAP patients (after complete polypectomy under colonoscopy) will be randomly assigned into Xiaoai Jiedu Decoction group and Xiaoai Jiedu Decoction mimetic agent group. Patients will receive 6-course treatments and a 2-year follow-up. Follow-up colonoscopy will be anticipated to perform in 1 and 2 years after the baseline examinations. The primary outcome measure is the new CAP occurrence in 1 and 2 years. The secondary outcome measure is the occurrence of advanced adenoma in 1 and 2 years. DISCUSSION: This study will provide objective evidences to evaluate the efficacy and safety of Xiaoai Jiedu Decoction as an adjuvant treatment for new CAP occurrence in post-polypectomy. TRIAL REGISTRATION: NCT03616444.
Subject(s)
Adenomatous Polyps/prevention & control , Colorectal Neoplasms/prevention & control , Drugs, Chinese Herbal/therapeutic use , Medicine, Chinese Traditional , Precancerous Conditions/prevention & control , Double-Blind Method , Humans , Multicenter Studies as Topic , Randomized Controlled Trials as TopicABSTRACT
BACKGROUND: Oral submucous fibrosis (OSF) is a premalignant and fibrosing disease, which is closely associated with the habit of chewing areca nut. Panax notoginseng Buck F. H. Chen is an often used antifibrotic and antitumor agent. To treat areca nut-induced OSF, we have developed a chewable tablet, in which one of the major medicines is total Panax notoginseng saponins (PNS). In this study, we have investigated the antifibrotic effect and mechanism of PNS on areca nut-induced OSF in vitro. METHODS: Through human procollagen gene promoter luciferase reporter plasmid, hydroxyproline assay, gelatin zymography, qRT-PCR, ELISA, and Western blot, the influences of PNS on areca nut extract (ANE)-induced cell growth, collagen accumulation, procollagen gene transcription, MMP-2/-9 activity, MMP-1/-13 and TIMP-1/-2 expression, cytokine secretion, and the activation of PI3K/AKT, ERK/JNK/p38 MAPK, and TGFß/Smads pathways were detected. RESULTS: Panax notoginseng saponins could inhibit the ANE-induced abnormal growth and collagen accumulation of oral mucosal fibroblasts in a concentration-dependent manner. PNS (25 µg/ml) could significantly inhibit the ANE-induced expression of Col1A1 and Col3A1, augment the ANE-induced decrease of MMP-2/-9 activity, inhibit the ANE-induced increase of TIMP-1/-2 expression, and decrease the ANE-induced transcription and release of CTGF, TGFß1, IL-6, and TNFα. PNS (25 µg/ml) also significantly inhibited the ANE-induced activation of AKT and ERK/JNK/p38 MAPK pathways in oral mucosal fibroblasts and the ANE-induced activation of TGFß/smad pathway in HaCaT cells. CONCLUSION: Panax notoginseng saponins possess excellent anti-OSF activity, and its mechanism may be related to its ability to inhibit the ANE-induced activation of PI3K/AKT, ERK/JNK/p38 MAPK, and TGFß/smad pathways.
