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.

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.

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.

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.

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.

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.

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.

Relationship of Microbial Profile With Airway Immune Response in Eosinophilic or Neutrophilic Inflammation of Asthmatics

PURPOSE: Different characteristics of airway microbiome in asthmatics may lead to differential immune responses, which in turn cause eosinophilic or neutrophilic airway inflammation. However, the relationships among these factors have yet to be fully elucidated.METHODS: Microbes in induced sputum samples were subjected to sequence analysis of 16S rRNA. Airway inflammatory phenotypes were defined as neutrophils (>60%) and eosinophils (>3%), and inflammation endotypes were defined by levels of T helper (Th) 1 (interferon-γ), Th2 (interleukin [IL]-5 and IL-13), Th-17 (IL-17), and innate Th2 (IL-25, IL-33, and thymic stromal lymphopoietin) cytokines, inflammasomes (IL-1β), epithelial activation markers (granulocyte-macrophage colony-stimulating factor and IL-8), and Inflammation (IL-6 and tumor necrosis factor-α) cytokines in sputum supernatants was assessed by enzyme-linked immunosorbent assay.RESULTS: The numbers of operational taxonomic units were significantly higher in the mixed (n = 21) and neutrophilic (n = 23) inflammation groups than in the paucigranulocytic inflammation group (n = 19; p < 0.05). At the species level, Granulicatella adiacens, Streptococcus parasanguinis, Streptococcus pneumoniae, Veillonella rogosae, Haemophilus parainfluenzae, and Neisseria perflava levels were significantly higher in the eosinophilic inflammation group (n = 20), whereas JYGU_s levels were significantly higher in the neutrophilic inflammation group compared to the other subtypes (P < 0.05). Additionally, IL-5 and IL-13 concentrations were correlated with the percentage of eosinophils (P < 0.05) and IL-13 levels were positively correlated with the read counts of Porphyromonas pasteri and V. rogosae (P < 0.05). IL-1β concentrations were correlated with the percentage of neutrophils (P < 0.05). had a tendency to be positively correlated with the read count of JYGU_s (P = 0.095), and was negatively correlated with that of S. pneumoniae (P < 0.05).CONCLUSIONS: Difference of microbial patterns in airways may induce distinctive endotypes of asthma, which is responsible for the neutrophilic or eosinophilic inflammation in asthma.

Effect of Baihu and Renshen Decoction on intestinal TLR4/NF-κB signaling pathway and intestinal barrier in type 2 diabetes MKR mice / 中草药

Objective: To observe the effect of Baihu and Renshen Decoction on intestinal barrier and TLR4/NF-κB inflammatory signaling pathway in type 2 diabetes MKR mice. Methods: A total of 32 MKR mice were randomly divided into model group, low dose Baihu and Renshen Decoction group, high dose Baihu and Renshen Decoction group and metformin group after 4 weeks with the high-fat diet. Another eight FVB/N mice were served as control group. After 4-week intragastric administration, fasting blood glucose (FBG), oral glucose tolerance test (OGTT), the levels of interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and lipopolysaccharide (LPS) in serum were examined. The morphological changes of colon tissue were observed by hematoxylin-eosin (HE) staining. The mRNA expression of TLR4, NF-κB, ZO-1, Claudin-1 and Occludin were measured by qRT-PCR method. The protein expression of ZO-1, Claudin-1 and Occludin in colon were measured by Western blotting. Results: Compared with model group, FBG in each administration group were significantly decreased (P < 0.01), serum IL-6, IL-1β, TNF-α, LPS and TLR4, NF-κB mRNA expressions were decreased (P < 0.05, P < 0.01) in high dose Baihu and Renshen Decoction group and metformin group. The mucosal epithelium integrity was improved. The mRNA and protein expression levels of ZO-1, Claudin-1, Occludin were increased (P < 0.05, P < 0.01) in high dose Baihu and Renshen Decoction group and metformin group; The mRNA expression levels of ZO-1, Claudin-1, Occludin were increased (P < 0.05) in low dose Baihu and Renshen Decoction group, while protein expressions showed no statistical difference. Conclusion: Baihu and Renshen Decoction can consequently reduce the levels of LPS and pro-inflammatory cytokines in MKR mice, regulate TLR4/NF-κB signaling pathway. It can significantly improve insulin resistance and reduce blood glucose levels by improving endotoxaemia, ameliorating intestinal injury and attenuating inflammation.

