Effects of Jianpi Yichang Powder on Expressions of IL-1β and IL-18 of NLRP3 Signaling Pathway in Ulcerative Colitis Rats / 中国中医药信息杂志

Objective To explore the effects of Jianpi Yichang Powder on the expressions of interleukin(IL)-1β and IL-18 of NLRP3 signaling pathway in ulcerative colitis(UC)model rats.Methods Ten rats were randomly selected from 40 SD rats as the normal group,and the other rats freely drank 5%dextran sulfate solution for 7 days to replicate UC rats model.The model rats were randomly divided into model group,sulfasalazine group and Jianpi Yichang Powder group,with 10 rats in each group.Jianpi Yichang Powder group and sulfasalazine group were given corresponding liquid medicine for gavage,and the normal and model groups were given equivalent volume distilled water for gavage for consecutive 14 d.The general status was observed,and the disease activity index(DAI)was scored,the contents of NLRP3,apoptosis-associated spotted proteins(ASC),and Caspase-1 in serum were detected by ELISA,the expressions of IL-1β and IL-18 protein and mRNA in colon tissue were detected by immunohistochemistry,Western blot and RT-PCR respectively.Results Compared with the normal group,the general status of the rats in model group was relatively worse,and DAI score significantly increased(P<0.01),the contents of NLRP3,ASC and Caspase-l in serum were significantly increased(P<0.01),the expressions of IL-1β and IL-18 protein and mRNA in colon tissue were significantly increased(P<0.01).Compared with the model group,the general status of the rats in Jianpi Yichang Powder group and sulfasalazine group were significantly improved,DAI score significantly decreased(P<0.01),the contents of NLRP3,ASC and Caspase-l in serum significantly reduced(P<0.05,P<0.01),and the expressions of IL-1β and IL-18 protein and mRNA in colon tissue significantly decreased(P<0.05,P<0.01).Conclusion Jianpi Yichang Powder can inhibit IL-1β and IL-18 expression of NLRP3 signaling pathway to reduce colon immune inflammatory damage,thus play a role in treating UC.

Molecular mechanism of adipose tissue inflammation induced by acute exposure to cooking oil fumes / 环境与职业医学

Background Cooking oil fumes are closely related to immune response, and adipose tissue also plays an important role in immune regulation. At present, the biological effect and mechanism of inflammation of adipose tissue induced by oil fume exposure are not clear yet. Objective To investigate the inflammatory effect of different exposure duration of cooking fumes on adipose tissue in mice and explore the role of Nod-like receptor pyrin domain 3 (NLRP3)/cysteinyl aspartate specific proteinase 1 (Caspase 1)/interleukin (IL)-1β signaling pathway. Methods Forty 8-week-old female C57BL/6J mice were randomly divided into 3-day control group (CON3 group), 7-day control group (CON7 group), 3-day oil fume exposure group (COF3 group), and 7-day oil fume exposure group (COF7 group), with 10 mice in each group. The mice were exposed to oil fumes in a cooking oil fume formation and exposure equipment (COFFEE) for 20 min, followed by a 10-min pause, 1 h a day for consecutive 3 d or 7 d. General condition of mice was observed and body weight was measured every day. After exposure, blood was sampled from the eyeball. Serum levels of IL-6, IL-27, and IL-1β were detected by enzyme-linked immunosorbent assay (ELISA). The adipose tissue of mice was collected and observed after hematoxylin-eosin (HE) staining. The percentages of CD4+ and CD8+T cells in adipose tissue were detected by flow cytometry. Real-time quantitative PCR (RT-qPCR) was used to detect the expression levels of nuclear factor-κB (NF-κB), NLRP3, Caspase 1, and IL-1β in adipose tissue. Western blot was used to detect the expression levels of NLRP3, Caspase 1, and IL-1β in adipose. Results Compared with the corresponding control group, serum IL-6, IL-27, and IL-1β contents in the COF3 group and the COF7 group were significantly increased (P<0.05) except IL-6 in the COF3 group, and the levels in the COF7 group were significantly higher than those in the COF3 group (P<0.05). Vacuolar lipid droplets in adipocytes decreased, cytoplasm shrank, and inflammatory cells infiltrated in the COF7 group after HE staining. The flow cytometry results showed that the proportions of CD4+ and CD8+T cells in adipocytes of the COF3 group and the COF7 group were increased compared to the corresponding control group, with a significant increase in the COF7 group (P<0.05), and the CD4+/CD8+T ratio also significantly increased progressively in the two groups (P<0.05). The results of RT-qPCR showed that compared with the corresponding control group, the mRNA expression levels of NF-κB, NLRP3, Caspase 1, and IL-1β in adipose tissue of mice in the COF3 group and the COF7 group were significantly increased (P<0.05, P<0.01). The mRNA expression levels of mice in each exposure group gradually increased over time. The Western blot results showed that compared with the corresponding control group, the protein expressions of NLRP3 and Caspase 1 in the COF3 group were significantly increased (P<0.01), and the expression of IL-1β protein also increased but without statistical significance. The protein expressions of NLRP3, Caspase 1, and IL-1β in the COF7 group were significantly higher than those in the CON7 group (P<0.05, P<0.01). Conclusion Acute exposure to cooking oil fumes can induce significant inflammatory response in adipose tissue, and the effect gradually increases with the extension of exposure time. The mechanism of action may be related to the activation of NLRP3 inflammasome signaling pathway.

