Gegen Qinlian decoction ameliorates TNBS-induced ulcerative colitis by regulating Th2/Th1 and Tregs/Th17 cells balance, inhibiting NLRP3 inflammasome activation and reshaping gut microbiota.

ETHNOPHARMACOLOGICAL RELEVANCE: Chinese herbal medicine Gegen Qinlian Decoction (GQD) has been clinically shown to be an effective treatment of ulcerative colitis (UC) in China. However, the underlying mechanism of GQD's anti-ulcerative colitis properties and its effect on gut microbiota still deserve further exploration. AIM OF THE STUDY: This study observed the regulatory effects of GQD on Th2/Th1 and Tregs/Th17 cells balance, the NOD-like receptor family pyrin domain containing 3 (NLRP3) infammasome and gut microbiota in TNBS-induced UC in BALB/c mice. MATERIALS AND METHODS: 61 main chemical compounds in the GQD were determined by UPLC-Q-TOF/MS. The UC BALB/c model was established by intrarectal administration of trinitrobenzene sulfonic acid (TNBS), and GQD was orally administered at low and high dosages of 2.96 and 11.83 g/kg/day, respectively. The anti-inflammatory effects of GQD for ulcerative colitis were evaluated by survival rate, body weight, disease activity index (DAI) score, colonic weight and index, spleen index, hematoxylin-eosin (HE) staining and histopathological scores. Flow cytometry was used to detect the percentage of CD4, Th1, Th2, Th17 and Tregs cells. The levels of Th1-/Th2-/Th17-/Tregs-related inflammatory cytokines and additional proinflammatory cytokines (IL-1ß, IL-18) were detected by CBA, ELISA, and RT-PCR. The expressions of GATA3, T-bet, NLRP3, Caspase-1, IL-Iß, Occludin and Zonula occludens-1 (ZO-1) on colon tissues were detected by Western blot and RT-PCR. Transcriptome sequencing was performed using colon tissue and 16S rRNA gene sequencing was performed on intestinal contents. Fecal microbiota transplantation (FMT) was employed to assess the contribution of intestinal microbiota and its correlation with CD4 T cells and the NLRP3 inflammasome. RESULTS: GQD increased the survival rate of TNBS-induced UC in BALB/c mice, and significantly improved their body weight, DAI score, colonic weight and index, spleen index, and histological characteristics. The intestinal barrier dysfunction was repaired after GQD administration through promoting the expression of tight junction proteins (Occludin and ZO-1). GQD restored the balance of Th2/Th1 and Tregs/Th17 cells immune response of colitis mice, primarily inhibiting the increase in Th2/Th1 ratio and their transcription factor production (GATA3 and T-bet). Morever, GQD changed the secretion of Th1-/Th2-/Th17-/Tregs-related cytokines (IL-2, IL-12, IL-5, IL-13, IL-6, IL-10, and IL-17A) and reduced the expressions of IL-1ß, IL-18. Transcriptome results suggested that GQD could also remodel the immune inflammatory response of colitis by inhibiting NOD-like receptor signaling pathway, and Western blot, immunohistochemistry and RT-PCR further revealed that GQD exerted anti-inflammatory effects by inhibiting the NLRP3 inflammasome, such as down-regulating the expression of NLRP3, Caspase-1 and IL-1ß. More interestingly, GQD regulated gut microbiota dysbiosis, suppressed the overgrowth of conditional pathogenic gut bacteria like Helicobacter, Proteobacteria, and Mucispirillum, while the probiotic gut microbiota, such as Lactobacillus, Muribaculaceae, Ruminiclostridium_6, Akkermansia, and Ruminococcaceae_unclassified were increased. We further confirmed that GQD-treated gut microbiota was sufficient to relieve TNBS-induced colitis by FMT, involving the modulation of Th2/Th1 and Tregs/Th17 balance, inhibition of NLRP3 inflammasome activation, and enhancement of colonic barrier function. CONCLUSIONS: GQD might alleviate TNBS-induced UC via regulating Th2/Th1 and Tregs/Th17 cells Balance, inhibiting NLRP3 inflammasome and reshaping gut microbiota, which may provide a novel strategy for patients with colitis.

Electroacupuncture improving obesity-induced insulin resistance via regulating intestinal SIRT1/TLR4 signaling pathway. / ç”µé’ˆè°ƒæŽ§å°è‚ æ²‰é»˜ä¿¡æ¯è°ƒèŠ‚å› å­1/Tollæ ·å—ä½“4ç‚Žæ€§é€šè·¯æ”¹å–„è‚¥èƒ–å¤§é¼ èƒ°å²›ç´ æŠµæŠ—çš„æœºåˆ¶ç ”ç©¶.

