Xiaoqinglong Decoction Enhances Autophagy to Antagonist Airway Inflammation Induced by Cold in Asthmatic Rats.

Asthma is a common chronic respiratory disease characterized by wheezing and shortness of breath. Its risk factors include genetic and acquired factors. The acquired factors are closely related to the environment, especially cold conditions. Autophagy plays a regulatory role in asthma. Therefore, we hypothesized that asthma can be controlled by drug intervention at the autophagy level under cold conditions. The Xiaoqinglong decoction (XQLT) was freeze-dried. The compounds in the freeze-dried powder were identified and quantified using reference standards via the high-performance liquid chromatography method. Ovalbumin (OVA)-sensitized rats were subjected to cold stimulation. The effect of cold stimulation on autophagy levels was determined, and it was confirmed that cold stimulation affected autophagy. The effects and mechanisms of XQLT in an asthmatic rat model (OVA-sensitized rats stimulated with cold) were explored. The concentrations of paeoniflorin, liquiritin, trans-cinnamic acid, glycyrrhizic acid, 6-gingerol, schisandrol A, and asarinin in XQLT freeze-dried powder were 14.45, 3.85, 1.03, 3.93, 0.59, 0.24, and 0.091 mg/g, respectively. Cold stimulation is an important cause of asthma. The inflammatory factors in bronchoalveolar lavage fluid and serum were increased in the model group, accompanied by a decline in autophagy level. The treatment with XQLT increased the expression of autophagy genes and decreased the expression of inflammatory factors. Histological studies showed that XQLT improved inflammatory infiltration and collagen fiber deposition in the lungs of rats. XQLT intervention increased autophagy in asthmatic rats. Autophagy plays a role in phagocytosis and reduces the accumulation of abnormal metabolites in the body to reduce airway inflammation and promote asthma recovery.

SIRT5-Related Desuccinylation Modification Contributes to Quercetin-Induced Protection against Heart Failure and High-Glucose-Prompted Cardiomyocytes Injured through Regulation of Mitochondrial Quality Surveillance.

Myocardial fibrosis represents the primary pathological change associated with diabetic cardiomyopathy and heart failure, and it leads to decreased myocardial compliance with impaired cardiac diastolic and systolic function. Quercetin, an active ingredient in various medicinal plants, exerts therapeutic effects against cardiovascular diseases. Here, we investigate whether SIRT5- and IDH2-related desuccinylation is involved in the underlying mechanism of myocardial fibrosis in heart failure while exploring related therapeutic drugs for mitochondrial quality surveillance. Mouse models of myocardial fibrosis and heart failure, established by transverse aortic constriction (TAC), were administered with quercetin (50 mg/kg) daily for 4 weeks. HL-1 cells were pretreated with quercetin and treated with high glucose (30 mM) in vitro. Cardiac function, western blotting, quantitative PCR, enzyme-linked immunosorbent assay, and immunofluorescence analysis were employed to analyze mitochondrial quality surveillance, oxidative stress, and inflammatory response in myocardial cells, whereas IDH2 succinylation levels were detected using immunoprecipitation. Myocardial fibrosis and heart failure incidence increased after TAC, with abnormal cardiac ejection function. Following high-glucose treatment, HL-1 cell activity was inhibited, causing excess production of reactive oxygen species and inhibition of mitochondrial respiratory complex I/III activity and mitochondrial antioxidant enzyme activity, as well as increased oxidative stress and inflammatory response, imbalanced mitochondrial quality surveillance and homeostasis, and increased apoptosis. Quercetin inhibited myocardial fibrosis and improved cardiac function by increasing mitochondrial energy metabolism and regulating mitochondrial fusion/fission and mitochondrial biosynthesis while inhibiting the inflammatory response and oxidative stress injury. Additionally, TAC inhibited SIRT5 expression at the mitochondrial level and increased IDH2 succinylation. However, quercetin promoted the desuccinylation of IDH2 by increasing SIRT5 expression. Moreover, treatment with si-SIRT5 abolished the protective effect of quercetin on cell viability. Hence, quercetin may promote the desuccinylation of IDH2 through SIRT5, maintain mitochondrial homeostasis, protect mouse cardiomyocytes under inflammatory conditions, and improve myocardial fibrosis, thereby reducing the incidence of heart failure.