Relationship between IL-1Ra gene polymorphism and the natural outcome of asymptomatic bacterial vaginosis / 安徽医科大学学报

Objective To investigate the relationship between interleukin 1 receptor a(IL-1Ra)gene polymorphism and different outcomes in asymptomatic bacterial vaginosis(aBV)patients with the aim of grouping and managing aBV patients.Methods In study on the natural attribution of aBV patients,all patients were enrolled and a sam-ple of venous blood and vaginal lavage fluid were separately frozen.After 4 months at the end of the clinical study,patients who completed the study were divided into three groups based on clinical outcomes:self-healing,progres-sive,and unchanged.The IL-1Ra gene polymorphism,the concentration of IL-1 β and IL-1Ra were tested,and the differences in the above indicators among three groups of patients with different outcomes were compared.Results 1 014 Chinese Han female patients were enrolled,and 984 patients completed clinical follow-up and obtained clini-cal outcome data.13 specimens were unusable during testing,with a total of 971 specimens completed the test.IL-1Ra gene was detected in all patients,with three genotypes:A1/A1,A1/A2,and A2/A2.Most population had a genotype of A1/A1,with the rarest genotype being A2/A2.No rare genotype of female was found.The frequency of A2 alleles in the progression group was significantly higher than that in the self-healing group(P＜0.05).IL-1 βand IL-1Ra were detected in all vaginal lavage fluid samples.Compared with the progression group,IL-1 β in the self-healing group was significantly lower(P＜0.05).When carrying the A2 allele,IL-1 β in progression group was relatively low,while the level of IL-1Ra was relatively high.The values of the unchanged group were middle.Con-clusion The polymorphism characteristics of the IL-1Ra gene in aBV patients are related to the IL-1Ra content in vaginal secretions.Carrying allele A2 is related to the elevation of IL-1Ra,the decrease of IL-1 β in vaginal secre-tions.Carrying allele A2 may affect the clinical outcome of aBV by some potential mechanism.

Up-regulation of Proinflammatory Cytokines in Rostral Ventromedial Medulla Contributes to Chronic Postsurgical Pain by Promoting 5-HT Release / 中山大学学报(医学科学版)

ObjectiveTo investigate the role of proinflammatory cytokines tumor necrosis factor alpha （TNFα） and interleukin-1β （IL-1β） in rostral ventromedial medulla （RVM） in chronic postsurgical pain （CPSP） induced by skin/muscle incision and retraction （SMIR）. MethodsSD rats were randomly divided into 5 groups： ① Sham group； ② SMIR group； ③ SMIR+TNFα/IL-1β neutralizing antibody group； ④ SMIR+TNFα/IL-1β group and ⑤ SMIR+vehicle group. 50% paw mechanical withdrawal threshold （MWT） was measured by the up-down method， immunofluroscence was used to detect the TNFα and IL-1β expression and ELISA for the 5-Hydroxytryptamine （5-HT） level. ResultsSMIR elicited persistent nociceptive sensitization， upregulated TNFα and IL-1β expression in RVM neurons and astrocytes. Microinjection of TNFα or IL-1β neutralizing antibody into RVM inhibited the development of nociceptive sensitization and decreased the level of 5-HT in both RVM and spinal dorsal horn. While microinjection of recombinant TNFα or IL-1β into RVM enhanced the development of nociceptive sensitization and increased the level of 5-HT in both RVM and spinal dorsal horn. ConclusionUp-regulation of proinflammatory cytokines in RVM may contribute to SMIR induced CPSP by promoting 5-HT release.

Changes in serum IL-1β, IFN-γ and IL-4 levels in children with sepsis and their values in differential diagnosis of Gram-positive/Gram-negative bacterial infection / 中华微生物学和免疫学杂志

Objective:To analyze the changes in serum IL-1β, IFN-γ and IL-4 levels in children with sepsis and analyze their values in the differential diagnosis of Gram-positive (G + )/Gram-negative (G -) bacterial infection. Methods:Clinical data of 195 children with sepsis admitted to Xuzhou Central Hospital from February 2020 to February 2023 were retrospectively analyzed. They were all confirmed to have G + /G - bacterial infection based on pathogen culturing and identification and enrolled as disease group. Another 180 healthy children taking physical examination in the same hospital during the same period were recruited as control group. The distribution of G + /G - bacterial infection in the disease group was analyzed after pathogen culturing. ELISA was used to detect serum IL-1β, IFN-γ and IL-4 levels in both groups. Differences in the levels of serum IL-1β, IFN-γ and IL-4 were compared between the disease group and the control group as well as between the patients with G + /G - bacterial infection. Receiver operating characteristic (ROC) curve was drawn to evaluate the values of serum IL-1β, IFN-γ and IL-4 levels used alone or in combination in the differential diagnosis of G + /G - bacterial infection. Results:A total of 211 strains were isolated from 195 blood culture samples in the disease group, including 181 strains (85.78%) of G + bacteria and 30 strains (14.22%) of G - bacteria. Coagulase-negative Staphylococcus and Salmonella were the predominant bacteria causing G + /G - bacterial infection, respectively. The levels of serum IL-1β and IFN-γ in the disease group were higher than those in the control group ( P<0.05), while the levels of serum IL-4 were similar in the two groups ( P>0.05). The levels of serum IL-1β, IFN-γ and IL-4 in the patients with G - bacterial infection were higher than those in the patients with G + bacterial infection ( P<0.05). The sensitivity and the area under the ROC curve of serum IL-1β, IFN-γ and IL-4 levels used in combination in the differential diagnosis of G + /G - bacterial infection were respectively 91.62% and 0.960, which were higher than those of each parameter used alone ( P<0.01, P<0.05). However, there was no significant difference in the specificity ( P>0.05). Conclusions:The levels of serum IL-1β and IFN-γ increased in children with sepsis, while the serum IL-4 level was basically normal in them. Children with G - bacterial infection tended to have higher serum IL-1β, IFN-γ and IL-4 levels. The combination use of all three parameters showed higher value in the differential diagnosis of G + /G - bacterial infection.