OBJECTIVES: To observe the effect of electroacupuncture (EA) in obese rats with insulin resistance (IR) through regulating intestinal silent information regulator 1 (SIRT1)/Toll-like receptor 4 (TLR4) signaling pathway, so as to explore the underlying mechanism of EA in improving obesity-induced IR. METHODS: A total of 40 Wistar rats were randomly divided into 4 groups, i.e. normal group, model group, EA group and EA combined with inhibitor group, with 10 rats in each group. The obesity-induced IR model was induced by feeding high-fat diet for 8 weeks. EA (2 Hz, 1mA) was applied at "Zhongwan"(CV12), "Guanyuan"(CV4), "Zusanli"(ST36) and "Fenglong" (ST40) for 10 min, 3 times a week for 8 weeks in both EA and EA combined with inhibitor groups. Sirtinol, an inhibitor of SIRT1 was injected into the tail vein (1 mg/kg), 3 times a week for 8 weeks in EA combined with inhibitor group. The body weight, glucose infusion rate (GIR) of rats in each group were recorded. The contents of serum C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and lipopolysaccharide (LPS) were detected by ELISA. Mucosal morphological changes in the small intestine was observed by HE staining and was graded using Chiu's score. The protein relative expression levels of SIRT1 and TLR4 and the co-labeling of SIRT1 with TLR4 in the small intestine was detected by Western blot and double immunofluorescence staining, separately. RESULTS: Compared with the normal group, the body weight, serum contents of CRP, TNF-α, IL-6, LPS, Chiu's score, TLR4 protein relative expression level and percentage of TLR4 positive expression area were increased (P<0.01, P<0.05), while the GIR, SIRT1 protein expression, percentage of SIRT1 positive expression area and SIRT1/TLR4 were decreased (P<0.01) in the model group. The pathological injury of small intestine mucosa was severe, accompanied with inflammatory cell infiltration in the model group. Following interventions, the body weight, serum contents of CRP, TNF-α and LPS, Chiu's score, TLR4 protein relative expression level and percentage of TLR4 positive expression area were decreased(P<0.01, P<0.05), and the GIR was increased (P<0.01), the pathological injury and inflammatory cell infiltration of small intestine mucosa were reduced in both EA and EA combined with inhibitor groups in contrast to the model group. Compared with the model group, the serum IL-6 content was significantly decreased (P<0.01), and the SIRT1 protein relative expression level and percentage of positive expression area, SIRT1/TLR4 were increased (P<0.01, P<0.05) in the EA group. Compared with the EA group, EA combined with inhibitor group showed the body weight, serum CRP, IL-6, LPS, Chiu's score, TLR4 protein relative expression level and TLR4 positive expression area were increased (P<0.01, P<0.05), and the GIR level , SIRT1 relative expression level, SIRT1/TLR4 ratio were decreased (P<0.05, P<0.01). CONCLUSIONS: EA can reduce the body weight and ameliorate peripheral insulin sensitivity in IR obese rats, which may be related with its function in regulating intestinal SIRT1/TLR4 signaling pathway to reduce inflammatory response.

Mechanisms of gut microbiota-immune-host interaction on glucose regulation in type 2 diabetes.

Intestinal absorption of food is one of the sources of glucose. Insulin resistance and impaired glucose tolerance caused by lifestyle and diet are the precursors of type 2 diabetes. Patients with type 2 diabetes have trouble controlling their blood sugar levels. For long-term health, strict glycemic management is necessary. Although it is thought to be well correlated with metabolic diseases like obesity, insulin resistance, and diabetes, its molecular mechanism is still not completely understood. Disturbed microbiota triggers the gut immune response to reshape the gut homeostasis. This interaction not only maintains the dynamic changes of intestinal flora, but also preserves the integrity of the intestinal barrier. Meanwhile, the microbiota establishes a systemic multiorgan dialog on the gut-brain and gut-liver axes, intestinal absorption of a high-fat diet affects the host's feeding preference and systemic metabolism. Intervention in the gut microbiota can combat the decreased glucose tolerance and insulin sensitivity linked to metabolic diseases both centrally and peripherally. Moreover, the pharmacokinetics of oral hypoglycemic medications are also influenced by gut microbiota. The accumulation of drugs in the gut microbiota not only affects the drug efficacy, but also changes the composition and function of them, thus may help to explain individual therapeutic variances in pharmacological efficacy. Regulating gut microbiota through healthy dietary patterns or supplementing pro/prebiotics can provide guidance for lifestyle interventions in people with poor glycemic control. Traditional Chinese medicine can also be used as complementary medicine to effectively regulate intestinal homeostasis. Intestinal microbiota is becoming a new target against metabolic diseases, so more evidence is needed to elucidate the intricate microbiota-immune-host relationship, and explore the therapeutic potential of targeting intestinal microbiota.

Relationship between acupuncture and transient receptor potential vanilloid: Current and future directions.