Efficacy of Shenling Baizhu San on stable chronic obstructive pulmonary disease patients: A systematic review and meta-analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Shenling Baizhu San (SBS) as a classic Chinese medicine prescription, has been extensively used in gastrointestinal diseases, such as ulcerative colitis and chronic diarrhea. In recent years, SBS has shown a beneficial effect on chronic obstructive pulmonary disease (COPD) patients. However, clinical trials had shown conflicting results of SBS on improving pulmonary function and other related indicators of patients with stable COPD. The efficacy of SBS on stable COPD patients has not been fully assessed. AIM OF THE STUDY: To determine whether the SBS used in the treatment of gastrointestinal disease was effective to treat COPD, we assessed the clinical evidence and efficacy of SBS supplemental treatment on stable COPD patients by a systematic review and meta-analysis of clinical trials. MATERIALS AND METHODS: Nine electronic databases were searched to include clinical trials (published until August 31, 2020) with SBS as a supplementation treatment on stable COPD. Mean difference (MD) was used to evaluate continuous variables, odds ratio (OR) was calculated to evaluate dichotomous. The Egger's test was applied for publication bias. RESULTS: A total of 770 COPD participants from 11 trials that met the inclusion criteria were included. The meta-analysis showed that modified SBS could improve the exercise endurance, life quality scores of stable COPD patients, and also showed the potential benefits to pulmonary function of COPD patients than original SBS. CONCLUSION: The methodological quality of included trials may limit the conclusions that indicate that modified SBS may have a promising treatment for improving FEV1/FVC and MVV, increasing exercise endurance and life quality scores on stable COPD patients.

Analysis of alkaloids from Peganum harmala L. sequential extracts by liquid chromatography coupled to ion mobility spectrometry.

An orthogonal two dimensional analysis method based on high performance liquid chromatography (HPLC) separation and electrospray ionization-ion mobility spectrometry (ESI-IMS) detection was developed for the analysis of alkaloid compounds from Peganum harmala L. seeds. Reverse phase (RP) and hydrophilic interaction chromatography (HILIC) were compared for the most optimal performance using three different chromatographic columns. The experimental results suggest that HILIC mode is a better option for combining with the ESI-IMS system for higher sensitivity and ease in hyphenating. Under optimized conditions, alkaloids from different extraction phases were determined by means of the established HPLC-IMS method. More compounds from Peganum harmala L. seed extracts were differentiated on the HPLC-ESI-IMS system by their retention time and drift time than by HPLC or ESI-IMS alone, and thirteen alkaloids were tentatively identified based on m/z and fragment ions using ultra-high-performance liquid chromatography tandem mass-spectrometry (UPLC-MS/MS). Hence, our results indicate that this method can be considered to be advantageous over traditional absorbance detection methods for resolving complex mixtures because of complementary separation steps, elevated peak capacity, and higher sensitivity.

Effect of parenteral glutamine supplementation combined with enteral nutrition on Hsp90 expression and Peyer's patch apoptosis in severely burned rats.