Study on the Mechanism of Liuling Jiedu Pills Inhibiting Acute Pharyngitis Induced by Staphylococcus Aureus in Rats / 中药新药与临床药理

Objective To investigate the effect and mechanism of Liuling Jiedu Pills on acute pharyngitis caused by Staphylococcus aureus in rats.Methods The rat model of acute pharyngitis was replicated using the method of injecting 1×109 CFU·mL-1 of Staphylococcus aureus solution into the pharynx of rats.SD rats were randomly divided into a blank group,a model group,a Lanqin Oral Solution group(5 mL·kg-1),and a low-,medium-,and high-dose group of Liuling Jiedu Pills(4.375,8.750,and 17.500 mg·kg-1),with 10 rats in each group.Rats in each group were administered the drug by gavage once a day for 7 days.The general conditions of the rats were observed and recorded every day during the modeling and drug administration periods,and the local inflammation in the pharynx was scored;histopathological changes in the pharynx of the rats were observed by hematoxylin-eosin(HE)staining;serum interleukin 1β(IL-1β),interleukin 6(IL-6),tumor necrosis factor α(TNF-α),and tumor necrosis factor-α(TNF-α)were detected by ELISA.Immunohistochemistry and Western Blot were used to detect the protein expression levels of IL-1β,IL-6 and TNF-α in rat pharyngeal tissue.Results Compared with the blank group,rats in the model group had significantly increased pharyngeal erythema,significantly higher inflammation scores(P＜0.01),significantly lower body mass on days 5-7 after modeling(P＜0.05,P＜0.01),significantly higher pathological scores(P＜0.01),significantly higher levels of the serum inflammatory factors IL-1β,IL-6,and TNF-α(P＜0.01),and significantly higher pharyngeal tissues showed significantly higher levels of IL-1β,IL-6,and TNF-α proteins(P＜0.01).Compared with the model group,the pharyngeal erythema was significantly reduced in the Lanqin Oral Solution group and the low-,medium-and high-dose groups of Liuling Jiedu Pills,and the inflammation scores were significantly reduced(P＜0.01),and the serum levels of IL-1β,IL-6,and TNF-α were significantly reduced(P＜0.01);the body mass of the rats in the Lanqin Oral Solution group,and in the medium-and high-dose groups of Liuling Jiedu Pills,were significantly increased on the seventh day of the modeling(P＜0.01);the histopathological scores and the levels of IL-1β,IL-6 and TNF-α proteins in pharyngeal tissue were significantly decreased(P＜0.05,P＜0.01).Conclusion Liuling Jiedu Pills can significantly improve the symptoms and inflammatory pathological changes of pharyngeal tissues in rats with acute pharyngitis,and its mechanism may be related to the down-regulation of the expression levels of inflammatory factors such as IL-1β,IL-6,and TNF-α.

MS4A6D promotes carrageenan-induced mouse foot pad swelling by activating NLRP3 inflammasome / 免疫学杂志

We here investigated the effect of MS4A6D(the membrane spanning 4-domain subfamily A(MS4A)superfamily protein 6D),one of the tetraspanins which specifically expresses on the membrane of macrophages,on carrageenan(CGN)-induced mouse foot pad swelling and its possible mechanism.Male C57BL/6 wild-type(WT)and various gene knockout(including Ms4a6d-/-,Nlrp3-/-,Casp-1-/-and IlR1-/-)mice were recruited and injected with 100 μl 1%CGN(w/v)and 20 μl CaCl2(50 mmol/L)to establish the foot pad swelling model,and then the severity of foot pad swelling in WT and gene knockout mice were compared.H&E staining was performed to investigate the pathological changes,and immunofluorescence was used to detect the infiltration of F4/80+ macrophages in the foot pad tissue.Finally,Western blot was used to determine the protein expression of NLRP3,IL-1 β,TNF-α and IL-6.Data showed that the combined injection of 100 μl 1%CGN(w/v)and 20 μl CaCl2(50 mmol/L)significantly promoted the swelling of foot pads,while the degree of foot pad swelling in Ms4a6d-/-mice was significantly lower than that in WT littermates(P<0.05);Significant tissue damage and inflammatory infiltration as well as necrotic lesions were observed in the WT foot pads,whereas,the degrees from Ms4a6d-/-foot pads showed significantly reduced;The protein levels of Caspase-1 and IL-1β in the foot pad tissue of the WT model were significantly higher than those of the Ms4a6d-/-groups;Compared with WT controls,the degree of foot pad swelling in Nlrp3-/-,Casp-1-/-,and IlR1-/-mice induced by 1%CGN(w/v)and CaCl2(50 mmol/L)were also significantly reduced(P<0.05).Taken together,MS4A6D promotes the activation of NLRP3 inflammasome in macrophages and induces IL-1β secretion,by thus deteriorates CGN-mediated swelling of mouse foot pads.

