Exploring the mechanism of electroacupuncture at different acupoints on acute colitis rats based on JAK2/STAT3/SOCS1 signaling pathway

Objective:To investigate the mechanism of JAK2/STAT3/SOCS1 signaling pathway in electroacupuncture of different acupoints on acute colitis rats.Methods:36 SPF SD rats were randomly divided into 6 groups,with 6 rats in each group.The rat model of acute colitis was prepared by enema with glacial acetic acid solution.After the model was established,electroacupuncture was given to each acupoint group,with density wave,frequency 2Hz-50 Hz,intensity 2 mA,muscle tremor as the degree 20 min/time,1 time/day,for 3 consecutive days.Observe the general condition of rats;the pathological changes of colonic mucosa in rats were observed by HE method.The contents of serum interleukin-4(IL-4)and interleukin-8(IL-8)were detected by ELISA.Western blot and RT-PCR were used to detect the expression of JAK2,STAT3,SOCS1 protein and mRNA in rat colon tissue.Results:In contrast to the normal group,the overall condition of the model group was worse,the colonic mucosa was severely damaged,even necrotic,and the ulcer surface was obvious.The content of IL-4 in serum was obviously reduced,and the content of IL-8 was obviously go up(P<0.01).The protein content of JAK2,STAT3 and the expression of JAK2,STAT3 mRNA in colon tissue of rats were obviously go up,while the protein content of SOCS1 and the expression of SOCS1 mRNA were obviously reduced(P<0.01).In contrast to the model group,the general condition of rats in each acupoint group was significantly improved,the damage and necrosis of colonic mucosa and ulcer surface were obviously alleviated,the content of IL-4 in serum was obviously go up,and the content of IL-8 was significantly decreased(P<0.01).The protein content of JAK2,STAT3 and the expression of JAK2,STAT3 mRNA in colon tissue of rats were obviously reduced,while the protein content of SOCS1 and the expression of SOCS1 mRNA were obviously go up(P<0.05,P<0.01).Comparison of different acupoint groups,the colonic mucosal injury in the Zusanli group was significantly reduced,the content of serum IL-4 was significantly increased,and the content of IL-8 was significantly decreased(P<0.05,P<0.01).The protein content and mRNA expression of JAK2 and STAT3 in colon tissue were significantly down-regulated,while the protein content and mRNA expression of SOCS1 were significantly go up(P<0.05,P<0.01).Conclusion:Electroacupuncture at each acupoint can improve the damage of colonic mucosa and reduce the inflammatory response.The therapeutic effect of Zusanli(ST36)is better than that of Tianshu(ST25),Dachangshu(BL25)and Shangjuxu(ST37).The mechanism may be related to the regulation of JAK2/STAT3/SOCS1 signaling pathway related proteins and inflammatory cytokines IL-4 and IL-8.

LncRNA Airsci increases the inflammatory response after spinal cord injury in rats through the nuclear factor kappa B signaling pathway

Spinal cord injury(SCI)is a serious traumatic event to the central nervous system.Studies show that long non-coding RNAs(lncRNAs)play an important role in regulating the inflammatory response in the acute stage of SCI.Here,we investigated a new lncRNA related to spinal cord injury and acute inflammation.We analyzed the expression profile of lncRNAs after SCI,and explored the role of lncRNA Airsci(acute inflammatory response in SCI)on recovery following acute SCI.The rats were divided into the control group,SCI group,and SCI+lncRNA Airsci-siRNA group.The expression of inflammatory factors,including nuclear factor kappa B[NF-κB(p65)],NF-κB inhibitor IκBαand phosphorylated IκBα(p-IκBα),and the p-IκBα/IκBαratio were examined 1–28 days after SCI in rats by western blot assay.The differential lncRNA expression profile after SCI was assessed by RNA sequencing.The differentially expressed lncRNAs were analyzed by bioinformatics technology.The differentially expressed lncRNA Airsci,which is involved in NF-κB signaling and associated with the acute inflammatory response,was verified by quantitative real-time PCR.Interleukin(IL-1β),IL-6 and tumor necrosis factor(TNF-α)at 3 days after SCI were measured by western blot assay and quantitative real-time PCR.The histopathology of the spinal cord was evaluated by hematoxylin-eosin and Nissl staining.Motor function was assessed with the Basso,Beattie and Bresnahan Locomotor Rating Scale.Numerous differentially expressed lncRNAs were detected after SCI,including 151 that were upregulated and 186 that were downregulated in the SCI 3 d group compared with the control group.LncRNA Airsci was the most significantly expressed among the five lncRNAs involved in the NF-κB signaling pathway.LncRNA Airsci-siRNA reduced the inflammatory response by inhibiting the NF-κB signaling pathway,alleviated spinal cord tissue injury,and promoted the recovery of motor function in SCI rats.These findings show that numerous lncRNAs are differentially expressed following SCI,and that inhibiting lncRNA Airsci reduces the inflammatory response through the NF-κB signaling pathway,thereby promoting functional recovery.All experimental procedures and protocols were approved by the approved by the Animal Ethics Committee of Jining Medical University(approval No.JNMC-2020-DW-RM-003)on January 18,2020.