OBJECTIVES: The aim of this study was to investigate the effects of parenteral glutamine (GLN) supplementation combined with enteral nutrition (EN) on heat shock protein (Hsp) 90 expression and Peyer's patch (PP) apoptosis in severely burned rats. METHODS: Male Sprague-Dawley (SD) rats were randomly assigned to four groups: Sham burn + EN + GLN-free amino acid (AA; n = 10), sham burn + EN + GLN (n = 10), burn + EN + AA (n = 10), and burn + EN + GLN (n = 10). Two hours after a 30% total body surface area (TBSA), full-thickness scald burn injury on the back, burned rats in two of the experimental groups (burn + EN + AA and burn + EN + GLN groups) were fed with a conventional EN solution by oral gavage for 7 d. Simultaneously, rats in the burn + EN + GLN group were given 0.35 g GLN/kg body weight/d once via a tail vein injection for 7 d and rats in the burn + EN + AA group were administered isocaloric/isonitrogenous GLN-free amino acid solution (Tyrosine) for comparison. Rats in two sham burn control groups (sham burn + EN + AA and sham burn + EN + GLN groups) were treated in the same manner except for the burn injury. All rats in the four groups were given 175 kcal/kg body wt/d. There was isonitrogenous, isovolumic, and isocaloric intake among the four groups. At the end of the seventh day after completion of the nutritional program, all rats were anesthetized and samples were collected for further analysis. PP apoptosis was measured by terminal deoxyuridine nick-end labeling (TUNEL). The expression of Hsp90 in PPs was analyzed by western blotting. Caspase-3 activity of PPs was also assessed. Levels of proinflammatory cytokines of gut tissues were evaluated by enzyme-linked immunosorbent assay (ELISA). The intestinal immunoglobulin A (IgA) content was also determined by ELISA. RESULTS: The results revealed that intestinal IgA content in rats of the burn + EN + GLN group were significantly increased compared with those in the burn + EN + AA group (P < 0.05). The expression of Hsp90 of PPs in rats in the burn + EN + GLN group was significantly upregulated compared with those in the burn + EN + AA group (P < 0.05). On the other hand, levels of proinflammatory cytokines of gut tissues, caspase-3 activity, and the number of TUNEL-stained cells of PPs in rats of the burn + EN + GLN group were markedly decreased compared with those of the burn + EN + AA group (P < 0.05). CONCLUSIONS: The results of this study show that parenteral glutamine supplementation combined with EN may upregulate the expression of Hsp90, reduce caspase-3 activity, lessen the release of proinflammatory cytokines, attenuate PP apoptosis, and improve intestinal IgA response in burned rats. Clinically, therapeutic efforts to improve intestinal immunity may contribute to a favorable outcome in severely burned patients.

Effect of parenteral glutamine supplementation combined with enteral nutrition on Hsp90 expression and lymphoid organ apoptosis in severely burned rats.

OBJECTIVE: The aim of this study is to investigate the effects of parenteral glutamine(GLN) supplementation combined with enteral nutrition (EN) on heat shock protein 90(Hsp90) expression, apoptosis of lymphoid organs and circulating lymphocytes, immunological function and survival in severely burned rats. METHODS: Male SD rats were randomly assigned into 4 groups: a sham burn+EN+GLN-free amino acid (AA) group (n=10), a sham burn+EN+GLN group (n=10), a burn+EN+AA group (n=10), and a burn +EN +GLN group (n=10). Two hours after a 30% total body surface area (TBSA), full-thickness scald burn injury on the back was made, the burned rats in two experimental groups (the burn+EN+AA group and the burn+EN +GLN group) were fed with a conventional enteral nutrition solution by oral gavage for 7 days. Simultaneously, the rats in the burn+EN+GLN group were given 0.35g GLN/kg body weight/day once via a tail vein injection for 7 days, whereas those in the burn+EN+AA group were administered isocaloric/isonitrogenous GLN-free amino acid solution (Tyrosine) for comparison. The rats in two sham burn control groups (the sham burn+EN+AA group and the sham burn+EN +GLN group) were treated in the same procedure as above, except for burn injury. All rats in each of the four groups were given 175kcal/kg body wt/day. There was isonitrogenous, isovolumic and isocaloric intake among four groups. At the end of the 7th day after nutritional programme were finished, all rats were anesthetized and samples were collected for further analysis. Serum immunoglobulin quantification was conducted by ELISA. Circulating lymphocyte numbers were counted by Coulter LH-750 Analyzer. The percentages and apoptotic ratio of CD4 and CD8T lymphocytes in circulation were determined by flow cytometry (FCM). The neutrophil phagocytosis index (NPI) was examined. The GLN concentrations in plasma, thymus, spleen and skeletal muscle were measured by high performance liquid chromatography (HPLC). The organ index evaluation and TUNEL analysis of thymus and spleen were carried out. The expression of Hsp90 in thymus and spleen was analyzed by western blotting. Moreover, the survival in burned rats was observed. RESULTS: The results revealed that parenteral GLN supplementation combined with EN significantly increased the GLN concentrations of plasma and tissues, the serum immunoglobulin content, the circulating lymphocyte number, the CD4/CD8 ratio, the indexes of thymus and spleen, NPI and survival as compared with the burn+EN+AA group (p<0.05). The expression of Hsp90 in thymus and spleen in the burn+EN+GLN group was significantly up-regulated as compared with the burn+EN+AA group (p<0.05). The apoptosis in circulating CD4 and CD8 lymphocytes, thymus and spleen in the burn+EN+GLN group was significantly decreased as compared with the burn+EN+AA group (p<0.05). CONCLUSION: The results of this study show that parenteral GLN supplementation combined with EN may increase the GLN concentrations of plasma and tissues, up-regulate the expression of Hsp90, attenuate apoptosis in lymphoid organ and circulating lymphocyte, enhance the immunological function and improve survival in severely burned rats. Clinically, therapeutic efforts at the modulation of the immune dysfunction may contribute to a favorable outcome in severely burned patients.

