1.Advances in research on cardiomyocyte pyroptosis mediated by NLRP3 inflammasome and traditional Chinese medicine intervention.
Zi-Shuo GE ; Zhi-Meng XU ; Ping LI
China Journal of Chinese Materia Medica 2021;46(20):5210-5217
Pyroptosis is a pro-inflammatory programmed cell death, and its role in cardiac inflammatory response has become a hot topic. The activation of nucleotide binding oligomerization domain like receptor protein 3(NLRP3) inflammasome is an important mechanism for pyroptosis induced by cysteinyl aspartate specific proteinase 1(caspase-1). The existing studies have shown that cardiomyocyte pyroptosis participates in the pathogenesis of different cardiovascular diseases and the NLRP3 inflammasome-mediated cardiomyocyte pyroptosis has been most widely studied. Also, the intervention in NLRP3 inflammasome activation and cardiomyocyte pyroptosis contributes to ameliorating myocardial injury, which may be the main mechanism of many traditional Chinese medicines in exerting the cardio-protective effects. Therefore, this paper reviewed the studies on cardiomyocyte pyroptosis mediated by NLRP3 inflammasome and put forward the importance of exploring traditional Chinese medicine intervention in the activation of NLRP3 inflammasome.
Inflammasomes
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Medicine, Chinese Traditional
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Myocytes, Cardiac
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NLR Family, Pyrin Domain-Containing 3 Protein/genetics*
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Pyroptosis
2.Therapeutic mechanism of Psammosilene tunicoides extract on rheumatoid arthritis based on NLRP3 inflammasome.
Zhi-Ming HE ; Zhi-Pu HUANG ; Ying-Chun WEI ; Meng CUI ; Zi-Gang QIAN ; Hai-Feng CHEN
China Journal of Chinese Materia Medica 2021;46(17):4504-4510
In this study, we investigated the mechanism of crude extract of Psammosilene tunicoides(CEPT) in the treatment of rheumatoid arthritis(RA) based on the Nod-like receptor protein 3(NLRP3) inflammasome. The collagen-induced arthritis(CIA) mouse model was established. On day 32 after the primary immunization, according to the arthritis score, the mice were randomly divided into model group, positive control(methotrexate) group, low-and high-dose CEPT groups, and normal group, with 10 mice in each group. According to the administration dose of each group, the mice were continuously administered for 21 days. Every four days during the administration, the paw edema degree, arthritis score, and spleen index of the mice were measured; histopathological examination was performed for the ankles of the mice; the contents of IL-1β and IL-18 in the serum were determined; the protein expression levels of NLRP3, caspase-1, and apoptosis-associated speck-like protein containing a CARD(ASC), as well as the mRNA expression levels of NLRP3 and caspase-1 in the ankle joints of the mice were detected. The results showed that compared with those in the model group, the mice in the positive control group and CEPT groups had significantly decreased the contents of IL-1β and IL-18 in the serum and spleen index(P<0.01), significantly lowered arthritis score and degree of paw edema(P<0.01), alleviated arthritic infiltration of the knee, and down-regulated protein and mRNA levels of NLRP3, ASC, and caspase-1 in the ankle joint(P<0.01). These results suggest that P. tunicoides may reduce the paw edema and arthritis score and alleviate the inflammatory response in CIA mice by inhibiting the expression of NLRP3. This study provides a basis for the study of immune regulation of P. tunicoides in RA.
Animals
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Arthritis, Experimental/genetics*
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Arthritis, Rheumatoid/genetics*
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Caspase 1/genetics*
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Inflammasomes/genetics*
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Mice
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NLR Family, Pyrin Domain-Containing 3 Protein/genetics*
3.Effect of pretreatment of acupuncture on learning-memory ability and related protein of NLRP3 inflammasome in hippocampus in Alzheimer's disease like rats.
Chuan HE ; Zhong-Sheng HUANG ; Hong-Ru CHEN ; Chao-Chao YU ; Xue-Song WANG ; Tao JIANG ; Li-Hong KONG
Chinese Acupuncture & Moxibustion 2020;40(12):1323-1327
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.
