Mechanism of Polygonum capitatum in treatment of Helicobacter Pylori associated gastritis by network pharmacology, microarray data analysis and molecular docking / 中国药理学通报

Aim To explore the mechanism of Polygonum capitatum(PC)in the treatment of Helicobacter Pylori associated gastritis(HAG). Methods The databases were used to identify the target of PC active compounds and HAG-related genes,and the intersection was taken to obtain the potential targets of PC treatment of HAG. The interaction network diagram of “drug-active compound-target-disease” and the protein-protein interaction(PPI)network of potential target protein interaction in HAG treated by PC were constructed by software Cytoscape 3.6.0. The important nodes in the network were screened by several topological indexes,and the GO and KEGG enrichment were analyzed by STRING database to obtain the potential signaling pathway of PC in the treatment of HAG. The binding ability of PC active components with key target proteins was observed by molecular docking method. On this basis,the related targets of PC in the treatment of HAG were verified in vivo and in vitro experiments. Results The PC active compounds and targets were identified through the database,and the “drug-active compound-target-disease” network diagram and the PPI network of potential target proteins were constructed. Combined with several topological indexes,the PPI network of potential target-protein interaction was analyzed,and 52 hub genes were screened. Further bioinformatics analysis and high-throughput sequencing revealed that PC exerted an effect on HAG through the Akt/NF-κB/NLRP3 pathway. Based on this,it was found that PC could reduce IL-18 and IL-1β in HAG GES-1 cells and HAG SD rats,up-regulate Akt and its phosphorylation level and reduce NF-κB expression,inhibit the activation of NLRP3 inflammatory body,so as to improve HAG inflammatory response. Conclusions PC could exert a therapeutic effect on HAG by activating Akt and its phosphorylation level,and inhibiting the expression of NF-κB and NLRP3 inflammasome related factors. This study provides a theoretical basis for explaining the mechanism of PC in the treatment of HAG.

Hydrogen intervenes early inflammation of silicosis through the NF-κB/NLRP3 signaling pathway / 中国职业医学

{L-End}Objective To study the intervention effect of hydrogen on early inflammation in a rat silicosis model and its mechanism. {L-End}Methods Wistar rats of specific pathogen free were randomly divided into the control group, model group, tetrandrine group, hydrogen group and combined intervention group, with 10 rats in each group. The rats in the last four groups were treated with a dose of l.00 mL silica suspension with a mass concentration of 50.0 g/L by a one-time non-exposed tracheal method. The rats in the control group were given 0.9% sodium chloride solution in equal volume. After 24 hours of dust exposure, rats of the tetrandrine group were given 30 mg/kg body mass tetrandrine by gavage daily, rats of the hydrogen group were given 66.6% hydrogen inhalation continuously for four hours daily, rats of the combined intervention group were given the same interventions as the rats in the tetrandrine group and the hydrogen group, rats in the control group and model group were given 0.9% sodium chloride solution in equal volume by gavage. After 14 days of treatment, the lung coefficient of rats was determined, and lung histopathology was performed. The level of malondialdehyde in serum was detected by colorimetry. The level of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in serum were detected by enzyme-linked immunosorbent assay. The relative expression of nucleotide-binding oligomerization domain-like receptor thermal protein domain associated protein 3 (NLRP3), nuclear factor-κB (NF-κB) p65, NF-κB phosphorylated p65 (NF-κB p-p65), cysteinyl aspartate specific proteinase1 (Caspase1) and apoptosis-associated speck-like protein containing a CARD (ASC) in serum were detected in lung tissues by Western blot. The relative expression of NLRP3 and NF-κB p65 in lung tissues were detected by immunohistochemistry. {L-End}Results The result of pulmonary histopathology showed that the model group had obvious alveolar rupture and fusion, interstitial lymphocyte and macrophage infiltration, and alveolar wall thickening, collagen fibre deposition, and mild fibrotic hyperplasia, compared with the control group. The pathological outcomes of lung tissues in the three treated groups were alleviated compared with the model group, and the alveolar structure was more complete and the alveolar wall was thinner and the fewer collagen fibres in the rats of combined intervention group, compared with tetrandrine group and hydrogen group. The lung coefficient and Szapiel score of rats of the tetrandrine group, hydrogen group and combined intervention group were lower than those of the model group (all P<0.05). The levels of serum malondialdehyde, TNF-α and IL-1β in lung tissues, and the relative expression of NLRP3, NF-κB p65, NF-κB P-p65, Caspase1 and ASC in lung tissues increased in the model group, compared with the control group (all P<0.05). The indexes above decreased in the three treated groups than those in the model group (all P<0.05). The indexes above decreased in the combined intervention group than those in the tetrandrine group and hydrogen group (all P<0.05), except for the level of malondialdehyde in serum and the relative expression of NF-κB p-p65 in lung tissue. {L-End}Conclusion Hydrogen can intervene the early inflammation of silicosis through NF-κB/ NLRP3 signaling pathway.