Mechanism of Mongolian drug Naru-3 in initiation of neuroinflammation of neuropathic pain from MMP9/IL-1β signaling pathway / 中国中药杂志

Neuropathic pain(NP) has similar phenotypes but different sequential neuroinflammatory mechanisms in the pathological process. It is of great significance to inhibit the initiation of neuroinflammation, which has become a new direction of NP treatment and drug development in recent years. Mongolian drug Naru-3 is clinically effective in the treatment of trigeminal neuralgia, sciatica, and other NPs in a short time, but its pharmacodynamic characteristics and mechanism of analgesia are still unclear. In this study, a spinal nerve ligation(SNL) model simulating clinical peripheral nerve injury was established and the efficacy and mechanism of Naru-3 in the treatment of NPs was discussed by means of behavioral detection, side effect evaluation, network analysis, and experimental verification. Pharmacodynamic results showed that Naru-3 increased the basic pain sensitivity threshold(mechanical hyperalgesia and thermal radiation hyperalgesia) in the initiation of SNL in animals and relieved spontaneous pain, however, there was no significant effect on the basic pain sensitivity threshold and motor coordination function of normal animals under physiological and pathological conditions. Meanwhile, the results of primary screening of target tissues showed that Naru-3 inhibited the second phase of injury-induced nociceptive response of formalin test in mice and reduced the expression of inflammatory factors in the spinal cord. Network analysis discovered that Naru-3 had synergy in the treatment of NP, and its mechanism was associated with core targets such as matrix metalloproteinase-9(MMP9) and interleukin-1β(IL-1β). The experiment further took the dorsal root ganglion(DRG) and the stage of patho-logical spinal cord as the research objects, focusing on the core targets of inducing microglial neuroinflammation. By means of Western blot, immunofluorescence, agonists, antagonists, behavior, etc., the mechanism of Naru-3 in exerting NP analgesia may be related to the negative regulation of the MMP9/IL-1β signaling pathway-mediated microglia p38/IL-1β inflammatory loop in the activation phase. The relevant research enriches the biological connotation of Naru-3 in the treatment of NP and provides references for clinical rational drug use.

Effects of moxibustion on serum levels of β-EP, SP and expression of IL-1β and COX-2 protein in brainstem in rats with migraine / 中国针灸

OBJECTIVE@#To observe the effects of moxibustion at "Baihui" (GV 20) and "Dazhui" (GV 14) at different time points on the serum level of β-endorphin (β-EP), substance P (SP) and expression of interleukin-1β (IL-1β) and cyclooxygenase-2 (COX-2) protein in brainstem in rats with migraine, and to explore the effect and mechanism of moxibustion in preventing and treating migraine.@*METHODS@#Forty male SD rats were randomly divided into a blank group, a model group, a prevention+treatment (PT) group and a treatment group, 10 rats in each group. Except the blank group, the rats in the remaining groups were injected with nitroglycerin subcutaneously to prepare migraine model. The rats in the PT group were treated with moxibustion 7 days before modeling (once a day) and 30 min after modeling, while the rats in the treatment group were treated with moxibustion 30 min after modeling. The "Baihui" (GV 20) and "Dazhui" (GV 14) were taken for 30 minutes each time. The behavioral scores in each group were observed before and after modeling. After intervention, ELISA method was used to detect the serum level of β-EP and SP; the immunohistochemistry method was used to detect the number of positive cells of IL-1β in brainstem; the Western blot method was used to detect the expression of COX-2 protein in brainstem.@*RESULTS@#Compared with the blank group, the behavioral scores in the model group were increased 0-30 min, 60-90 min and 90-120 min after modeling (P<0.01); compared with the model group, in the treatment group and the PT group, the behavioral scores were decreased 60-90 min and 90-120 min after modeling (P<0.01). Compared with the blank group, in the model group, the serum level of β-EP was decreased (P<0.01), while the serum level of SP, the number of positive cells of IL-1β in brainstem and the expression of COX-2 protein were increased (P<0.01). Compared with the model group, in the PT group and and the treatment group, the serum level of β-EP was increased (P<0.01), while the serum level of SP, the number of positive cells of IL-1β and the expression of COX-2 protein in brainstem were decreased (P<0.01, P<0.05). Compared with the treatment group, in the PT group, the serum level of β-EP was increased and COX-2 protein expression was decreased (P<0.05).@*CONCLUSION@#Moxibustion could effectively relieve migraine. The mechanism may be related to reduce the serum level of SP, IL-1β and COX-2 protein expression in brainstem, and increase the serum level of β-EP, and the optimal effect is observed in the PT group.

Study of the improvement effect of "DuZhong-DangGui" on osteoarthritis rats via regulating NLPR3 inflammasome / 中国临床药理学与治疗学