Resveratrol and its derivates improve inflammatory bowel disease by targeting gut microbiota and inflammatory signaling pathways

Inflammatory bowel disease(IBD)is a chronic inflammatory lesion of the intestine,mainly manifested by infiltration of intestinal inflammatory cells and imbalance of gut microbiota.Conventional treatments for IBD include antibiotics,immunosuppressive agents,5-aminosalicylic acid,steroids and surgery,which have high toxic side effects.Resveratrol is a natural polyphenol,and its various derivatives have anti-oxidation and anti-inflammatory properties.In this paper,we comprehensively review the mechanism of resveratrol and its derivates to alleviate IBD by improving intestinal barrier,regulating the unbalanced gut microbiota,and targeting various inflammatory signaling pathways.

Nucleolin Mediates LPS-induced Expression of Inflammatory Mediators and Activation of Signaling Pathways

Summary:In this study,we investigated the effects of nucleolin on lipopolysaccharide(LPS)-induced activation of MAPK and NF-KappaB(NF-kB)signaling pathways and secretion of TNF-a,IL-1βand HMGB1 in THP-1 monocytes.Immunofluorescence assay and Western blotting were used to identify the nucleolin expression in cell membrane,cytoplasm and nucleus of THP-1 monocytes.Inactivation of nucleolin was induced by neutralizing antibody against nucleolin.THP-1 monocytes were pretreated with anti-nucleolin antibody for 1 h prior to LPS challenge.The irrelevant IgG group was used as control.Secretion of inflammatory mediators(TNF-a,IL-1β and HMGB1)and activation of MAPK and NF-kB/I-kB signaling pathways were examined to assess the effects of nucleolin on LPS-mediated inflammatory response.Nucleolin existed in cell membrane,cytoplasm and nucleus of THP-1 monocytes.Pretreatment of anti-nucleolin antibody significantly inhibited the LPS-induced secretion of TNF-a,IL-1β and HMGB1.P38,JNK,ERK and NF-κB subunit p65 inhibitors could significantly inhibit the secretion of IL-1β,TNF-a and HMGB1 induced by LPS.Moreover,the phosphorylation of p38,JNK,ERK and p65(or nuclear translocation of p65)was significantly increased after LPS challenge.In contrast,pretreatment of anti-nucleolin antibody could significantly inhibit the LPS-induced phosphorylation of p38,JNK,ERK and p65(or nuclear translocation of p65).However,the irrelevant IgG,as a negative control,had no effect on LPS-induced secretion of TNF-a and IL-Iβ and phosphorylation of p38,JNK,ERK and p65(or nuclear translocation of p65).We demonstrated that nucleolin mediated the LPS-induced activation of MAPK and NF-κB signaling pathways,and regulated the secretion of inflammatory mediators(TNF-a,IL-1β and HMGB1).

Effects of the Nocth signaling pathway on expression of inflammatory factors and transforming growth factors in diabetic foot ulcers

Objective:persistent hyperinflammation is an important reason for the development of diabetic foot ulcer.Notch signaling is an important signaling pathway involved in the inflammatory response and cell proliferation in diabetic foot ulcer rats.This paper aims to explore the effect of Notch signaling on inflammatory factors,chemokines and growth factors through the intervention of Notch signaling in diabetic foot ulcer rats.Methods:the experimental model was made by using high-fat feed combined with streptozotocin(STZ)to cause diabetes,and the experimental model of diabetic foot ulcer was established by constant temperature and constant pressure scald apparatus.The normal ulcer model was used as a control.The intervention controls of the experimental model included normal saline,western medicine growth factor,Notch agonist Jagged1,Notch signaling inhibitor ly-411575,and the intervention of traditional Chinese medicine Zizhu ointment for 7 days.Serum il-1,il-6,TNF-radiation,and il-17 were detected by ELISA.Real-time PCR was used to detect the inflammatory factors,chemokines,and growth factors associated with Notch signaling in wound tissues:tnf-uum,il-1,il-6,il-17,interleukin-8,ip-10,McP-1,TGF-uum,TGF-livelihood.Results:serum levels of il-1,il-6,TNF-radiation and il-17 in diabetic foot ulcer rats were significantly higher than that in normal ulcer rats.The contents of il-1,il-6,TNF-radiation and il-17a in ly-411575 group and Zizhu ointment group were significantly reduced.Real-time PCR results of wound tissue showed that the levels of inflammatory cytokines il-1,il-6,TNF-radiation,il-17 and chemokines ip-10,il-8 and McP-1 in the wound tissue of diabetic foot ulcer rat model were significantly higher than that of normal ulcer model,and the levels of growth factor TGF-exposure were lower than that of normal ulcer model.LY-411575 significantly reduced il-1,il-6,TNF-maxima,il-17,and the chemokines ip-10,il-8,and McP-1 in diabetic foot ulcer rats,and reduced the expression of TGF-,TGF-earth.Jagged1 can increase the expression of TGF--,TGF---,suggesting that inhibition of the Notch signaling pathway can reduce the expression of the inflammatory factors il-1,il-6,TNF--,il-17a,il-8,and the growth factors TGF--,TGF---.Zizhu ointment can reduce the levels of il-1,il-6,TNF-benand,il-17,and the chemokines ip-10,il-8,and McP-1 on the wound surface of diabetic foot rats,and improve the expression of TGF-benand TGF-SUNS.Ly-411575 inhibited the expression of TGF-bento and TGF-promoting of Zizhu ointment.Conclusion:the expression of inflammatory cytokines and chemokines was higher and the expression of growth factors was lower in diabetic foot ulcer rats than in normal ulcer rats.Inhibition of Notch signaling pathway can reduce the expression of inflammatory factors,chemokines and growth factors in experimental model rats,and Notch signaling pathway can promote inflammation and cell proliferation.Zizhu ointment can reduce the levels of inflammatory cytokines and chemokines in diabetic foot ulcer rats,improve the expression of growth factors,and reduce wound inflammation,which may be related to the inhibition of Nocth signal expression.