Acupuncture is a common complementary and alternative therapy around the world, but its mechanism remains still unclear. In the past decade, some studies indicated that transient receptor potential vanilloid (TRPV) channels play a great role in the response of acupuncture stimulation. In this article, we discussed the relationship between acupuncture and TRPV channels. Different from inhibitors and agonists, the regulation of acupuncture on TRPV channels is multi-targeted and biphasic control. Acupuncture stimulation shows significant modulation on TRPV1 and TRPV4 at the autonomic nervous system (ANS) including central and peripheral nervous systems. On the contrary, the abundant expression and functional participation of TRPV1 and TRPV4 were specific to acupuncture stimulation at acupoints. The enhancement or inhibition of TRPV channels at different anatomical levels will affect the therapeutic effect of acupuncture. In conclusion, TRPV channels help to understand the principle of acupuncture stimulation, and acupuncture also provides a potential approach to TRPV-related trials.

[Mechanisms of electroacupuncture in improving obesity-induced insulin resistance via TLR4/NF-κB inflammatory pathway in liver].

OBJECTIVE: To investigate the effect of electroacupuncture (EA) on the Toll-like receptor 4 (TLR4)/nuclear factor kappa B (NF-κB) inflammatory pathway in the liver of obese rats with insulin resistance, and explore its mechanism. METHODS: Male Wistar rats were randomly divided into a normal group (n=15) and an experimental group (n=30). The obesity-induced insulin resistance model was induced by the high-fat diet (HFD) in rats of the experimental group for 8 weeks. Subsequently, the model rats were further divided into a model group (n=15) and an EA group (n=15). EA was applied at "Zhongwan "(CV12), "Guanyuan" (CV4), "Zusanli "(ST36) and "Fenglong "(ST40) in the EA group for 10 min, three times a week for 8 weeks. The body weight of rats in each group was measured before intervention and at the 2nd, 4th, 6th, and 8th weeks during the intervention. Glucose infusion rate (GIR) was measured by glucose clamp test before and after treatment. After treatment, fast blood glucose (FBG) was detected by the glucometer, and homeostasis model assessment of insulin resistance (HOMA-IR) was calculated. The contents of fasting insulin (FINS), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) were determined by ELISA. The protein expressions of TLR4, IκB kinase ß (IKKß), phosphorylated IKKß (p-IKKß), NF-κB p65, and TNF-α related to the TLR4/NF-κB signaling pathway in the liver of rats were detected by Western blot. RESULTS: Compared with the normal group, the body weight, HOMA-IR levels, serum levels of FINS, TNF-α, and IL-6 were up-regulated (P<0.01), and the GIR level was down-regulated (P<0.01), the protein expressions of TLR4, IKKß, p-IKKß, NF-κB p65 and TNF-α in liver tissues were increased(P<0.05) in the model group. Compared with the model group, the EA group showed weight loss from the 6th week, and the HOMA-IR levels，serum levels of FINS, TNF-α, and IL-6 were decreased(P<0.01, P<0.05), the GIR level was up-regulated (P<0.01), the protein expressions of TLR4, IKKß, p-IKKß, NF-κB p65 and TNF-α in liver tissues were down-regulated (P<0.05). CONCLUSION: EA can reduce the inflammatory response and improve peripheral insulin sensitivity by inhibiting the TLR4/NF-κB pathway in liver tissues of obese rats with insulin resistance, showing a good regulatory effect on insulin resistance induced by obesity.

Electroacupuncture Pretreatment Exhibits Lung Protective and Anti-Inflammation Effects in Lipopolysaccharide-Induced Acute Lung Injury via SIRT1-Dependent Pathways.

To investigate the effect of electroacupuncture (EA) on acute lung injury (ALI), a lipopolysaccharide (LPS) induced ALI mouse model was used in this study. Before receiving intratracheal LPS instillation, mice were given EA at ST36 for 7 days as a long-term treatment or one time as a short-term treatment. Lung histopathological examination, lung injury scores, lung wet/dry (W/D) ratio, and inflammatory cytokines included proinflammation factors such as TNF-α, IL-1ß, and IL-6 and anti-inflammation factors such as IL-4 and IL-10 in serum and bronchoalveolar lavage fluid (BALF) were detected at the end of experiment. The results show that EA pretreatment ameliorated the lung damage and inflammatory response by LPS. In addition, we found that SIRT1 and its deacetylation of NF-κB were promoted after EA pretreatment in lung tissues. Meanwhile, the expression of angiotensin-converting enzyme 2 (ACE2) is also enhanced by EA pretreatment. Thus, the present findings suggest that EA could be a potential therapy of ALI.