Effects of enteral nutrition with parenteral glutamine supplementation on the immunological function in septic rats.

The aim of the present study was to investigate the effects of enteral nutrition (EN) with parenteral glutamine (GLN) supplementation on inflammatory response, lymphatic organ apoptosis, immunological function and survival in septic rats by caecal ligation and puncture (CLP). Male rats were randomly assigned into two experimental groups and two sham CLP control groups (n 10 per group). After CLP or sham CLP model and nutrition programme were completed, the GLN concentrations of plasma and tissues and several indices of immunological function including serum Ig content, circulating lymphocyte number, the CD4:CD8 ratio, the neutrophil phagocytosis index (NPI), the organ index and apoptosis of thymus and spleen, and plasma cytokine levels were determined. Moreover, the survival in septic rats was observed. The results revealed that EN with parenteral GLN supplementation remarkably increased the GLN concentrations of plasma and tissues, serum Ig content, the circulating lymphocyte number, the CD4:CD8 ratio, the indexes of thymus and spleen, NPI and survival compared with the control group (P< 0·05). In contrast, the apoptosis of thymus and spleen and the levels of TNF-α, IL-1ß and IL-6 in plasma were obviously decreased compared with the control group (P< 0·05). These results show that EN with parenteral GLN supplementation diminished the release of inflammatory cytokines, attenuated lymphatic organ apoptosis, enhanced the immunological function and improved survival in septic rats.

Parenteral glutamine supplementation in combination with enteral nutrition improves intestinal immunity in septic rats.