Effects of Liancao-Xieli capsule on intestinal mucosal inflammatory factors and TLR4/PI3K/Akt/mTOR signaling pathway in mice with ulcerative colitis

Objective:To observe the effect of Liancao-Xieli capsule on intestinal mucosal inflammatory factors and TLR4/PI3K/Akt/mTOR signaling pathway in mice with ulcerative colitis(UC);Methods:40 male C57BL/6 mice were randomly divided into the control group,model group,Liancao-Xieli group and mesalazine group,with 10 mice in each group.In addition to the control group,the remaining three groups of mice were induced by 3%dextran sulfate sodium(DSS)to induce acute UC model.During the modeling period,mice in each group were given corresponding drugs and normal saline by gavage.At the end of the experiment,HE staining was used to observe the pathological changes of colonic tissue in each group,and ELISA was used to detect the inflammatory factors(TNF-α,IL-6,IL-1β,IL-8,IL-17,and INF-γ)in serum and colonic tissue.The expression levels of TLR4/PI3K/Akt/mTOR signaling pathway related proteins were also detected by Western blot;Results:Compared with the model group,Liancao-Xieli capsule could significantly increase the colon length and decrease the score of colon histopathology in UC mice(P<0.01).In addition,the levels of TNF-α,IL-6,IL1β,IL-8,IL-17,and INF-γwere significantly reduced in serum and colon tissue,and the expressions of TLR4,PI3K,p-Akt and p-mTOR were significantly down-regulated in LiancaoXieyi group when compared with the model group(P<0.01).While the expressions of Akt and mTOR were not significantly affected in Liancao-Xieyi group(P>0.05);Conclusion:LiancaoXieli capsule can reduce the secretion of inflammatory factors,improve the intestinal mucosal damage and inflammatory response in UC by inhibiting the activation of TLR4/PI3K/Akt/mTOR signaling pathway。

Value of spiral CT perfusion parameters for evaluating acute pancreatitis and their correlation with inflammatory factor and JAK2/STAT3 signaling pathway

Objective: To study the value of spiral CT perfusion parameters for evaluating acute pancreatitis and their correlation with inflammatory factor and JAK2/STAT3 signaling pathway. Methods: Patients with acute pancreatitis and patients with pancreatic trauma who underwent surgical resection in Liaocheng Dongchangfu People's Hospital between May 2014 and March 2017 were selected and enrolled in the AP group and the control group of the research respectively;spiral CT perfusion scanning was conducted before surgery to measure the blood flow (BF), blood volume (BV), and mean transit time (MTT), and the serum was collected to determine the contents of inflammatory factors;pancreatitis tissue and normal pancreatic tissue were collected after surgical resection to determine the expression of JAK2/STAT3 signal molecules. Results: pancreatic tissue BF and BV levels of AP group were significantly lower than those of control group while MTT level was not different from that of control group;CRP, PCT, HMGB-1, Ghrelin and sTREM-1 contents in serum as well as JAK2, STAT3, Bcl-2 and Bcl-xL mRNA expression in pancreatic tissue of AP group were significantly higher than those of control group and negatively correlated with BF and BV levels in pancreatic tissue. Conclusion: Spiral CT perfusion parameters BF and BV can reflect the microcirculatory disorder of acute pancreatitis and are associated with the increased secretion of inflammatory factors and the activation of JAK2/STAT3 signaling pathway in the course of disease.