AIM: To investigate the protective effect of "DuZhong-DangGui" (DZ-DG) on the knee tissue of rats with osteoarthritis (OA) and its regulation role on NLPR3 inflammasome. METHODS: Twenty-four SPF male SD rats, eighteen rats were randomly selected to establish OA model by anteri- or cruciate ligament amputation (ACLT) method, and rats were divided into control group, OA model group, DZ-DG group and positive drug group, 6 in each group, treatment for 8 weeks. The peripheral blood were collected, ELISA was used to detect the levels of IL-1β, IL-6, IL-18, and TNF-α; cartilage tissue of knee joint were collected, HE staining was used to observe pathology changes, OARSI staining was used to observe cartilage calcification and preform quantitative OARSI scoring; immunohistochemistry and TUNEL staining were used to detect the contents of Caspase1 and Collagen II and the number of apoptosis in the tissue, respectively, and western blot was used to detect the protein expressions of matrix metalloproteinase-13 (MMP-13), tissue inhibitor of metalloproteinase-1 (TIMP-1), p53, p21, NLPR3, apoptosis-associated speck-like protein containing a CARD (ASC) and Pro-Caspase-1. RESULTS: Compared to control group, OA model group rats serum levels of IL-1β, IL-6, IL-18, and TNF - α significantly increased (P<0.01), OARSI scores significantly increased (P<0.01), chondrocyte apoptosis was increased (P<0.01), Caspase-1 content and MMP-13, p53, p21, NLPR3, ASC, Pro-Caspase-1 protein expressions significantly increased (P <0.01), while Collagen II content and TIMP-1 protein expression significantly decreased (P<0.01). Compared with the OA model group, DZ-DG group and positive drug group rats serum levels of IL-1β, IL-6, IL -18 and TNF - α significantly decreased (P< 0.05), chondrocyte apoptosis were significantly decreased (P<0.01), Caspase1 content, MMP-13, p53, NLPR3, Pro-Caspase-1 significantly decreased (P< 0.05), Collagen Ⅱ content and TIMP- 1 protein expression (P<0.01); DZ-DG group rats protein expression of p21 and ASC were decreased (P<0.01). CONCLUSION: The DZ-DG have protection role on cartilage of OA rats, its effect may related to mediation of NLPR3/ASC/Pro-Caspase-1 pathway, to decrease of IL-1β, and inhibition of cell apoptosis.

Mechanism of Qingre Lishi Huazhuo Method（Modified Simiaosan） on Chronic Gouty Arthritis of Dampness-heat Obstruction Syndrome Based on NLRP3/IL-1β Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo study the clinical efficacy of the Qingre Lishi Huazhuo method on patients with chronic gouty arthritis of dampness-heat obstruction syndrome and the effect on nucleotide-binding oligomerization domain-like receptor 3（NLRP3）/interleukin-1β （IL-1β） signaling pathway to preliminarily explore its mechanism. MethodSixty patients with chronic gouty arthritis of dampness-heat obstruction syndrome were enrolled and divided into a treatment group （30 cases） and a control group （30 cases） according to the random number table method. Thirty people were assigned to the healthy group. Patients in the control group were treated with oral Febuxostat， while those in the treatment group were treated with modified Simiaosan combined with Febuxostat. Treatment lasted four weeks. The general clinical data， traditional Chinese medicine （TCM） syndrome scores， serum uric acid （UA）， serum creatinine （SCr）， blood urea nitrogen （BUN）， fasting blood glucose （FPG）， low-density lipoprotein （LDL）， triglyceride （TG）， total cholesterol （TC）， erythrocyte sedimentation rate （ESR）， and C-reactive protein （CRP） of patients were recorded. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the levels of IL-1β，TNF-α， and IL-6，and the levels of NLRP3，cysteinyl aspartate-specific protease-1 （Caspase-1）， and apoptosis-associated speck-like protein containing a CARD （ASC） were detected by Western blot. ResultBefore treatment， the levels of body mass index （BMI）， systolic blood pressure （SBP），diastolic blood pressure （DBP），UA，SCr，BUN，FPG，LDL，TG，and TC in both groups significantly increased （P<0.05，P<0.01），and the levels of HDL significantly decreased as compared with those in the healthy group（P<0.05）. Additionally， the levels of IL-1β， TNF-α， and IL-6 in both groups significantly increased before treatment （P<0.01）. Compared with the results before treatment， patients in the two groups had significant reductions in tube pain， joint tenderness， joint swelling，joint fever， activity disorders， body fatigue， sliminess， bitter mouth， yellow and red urine， and tongue manifestation scores （P<0.05，P<0.01）. Compared with patients in the control group after treatment， those in the treatment group had a significant decrease in joint fever， body fatigue， sliminess， bitter mouth，sticky stool，yellow and red urine， tongue manifestation score， and pulse score （P<0.05）. The total effective rate in the treatment group was 80.0% （24/30）， higher than 56.7% （17/30）in the control group（χ2=11.916，P<0.05）. Compared with the results before treatment， BMI， SBP， DBP， UA， SCr， BUN， FPG， LDL， TG， TC， ESR，CRP， IL-1β， TNF-α， IL-6 levels， and VAS score in both groups significantly decreased （P<0.05，P<0.01）. Compared with patients in the control group after treatment， those in the treatment group had decreased DBP，ESR， IL-1β levels， and VAS score （P<0.05）. Western blot results showed that before treatment， the protein expression of NLRP3， Caspase-1， and ASC in peripheral blood mononuclear cells （PBMCs） of patients in both groups were higher than those in the healthy group （P<0.01）. Compared with the results before treatment， the protein expression of NLRP3， Caspase-1， and ASC in PBMCs in patients of both groups after treatment decreased （P<0.05，P<0.01）. Compared with the control group after treatment， the treatment group showed decreased expression levels of NLRP3 and Caspase-1（P<0.05）. ConclusionThe Qingre Lishi Huazhuo method can effectively improve the clinical symptoms and reduce inflammation of chronic gouty arthritis of dampness-heat obstruction syndrome with good safety. The mechanism may be related to the inhibition of the NLRP3/IL-1β signaling pathway.