OBJECTIVES: The gut-associated lymphoid tissue is continuously exposed to antigens in the gut lumen and becomes the first line of defense against enteric bacteria and associated toxin. The aim of this study was to investigate the effects of parenteral glutamine (GLN) supplementation in combination with enteral nutrition (EN) on intestinal mucosal immunity in septic rats by cecal ligation and puncture (CLP). METHODS: Male Sprague-Dawley rats were randomly assigned into four groups: A sham CLP + EN + saline group (n = 10), a sham CLP + EN + GLN group (n = 10), a CLP + EN + saline group (n = 10), and a CLP + EN + GLN group (n = 10). At 2 h after CLP or sham CLP, all rats in each of the four groups received an identical enteral nutrition solution as their base formula. Then, the rats in the sham CLP + EN + GLN group and CLP + EN + GLN group were given 0.35 g GLN/kg body weight daily for 7 d, all at the same time, via a tail vein injection; whereas those in the sham CLP + EN + saline group and CLP + EN + saline group were daily administered isovolumic sterile 0.9% saline for comparison. All rats in each of the four groups were given 290 kcal/kg body wt/d for 7 d. At the end of the seventh day after the nutritional program was finished, all rats were euthanized and the entire intestine was collected. Total Peyer's patches (PP) cell yield was counted by a hemocytometer. The percentage of PP lymphocyte subsets was analyzed by flow cytometry. The number of intestinal lamina propria IgA plasma cells was determined by the immunohistochemistry technique. The intestinal immunoglobulin A (IgA) levels were assessed by ELISA. PP apoptosis was evaluated by terminal deoxyuridine nick-end labeling. RESULTS: The results revealed total PP cell yield, the numbers of PP lymphocyte subsets, intestinal lamina propria IgA plasma cells, and intestinal IgA levels in the CLP + EN + GLN group were significantly increased when compared with the CLP + EN + saline group (P < 0.05). On the other hand, the number of TUNEL-stained cells within PPs in the CLP + EN + GLN group was markedly decreased as compared with the CLP + EN + saline group (P < 0.05). CONCLUSION: The results of this study show that parenteral glutamine supplementation in combination with enteral nutrition may attenuate PP apoptosis, increase PP cell yield and intestinal lamina propria IgA plasma cells, and subsequently improve intestinal mucosal immunity. Clinically, these results suggest therapeutic efforts at improving intestinal immunity may contribute to the prevention and treatment of sepsis.

Effects of early enteral nutrition supplemented with arginine on intestinal mucosal immunity in severely burned mice.

BACKGROUND: To investigate the effects of early enteral nutrition (EN) supplemented with Arginine (Arg) on intestinal mucosal immunity in severely burned mice. METHODS: Forty-four mice were randomly assigned into four groups: a sham injury+EN group (n=10), a sham injury+EN+Arg group (n=10), a burn+EN group (n=12), and a burn+EN+Arg group (n=12) and the mice in two experimental groups received a 20% total body surface area (TBSA), full-thickness scald burn on the back. Then, the burned mice were given a 175 kcal/kg body wt/day of conventional enteral nutrition or an isonitrogenous and isocaloric enteral nutrition supplemented with Arg by gastric gavage for 7 days. There was isonitrogenous and isocaloric intake in two experimental groups. The mice in two control groups received the same procedures as above, except for burn injury. On day 7 after injury, all mice among four groups were euthanized and the entire intestine was harvested. Intestinal immunoglobulin A (IgA) levels, total lymphocyte yield, and lymphocyte subpopulations in Peyer's patches were analyzed. Levels of IFN-gamma, IL-2, IL-4 and IL-10 in gut homogenates were also measured by ELISA. RESULTS: Total lymphocyte yield, numbers of lymphocyte subpopulations, and intestinal IgA levels in the EN+ARG group were higher than those in the EN group (p<0.05). Levels of gut tissue cytokines were significantly altered with enteral Arg supplementation: levels of IL-4 and IL-10 were increased, and levels of IFN-gamma and IL-2 declined, when compared with the EN-fed mice (p<0.05). CONCLUSIONS: The results of this study suggested that enteral nutrition supplemented with Arg has changed the cytokine concentrations in intestinal homogenates from a pro- to an anti-inflammatory profile, increased sIgA levels and changed lymphocytes in severely burned mice.

Effects of glutamine added to enteral nutrition on Peyer's patch apoptosis in severely burned mice.