Subject(s)
Areca/adverse effects , Mouth Mucosa/drug effects , Nuts/adverse effects , Oral Submucous Fibrosis/pathology , Panax notoginseng , Plant Extracts/pharmacology , Saponins/pharmacology , Animals , Cell Culture Techniques , Cell Line , Collagen Type I/drug effects , Collagen Type I, alpha 1 Chain , Collagen Type III/drug effects , Connective Tissue Growth Factor/drug effects , Fibroblasts/drug effects , Humans , Hydroxyproline/analysis , Interleukin-6/analysis , MAP Kinase Signaling System/drug effects , Matrix Metalloproteinase 2/drug effects , Matrix Metalloproteinase 9/drug effects , Mice , Mice, Inbred BALB C , Mouth Mucosa/cytology , Oral Submucous Fibrosis/etiology , Phosphatidylinositol 3-Kinases/drug effects , Plant Extracts/adverse effects , Proto-Oncogene Proteins c-akt/drug effects , Smad Proteins/drug effects , Tissue Inhibitor of Metalloproteinase-1/drug effects , Tissue Inhibitor of Metalloproteinase-2/drug effects , Transforming Growth Factor beta1/drug effects , Tumor Necrosis Factor-alpha/drug effectsABSTRACT
It has been reported that autophagy is involved in the replication of many viruses. In this study, we screened 89 medicinal plants, using an assay based on the inhibition of the formation of the Atg12-Atg5/Atg16 heterotrimer, an important regulator of autophagy, and selected Silybum marianum L. for further study. An antiviral assay indicated that silybin (S0), the major active compound of S. marianum L., can inhibit influenza A virus (IAV) infection. We later synthesized 5 silybin derivatives (S1 through S5) and found that 23-(S)-2-amino-3-phenylpropanoyl-silybin (S3) had the best activity. When we compared the polarities of the substituent groups, we found that the hydrophobicity of the substituent groups was positively correlated with their activities. We further studied the mechanisms of action of these compounds and determined that S0 and S3 also inhibited both the formation of the Atg12-Atg5/Atg16 heterotrimer and the elevated autophagy induced by IAV infection. In addition, we found that S0 and S3 could inhibit several components induced by IAV infection, including oxidative stress, the activation of extracellular signal-regulated kinase (ERK)/p38 mitogen-activated protein kinase (MAPK) and IκB kinase (IKK) pathways, and the expression of autophagic genes, especially Atg7 and Atg3. All of these components have been reported to be related to the formation of the Atg12-Atg5/Atg16 heterotrimer, which might validate our screening strategy. Finally, we demonstrated that S3 can significantly reduce influenza virus replication and the associated mortality in infected mice. In conclusion, we identified 23-(S)-2-amino-3-phenylpropanoyl-silybin as a promising inhibitor of IAV infection.
Subject(s)
Antiviral Agents/pharmacology , Influenza A Virus, H1N1 Subtype/drug effects , Plant Extracts/chemistry , Silybum marianum/chemistry , Silymarin/analogs & derivatives , Animals , Antiviral Agents/chemical synthesis , Antiviral Agents/isolation & purification , Autophagy/drug effects , Autophagy-Related Protein 12 , Autophagy-Related Protein 5 , Autophagy-Related Proteins , Carrier Proteins/antagonists & inhibitors , Carrier Proteins/genetics , Carrier Proteins/metabolism , Chlorocebus aethiops , Dogs , Gene Expression Regulation , High-Throughput Screening Assays , Humans , Influenza A Virus, H1N1 Subtype/growth & development , Madin Darby Canine Kidney Cells , Microtubule-Associated Proteins/antagonists & inhibitors , Microtubule-Associated Proteins/genetics , Microtubule-Associated Proteins/metabolism , Mitogen-Activated Protein Kinases/antagonists & inhibitors , Mitogen-Activated Protein Kinases/genetics , Mitogen-Activated Protein Kinases/metabolism , Oxidative Stress/drug effects , Plasmids , Protein Multimerization/drug effects , RNA, Small Interfering/genetics , RNA, Small Interfering/metabolism , Signal Transduction , Silymarin/chemical synthesis , Silymarin/isolation & purification , Silymarin/pharmacology , Small Ubiquitin-Related Modifier Proteins/antagonists & inhibitors , Small Ubiquitin-Related Modifier Proteins/genetics , Small Ubiquitin-Related Modifier Proteins/metabolism , Vero CellsABSTRACT