Acacetin protects against cerebral ischemia-reperfusion injury via the NLRP3 signaling pathway

Acacetin(5,7-dihydroxy-4′-methoxyflavone), a potential neuroprotective agent, has an inhibitory effect on lipopolysaccharide-induced neuroinflammatory reactions. However, whether acacetin has an effect on inflammatory corpuscle 3(NLRP3) after cerebral ischemia-reperfusion injury has not been fully determined. This study used an improved suture method to establish a cerebral ischemia-reperfusion injury model in C57BL/6 mice. After ischemia with middle cerebral artery occlusion for 1 hour, reperfusion with intraperitoneal injection of 25 mg/kg of acacetin(acacetin group) or an equal volume of saline(0.1 mL/10 g, middle cerebral artery occlusion group) was used to investigate the effect of acacetin on cerebral ischemia-reperfusion injury. Infarct volume and neurological function scores were determined by 2,3,5-triphenyltetrazolium chloride staining and the Zea-Longa scoring method. Compared with the middle cerebral artery occlusion group, neurological function scores and cerebral infarction volumes were significantly reduced in the acacetin group. To understand the effect of acacetin on microglia-mediated inflammatory response after cerebral ischemia-reperfusion injury, immunohistochemistry for the microglia marker calcium adapter protein ionized calcium-binding adaptor molecule 1(Iba1) was examined in the hippocampus of ischemic brain tissue. In addition, tumor necrosis factor-α, interleukin-1β, and interleukin-6 expression in ischemic brain tissue of mice was quantified by enzyme-linked immunosorbent assay. Expression of Iba1, tumor necrosis factor-α, interleukin-1β and interleukin-6 was significantly lower in the acacetin group compared with the middle cerebral artery occlusion group. Western blot assay results showed that expression of Toll-like receptor 4, nuclear factor kappa B, NLRP3, procaspase-1, caspase-1, pro-interleukin-1β, and interleukin-1β were significantly lower in the acacetin group compared with the middle cerebral artery occlusion group. Our findings indicate that acacetin has a protective effect on cerebral ischemia-reperfusion injury, and its mechanism of action is associated with inhibition of microglia-mediated inflammation and the NLRP3 signaling pathway.

Oxymatrine reduces neuroinflammation in rat brain A signaling pathway

Cerebral neuroinflammation models were established by injecting 10μg lipopolysaccharide into the hippocampus of male Sprague-Dawley rats. The rats were treated with an intraperitoneal injection of 120, 90, or 60 mg/kg oxymatrine daily for three days prior to the lipopolysaccharide injection. Twenty-four hours after model induction, the hippocampus was analyzed by real-time quantitative PCR, and the cerebral cortex was analyzed by enzyme-linked immunosorbent assay and western blot assay. The results of the enzyme-linked immunosorbent assay and the real-time quantitative PCR showed that the secretion and mRNA expression of the pro-inflammatory cytokines interleukin-113 and tumor necrosis factor-a were significantly decreased in the hippocampus and cerebral cortex of model rats treated with oxymatrine. Western blot assay and real-time quantitative PCR analysis indicated that toll-like receptor 4 mRNA and protein expression were significantly decreased in the groups receiving different doses of oxymatrine. Additionally, 120 and 90 mg/kg oxymatrine were shown to reduce protein levels of nuclear factor-KB p65 in the nucleus and of phosphorylated IKBa in the cytoplasm of brain cells, as detected by western blot assay. Experimental findings indicate that oxymatrine may inhibit neuroinflammation in rat brain via downregulating the expression of molecules in the toll-like receptor 4/nuclear factor-KB signaling Dathwav.

Neuroprotection mediated by the Wnt/Frizzled signaling pathway in early brain injury induced by subarachnoid hemorrhage

The Wnt/Frizzled signaling pathway participates in many inflammation-linked diseases. However, the inflammatory response mediated by the Wnt/Frizzled signaling pathway in experimental subarachnoid hemorrhage has not been thoroughly investigated. Consequently, in this study, we examined the potential role of the Wnt/Frizzled signaling pathway in early brain injury in rat models of subarachnoid hemorrhage.Simultaneously, possible neuroprotective mechanisms were also investigated. Experimental subarachnoid hemorrhage rat models were induced by injecting autologous blood into the prechiasmatic cistern. Experiment 1 was designed to examine expression of the Wnt/Frizzled signaling pathway in early brain injury induced by subarachnoid hemorrhage. In total, 42 adult rats were divided into sham(injection of equivalent volume of saline), 6-, 12-, 24-, 48-, 72-hour, and 1-week subarachnoid hemorrhage groups. Experiment 2 was designed to examine neuroprotective mechanisms of the Wnt/Frizzled signaling pathway in early brain injury induced by subarachnoid hemorrhage. Rats were treated with recombinant human Wnt1(rhwnt1), small interfering Wnt1(siwnt1) RNA, and monoclonal antibody of Frizzled1(anti-Frizzled1) at 48 hours after subarachnoid hemorrhage. Expression levels of Wnt1, Frizzled1, β-catenin, peroxisome proliferator-activated receptor-γ, CD36, and active nuclear factor-κB were examined by western blot assay and immunofluorescence staining. Microglia type conversion and inflammatory cytokine levels in brain tissue were examined by immunofluorescence staining and enzyme-linked immunosorbent assay. Our results show that compared with the sham group, expression levels of Wnt1, Frizzled1, and β-catenin were low and reduced to a minimum at 48 hours, gradually returning to baseline at 1 week after subarachnoid hemorrhage. rhwnt1 treatment markedly increased Wnt1 expression and alleviated subarachnoid hemorrhage-induced early brain injury(within 72 hours), including cortical cell apoptosis, brain edema, and neurobehavioral deficits, accompanied by increasing protein levels of β-catenin, CD36, and peroxisome proliferator-activated receptor-γ and decreasing protein levels of nuclear factor-κB. Of note, rhwnt1 promoted M2-type microglia conversion and inhibited release of inflammatory cytokines(interleukin-1β, interleukin-6, and tumor necrosis factor-α). In contrast, siwnt1 RNA and anti-Frizzled1 treatment both resulted in an opposite effect. In conclusion, the Wnt/Frizzled1 signaling pathway may participate in subarachnoid hemorrhage-induced early brain injury via inhibiting the inflammatory response, including regulating microglia type conversion and decreasing inflammatory cytokine release. The study was approved by the Animal Ethics Committee of Anhui Medical University and First Affiliated Hospital of USTC,Division of Life Sciences and Medicine, University of Science and Technology of China(approval No. LLSC-20180202) in May 2017.