[Effect of electroacupuncture at different acupoint combination on intestinal inflammatory response and intestinal flora in obese rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) at different acupoint combination on intestinal inflammatory response, intestinal flora structure and metabolic function in obese rats. METHODS: Ninety male Wistar rats aged 8 weeks were collected. Ten rats were randomly collected from 15 rats fed with regular forage in a normal group and the rest 75 rats were fed with high-fat forage to induce obesity models. Forty rats were modeled successfully and randomized into a model group, a lower-limb EA group, an abdomen EA group and a biaoben acupoints group, 10 rats in each one. "Zusanli" (ST 36) and "Fenglong" (ST 40) were selected in the lower-limb EA group, "Zhongwan "(CV 12), "Tianshu "(ST 25) and "Guanyuan" (CV 4) were selected in the abdomen EA group. The acupoint prescriptions in the above two groups were combined in the biaoben acupoints group. EA was delivered in all of the intervention groups, with continuous wave, 2 Hz in frequency and 1 mA in current intensity. The intervention was administered three times weekly (on Monday, Wednesday and Friday), for consecutive 8 weeks. Before intervention and on the last day of the 8th week of intervention, the body weight and 24 h food intake were observed. After intervention, using Western blot method, the proteins expressions of interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) were detected in the tissue of the small intestine; 16S rRNA sequencing technology was adopted to detect the distribution structure and metabolic function of intestinal flora. RESULTS: Compared with the normal group, the body weight, the food intake and the proteins expressions of IL-6 and TNF-α in small intestine were all increased in the model group (P<0.01). The above indexes of each EA intervention group were all decreased (P<0.01) when compared with the model group. The proteins expressions of IL-6 and TNF-α in the small intestine tissue of rats in biaoben acupoints group were lower than those in the other two EA intervention groups (P<0.01). Compared with the normal group, the ratio of Firmicutes/Bacteroidetes was elevated (P<0.01), while the abundance of Lactobacillus, Muribaculaceae and Bifidobacteria decreased in the model group (P<0.01). When compared with the model group, the ratio of Firmicutes/Bacteroidetes in each EA intervention group was reduced (P<0.01) and the abundance of Lactobacillus, Bifidobacteria and Bacteroidetes increased (P<0.01). Compared with the other two EA intervention groups, the abundance of Lactobacillus and Muribaculaceae was increased (P<0.01), while the abundance of Collinsella and Ruminococcus Gauvreauii reduced (P<0.01) in the biaoben acupoints group. In the model group, the abundance of clusters of orthologous groups of proteins (COG) function of intestinal flora in the transportation and metabolism of carbohydrate, amino acid and lipid, as well as in the signal transduction mechanisms were reduced when compared with the normal group (P<0.01). In comparison with the model group, the abundance of the above COG function was increased in each EA intervention group (P<0.01, P<0.05). CONCLUSION: Electroacupuncture at biaoben acupoint combination may attenuate intestinal inflammatory response and effectively improve the structure and function of intestinal flora. The effect is superior to the intervention at the acupoints on the lower limbs and those on the abdomen, better regulating the abundance of specific intestinal flora.

[Effect of electroacupuncture stimulation of different acupoint groups on lipid metabolism and liver TLR4/NF-κB signaling in obese rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of different acupoint combinations on lipid metabolism and liver Toll-like receptor 4 (TLR4)/nuclear factor kappa B (NF-κB) signaling in obese rats, so as to explore the specificity of regulatory effects of different acupoints for obesity. METHODS: Male SD rats were randomly divided into normal control, model, hindlimb acupoint EA ï¼»"Fenglong" (ST40)-"Zusanli" (ST36), hindlimb EAï¼½, abdominal acupoint EA ï¼»("Zhongwan" (CV12)-"Tianshu" (ST25), "Guanyuan" (CV4) - the opposite ST25, abdominal EAï¼½, BiaoBen-acupoint EA ï¼»ST40-ST36, CV12 -CV4, ST25, BiaoBen EAï¼½ groups, with 10 rats in each group. The obesity model was induced by feeding the rats with high fat diet. EA (2 Hz, 1 mA) was applied to the acupoint groups mentioned above for 10 min (per acupoint group), 3 times a week for 8 weeks. After the treatment, the body weight and food intake in each group were recorded. The contents of serum total cholesterol (TC), triglyceride (TG), non-esterified fatty acid (NEFA) were detected by using an automatic biochemical analyzer. The expression levels of TLR4 and NF-κB p65 mRNAs and proteins in the liver tissue were detected by quantitative real-time PCR and Western blot, separately. RESULTS: Compared with the normal group, the body weight and food intake, serum TC, TG and NEFA contents, hepatic TLR4 and NF-κB p65 protein and gene expression were significantly increased in the model group (P<0.05, P<0.01). After the treatment, compared with the model group, the body weight, food intake, TC, TG, NEFA, TLR4 and NF-κB p65 protein and mRNA expression were significantly decreased in the hindlimb EA, abdominal EA and BiaoBen EA groups (P<0.05, P<0.01). Comparison among the 3 treatment groups showed that the serum TC, TG and NEFA contents were significantly lower in the hindlimb EA and BiaoBen EA groups than in the abdominal EA group (P<0.01，P<0.05). The expression levels of TLR4 and NF-κB p65 mRNAs and proteins were significantly lower in the BiaoBen EA group than in the hindlimb EA and abdominal EA groups (P<0.01, P<0.05). CONCLUSION: BiaoBen EA is superior to abdominal EA in improving lipid metabolism in obesity rats, which may be related to its stronger effectiveness in down-regulating hepatic TLR4/NF-κB p65 signaling.