Mechanism of lung and intestine combination therapy in treatment of acute lung injury by inhibiting inflammatory response based on NF-κB/NLRP3 signaling pathway and alveolar macrophage activation / 中国中药杂志

Lung and intestine combination therapy(LICT) is effective in the treatment of acute lung injury(ALI). In this study, the combination of Mahuang Decoction and Dachengqi Decoction(hereinafter referred to as the combination), a manifestation of LICT, was employed to explore the effect of nuclear factor kappaB(NF-κB)/nucleotide binding oligomerization domain-like receptors-3(NLRP3) pathway and alveolar macrophage activation on the lung inflammation in rats with ALI, for the purpose of elucidating the mechanism of LICT in treating ALI. After the modeling of ALI with limpolysaccharide(LPS, ip), rats were respectively given(ig) the combination at 10, 7.5, and 5 g·kg~(-1)(high-dose, medium-dose, and low-dose LICT groups, separately), once every 8 h for 3 times. Haematoxylin-eosin(HE) staining was used to observe the histopathological changes of lung tissue, followed by the scoring of inflammation. Immunohistochemistry was applied to detect alveolar macrophage activation, enzyme-linked immunosorbent assay(ELISA) was applied to detect the serum content of tumor necrosis factor-α(TNF-α) and interleukin-18(IL-18), Western blot was applied to detect the protein expression of phosphorylated-nuclear factor kappaB p65(p-NF-κB p65), nuclear factor kappaB p65(NF-κB p65), phosphorylated-inhibitor kappaB alpha(p-IκBα), inhibitor kappaB alpha(IκBα), and NLRP3 in lung tissue, and quantitative reverse transcription-PCR(qRT-PCR) was applied to detect the mRNA expression of TNF-α, IL-18, NLRP3, and NF-κB p65 in lung tissue. The results showed that LICT groups demonstrated lung injury relief, decrease in inflammation score, alleviation of alveolar macrophage activation, significant decline in serum content of inflammatory factors TNF-α and IL-18, and decrease of the protein expression of p-NF-κB p65/NF-κB p65, p-IκBα/IκBα, and NLRP3, and mRNA expression of TNF-α, IL-18, NLRP3, and NF-κB p65 in lung tissue. In summary, LICT has definite therapeutic effect on ALI. The mechanism is that it inhibits alveolar macrophage activation by suppressing NF-κB/NLRP3 signaling pathway, thereby reducing the activation and release of inflammatory factors and finally inhibiting inflammation.

Rehmanniae Radix and Rehmanniae Radix Praeparata improve diabetes induced by high-fat diet coupled with streptozotocin in mice through AMPK-mediated NF-κB/NLRP3 signaling pathway / 中国中药杂志

This study investigated the differential mechanisms of Rehmanniae Radix and Rehmanniae Radix Praeparata in improving diabetes in mice through AMPK-mediated NF-κB/NLRP3 signaling pathway. The diabetic mouse model was established with high-fat diet coupled with streptozotocin(STZ, intraperitoneal injection, 100 mg·kg~(-1), once a day for three consecutive days), after which the mice were randomly divided into model group, low-dose(5 g·kg~(-1)) and high-dose(15 g·kg~(-1)) Rehmanniae Radix groups, low-dose(5 g·kg~(-1)) and high-dose(15 g·kg~(-1)) Rehmanniae Radix Praeparata groups, catalpol group(250 mg·kg~(-1)), 5-hydroxymethylfurfural(5-HMF) group(250 mg·kg~(-1)), metformin group(250 mg·kg~(-1)), with the normal group also set. The organ indexes of heart,liver, spleen, lung, kidney and pancreas were calculated after four weeks of administration. The pathological changes and fibrosis of pancreas, kidney and liver in mice were observed by hematoxylin-eosin(HE) staining and Masson staining. Western blot was used to determine the expression levels of Toll-like receptor-4(TLR4), nuclear factor-κB(NF-κB), Nod-like receptor protein 3(NLRP3),interleukin-1β(IL-1β), adenosine monophosphate-activated protein kinase(AMPK), phosphorylated AMPK(p-AMPK) in the pancreas, kidney and liver of mice. Compared with the model group, the administration groups witnessed significant decrease in the liver,spleen, kidney, pancreas and fat indexes of diabetic mice, and there was no significant difference in heart and lung indexes. The pathological states and fibrosis of pancreatic, kidney and liver tissues were significantly improved after administration. Additionally, the expression levels of TLR4, NF-κB and NLRP3 in pancreas, kidney and liver of diabetic mice were significantly lowered. The expression levels of p-AMPK/AMPK were enhanced significantly in kidney and liver of mice in Rehmanniae Radix group while in pancreas, kidney and liver in Rehmanniae Radix Praeparata group. This suggests that Rehmanniae Radix and Rehmanniae Radix Praeparata differ in the mechanism of regulating energy metabolism of multiple organs and thereby exerting anti-inflammatory effects to alleviate symptoms of diabetic mice.