Autophagy is involved in many human diseases, such as cancer, cardiovascular disease and virus infection, including human immunodeficiency virus (HIV), hepatitis C virus (HCV), influenza A virus (IAV) and coxsackievirus B3/B4 (CVB3/B4), so a drug screening model targeting autophagy may be very useful for the therapy of these diseases. In our study, we established a drug screening model based on the inhibition of the dissociation of Beclin1-Bcl2 heterodimer, an important negative regulator of autophagy, using bimolecular fluorescence complementation (BiFC) technique for developing novel autophagy inhibitors and anti-IAV agents. From 86 examples of traditional Chinese medicines, we found Syzygium aromaticum L. had the best activity. We then determined the anti-autophagy and anti-IAV activity of eugenol, the major active compound of Syzygium aromaticum L., and explored its mechanism of action. Eugenol could inhibit autophagy and IAV replication, inhibited the activation of ERK, p38MAPK and IKK/NF-κB signal pathways and antagonized the effects of the activators of these pathways. Eugenol also ameliorated the oxidative stress and inhibited the expressions of autophagic genes. We speculated that the mechanism underlying might be that eugenol inhibited the oxidative stress and the activation of ERK1/2, p38MAPK and IKK/NF-κB pathways, subsequently inhibited the dissociation of Beclin1-Bcl2 heterodimer and autophagy, and finally impaired IAV replication. These results might conversely display the reasonableness of the design of our screening model. In conclusion, we have established a drug screening model for developing novel autophagy inhibitor, and find eugenol as a promising inhibitor for autophagy and IAV infection.
Subject(s)
Antiviral Agents/pharmacology , Autophagy/drug effects , Drug Evaluation, Preclinical/methods , Eugenol/pharmacology , Influenza A virus/drug effects , Cell Line , Drugs, Chinese Herbal/pharmacology , Humans , Syzygium/chemistryABSTRACT
<p><b>OBJECTIVE</b>To examine whether acupuncture treatment would improve outcome in chronic Achilles tendinopathy.</p><p><b>METHODS</b>A randomized, controlled trial at two centers of 64 randomized patients aged 18 to 70 years with chronic Achilles tendinopathy was conducted from July 2007 to April 2010, with follow-up until October, 2010. These patients were randomly allocated into an acupuncture treatment group (acupuncture group) and an eccentric exercises group (control group). The validated Victorian Institute of Sports Assessment-Achilles (VISA-A) questionnaire was completed at baseline and 8, 16, and 24 weeks. The pain at rest and after activity was accessed at baseline and 8 weeks with Visual Analogue Scale (VAS).</p><p><b>RESULTS</b>After randomization into the acupuncture group or control group, one patient was loss of follow-up. The mean VISA-A score improved signifificantly after 8 weeks in the acupuncture group to 67.1 points [95% confifidence interval (CI), 64.1-70.2] and in the control group to 48.5 points (95% CI, 45.5-51.6) with an additional 18.6 points increase in acupuncture treatment patients (P=0.0000). Acupuncture treatment resulted in a significant increase from baseline in VISA-A of 25.8 after 16 weeks and 28.4 after 24 weeks. Whereas, in the control group the increase from baseline in VISA-A were 10.0 and 16.6 after 16 and 24 weeks, respectively (P=0.0000). The VAS diminished by 2.0 cm after activity, and by 1.5 cm at rest after 8 weeks in the control group. In the acupuncture group, the pain scores diminished significantly more than in the control group, with pain reduction of 3.7 cm after activity (P=0.0000) and 3.2 cm at rest (P =0.0000).</p><p><b>CONCLUSION</b>Acupuncture intervention could improve pain and activity in patients with chronic Achilles tendinopathy compared with eccentric exercises.</p>
Subject(s)
Female , Humans , Male , Middle Aged , Achilles Tendon , Pathology , Acupuncture , Chronic Disease , Tendinopathy , Therapeutics , Treatment Outcome , Visual Analog ScaleABSTRACT
In this research, we have established a drug screening method based on the autophagy signal pathway using the bimolecular fluorescence complementation-fluorescence resonance energy transfer (BiFC-FRET) technique to develop novel anti-influenza A virus (IAV) drugs. We selected Evodia rutaecarpa Benth out of 83 examples of traditional Chinese medicine and explored the mechanisms of evodiamine, the major active component of Evodia rutaecarpa Benth, on anti-IAV activity. Our results showed that evodiamine could significantly inhibit IAV replication, as determined by a plaque inhibition assay, an IAV vRNA promoter luciferase reporter assay and the Sulforhodamine B method using cytopathic effect (CPE) reduction. Additionally, evodiamine could significantly inhibit the accumulation of LC3-II and p62, and the dot-like aggregation of EGFP-LC3. This compound also inhibited the formation of the Atg5-Atg12/Atg16 heterotrimer, the expressions of Atg5, Atg7 and Atg12, and the cytokine release of TNF-α, IL-1ß, IL-6 and IL-8 after IAV infection. Evodiamine inhibited IAV-induced autophagy was also dependent on its action on the AMPK/TSC2/mTOR signal pathway. In conclusion, we have established a new drug screening method, and selected evodiamine as a promising anti-IAV compound.