Atsttrin reduces lipopolysaccharide-induced neuroinflammation by inhibiting the nuclear factor kappa B signaling pathway

Progranulin is closely related to neuronal survival in a neuroinflammatory mouse model and attenuates inflammatory reactions. Atsttrin is an engineered protein composed of three progranulin fragments and has been shown to have an effect similar to that of progranulin. Atsttrin has anti-inflammatory actions in multiple arthritis mouse models, and it protects against further arthritis development. However, whether Atsttrin has a role in neuroinflammation remains to be elucidated. In this study, we produced a neuroinflammatory mouse model by intracerebroventricular injection of 1 μL lipopolysaccharide(10 μg/μL). Atsttrin(2.5 mg/kg) was administered via intraperitoneal injection every 3 days over a period of 7 days before intracerebroventricular injection of 1 μL lipopolysaccharide(10 μg/μL). In addition, astrocyte cultures were treated with 0, 100 or 300 ng/mL lipopolysaccharide, with 200 ng/mL Atsttrin simultaneously. Immunohistochemistry, enzyme-linked immunosorbent assay and real-time reverse transcription-polymerase chain reaction were performed to examine the protein and mRNA levels of inflammatory mediators and to assess activation of the nuclear factor kappa B signaling pathway. Progranulin expression in the brain of wild-type mice and in astrocyte cultures was increased after lipopolysaccharide administration. The protein and mRNA expression levels of tumor necrosis factor-α, interleukin-1β and inducible nitric oxide synthase were increased in the brain of progranulin knockout mice after lipopolysaccharide administration. Atsttrin treatment reduced the lipopolysaccharide-induced increase in the protein and mRNA levels of tumor necrosis factor-α, interleukin-1β, matrix metalloproteinase-3 and inducible nitric oxide synthase in the brain of progranulin knockout mice. Atsttrin also reduced the expression of cyclooxygenase-2, inducible nitric oxide synthase and matrix metalloproteinase 3 mRNA in lipopolysaccharide-treated astrocytes in vitro, and decreased the concentration of tumor necrosis factor α and interleukin-1β in the supernatant. Furthermore, Atsttrin significantly reduced the levels of phospho-nuclear factor kappa B inhibitor α in the brain of lipopolysaccharide-treated progranulin knockout mice and astrocytes, and it decreased the expression of nuclear factor kappa B2 in astrocytes. Collectively, our findings show that the anti-neuroinflammatory effect of Atsttrin involves inhibiton of the nuclear factor kappa B signaling pathway, and they suggest that Atsttrin may have clinical potential in neuroinflammatory therapy.

Effect of Catechins on Myocardial Injury and Inflammatory Factors in Rats with Coronary Heart Disease under PI3K/Akt/eNOS Signaling Pathway

Objective:To discuss the effect of catechins on myocardial injury and inflammatory factors in rats with coronary heart disease under PI3K/Akt/eNOS signaling pathway.Methods:A total of 50 healthy adult pathogen-free(SPF)-grade SD rats were divided into five groups by random number table method.Except for the blank group,the other four groups were fed with high fat to construct a rat model of coronary artery disease.After the model was successfully constructed,the blank group and the model group were given saline by gavage,the positive group was given 25 mg/kg aspirin by gavage,the low-dose group was given 20 mg/kg catechin by gavage,and the high-dose group was given 60 mg/kg catechin by gavage.The expression levels of PI3K/Akt/eNOS signaling pathway-related proteins,myocardial injury markers,myocardial infarction and myocardial inflammatory factors were observed and compared in the five groups.Results:Overall,there were significant differences in the expression levels of PI3K,p-Akt/Akt,and p-eNOS/eNOS in the five groups of rats(P<0.05);there were significant differences in the expression levels of CK-MB and c Tn I in the five groups of rats(P<0.05);there were significant differences in ischemic area,infarct area,and myocardial infarction range in the four groups of rats,except for the blank group(P<0.05);there were significant differences in the expression levels of IL-1β,IL-18,TNF-α,and ET-1 in the five groups of rats(P<0.05).Conclusion:Catechins can reduce the severity of myocardial injury,reduce the range of myocardial infarction,and reduce myocardial inflammation in rats with coronary heart disease by up-regulating expression level of PI3K/Akt/eNOS signaling pathway-related proteins.Compared with aspirin,high-dose catechins have a more prominent protective effect on the myocardium of rats with coronary heart disease.