Research of Influence on Inflammation of Xieheyin on PCOS-IR Mice Based on Intestinal Mucosal Barrier and TLR4/NF-κB/NLRP3 Pathway / 中国实验方剂学杂志

Objective:To investigate the possible mechanism of Xieheyin in alleviating obese polycystic ovary syndrome with insulin resistance（PCOS-IR）and reducing inflammatory response. Method:Ten of sixty SPF femlae C57BL/6J mice were randomly selected as the normal group，and the rest mice were given letrozole 0.002 g·kg^-1 combined with fecal suspension 2 g·kg^-1 for 28 consecutive days to establish model of PCOS-IR.The mice that were successfully modeled were randomized into the model group，metformin group（0.25 g·kg^-1），and low（10 g·kg^-1），medium（20 g·kg^-1），and high-dose（40 g·kg^-1）Xieheyin groups，and administered with the corresponding drugs by gavage，once a day，for four consecutive weeks. Except the normal control group， the mice in the other groups were continuously given fecal suspension combined with letrozole solution to maintain the model during the treatment. The mice were weighed once a week.Levels of fasting blood glucose （FBG） were detected by blood glucose test strips.And enzyme-linked immunosorbent assay （ELISA） method was used to detect serum testosterone（T），follicle stimulating hormone（FSH），luteinizing hormone（LH），fasting insulin（FINS）level，and LH/FSH and Homeostasis model assesment of insulin resistance （HOMA-IR） were calculated．The uterus and ovaries were weighed and fixed.Hematoxylin-eosin（HE）staining was used to observe ovarian tissue pathology morphology. Western blot was used to detect the expression levels of tight junction key molecular zonula occludens 1（ZO-1），occludin in colon tissues，and the expression levels of Toll-like receptor 4/nuclear factor kappa B/Nod-like receptor protein 3（TLR4/NF-<italic>κ</italic>B/NLRP3）signaling pathway and inflammation associated proteins cysteinyl aspartate specific proteinase-1（Caspase-1） and interleukin-1<italic>β</italic>（IL-1<italic>β</italic>） in colon tissues. Result:Compared with normal control group，the body weight of mice in the model control group increased significantly（<italic>P</italic><0.01）. Serum FINS，FBG，HOMA-IR，T，LH/FSH were significantly increased（<italic>P</italic><0.05，<italic>P</italic><0.01）. The uterine organ ratio were decreased significantly（<italic>P</italic><0.01），while the ovarian organ ratio were significantly increased（<italic>P</italic><0.01）. The number of atresia follicles and cystic dilatation follicles increased significantly，and the number of corpus luteum significantly decreased，the thickness of follicular granulosa cells also decreased，while the white membrane thickness of the ovary increased. Tight junction related ZO-1，occludin proteins in colon tissues were all decreased（<italic>P</italic><0.01）.The relative expression levels of inflammation-related protein IL-1<italic>β</italic>，Caspase-1 and TLR4/NF-<italic>κ</italic>B/NLRP3 target protein signaling pathway were significantly increased（<italic>P</italic><0.05）.Compared with model control group， the body weight of mice in the low，middle and high dose Xieheyin group decreased significantly（<italic>P</italic><0.01）. The serum T，LH/FSH，FINS，FBG，HOMA-IR were significantly decreased（<italic>P</italic><0.05，<italic>P</italic><0.01）. The uterine organ ratio were increased（<italic>P</italic><0.05），while the ovarian organ ratio were decreased（<italic>P</italic><0.05）. The number of cystic follicles decreased and corpus luteum increased，the thickness of follicular granulosa cells increased and be arranged normally，while the white membrane thickness of the ovary increased slightly. The expressions of ZO-1，occludin proteins were increased（<italic>P</italic><0.01）. The expression levels of IL-1<italic>β</italic>，Caspase-1 and TLR4/NF-<italic>κ</italic>B/NLRP3 target protein in the high dose group were significantly decreased（<italic>P</italic><0.01）. Conclusion:Xieheyin could activate intestinal TLR4/NF-<italic>κ</italic>B/NLRP3 signaling pathway，inhibit pro-inflammatory factor secretion，improve obesity and IR，which was correlated with rebuilding intestinal mucosal barrier and inhibiting intestinal inflammation．