Alzheimer Disease/therapy*
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Animals
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Electroacupuncture
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Hippocampus
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Inflammasomes
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NLR Family, Pyrin Domain-Containing 3 Protein/genetics*
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Rats
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Rats, Sprague-Dawley
4.Pathogenic role of NDUFA13 inactivation in spontaneous hepatitis in mice and the mechanism.
Xiaohui XU ; Rui LI ; Xin ZENG ; Xin WANG ; Bingqian XUE ; Daochao HUANG ; Yi HUANG
Journal of Southern Medical University 2021;41(1):55-63
OBJECTIVE:
To investigate the role of NDUFA13 inactivation in the pathogenesis of spontaneous hepatitis in mice and explore the possible mechanisms.
METHODS:
Hepatocyte-specific NDUFA13 knockout (NDUFA13
RESULTS:
Liver-specific NDUFA13 heterozygous knockout mice were successfully constructed as verified by PCR results. HE staining revealed severe liver damage in both 4- week-old and 2-year-old NDUFA13
CONCLUSIONS
Hepatocytes-specific NDUFA13 ablation can trigger spontaneous hepatitis in mice possibly mediated by the activation of ROS/NF-κB/NLRP3 signaling.
Animals
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Hepatitis
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Inflammasomes
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Mice
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NF-kappa B/metabolism*
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NLR Family, Pyrin Domain-Containing 3 Protein/genetics*
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Signal Transduction
5.Effect of extract of Quzhou Aurantii Fructus on hepatic inflammation and NF-κB/NLRP3 inflammasome pathway in CCl_4-induced liver fibrosis mice.
Si-Wei WANG ; Tian LAN ; Fang ZHENG ; Mei-Kang LEI ; Feng ZHANG
China Journal of Chinese Materia Medica 2021;46(6):1474-1479
To study the effect and mechanism of extract of Quzhou Aurantii Fructus(QAF) on liver inflammation in CCl_4-induced liver fibrosis mice. Totally 60 C57 BL/6 male mice were randomly divided into control group(distilled water, oral), model group(distilled water, oral), colchicines group(Col, colchicines 2 mg·kg~(-1)·d~(-1), oral), low-dose QAF group(QAF-L, QAF 100 mg·kg~(-1)·d~(-1), oral) and high-dose QAF group(QAF-H, QAF 300 mg·kg~(-1)·d~(-1), oral) by random number table method. The model group and each administration group were injected with carbon tetrachloride(CCl_4) 1 mL·kg~(-1)(CCl_4-olive oil 1∶4), twice a week, totally 6 weeks. After the last administration, the mice were sacrificed, and serum and liver tissue were collected. Serum ALT and AST levels were measured in each group to observe the liver function of mice. The pathological changes and inflammatory cell infiltration in liver were observed by HE staining and F4/80 immunohistochemical staining. The mRNA expressions of TNF-α, IL-18 and IL-1β were detected by RT-PCR. The protein expressions of IκBα, p-IKKα/β, p-p65, NLRP3, caspase-1 and cleaved caspase-1 were analyzed by Western blot. The results showed that QAF significantly reduced serum ALT and AST levels, and alleviated the degree of liver damage.The results of immunohistochemistry showed that QAF significantly reduced liver inflammatory cell infiltration in liver fibrosis mice. The results of RT-PCR and Western blot showed that QAF significantly inhibited mRNA expressions of TNF-α, IL-18 and IL-1β in liver of fibrosis mice. QAF also suppressed the degradation of IκBα protein and reduced p-IKKα/β, p-p65, NLRP3 and cleaved caspase-1 protein expressions. In conclusion, QAF improves CCl_4-induced liver fibrosis in mice. The mechanism may be related to the inhibition of NF-κB/NLRP3 inflammasome-mediated inflammation signaling pathway.
Animals
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Inflammasomes/genetics*
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Inflammation
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Liver/pathology*
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Liver Cirrhosis/genetics*
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Male
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Mice
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NF-kappa B/genetics*
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NLR Family, Pyrin Domain-Containing 3 Protein/genetics*
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Plant Extracts
6.Investigation of inflammasome during excitation of IgG-HepG2 cells for evaluation of allergenic ingredients.