Mechanism of Chaihu Qinggantang in Intervening NLRP3/IL-1β Pathway to Treat Granulomatous Lobular Mastitis in Rat Model / 中国实验方剂学杂志

ObjectiveTo investigate the mechanism of Chaihu Qinggantang （CHQGT） in the treatment of granulomatous lobular mastitis （GLM） in the rat model. MethodSixty female rats were divided into a normal group， a model group， a prednisolone group （0.001 8 g·kg-1）， and three CHQGT low-dose， medium-dose， and high-dose groups （4.5， 8.9， 17.8 g·kg-1）. The tissue homogenates mixed with GLM lesion tissue and Fritner's reagent were used for modeling. After modeling， the treatment groups were given corresponding treatment factors， and the normal group and the model group were given the equal volume of normal saline. The changes in mammary gland of rats were observed after 14 d. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes in breast samples. The mRNA expressions of NOD-like receptor thermal protein domain associated protein 3 （NLRP3） inflammasome， Caspase-1， and interleukin-1β （IL-1β） were detected by real-time quantitative fluorescence polymerase chain reaction （Real-time PCR）. The protein expressions of NLRP3， Caspase-1， IL-1β， and IL-18 were detected by Western bolt. ResultAs compared with the normal group， the breasts of rats in the model group were obviously swelling， and mammary gland inflammation index was significantly increased （P<0.01）. Pathological changes included the formation of granuloma centered on the lobule of mammary gland with a large number of inflammatory cells such as lymphocytes and plasma cells. The mRNA expressions of NLRP3， Caspase-1， and IL-1β， and the protein expressions of NLRP3， Caspase-1， IL-1β， and IL18 in the model group were significantly increased （P<0.01）. Compared with the model group， the treatment groups improved breast swelling， and the CHQGT medium and high-dose groups and the prednisolone group reduced inflammation index to some extent after treatment （P<0.05， P<0.01）. The inflammation degree of mammary gland was significantly improved， and inflammatory cells such as macrophages， lymphocytes， and plasma cells were reduced to varying degrees in pathological aspects. The mRNA expressions of NLRP3， Caspase-1， and IL-1β， and the protein expressions of NLRP3， Caspase-1， IL-1β， and IL-18 in the CHQGT high-dose group and the prednisolone group were significantly down-regulated （P<0.05， P<0.01）. ConclusionCHQGT inhibits inflammation and treats GLM in rats. The mechanism is possibly related to the inhibition of NLRP3/IL-1β signaling pathway， which provides a new target for the prevention and treatment of GLM by Qingxiao method.

Effect mechanism of Ma Xing Gan Shi Decoction against influenza virus from JAK2/STAT3 signaling pathway by combining network pharmacology / 中国药理学通报

Aim To explore the effect of JAK2/STAT3 signaling pathway on lung tissue injury induced by influenza A virus in combination with network pharmacology and to further explore the intervention effect of Ma Xing Gan Shi Decoction.Methods Network pharmacological method was used to screen the signal pathway enriched by Ma Xing Gan Shi Decoction on the potential target of influenza virus.BLAB/c mice were intranasally infected with influenza A virus.The mice were divided into normal control group, model control group, oseltamivir group, antiviral granule group and Ma Xing Shigan decoction group.The animals were treated with corresponding drugs for 3 and 7 days.Body weight and lung index were detected by HE for observation of the pathological changes of lung tissues.Real-time PCR(RT-PCR)was used to detect the mRNA expression levels of JAK2, STAT3, IL-1β and IL-4 in lung tissues.Western blot and ELISA were used to detect the protein expression levels of JAK2, STAT3, IL-1β and IL-4 in lung tissues.AutoDock Vina software was used to conduct molecular docking between STAT3 and target compounds.Results The main active components of Ma Xing Gan Shi Decoction had 110 intersection targets with influenza virus and were enriched in 170 signaling pathways.Ma Xing Shigan decoction could up-regulate the body weight of mice infected with influenza A virus, improve the pathological injury of lung tissues, down-regulate the lung index and the expression levels of JAK2, STAT3, IL-1β mRNA and protein in lung tissues, and up-regulate the expression levels of IL-4 mRNA and protein.STAT3 had better binding activity with glycyrrhiza chalcone A, an active compound in Ma Xing Gan Shi Decoction.Conclusions Ma Xing Gan Shi Decoction, as an effective compound prescription of traditional Chinese medicine against influenza virus, can effectively reduce pulmonary inflammation and regulate the balance of cytokines.The possible mechanism is to alleviate the lung injury caused by influenza A virus infection in mice by inhibiting the activation of JAK2/STAT3 signal pathway.

Enphorbia lunulata Bge extract inhibits interleukin-1β-induced colorectal cancer cell proliferation, migration and invasion mechanism through miR-30a-5p/NFκB / 中国药理学通报