Effect of pretreatment of acupuncture on learning-memory ability and related protein of NLRP3 inflammasome in hippocampus in Alzheimer's disease like rats / 中国针灸

OBJECTIVE@#To observe the effect of pretreatment of acupuncture on the expression of nucleotide-binding oligomerization domain-like receptor 3(NLRP3), Caspase-1, interleukin1β(IL-1β) and the number of activated microglia (MG) in the hippocampus in Alzheimer's disease (AD) like rats, so as to explore the mechanism of pretreatment of acupuncture in preventing and treating AD.@*METHODS@#A total of 36 SD rats were randomly divided into a blank group, a model group and an electroacupuncture (EA) group, 12 rats in each group. The AD like rat model was established by 8-week continuous intraperitoneal injection of D-galactose (120 mg·kg@*RESULTS@#Compared with the blank group, the average escape latency was prolonged (@*CONCLUSION@#Pretreatment of acupuncture could prevent and treat the learning-memory dysfunction in AD like rats, and its mechanism may be related to the inhibition of NLRP3 inflammatsome related protein and MG activation.

Comparasion on Effect of Xuefu Zhuyutang and Modified Erchentang on Nonalcoholic Fatty Liver in Mice / 中国实验方剂学杂志

Objective:To compare the effect and mechanisms of modified Erchentang and Xuefu Zhuyutang on high-fat diet-induced apolipoprotein-E knockout (ApoE-/-) mice nonalcoholic fatty liver disease (NAFLD). Method:Ten C57/BL6J mice were taken as normal control group and fed with normal feed. Totally 30 ApoE-/- mice were fed with high-fat diet to establish a disease model for 4 weeks. After 4 weeks, the 30 ApoE-/- mice were divided into model group, Xuefu Zhuyutang group (hereinafter referred to as Huoxue group) and modified Erchentang group (hereinafter referred to as Huatan group) by random number table method, with 10 in each group. The normal group and the model group were intragastrically administered with normal saline. The drug-administered group was intragastrically administered at a dosage that was ten times of the adult dose, once a day, for 8 weeks. Serum and liver were collected after the end of the 12-week experiment. The serum lipid and liver function levels of each group were measured, and the liver pathological morphology was observed. Protein and mRNA expressions of liver inflammatory mediators interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), matrix metalloproteinase-9 (MMP-9), monocyte chemotactic factor-1 (MCP-1) were detected by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) and Western blot. Result:The results of serum lipids and liver function showed that compared with the normal group, serum total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) in the model group were significantly increased, while serum high-density lipoprotein (HDL) was significantly reduced (P<0.01). Compared with the model group, serum TG ，LDL and ALT were significantly decreased, HDL was significantly increased in the Huoxue group (P<0.05). The serum levels of TC, TG， LDL, AST and ALT in the Huatan group were significantly decreased，HDL was significantly increased (P<0.05，P<0.01), and TG was decreased. The mice serum HDL in the Huatan group was higher than that in the Huoxue group. The serum ALT in the Huoxue group was lower than that in the Huatan group. The pathological observation showed that compared with the normal group, hepatocytes in the model group had severe steatosis with many lipid droplet vacuoles, suggesting that the mouse NAFLD model was successful. Compared with the model group, each administration group alleviated hepatocyte steatosis, with no significant difference between the two administration groups. Western blot and Real-time PCR results showed that compared with the normal group, protein and mRNA expression levels of TNF-α, IL-1β, MCP-1, and MMP-9 in the model group were significantly increased (P<0.05，P<0.01). Compared with the model group, the Huoxue group significantly down-regulated the expressions of IL-1β, MCP-1 protein and MCP-1 mRNA(P<0.05，P<0.01). The Huatan group significantly reduced the expressions of IL-1β, TNF-α, MMP-9, MCP-1 protein, TNF-α and MMP-9 mRNA(P<0.05，P<0.01). Conclusion:Modified Erchentang and Xuefu Zhuyutang can alleviate the therapeutic effect of NAFLD mice to a certain extent, modified Erchentang has a better therapeutic effect.