Subject(s)
Antiviral Agents/pharmacology , Autophagy/drug effects , Drug Evaluation, Preclinical/methods , Influenza A virus/drug effects , Quinazolines/pharmacology , Signal Transduction/drug effects , Adenylate Kinase/metabolism , Animals , Autophagy/genetics , Carrier Proteins/genetics , Carrier Proteins/metabolism , Cell Line , Cytokines/biosynthesis , Gene Expression/drug effects , Humans , Microtubule-Associated Proteins/genetics , Microtubule-Associated Proteins/metabolism , Protein Binding/drug effects , Protein Multimerization/drug effects , Small Ubiquitin-Related Modifier Proteins/genetics , Small Ubiquitin-Related Modifier Proteins/metabolism , TOR Serine-Threonine Kinases/metabolism , Tuberous Sclerosis Complex 2 Protein , Tumor Suppressor Proteins/metabolism , Virus Replication/drug effectsABSTRACT
OBJECTIVE: To investigate the intestinal absorption and metabolism of daidzein in Caco-2 cell model. METHODS: The damage of daidzein to Caco-2 cell was evaluated by MTT value and alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) activity. With the reference to propranolol, the apparent permeability coefficient (Papp) of daidzein was measured through the monolayer under the different concentrations and pH media. The metabolites were also measured by enzyme hydrolysis. RESULTS: Daidzein had no damage to the growth of the Caco-2 cells in the concentration of 1-50 microg/mL. The apparent permeability coefficient (Papp) of daidzein across the monolayer showed concentration- and pH- independent manner, which was similar to that of transcellcular marker-propranolol, suggesting the good absorption of daidzein in vivo. By hydrolysis with beta-glucuronidase, low metabolites were detected in monolayer and transport medium, verifying the existence of glucuronides and sulfates. CONCLUSION: The daidzein absorption in Caco-2 cell model is a passive and transcellular transport and quite good in the intestines.
Subject(s)
Glycine max , Isoflavones/pharmacokinetics , Phytoestrogens/pharmacokinetics , Absorption , Biological Transport , Caco-2 Cells , Cell Membrane Permeability , Humans , Hydrogen-Ion Concentration , Isoflavones/chemistry , Models, Biological , Phytoestrogens/chemistry , Plants, Medicinal/chemistry , Glycine max/chemistryABSTRACT
OBJECTIVE: To prepare tetrandrine alginate calcium sustained release gel pellets twice daily with Eudragit RS 30D and Eudragit RL 30D. METHODS: The sustained release gel pellets were prepared by fluid bed technique and release in vitro was selected as the evaluate index. The formulation was optimized by full design test based on the studies of coating factors. RESULTS: The optimal coating formulation was shown at the ratio of Eudragit RS 30D and Eudragit RL 30D to 5:1, the loading weight of polymers of 45%, the plasticizer concentration of 20% and 35% talcum powder. CONCLUSION: The perfect sustained release of tetrandrine pellets can be obtained by adjusting the ratio of Eudragit RS 30D and Eudragit RL 30D and the loading weight of polymers.