Aim To investigate whether the Enphorbia lunulata Bge extract regulates the proliferation, migration and invasion of colorectal cancer cells induced by interleukin-1β(IL-1β)through miR-30a-5p/nuclear factor κB(NF-κB). Methods HT29 cells were divided into NC group, IL-1β group, low-dose(2.5 mg·L-1)+IL-1β group, middle-dose(5 mg·L-1)+IL-1β group, high-dose(10 mg·L-1)+IL-1β group, miR-NC+IL-1β group, miR-30a-5p+IL-1β group, anti-miR-NC+high-dose+IL-1β group, anti-miR-30a-5p+high-dose+IL-1β group. Cell counting kit-8 method was used to detect cell proliferation activity in each group, clone formation experiment was applied to assess cell clones, Transwell method was employed to monitor cell migration and invasion, Western blot was performed to determine the protein expression level, and qRT-PCR was employed to detect the expression level of miR-30a-5p. Results Compared with the NC group, the proliferation activity, cell clone number, migration and invasion number of colorectal cancer cell HT29 in IL-1β group increased, and the expression level of miR-30a-5p decreased(all P<0.01). Compared with the IL-1β group, the proliferation activity, the number of cell clones, the number of migration and invasion of colorectal cancer cell HT29 decreased, and the expression level of miR-30a-5p increased(all P<0.01); The expression level of p-p65 in the high-dose+IL-1β group was lower than that in the IL-1β group(P<0.01). The proliferation activity, cell clone number, migration and invasion number of colorectal cancer cell HT29 in the miR-30a-5p+IL-1β group were lower than those in the miR-NC+IL-1β group(all P<0.01). The proliferation activity, cell clone number, migration and invasion number of colorectal cancer cell HT29 in the anti-miR-30a-5p+high-dose+IL-1β group were higher than those of anti-miR-NC+high-dose+IL-1β group(all P<0.01). Conclusions Enphorbia lunulata Bge extract can inhibit the proliferation, migration and invasion of colorectal cancer cells induced by IL-1β by up-regulating miR-30a-5p and down-regulating the activity of NF-κB signaling pathway.

PM / 中国生物化学与分子生物学报

Particulate matter (PM) can damage respiratory system, cardiovascular system, nervous system and immune system, but there are few researches on reproductive damage of particulate matter. The objectives of this study were to investigate the effect of short-term particulate matter 2.5 (PM

Bavachin enhances NLRP3 inflammasome activation induced by ATP or nigericin and causes idiosyncratic hepatotoxicity / 医学前沿

Psoraleae Fructus (PF) is a well-known traditional herbal medicine in China, and it is widely used for osteoporosis, vitiligo, and other diseases in clinical settings. However, liver injury caused by PF and its preparations has been frequently reported in recent years. Our previous studies have demonstrated that PF could cause idiosyncratic drug-induced liver injury (IDILI), but the mechanism underlying its hepatotoxicity remains unclear. This paper reports that bavachin isolated from PF enhances the specific stimuli-induced activation of the NLRP3 inflammasome and leads to hepatotoxicity. Bavachin boosts the secretion of IL-1β and caspase-1 caused by ATP or nigericin but not those induced by poly(I:C), monosodium urate crystal, or intracellular lipopolysaccharide. Bavachin does not affect AIM2 or NLRC4 inflammasome activation. Mechanistically, bavachin specifically increases the production of nigericin-induced mitochondrial reactive oxygen species among the most important upstream events in the activation of the NLRP3 inflammasome. Bavachin increases the levels of aspartate transaminase and alanine aminotransferase in serum and hepatocyte injury accompanied by the secretion of IL-1β via a mouse model of lipopolysaccharide-mediated susceptibility to IDILI. These results suggest that bavachin specifically enhances the ATP- or nigericin-induced activation of the NLRP3 inflammasome. Bavachin also potentially contributes to PF-induced idiosyncratic hepatotoxicity. Moreover, bavachin and PF should be evaded among patients with diseases linked to the ATP- or nigericin-mediated activation of the NLRP3 inflammasome, which may be a dangerous factor for liver injury.

Mangiferin attenuates IL / 中南大学学报(医学版)

OBJECTIVES@#Chondrocyte apoptosis is an important process in the pathogenesis of osteoarthritis. Mangiferin exerts multiple pharmacological effects such as anti-inflammatory and anti-apoptosis. However, the role of mangiferin in chondrocyte apoptosis is not clear. In this study, we aimed to explore the role of mangiferin in IL-1β-induced chondrocyte apoptosis.@*METHODS@#ATDC5 cells were randomly divided into a control group, a IL-1β group, a MFN-L group, a MFN-M group, a MFN-H group and a MFN+LY294002 group. Cells in the control group were treated with IL-1β (10 ng/mL) for 24 h; cells in the MFN-L group, the MFN-M group and the MFN-H group were pretreated with 5, 10 and 20 μmol/L mangiferin for 1 h respectively, and then they were treated with IL-1β (10 ng/mL) for 24 h; cells in the MFN+LY294002 group were treated with LY294002 (25 μmol/L) for 1 h, then mangiferin (20 μmol/L) and IL-1β (10 ng/mL) for 1 h and 24 h, respectively. Cell viability was detected by CCK-8 assay and cell apoptosis was measured by flow cytometry. Colorimetric assay was conducted to measure the caspase-3 activity. The protein levels of Bcl-2, Bax, and phosphoinositide 3-kinase (PI3K)/Akt signaling pathway related proteins were detected by Western blotting.@*RESULTS@#Compared to the control group, cell viability was significantly decreased; cell apoptosis, caspase-3 activity and Bax protein expression were significantly increased; the protein levels of Bcl-2, p-PI3K, and p-Akt were significantly decreased in the IL-1β group (all @*CONCLUSIONS@#Mangiferin could attenuate IL-1β-induced apoptosis of the mice chondrocytes, which is mediated by the activation of PI3K/Akt signaling pathway.