Effect of Modified Buwangsan on Expression of Related Molecules in NLRP3 Inflammatory Signaling Pathway in Hippocampus of AD Rats / 中国实验方剂学杂志

Objective::To investigate the effects of modified Buwangsan on the learning and memory ability of Alzheimer's disease (AD) model rats and the expression of NOD-like receptor 3 (NLRP3), cysteine-containing aspartate-specific proteases 1 (Caspase-1) and interleukin-1 beta (IL-1β) in NLRP3 inflammatory pathway in hippocampus of AD model rats, and exploring the underlying mechanism of modified Buwangsan. Method::The 52 eligible rats were randomly divided into sham control group, AD model group, low-dose modified Buwangsan group (1.5 g·kg-1) and high-dose modified Buwangsan group (3 g·kg-1). AD mouse model was established by bilateral hippocampus injection of Aβ1-425 μL (2 g·L-1). The rats in low-dose and high-dose modified Buwangsan group received low and high dose modified Buwangsan respectively within the next 4 weeks, once daily. The learning and memory ability was tested by Morris water maze. The expression of NLRP3, Caspase-1 and IL-1β mRNA was tested by quantitative PCR(Real-time PCR) and Western blot. Result::As compared with the sham group, the learning and memory ability of the rats were significantly impaired (P<0.05). Compared with AD model group, the learning and memory ability and the expression levels of NLRP3, Caspase-1, and IL-1β mRNA and protein were all no statistical differences in low-dose modified Buwangsan group, while the learning and memory ability of the rats were significantly improved and the expression of NLRP3, Caspase-1 and IL-1β mRNA in hippocampus of rats was significantly decreased in high-dose modified Buwangsan group (P<0.05). Conclusion::High-dose modified Buwangsan could attenuate neuroinflammation in the hippocampus of AD mouse model via inhibiting the expression of NLRP3, Caspase-1 and IL-1β, which may be the mechanisms of modified Buwangsan could be used to ameliorate the learning and memory ability of AD mouse model.

Effect of Huoxue Ointment on Osteoarthritis Caused by Knee Joint Injury in Rabbits / 中国实验方剂学杂志

Objective:To investigate the efficacy and anti-inflammatory and analgesic effect of Huoxue ointment on osteoarthritis of the knee caused by knee joint injury in rabbits. Method:A total of 60 big ear white rabbits were selected, with half male and half female. Ten rabbits were randomly selected by weight and set as normal control group. For the remaining rabbits, the injury of articular cartilage and anterior cruciate ligament were caused in the femoral ankle joint surface of the right hind knee joint, so as to establish an animal model of knee osteoarthritis in rabbits. For the normal control group, the skin was cut open and immediately sutured. Two weeks after the operation, rabbits were randomly divided into model control group according to body weight，positive voltaren group (1 cm·kg-1·d-1), and high-dose (6 g·kg-1·d-1), medium-dose (3 g·kg-1·d-1) and low-dose(1.5 g·kg-1·d-1) Huoxue ointment groups, with 10 animals in each group. Interleukinin-1β （IL-1β） in rabbit articular cavity fluid and peripheral blood were measured, contents of tumor necrosis factor-α （TNF-α） and free radical nitric oxide（NO）, rabbit articular cartilage was examined by histopathology, in order to study the efficacy of Huoxue ointment on rabbit knee arthritis. The model of increased permeability of capillaries in abdominal cavity of mice induced by acetic acid and the model of foot swelling of rats induced by carrageenan were used to determine the absorbance of mice peritoneal fluid and the rat foot swelling value, and study the anti-inflammatory effect of huoxue ointment. A total of 120 SPF ICR mice and 60 SD rats were selected, with half male and half female. The mice were randomly divided according to body weight into model control groups, positive Voltaren group (3 cm·kg-1·d-1), and high-dose (16 g·kg-1·d-1), medium-dose (8 g·kg-1·d-1), low-dose (4 g·kg-1·d-1) Huoxue ointment groups，with 12 animals in each group. The rats were randomly divided into the model control group, positive voltaren group (2 cm·kg-1·d-1), and high-dose (8 g·kg-1·d-1), medium-dose (4 g·kg-1·d-1), low-dose (2 g·kg-1·d-1) Huoxue ointment groups，with 10 animals in each group. Result:The tested dose of Huoxue ointment，the levels of IL-1β, TNF-α and NO in rabbit serum and joint fluid were decreased to varying degrees, and the effect was significant in the high-dose group (P<0.05). According to the pathological examination, the tested dose of Huoxue ointment, knee cartilage and synovial tissue lesions were significantly lower than those of the model control group (P<0.05)，the value of peritoneal fluid concentration and the foot swelling in medium and high-dose Huoxue ointment groups were significantly lower than those in the model control group (P<0.05). The analgesic effect of tail shaking in mice, and the pain threshold rate of the high-dose Huoxue ointment group were significantly higher than those of the model control group (P<0.05)， the analgesic effect lasted for 4 hours, the foot tenderness of rats in the large and medium-dose Huoxue ointment groups was significantly higher than that in the model control group (P<0.05). Conclusion:Huoxue ointment has obvious joint repair and anti-inflammatory and analgesic effects on osteoarthritis of knee caused by knee joint injury in rabbits.