[Mechanism of electroacupuncture for regulation of lipid production and improvement in obesity by mediating Wnt/ ß-catenin pathway through activating SIRT1].

OBJECTIVE: To explore the mechanism of electroacupuncture (EA) for the regulation of lipid production and improvement in obesity by mediating Wnt/ß-catenin pathway through activating silent information regulator 1 (SIRT1). METHODS: Of 75 Wistar male rats, 10 rats were selected randomly as the normal group and fed with standard diet. The rest rats were fed with high-fat diet for 8 weeks to establish the obesity model. Forty rats of successful modeling were randomized into a model group, an EA group, an EA plus inhibitor group (EA+I group) and an agonist group, 10 rats in each one. In the EA group, EA was applied at "Guanyuan" (CV 4), "Zhongwan" (CV 12), "Zusanli" (ST 36) and "Fenglong" (ST 40), with continuous wave, 2 Hz in frequency and around 1 mA in intensity. The needles were retained for 20 min. In the EA+I group, sirtinol solution was injected from caudal vein and EA was exerted simultaneously. In the agonist group, resveratrol solution was given by intragastric administration. The intervention of the above three groups was given once every two days, 3 times a week, consecutively for 8 weeks. Before and after intervention, body mass and Lee's index were recorded in the rats of each group. After intervention, the levels of serum total cholesterol (TC), triglyceride (TG) and free fatty acid (FFA) were detected in the rats of each group. After intervention, the mass of white adipose tissue (WAT) and the area of adipocytes were compared in the rats among the 5 groups. Using Western blot method, the protein expressions of SIRT1, glycogen synthase kinase-3ß (GSK3ß), ß-catenin, cyclin D1 and peroxisome proliferators-activated receptor γ (PPARγ) were detected in WAT in the rats of each group. RESULTS: After intervention, compared with the model group, the body mass and Lee's index were reduced in the rats of the EA group and the agonist group (P<0.01, P<0.05), the body mass was reduced in the rats of the EA+I group (P<0.05). Compared with the normal group, the levels of serum TC, TG and FFA, as well as WAT mass were increased in the rats of the model group (P<0.01), as well as the area of adipocytes (P<0.01). Compared with the model group, the levels of serum TC and TG (except in the EA+I group), the levels of FFA and WAT mass were all decreased (P<0.01, P<0.05) and the area of adipocytes was reduced (P<0.01, P<0.05) in the EA group, the agonist group and the EA+I group. Compared with the EA group, the area of adipocytes was increased in the EA+I group (P<0.05). Compared with the normal group, the protein expressions of SIRT1, ß-catenin and cyclin D1 in WAT were down-regulated (P<0.01) and the protein expressions of GSK3ß and PPARγ in WAT were up-regulated (P<0.01) in the model group. Compared with the model group, the protein expressions of SIRT1, ß-catenin and cyclin D1 in WAT were up-regulated (P<0.05, P<0.01) and the expressions of GSK3ß and PPARγ in WAT were down-regulated (P<0.01, P<0.05) in the EA group and the agonist group, and in the EA+I group, GSK3ß protein expression was down-regulated andß-catenin protein expression was up-regulated (P<0.05). CONCLUSION: Electroacupuncture remarkably improves the body mass, Lee's index and blood lipid metabolism and reduces WAT mass and adipocyte size in obesity model rats, which is probably related to up-regulating the protein expression of SIRT1 in WAT, activating Wnt/ß-catenin pathway and inhibiting the expression of PPARγ of downstream lipogenic gene so as to affect lipid production.

Ketone Body 3-Hydroxybutyrate Ameliorates Atherosclerosis via Receptor Gpr109a-Mediated Calcium Influx.

Atherosclerosis is a chronic inflammatory disease that can cause acute cardiovascular events. Activation of the NOD-like receptor family, pyrin domain containing protein 3 (NLRP3) inflammasome enhances atherogenesis, which links lipid metabolism to sterile inflammation. This study examines the impact of an endogenous metabolite, namely ketone body 3-hydroxybutyrate (3-HB), on a mouse model of atherosclerosis. It is found that daily oral administration of 3-HB can significantly ameliorate atherosclerosis. Mechanistically, 3-HB is found to reduce the M1 macrophage proportion and promote cholesterol efflux by acting on macrophages through its receptor G-protein-coupled receptor 109a (Gpr109a). 3-HB-Gpr109a signaling promotes extracellular calcium (Ca2+) influx. The elevation of intracellular Ca2+ level reduces the release of Ca2+ from the endothelium reticulum (ER) to mitochondria, thus inhibits ER stress triggered by ER Ca2+ store depletion. As NLRP3 inflammasome can be activated by ER stress, 3-HB can inhibit the activation of NLRP3 inflammasome, which triggers the increase of M1 macrophage proportion and the inhibition of cholesterol efflux. It is concluded that daily nutritional supplementation of 3-HB attenuates atherosclerosis in mice.