Effect of phosphoglycerate mutase 5 mediated pyroptosis on liver ischemia-reperfusion injury / 器官移植

Objective To investigate the effect and its molecular mechanism of phosphoglycerate mutase 5 (PGAM5) mediated pyroptosis on liver ischemia-reperfusion injury (IRI). Methods C57 mouse models of liver IRI were established and randomly divided into the 6 h reperfusion (6 h group) and 12 h reperfusion (12 h group), and sham operation group (sham group) was established too, 10 rats in each group. The effect of IRI on the parameters in the liver tissues and serum samples was evaluated. The expression levels of PGAM5 and cysteinyl aspartate specific proteinase (Caspase)-1 in the liver tissues during IRI were quantitatively detected. The IRI models of liver cells were established (IRI group). The IRI models of liver cells were established after pretreatment with Caspase-1 inhibitor Z-YVAD-FMK (inhibitor group). The untreated AML12 cells were allocated into the control group. The effect of inhibiting Caspase-1 activity on pyroptosis was analyzed. AML12 cells were transfected with PGAM5 small interfering ribonucleic acid (siRNA) (siRNA group) and siRNA-negative control (siRNA-NC) (siRNA-NC group) by liposome 3000, and then IRI models of liver cells were established. The untreated AML12 cells were assigned into the control group. The effect of PGAM5 mediated pyroptosis on IRI of liver cells was assessed. Results In the 6 h and 12 h groups, partial liver cell edema, hepatic sinusoid narrowing, central vein congestion and occasional spot necrosis were observed in the mouse liver tissues, and these changes in the 12 h group were more aggravated than those in the 6 h group. The serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the 6 h and 12 h groups were higher than those in the sham group, and the values in the 12 h group were higher than those in the 6 h group. The levels of tumor necrosis factor (TNF)-α and interleukin (IL)-1β were increased in the 6 h and 12 h groups, and the values in the 12 h group were lower than those in the 6 h group. The relative expression levels of IL-1β messenger ribonucleic acid (mRNA) in the mouse liver tissues in the 6 h and 12 h groups were up-regulated, and the value in the 12 h group was lower than that in the 6 h group. The cell apoptosis rates in the liver tissues were significantly increased in the 6 h and 12 h groups, and the value in the 12 h group was remarkably lower than that in the 6 h group (P < 0.01-0.05). Compared with the sham group, the relative expression levels of PGAM5 mRNA and protein in the mouse liver tissues in the 6 h and 12 h groups were significantly up-regulated, and the values in the 12 h group were significantly higher than those in the 6 h group (P < 0.01-0.05). The protein expression levels of PGAM5 and Caspase-1 in the liver tissues were up-regulated in the 6 h and 12 h groups. Compared with the control group, the relative expression levels of NOD-like receptor protein 3 (NLRP3), cleaved Caspase-1 and Gasdermin D (GSDMD) proteins were up-regulated and the fluorescence intensity of GSDMD was increased in the IRI group. Compared with the IRI group, the relative expression levels of NLRP3, cleaved Caspase-1 and GSDMD proteins were significantly down-regulated and the fluorescence intensity of GSDMD was considerably decreased in the inhibitor group (P < 0.01-0.05). Compared with the control group, the cell survival rate was significantly decreased, and the relative expression levels of PGAM5, NLRP3, cleaved Caspase-1 and GSDMD proteins were significantly up-regulated in the siRNA-NC group (P < 0.01-0.05). Compared with the siRNA-NC group, the cell survival rate was remarkably increased, whereas the relative expression levels of PGAM5, NLRP3, cleaved Caspase-1 and GSDMD proteins were significantly down-regulated in the siRNA group (P < 0.01-0.05). Conclusions PGAM5 may aggravate the liver IRI in mouse models probably by mediating pyroptosis via PGAM5/Caspase-1/GSDMD signaling pathway and aggravating liver cell injury.

p-Coumaric acid alleviates adriamycin-induced hepatotoxicity in rats

Objective: To evaluate the effect of p-coumaric acid against adriamycin-induced hepatotoxicity in rats. Methods: The rats were divided into 4 groups. The control group received solvent; the p-coumaric acid group was treated with 100 mg/kg of p-coumaric acid orally for five consecutive days; the adriamycin group was administered with a single dose of adriamycin (15 mg/kg, i.p.), and the p-coumaric acid + adriamycin group was given p-coumaric acid five days before adriamycin administration. Twenty-four hours after the last administration, blood samples were collected for biochemical analysis, and liver tissues were removed for histopathological and immunohistochemistrical studies. Moreover, the levels of tissue lipid peroxidation and enzyme activities of glutathione peroxidase, superoxide dismutase, and catalase in liver tissue were measured. Results: Treatment with p-coumaric acid protected the liver from the toxicity of adriamycin by attenuating the increase in alkaline phosphatase, alanine transaminase, aspartate transaminase, total bilirubin, total cholesterol, triglyceride, and low-density lipoprotein cholesterol and lessening the decrease in high-density lipoprotein cholesterol and albumin. p-Coumaric acid also raised the levels of glutathione peroxidase, superoxide dismutase, and catalase, as well as decreased lipid peroxidation in liver tissue and hepatic IL-1β expression. Additionally, histopathological study confirmed the protective effect of p-coumaric acid against liver damage. Conclusions: p-Coumaric acid can alleviate adriamycin-induced hepatotoxicity.

Aryl Hydrocarbon Receptor is Involved in the Proinflammatory Cytokine Response to Cadmium / 生物医学与环境科学（英文）

Objective@#To investigate involvement of the aryl hydrocarbon receptor (AhR) in the immunomodulatory effects of cadmium (Cd).@*Methods@#The effect of Cd on AhR activation ( @*Results@#Cd increased @*Conclusion@#AhR signaling is involved in the lung leukocyte proinflammatory cytokine response to Cd. The relevance of the AhR to the cytokine response to Cd provides new insight into the mechanisms of Cd immunotoxicity.