Effect of miR-124 Overexpression under Wnt/β-catenin Signaling Pathway on Inflammatory Factor and Epidermal Growth Factor in Rats with Precancerous Gastric Lesions

Objective:To investigate the effects of miR-124 overexpression on inflammatory factors and epidermal growth factor in rats with gastric precancerous lesions by modulating the Wnt/β-catenin signaling pathway.Methods:Eighty SPF-grade Wistar rats were selected as the research subjects.After adaptive feeding,they were divided into 4 groups using a random number table method.The control group was fed a regular diet,while the model group,low expression group,and overexpression group of rats were treated with N-methyl-N’-nitro-N-nitrosoguanidine to establish a model of gastric cancer precancerous lesions.After successful modeling,the control group and model group rats were injected intraperitoneally with 0.9%sodium chloride solution,the low-expression group rats were injected intraperitoneally with miR-124-5p inhibitor,and the over-expression group rats were injected intraperitoneally with miR-124-5p mimic.After 8 weeks of continuous injection,the levels of miR-124,Wnt/β-catenin signaling pathway-related proteins,inflammatory factors,and epidermal growth factor were observed and compared among the four groups of rats.Results:The differences in the relative expression of miR-124 among the four groups of rats were statistically significant(P<0.05);the differences in the expression of Wnt protein andβ-catenin protein among the four groups of rats were statistically significant(P<0.05),and the expression of Wnt protein andβ-catenin protein in the overexpression group was significantly higher than that of the control group,and significantly lower than that in the model group and the low-expression group;the levels of IL-1β,IL-6,and TNF-αof rats in the four groups of rats IL-1β,IL-6 and TNF-αlevels were statistically different(P<0.05),and the levels of IL-1β,IL-6 and TNF-αin rats in the overexpression group were higher than those in the model group and significantly lower than those in the model group and the low-expression group(P<0.05);the differences in the levels of EGF,HER1 and HER2 among the four groups of rats were statistically different(P<0.05),and the levels of EGF levels were higher than those of the control group,and EGF and HER1 were significantly lower than those of the model and low expression groups(P<0.05).Conclusion:Mi R-124 overexpression helps to reduce gastric mucosal damage and inflammatory response in rats with gastric precancerous lesions,and its mechanism of action may be linked to the inhibition of the Wnt/β-catenin signaling pathway.

Study on the Hepatoprotective Effect of Oxalis coriniculata L. and Related Mechanism by Regulating Oxidative Stress and TLR-2/NF-κB Signaling Pathway

[Objectives]This study aimed to explore the protective effect of Oxalis coriniculata L.on rats with acute liver injury induced by carbon tetrachloride(CCl4)and related mechanism by regulating oxidative stress and the TLR-2 TLR-2/NF-κB signaling pathway.[Methods]A total of 48 female rats were randomly and evenly divided into normal group,model group,silymarin group(0.12 g/kg),and high(16 g/kg),middle(8 g/kg)and low-dose(4 g/kg)O.coriniculata L.groups.The rats in the groups were intragastrically administered with 5 mL/kg of corresponding drugs(equal-volume distilled water for normal group and control group),respectively.The administration was conducted twice a day,for 10 consecutive days.After 2 h of the last administration,the rats in all the groups except the normal group were intraperitoneally injected with 12%carbon tetrachloride(CCl4)olive oil solution(5 mL/kg),respectively to establish liver injury rat models.After 16 h,the eyeball blood of the rats was collected,and their liver tissues were collected for preparation of HE sections.The biochemical indicators detected included aspartate aminotransferase(AST),alanine aminotransferase(ALT),total superoxide dismutase(T-SOD)and glutathione peroxidase(GSH-Px)activity and malondialdehyde(MDA)content in the serum.The contents of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-6(IL-6)in the serum were detected by ELISA.The expression of Toll-like receptor-2(TLR-2)and nuclear factor-κB(NF-κB)in liver tissue was detected using Western blotting.The pathological changes of liver were observed under light microscope.[Results]Compared with the normal group,the ALT,AST activity and MDA,IL-1β,IL-6,TNF-αlevels in rat serum significantly increased(P<0.01),the GSH-Px,T-SOD activity in rat serum significantly decreased(P<0.01),and the expression of TLR-2 and NF-κB in liver tissue was up-regulated(P<0.01)in the model group.Compared with the model group,the ALT,AST activity and MDA,IL-1β,IL-6 and TNF-αlevels in rat serum reduced(P<0.05,P<0.01),the GSH-Px and T-SOD activity in rat serum increased(P<0.05,P<0.01),and the expression of TLR-2 and NF-κB in liver tissue was down-regulated(P<0.05,P<0.01)in the O.coriniculata L.administration groups.Pathological sections show that O.coriniculata L.had an improving effect on rats with acute liver injury induced by CCl4.[Conclusions]O.coriniculata L.has a good protective effect on rats with acute liver injury induced by CCl4.Its mechanism may be related to inhibition of oxidative stress,inhibition of inflammatory response and regulation of the TLR-2/NF-κB signaling pathway.