Gegen Qinlian Decoction Ameliorates Nonalcoholic Fatty Liver Disease in Rats via Oxidative Stress, Inflammation, and the NLRP3 Signal Axis.

Gegen Qinlian Decoction (GQD), a classic Chinese herbal formula, has been widely used in Chinese clinic for centuries and is well defined in treating nonalcoholic fatty liver disease (NAFLD). However, the mechanism action of GQD on NAFLD is still rarely evaluated. The present study aims to investigate the effect of GQD on treatment of NAFLD in rats and to further explore the underlying mechanism. The rat NAFLD model established by high-fat-diet feeding was used in the research. Our results exhibited the liver lesions and steatosis was significantly alleviated in NAFLD rats treated with GQD via Oil Red O and H&E staining. Body weight and liver index in GQD groups were reduced significantly (P < 0.05). Moreover, the biochemical analyzer test results showed that GQD significantly decreased blood lipid levels total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and liver injury indicators alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP), while it increased the level of high-density lipoprotein cholesterol (HDL-C) (P < 0.05). The levels of interferon-ß (IFN-ß), tumor necrosis factor-α (TNF-α), and malondialdehyde (MDA) after the GQD treatment were significantly lower, and then interleukin-2 (IL-2), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) levels were lifted significantly (P < 0.05). Further, GQD blocked the expression of NLRP3, ASC, caspase-1 mRNA, and proteins in the liver tissues significantly (P < 0.05). These findings indicated that GQD can ameliorate the hepatic steatosis and injury of NAFLD. Its possible mechanism involves the modulation of inflammatory cytokines and antioxidative stress and the inhibition of NLRP3 signal axis activation. The results support that GQD may be a promising candidate in the treatment of NAFLD.

Mechanism of electroacupuncture for regulation of lipid production and improvement in obesity by mediating Wnt/ β-catenin pathway through activating SIRT1 / 中国针灸

OBJECTIVE@#To explore the mechanism of electroacupuncture (EA) for the regulation of lipid production and improvement in obesity by mediating Wnt/β-catenin pathway through activating silent information regulator 1 (SIRT1).@*METHODS@#Of 75 Wistar male rats, 10 rats were selected randomly as the normal group and fed with standard diet. The rest rats were fed with high-fat diet for 8 weeks to establish the obesity model. Forty rats of successful modeling were randomized into a model group, an EA group, an EA plus inhibitor group (EA+I group) and an agonist group, 10 rats in each one. In the EA group, EA was applied at "Guanyuan" (CV 4), "Zhongwan" (CV 12), "Zusanli" (ST 36) and "Fenglong" (ST 40), with continuous wave, 2 Hz in frequency and around 1 mA in intensity. The needles were retained for 20 min. In the EA+I group, sirtinol solution was injected from caudal vein and EA was exerted simultaneously. In the agonist group, resveratrol solution was given by intragastric administration. The intervention of the above three groups was given once every two days, 3 times a week, consecutively for 8 weeks. Before and after intervention, body mass and Lee's index were recorded in the rats of each group. After intervention, the levels of serum total cholesterol (TC), triglyceride (TG) and free fatty acid (FFA) were detected in the rats of each group. After intervention, the mass of white adipose tissue (WAT) and the area of adipocytes were compared in the rats among the 5 groups. Using Western blot method, the protein expressions of SIRT1, glycogen synthase kinase-3β (GSK3β), β-catenin, cyclin D1 and peroxisome proliferators-activated receptor γ (PPARγ) were detected in WAT in the rats of each group.@*RESULTS@#After intervention, compared with the model group, the body mass and Lee's index were reduced in the rats of the EA group and the agonist group (@*CONCLUSION@#Electroacupuncture remarkably improves the body mass, Lee's index and blood lipid metabolism and reduces WAT mass and adipocyte size in obesity model rats, which is probably related to up-regulating the protein expression of SIRT1 in WAT, activating Wnt/β-catenin pathway and inhibiting the expression of PPARγ of downstream lipogenic gene so as to affect lipid production.

[Electroacupuncture improves obesity by lowering gastrointestinal motility, blood lipids and expression of intestinal leptin and cholecystokinin in obese rats].