BACKGROUND AND AIMS: This study aimed to investigate the influence of enteral glutamine (GLN) supplementation on Peyer's patch apoptosis in severely burned mice. METHODS: Thirty-four mice were randomly assigned to a normal group (n=10), an EN group (n=12) and an EN supplemented with GLN (EN+GLN) group (n=12) and the mice in the EN and EN+GLN groups received a full-thickness scald burn over 20% total body surface area (TBSA) on the back. The burned mice then were fed orally with a common EN or an isonitrogenous and isocaloric EN supplemented with GLN for 7 days. On day 7 after injury, all surviving mice were euthanised and the entire intestine was collected. The percentage of apoptotic cells and cell percentage of phenotype in Peyer's patches were determined by flow cytometry (FCM). The FasL expression in Peyer's patches was analysed by reverse transcription polymer chain reaction (RT-PCR) and FCM. Both TNF-alpha levels and caspase-3 activity in Peyer's patches were also assessed. RESULTS: The results revealed that the percentage of lymphocyte subsets in Peyer's patches after burn injury significantly altered: the percentage of CD4 and CD19 cells declined and the percentage of CD8 cells correspondingly increased, when compared with the normal control mice (p<0.05). On the other hand, the total apoptotic ratio and all lymphocytes subset apoptosis in Peyer's patches were markedly increased (p<0.05), which were consistent with up-regulation in the FasL expression at the levels of both mRNA and protein, TNF-alpha levels and caspase-3 activity in Peyer's patches. Enteral GLN supplementation partially reversed these changes: the total apoptotic ratio and all lymphocytes subpopulation apoptosis in Peyer's patches were markedly decreased when compared with the EN group (p<0.05). The percentage of lymphocyte subsets within Peyer's patches also restored the condition prior to injury. However, no significant differences in the FasL expression, including mRNA and protein, were observed between the EN and EN+GLN groups. Although, both TNF-alpha levels and caspase-3 activity in Peyer's patches were lower in the EN+GLN group than in the EN group (p<0.05). CONCLUSIONS: This study suggests that enteral GLN supplementation is superior to a common enteral nutrition with respect to attenuating apoptosis in Peyer's patches, which might be more effective in decreasing TNF-alpha levels and down-regulating caspase-3 activity in Peyer's patches.

Effects of enteral nutrition supplemented with glutamine on intestinal mucosal immunity in burned mice.

OBJECTIVE: The aim of this study was to investigate the effects of enteral nutrition (EN) supplemented with glutamine (GLN) on Peyer's patches and intestinal immunoglobulin A (IgA) response in burned mice. METHODS: Thirty-four mice were randomly assigned to a normal control group (n = 10), an EN group (n = 12), and an EN supplemented with GLN (EN + GLN) group (n = 12) and mice in the EN and EN + GLN groups received a 20% total body surface area, full-thickness scald burn on the back. Then the burned mice were fed with conventional EN or EN + GLN for 7 d. There was isonitrogenous and isocaloric intake in the EN and EN + GLN groups. On day 7 after injury, entire intestines were collected and intestinal IgA levels, total lymphocyte yield, lymphocyte subpopulations, and total apoptotic ratio in Peyer's patches were analyzed. RESULTS: Total lymphocyte yield, numbers of lymphocyte subpopulations, and intestinal IgA levels in the EN + GLN group were significantly higher than those in the EN group (P < 0.05). The total apoptotic ratio in Peyer's patches was markedly decreased in the EN + GLN group compared with that in the EN group (P < 0.05). CONCLUSION: The results indicated that EN supplemented with GLN is superior to conventional EN with respect to improvement of intestinal immunity in burned mice.

[Determination of plumbagin in different parts of Plumbago zeylanica by RP-HPLC].

OBJECTIVE: To develop a RP-HPLC method to determine plumbagin in Plumbago zeylanica, and to investigate contents of plumbagin in different parts of. P. zeylanica. METHOD: The analysis was carried out at 30 degrees C on a Kromasil C18, column eluted with a mobile phase consisting of a mixture of methanol-water (65: 35). The flow rate was 1 mL x min(-1), the detector wavelength was 213 nm. RESULT: The calibration curve was linear within the concentration ranges of 0.020 8-0. 104 microg (r = 0. 9999). The average recovery was 98.7%. The contents in the root, stem and leaf were 0.394 5%, 0.050 8%, 0.031 4% respectively. CONCLUSION: This method is simple, accurate, replicate and suitable for the determination of plumbagin in P. zeylanica.