Fangji Fuling Decoction Alleviates Sepsis by Blocking MAPK14/FOXO3A Signaling Pathway

Objective:To examine the therapeutic effect of Fangji Fuling Decoction(FFD) on sepsis through network pharmacological analysis combined with in vitro and in vivo experiments.Methods:A sepsis mouse model was constructed through intraperitoneal injection of 20 mg/kg lipopolysaccharide(LPS).RAW264.7 cells were stimulated by 250 ng/m L LPS to establish an in vitro cell model.Network pharmacology analysis identified the key molecular pathway associated with FFD in sepsis.Through ectopic expression and depletion experiments,the effect of FFD on multiple organ damage in septic mice,as well as on cell proliferation and apoptosis in relation to the mitogen-activated protein kinase 14/Forkhead Box O 3A(MAPK14/FOXO3A) signaling pathway,was analyzed.Results:FFD reduced organ damage and inflammation in LPS-induced septic mice and suppressed LPS-induced macrophage apoptosis and inflammation in vitro(P<0.05).Network pharmacology analysis showed that FFD could regulate the MAPK14/FOXO signaling pathway during sepsis.As confirmed by in vitro cell experiments,FFD inhibited the MAPK14 signaling pathway or FOXO3A expression to relieve LPS-induced macrophage apoptosis and inflammation(P<0.05).Furthermore,FFD inhibited the MAPK14/FOXO3A signaling pathway to inhibit LPS-induced macrophage apoptosis in the lung tissue of septic mice(P<0.05).Conclusion:FFD could ameliorate the LPS-induced inflammatory response in septic mice by inhibiting the MAPK14/FOXO3A signaling pathway.

Differential regulation of JAK/STAT-signaling in patients with ulcerative colitis and Crohn’s disease

In 2018,the pan-Janus kinase(JAK)inhibitor tofacitinib was launched for the treatment of ulcerative colitis(UC).Although tofacitinib has proven efficacious in patients with active UC,it failed in patients with Crohn’s disease(CD).This finding strongly hints at a different contribution of JAK signaling in both entities.Here,we review the current knowledge on the interplay between the JAK/signal transducer and activator of transcription(STAT)pathway and inflammatory bowel diseases(IBD).In particular,we provide a detailed overview of the differences and similarities of JAK/STAT-signaling in UC and CD,highlight the impact of the JAK/STAT pathway in experimental colitis models and summarize the published evidence on JAK/STAT-signaling in immune cells of IBD as well as the genetic association between the JAK/STAT pathway and IBD.Finally,we describe novel treatment strategies targeting JAK/STAT inhibition in UC and CD and comment on the limitations and challenges of the new drug class.

Changes and correlation of inflammatory polarization and Notch pathway in rats with adjuvant arthritis

Objective:To investigate the relationship between inflammatory polarization and Notch pathway in rats with adjuvant arthritis.Methods:Twelve rats were randomly divided into normal(NC)group(n=6)and model(MC)group(n=6).In the model group,complete Freund's adjuvant(0.1ml/rat)was injected into the right hindfoot to induce inflammation.On the 12th day after inflammation,the changes of plantar swelling degree(E)and arthritis index(AI)were observed,and the expressions of inflammatory polarization markers CD68 and CD206 in peripheral blood were detected by flow cytometry.PCR was used to detect the expression of factors related to Notch signal pathway in peripheral blood.Results:Compared with the normal group,the expression of E,AI,CD68,Notch2,Notch3,Notch4 and Delta1 in the model group increased significantly,while the expression of CD206,Notch1,Jagged1 and Jagged2 decreased(P<0.01or P<0.05).The results showed that CD68,toe swelling degree and arthritis index were negatively correlated with Notch1,Jagged1 and Jagged2,CD68,toe swelling degree and arthritis index were positively correlated with Notch2,Notch4 and Delta1,CD206 was positively correlated with Notch1 and Jagged1,Jagged2 and CD206 was negatively correlated with Notch2,Notch4 and Delta1.Conclusion:Notch signal pathway may promote the occurrence and development of AA by regulating inflammatory polarization of macrophages.

Inhibitory Effect of Bergenin on TLR-4/NF-κB Signal Pathway in Reducing Allergic Rhinitis in Mice

[Objectives]To explore the effect and possible mechanism of bergenin in relieving allergic rhinitis(AR)in mice.[Methods]50 C57/BL6 mice were randomly divided into blank group(n=10),model group(n=10)and high(100 mg/kg),medium(50 mg/kg)and low(25 mg/kg)dose bergenin groups with 10 mice in each group.Except for the blank group,the other mice were sensitized by basic ways combined with attack to replicate the AR model.From the 15th d of modeling(from the second d after the end of the basic modeling),the drug group was given bergenin orally for 15 d,and the blank group and model group were given the same volume of normal saline once a day.24 h after the last establishment of the model,the content of interleukin 4(IL-4),IL-6,TNF-αand IL-1βin nasal lavage fluid and serum of mice in each group was detected by ELISA.The expression of TLR-4,NF-κB and p-NF-κB in nasal mucosa of mice was detected by Western blot.[Results]Compared with the blank group,the content of inflammatory factors IL-4,IL-6,TNF-αand IL-1βin nasal lavage fluid and serum of model group was significantly increased,and the protein expression of TLR-4 and p-NF-κB was significantly increased.After the intervention of bergenin,the content of IL-4,IL-6,TNF-αand IL-1βin nasal lavage fluid and serum and TLR-4 and p-NF-κB protein in tissue was significantly inhibited in bergenin group.[Conclusions]Bergenin can effectively reduce allergic inflammation in AR model mice,and its mechanism may be related to inhibition of inflammation and down-regulation of TLR-4/NF-κB signal pathway.