OBJECTIVE: To investigate the effect of electroacupuncture (EA) on gastrointestinal motility and expression of leptin(LEP) and cholecystokinin(CCK) in the small intestine in obese rats，so as to explore the mechanism of EA underlying improvement of obesity. METHODS: Male Wistar rats were randomized into 5 groups: normal control, obesity model, abdominal acupoints ［"Guanyuan" (CV4), "Zhongwan" (CV12) and bilateral "Tianshu" (ST25)］, lower-leg acupoints ［bilateral "Zusanli" (ST36) and bilateral "Fenglong" (ST40)］, and abdominal+ lower-leg acupoints (n=10 rats in each group). The obesity model was established by feeding the animals with high-fat diet for 8 weeks. EA was applied to the abovementioned acupoints for 20 min every time, 3 times a week for 8 weeks. The food intake and body mass were recorded. The white adipose tissue around the testicles and in the abdominal region was weighed. The serum total cholesterol (TC), triglyceride (TG) and non-esterified fatty acid(NEFA) were detected by using automatic biochemical analyzer. The gastric empty rate and intestinal propulsive rate were calculated. The contents of serum CCK and LEP were detected by using ELISA, and the expression levels of CCK and LEP proteins in the small intestine were detected by using Western blot. RESULTS: Following modeling, the food intake, body mass, weight of white adipose around the testicles and abdomen, the gastric empty rate, and serum TC, TG, NEFA and LEP contents as well as intestinal LEP expression were significantly increased (P<0.05, P<0.01), while the intestinal propulsive rate, serum CCK content and intestinal CCK expression were evidently decreased (P<0.01) in the model group relevant to the normal control group. After EA interventions and compared with the model group, the increased levels of food intake, body mass, white adipose weight, gastric empty rate, TC, TG, and LEP in serum and small intestine, and the decreased levels of intestinal propulsive rate, CCK in the serum and intestine were reversed in the abdominal acupoints, lower-leg acupoints and abdominal+lower-leg acupoints groups (P<0.05, P<0.01). No significant differences were found in the effects of the three EA groups in down-regulating food intake, body mass, white adipose weight, gastric empty rate, serum TC, TG and LEP levels (except NEFA) and in up-regulating intestinal propulsive rate and CCK level (P>0.05). CONCLUSION: EA stimulation of the abdominal and lower-leg acupoints or both can reduce body weight on obesity rats, which is associated with its functions in regulating intestinal motility, food intake, and secretion of LEP and CCK.

The Prognostic Value of Autophagy-Related Markers Bclin-1 and LC-3 in Colorectal Cancers: A Systematic Review and Meta-analysis.

OBJECTIVE: At present, the relationship between autophagosomes and the prognosis of various cancers has become a subject of active investigation. A series of studies have demonstrated the correlation between autophagy microtubule-associated protein light chain 3 (LC-3), Beclin-1, and colorectal cancer (CRC). Since autophagy has dual regulatory roles in tumors, the results of this correlation are also uncertain. Hence, we summarized the relationship between Beclin-1, LC-3, and CRC using systematic reviews and meta-analysis to clarify their prognostic significance in it. METHODS: PubMed, EMBASE, Cochrane Library, and Web of Science databases were searched online up to April 1, 2019. The quality of the involving studies was assessed against the Newcastle-Ottawa Scale (NOS). Pooled hazard ratio (HR) and 95% confidence interval (CI) in a fixed or random effects model were used to assess the strength of correlation between Beclin-1, LC-3, and CRC. RESULTS: A total of 9 articles were collected, involving 2,297 patients. Most literatures scored more than 6 points, suggesting that the quality of our including research was acceptable. Our finding suggested that the expression of Beclin-1 was not associated with overall survival (HR = 0.68, 95% CI (0.31-1.52), P=0.351). Nonetheless, LC-3 expression exerted significant impact on OS (HR = 0.51, 95% CI (0.35-0.74), P < 0.05). Subgroup analysis exhibited that Beclin-1 expression was associated with OS at TNM stage III (HR = 0.04, 95% CI = 0.02-0.08, P < 0.05), surgical treatment (HR = 1.53, 95% CI (1.15-2.02), P=0.003), and comprehensive treatment (HR = 0.27 95% CI (0.08-0.92), P=0.036), respectively. Similarly, the results showed the increased LC-3 expression in CRC was related to OS in multivariate analyses (HR = 0.44, 95% CI (0.34-0.57), P < 0.05), stages (HR = 0.51, 95% CI (0.35-0.74), P < 0.05), and comprehensive treatment (HR = 0.44, 95% CI (0.34-0.57), P < 0.05). CONCLUSIONS: Autophagy-related proteins of LC-3 might be an important marker of CRC progression. However, since the number of the original studies was limited, more well-designed, large-scale, high-quality studies are warranted to provide more convincing and reliable information.

[Relationship between vegetation C, N, P stoichiometry and species diversity in sand land.] / æ²™åœ°æ¤è¢«ç¢³æ°®ç£·åŒ–å­¦è®¡é‡ç‰¹å¾ä¸Žç‰©ç§å¤šæ ·æ€§çš„å ³ç³».