Licochalcone A Inhibits Proliferation and Induces Apoptosis of MH7A Cells by Regulating MAPK Signaling Pathway / 中国实验方剂学杂志

Objective:To study the effects of licochalcone A （LCA） on the proliferation and apoptosis of rheumatoid arthritis fibroblast-like synoviocytes （MH7A） as well as the related inflammatory factors， also to reveal the relevance between mitogen activated protein kinase （MAPK） signaling pathway and LCA regulation of MH7A cell proliferation and apoptosis. Method:MH7A cells were cultured and divided into blank group， LCA groups （10，20，40 μmol·L-1）. The proliferation of MH7A cells was detected by methylthiazolyldiphenyl-tetrazolium bromide（MTT）and immunofluorescence staining. The cell cycle of MH7A cells was determined by flow cytometry after PI staining and apoptosis was detected by flow cytometry after Annexin V/PI staining. The effect of LCA on interleukin-1β（IL-1β） mRNA was detected by Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）. Western blot was used to detect the effect of LCA on the key proteins of MAPK signaling pathway， meanwhile， PD98059， a specific ERK inhibitor， was used to observe the expression levels of p-ERK and IL-1β. Result:Compared with blank group， LCA could inhibit the proliferation of MH7A cells in a dose-dependent manner， and the number of living cells decreased significantly（P<0.01）， while the number of early apoptotic cells increased significantly（P<0.01）. Compared with the tumor necrosis factor-α （TNF-α，10 μg·L-1）group， LCA could reverse the expression of IL-1β mRNA induced by TNF-α（P<0.01）. and compared with the blank group， LCA also promoted the phosphorylation of ERK， JNK and p38 in a dose-dependent manner（P<0.01）. After ERK inhibitor PD98059 inhibited ERK phosphorylation， the inhibitory effect of LCA 10， 20 μmol·L-1 on IL-1β disappeared. Conclusion:LCA can inhibit the proliferation and induce apoptosis of MH7A cells， which may be related to the phosphorylation of MAPK pathway related proteins， and then inhibit the expression of inflammatory factors.

Effects of electroacupuncture pretreatment on cardiac function and immune inflammatory response in mice with acute myocardial ischemia / 中国针灸

OBJECTIVE@#To observe the effects of electroacupuncture (EA) pretreatment on the cardiac ejection fraction (EF), the number of macrophages in spleen and heart, and the expression of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) and interleukin-1β (IL-1β) in myocardium in mice with acute myocardial ischemia, and to explore the possible mechanism of EA pretreatment on promoting myocardial protection.@*METHODS@#A total of 30 male C57BL/6J mice were randomly divided into a control group, a model group and an EA pretreatment group, 10 rats in each group. The acute myocardial ischemia model was established by ligating the left anterior descending branch of the coronary artery in the model group and EA pretreatment group, while threading but no ligating at left anterior descending branch of the coronary artery was applied in the control group. In the EA pretreatment group, mice were intervented with EA at bilateral "Neiguan" (PC 6), disperse-dense wave, frequency of 2 Hz/15 Hz, intensity of 2 mA; each EA treatment last for 20 min, once a day, and 3-day treatment was given before model establishment. The EF value was evaluated by ultrasonic cardiogram; the number of macrophages in spleen and heart was measured by flow cytometry; the expression level of NLRP3 and IL-1β in myocardium was measured by Western blot.@*RESULTS@#Compared with the control group, the EF value was decreased in the model group (<0.001), the number of macrophages in the heart and spleen was increased (<0.001), and the expression level of NLRP3 and IL-1β in the myocardium was increased (<0.001, <0.01). Compared with the model group, the EF value was increased in the EA pretreatment group (<0.01), the number of macrophages in the heart and spleen was decreased (<0.01), and the expression level of NLRP3 and IL-1β in the myocardium was decreased (<0.01, <0.05).@*CONCLUSION@#EA pretreatment could reduce the number of macrophages in spleen and heart, down-regulate the expression of NLRP3 and IL-1β in myocardial tissue in mice with acute myocardial ischemia, which could relieve the local inflammatory response and achieve the myocardial protective effect.