Correlation of MyD88 expression in the peripheral blood and placenta with the inflammatory response and insulin signal transduction in the placenta of patients with GDM

Objective: To study the correlation of MyD88 expression in peripheral blood and placenta with the inflammatory response and insulin signal transduction in the placenta of patients with gestational diabetes mellitus (GDM). Methods: The patients with GDM and healthy pregnant women who accepted antenatal care and gave birth in Guangyuan First People's Hospital between March 2015 and June 2017 were selected as the research subjects and enrolled in the GDM group and control group for the study respectively;the peripheral blood was collected before delivery to determine the MyD88 mRNA expression, and the placental tissue was collected after delivery to determine the mRNA expression of MyD88, inflammatory response molecules and insulin signal transduction molecules. Results: MyD88 mRNA expression levels in the peripheral blood and placenta of GDM group were significantly higher than those of control group, and the MyD88 mRNA expression in the peripheral blood was positively correlated with the MyD88 mRNA expression in the placenta;IL-1β, IL-6, RBP4, Chemerin, Resistin and PTP1B mRNA expression levels in the placenta of GDM group were significantly higher than those of control group whereas IRS1, ISR2, p-PI3K and GLUT4 protein expression levels were significantly lower than those of control group;IL-1β, IL-6, RBP4, Chemerin, Resistin and PTP1B mRNA expression levels in the placenta of GDM group of patients with high MyD88 expression were significantly higher than those of patients with low MyD88 expression whereas IRS1, ISR2, p-PI3K and GLUT4 protein expression levels were significantly lower than those of patients with low MyD88 expression. Conclusion:The expression of MyD88 in peripheral blood and placenta increase in patients with GDM and the change of MyD88 expression in peripheral blood could reflect the abnormality of inflammatory response and insulin signal transduction in the placenta.

Comparative transcriptomic analysis reveals the molecular changes of acute pancreatitis in experimental models

BACKGROUND Acute pancreatitis(AP)encompasses a spectrum of pancreatic inflammatory conditions,ranging from mild inflammation to severe pancreatic necrosis and multisystem organ failure.Given the challenges associated with obtaining human pancreatic samples,research on AP predominantly relies on animal models.In this study,we aimed to elucidate the fundamental molecular mechanisms underlying AP using various AP models.AIM To investigate the shared molecular changes underlying the development of AP across varying severity levels.METHODS AP was induced in animal models through treatment with caerulein alone or in combination with lipopolysaccharide(LPS).Additionally,using Ptf1αto drive the specific expression of the hM3 promoter in pancreatic acinar cells transgenic C57BL/6J-hM3/Ptf1α(cre)mice were administered Clozapine N-oxide to induce AP.Subsequently,we conducted RNA sequencing of pancreatic tissues and validated the expression of significantly different genes using the Gene Expression Omnibus(GEO)database.RESULTS Caerulein-induced AP showed severe inflammation and edema,which were exacerbated when combined with LPS and accompanied by partial pancreatic tissue necrosis.Compared with the control group,RNA sequencing analysis revealed 880 significantly differentially expressed genes in the caerulein model and 885 in the caerulein combined with the LPS model.Kyoto Encyclopedia of Genes and Genomes enrichment analysis and Gene Set Enrichment Analysis indicated substantial enrichment of the TLR and NOD-like receptor signaling pathway,TLR signaling pathway,and NF-κB signaling pathway,alongside elevated levels of apoptosis-related pathways,such as apoptosis,P53 pathway,and phagosome pathway.The significantly elevated genes in the TLR and NOD-like receptor signaling pathways,as well as in the apoptosis pathway,were validated through quantitative real-time PCR experiments in animal models.Validation from the GEO database revealed that only MYD88 concurred in both mouse pancreatic tissue and human AP peripheral blood,while TLR1,TLR7,RIPK3,and OAS2 genes exhibited marked elevation in human AP.The genes TUBA1A and GADD45A played significant roles in apoptosis within human AP.The transgenic mouse model hM3/Ptf1α(cre)successfully validated significant differential genes in the TLR and NOD-like receptor signaling pathways as well as the apoptosis pathway,indicating that these pathways represent shared pathological processes in AP across different models.CONCLUSION The TLR and NOD receptor signaling pathways play crucial roles in the inflammatory progression of AP,notably the MYD88 gene.Apoptosis holds a central position in the necrotic processes of AP,with TUBA1A and GADD45A genes exhibiting prominence in human AP.