Vegetation stoichiometry represents plant nutrition limitation status. Whether it can affect the species diversity of plant community remains to be explored. In this study, we examined plant carbon (C), nitrogen (N), phosphorus (P) concentrations and stoichiometry, and further analyzed the relationship between stoichiometry and species diversity in Artemisia ordosica community and Salix psammophila community in sand land of the Ningxia Habahu National Nature Reserve. The results showed that, for the S. psammophila community growing in mobile and semi-fixed sand dunes, there was a significant negative correlation between Simpson index and vegetation C/N ratio, but no significant correlation between Simpson index and vegetation N/P ratio. For A. ordosica community distributed in semi-fixed and fixed dunes, the Shannon index had a significant positive correlation with vegetation N/P ratio and a significant negative correlation with vegetation C/N ratio. Combined the vegetation stoichiometry with the results of regression analysis (RDA), P concentration and thus N/P ratio had different influences on the species diversity for A. ordosica community and S. psammophila community. Our results showed that vegetation stoichiometry has an important influence on species diversity of plant community in sand land.

Androgen Deprivation Accelerates the Prostatic Urethra Wound Healing After Thulium Laser Resection of the Prostate by Promoting Re-Epithelialization and Regulating the Macrophage Polarization.

BACKGROUND: Complications after a thulium laser resection of the prostate (TmLRP) are related to re-epithelialization of the prostatic urethra. Since prostate growth and development are induced by androgen, the aim of this study was to determine the role and explore the mechanism of androgen in wound healing of the prostatic urethra. METHODS: Beagles that received TmLRPs were randomly distributed into a castration group, a testosterone undecanoate (TU) group, and a control group. The prostate wound was assessed once a week using a cystoscope. Histological analysis was then carried out to study the re-epithelialization of the prostatic urethra in each group. The inflammatory response in the wound tissue and urine was also investigated. RESULTS: The healing of the prostatic urethra after a TmLRP was more rapid in the castration group and slower in the TU group than that in the control group. Castration accelerated re-epithelialization by promoting basal cell proliferation in the wound surface and beneath the wound and by accelerating the differentiation of basal cells into urothelial cells. Castration reduced the duration of the inflammatory phase and induced the conversion of M1 macrophages to M2 macrophages, thus accelerating the maturation of the wound. By contrast, androgen supplementation enhanced the inflammatory response and prolonged the inflammatory phase. Moreover, the anti-inflammatory phase was delayed and weakened. CONCLUSION: Androgen deprivation promotes re-epithelialization of the wound, regulates the inflammatory response, and accelerates wound healing of the prostatic urethra after a TmLRP. Prostate 77:708-717, 2017. © 2017 Wiley Periodicals, Inc.

[Impact of CCND1 A870G polymorphism on acute adverse events in postoperative rectal cancer patients treated with adjuvant concurrent chemoradiotherapy].

OBJECTIVE: The purpose of this study was to investigate the association between single nucleotide polymorphism (SNP) of CCND1 A870G and acute adverse events (AEs) in postoperative rectal cancer patients who received capecitabine-based postoperative chemoradiotherapy (CRT). METHODS: Four hundred patients with stage II and III rectal cancer received postoperative CRT of capecitabine with or without oxaliplatin were accumulated and prostectively studied in this study. The patients were randomly divided into two groups. Two hundred and twenty-eight patients were treated with concurrent capecitabine and radiotherapy (Cap-CRT), and 172 patients were treated with capecitabine and oxaliplatin plus radiotherapy (Cap-Oxa-CRT). Adverse events were graded according to the Common Terminology Criteria for Adverse Events, v. 3.0 (CTCAE v3.0). The genotype of CCND1 A870G in the patients was detected by polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) analysis. The associations between the SNP and acute AEs were indicated by odds ratios (ORs) and 95% confidence intervals (CIs), which were computed with logistic regression model. RESULTS: A total of 136 patients presented severe AEs. Among them the frequencies of the three genotypes GG, GA and AA were 16.9%, 50.7% and 32.4%, compared with 24.6%, 48.1% and 27.3%, respectively, among the patients without severe AEs. Diarrhea was the most common AE, and severe diarrhea occurred in 109 patients. The frequencies of the three genotypes GG, GA and AA were 15.6%, 47.7% and 36.7% among these patients, compared with 24.4%, 49.5% and 26.1%, respectively, among patients without severe diarrhea. Multivariate logistic regression analysis showed a 1.66-fold increased risk for severe diarrhea in patients with AA genotype (95%CI 1.03 - 2.67, P = 0.038) compared with the cases with GG or GA genotypes. Stratified analysis showed that in the Cap-Oxa-CRT group, patients with AA genotype showed a 2.34-fold increased risk for severe diarrhea (95%CI 1.16 - 4.76, P = 0.018) compared with those with GG or GA genotypes, but in the Cap-CRT group, the SNP was not associated with the risk of severe diarrhea. CONCLUSIONS: The genetic polymorphism of CCND1 A870G might be a potential biomarker for predicting acute AEs in postoperative stage II and III rectal cancer patients treated with adjuvant concurrent chemoradiotherapy of capecitabine and oxaliplatin.