Effect of acupuncture on NLRP3 inflammatory corpuscle in rats with intracerebral hemorrhage / 中国针灸

OBJECTIVE@#To observe the effect of acupuncture at "Baihui" (GV 20) through "Qubin" (GB 7) on NLRP3 inflammatory corpuscle in rats with intracerebral hemorrhage (ICH), and to explore the action mechanism of acupuncture on promoting the recovery of neural function in rats with ICH.@*METHODS@#Forty SPF six-week-old male SD rats were randomly divided into a sham operation group, a model group, a non-acupoint group and an acupuncture group, 10 rats in each group. The rats in the model group, non-acupoint group and acupuncture group were intervened with autologous blood injection to prepare ICH model, while the rats in the sham operation group were only intervened with operation but not injection with autologous blood. About 3 hours after the establishment of the model, the rats in the acupuncture group were intervened with acupuncture at "Baihui" (GV 20) through "Qubin" (GB 7), once every 12 hours, for 7 days; the rats in the non-acupoint group were intervened with acupuncture at the non-acupoint [parallel to the "Baihui" (GV 20), 1 cm next to the midline] on the affected side, and other treatment was the same as the acupuncture group. At the end of the intervention, the composite nerve function score of each group was evaluated; the histomorphology of the hemorrhage penumbra was observed by HE staining; the expression of NLRP3 inflammatory corpuscle in the brain was detected by immunohistochemistry; the relative protein expression levels of NLRP3, interleukin-1β (IL-1β) and interleukin-18 (IL-18) in brain were detected by the method of Western blot.@*RESULTS@#Seven days into intervention, compared with the sham operation group, each item score and total score of composite nerve function in the model group were significantly reduced (<0.01, <0.05). There was edema and karyopyknosis in brain neuron as well as necrocytosis and inflammatory cell infiltration in the model group. Compared with the model group and the non-acupoint group, the total score of composite nerve function and the scores of symmetrical movement of limbs (LS) and proprioception of tentacles (VP) in the acupuncture group were increased (<0.01, <0.05), and the cell necrosis and inflammatory cell infiltration were relieved. Compared with the sham operation group, NLRP3 inflammatory corpuscle expression and the relative protein expression levels of NLRP3, IL-1β and IL-18 in brain tissue in the model group were increased (<0.01); compared with the model group and the non-acupoint group, NLRP3 inflammatory corpuscle expression and the relative protein expression levels of NLRP3, IL-1β and IL-18 in brain tissue in the acupuncture group were reduced (<0.01).@*CONCLUSION@#Acupuncture at "Baihui" (GV 20) through "Qubin" (GB 7) could downregulate the expression of NLRP3, IL-1β and IL-18 in the brain tissue of ICH rats, inhibit the inflammatory response, and promote the recovery of neural function.

Anti-inflammatory dietary supplements in prevention of diseases in geriatric people

As our age advances, many changes are seen in our body, such as cellular changes, cardiovascular problems, cerebrovascular diseases, including cancer mediated by inflammation and their mediators such as free radicals (ROS, RNS), cytokines, transcription factors (NF-KB, STAT3) due to altered dietary patterns and digestive disorders. The disease pattern can be suppressed by including anti-inflammatory dietary supplements in our diet to prevent various diseases in geriatric peoples associated with inflammation. Chronic activation of the inflammatory response, defined as inflammation, is the key physio-pathological substrate for anabolic resistance, sarcopenia and frailty in older individuals. Nutrients can theoretically modulate this phenomenon. This article briefs about anti-inflammatory dietary supplements in prevention of diseases associated with inflammation in geriatric people.

Evaluation of Antidiabetic Potentiality of Truffles and Balanites Aegyptiaca among Streptozotocin Induced Diabetic Rats

The potential application of Truffle (Terfeziaceae) and Desert Date tree (Balanites aegyptiaca) for modulating the diabetesmellitus related symptoms has gained much interest. However, less firm evidence has come from data to increase theunderstanding of the mechanism by which Truffle and Balanites protect pancreatic β-cells. The present study aimed toevaluate the effect of methanolic extract of Truffle and Balanites aegyptiaca on the fasting blood glucose (FBG) level; thechanges in pancreatic histology as well as the changes in iNOS and IL-1β genes expression level among STZ induceddiabetic rats which might help in better clarification of possible mechanisms beside the beneficial effects of the studiedplants on diabetes. The elevation of the FBG level, pathological pancreatic changes and the level of both IL-1β and iNOSgene expressions in diabetic rats were observed in comparison with negative control rats. This increase was declinedsignificantly due to the administration of Truffle and Balanites extracts. All of the studied parameters did not completelyreverse to the normal levels as compared with negative control rats. The obtained results concluded that the beneficial effectof Truffle and Balanites aegyptiaca on STZ-induced diabetes was at least partly due to the reduction of IL-1β and iNOS geneover the expression which can have a protective effect on β cell.