Xi LU ; Mingwei WANG ; Fan LEI ; Yugang WANG ; Zhiyi YUAN ; Shuang ZHAO ; Huiyu LI ; Dongming XING ; Lijun DU
China Journal of Chinese Materia Medica 2011;36(14):1855-1859
OBJECTIVETo investigate the alteration of inflammasome and receptor during IgG promoter transfected to HepG2 cells.
METHODBy assay of Elisa to evaluate the secretion of IL-1 beta, IL-8, TNF-alpha and MCP-1 after puerarine and LPS administration, and by assay of real time PCR to evaluate the expression of mRNA of IL-1 beta, IL-8,TNF-alpha and MCP-1, as well as the receptors of TLR2, 4 and NOD2, MyD88.
RESULTIgG promoter did not active innate immunity and enhance the expression and secretion of inflammasome in HepG2. Puerarine did not active the inflammasome either. LPS activated the innate immunity and increased the secretion of IL-8, TNF-alpha and MCP-1.
CONCLUSIONIgG-HepG2 cells could be used specifically as the model of allergy type II for ingredients screening. It is suggested that puerarine was suite for the activator for this type of allergy as positive control.
Allergens ; analysis ; immunology ; Drugs, Chinese Herbal ; chemistry ; Gene Expression Regulation ; immunology ; Hep G2 Cells ; Humans ; Immunity, Innate ; immunology ; Immunoglobulin G ; genetics ; Inflammasomes ; immunology ; Promoter Regions, Genetic ; genetics ; Transfection
7.Effect of polysaccharides from seeds of Vaccaria segetalis in alleviating urinary tract infection induced bladder injury by inhibiting NLRP3 inflammasome.
Xin MAO ; Rong-Mei YAO ; Yan-Yan BAO ; Jing SUN ; Xiao-Lan CUI ; Hai-Jiang ZHANG ; Yu-Jing SHI
China Journal of Chinese Materia Medica 2021;46(13):3388-3393
To study the mechanism of polysaccharides from seeds of Vaccaria segetalis( PSV) in the treatment of bacterial cystitis through the NLRP3 inflammasome pathway. The rat model of urinary tract infection was used and treated with PSV,and the urine and bladders were collected. The level of interleukin-10( IL-10) in rat urine was detected by enzyme linked immunosorbent assay( ELISA). Western blot and immunofluorescence staining were used to detect the expressions of sonic hedgehog( SHH) and NLRP3 inflammasome [NOD-like receptor thermoprotein domain 3( NLRP3),apoptosis associated speck like protein( ASC) and pro-caspase-1]. The expression of Toll-like receptor pathway was detected by RT-PCR. The death of 5637 cells induced by uropathogenic Escherichia coli( UPEC) and lactate dehydrogenase( LDH) release were evaluated using live/dead staining. The results showed that in the rat bladder,the expressions of SHH,NLRP3 inflammasomes and Toll-like receptors were significantly up-regulated,and NLRP3 inflammasomes were significantly activated by UPEC infection. The administration with PSV could significantly increase the concentration of IL-10 in urine,inhibit the expressions of SHH,NLRP3 inflammasomes and Toll-like receptors in bladder,and inhibit the activation of NLRP3 inflammasomes. A large number of 5637 cells were dead after UPEC infection and caused LDH production. PSV could significantly inhibit the death of 5637 cells and the release of LDH. In conclusion,PSV could inhibit the expression and activation of NLRP3 inflammasomes by inhibiting the Toll-like receptor pathway,thereby mitigating the bladder injury.
Animals
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Hedgehog Proteins
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Inflammasomes/genetics*
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Interleukin-1beta
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NLR Family, Pyrin Domain-Containing 3 Protein/genetics*
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Polysaccharides/pharmacology*
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Rats
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Seeds
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Urinary Bladder
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Urinary Tract Infections/drug therapy*
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Vaccaria
8.10,11-Dehydrocurvularin attenuates inflammation by suppressing NLRP3 inflammasome activation.
Qun ZHAO ; Mengyuan FENG ; Shu JIN ; Xiaobo LIU ; Shengbao LI ; Jian GUO ; Xinran CHENG ; Guangbiao ZHOU ; Xianjun YU
Chinese Journal of Natural Medicines (English Ed.) 2023;21(3):163-171
10,11-Dehydrocurvularin (DCV) is a natural-product macrolide that has been shown to exert anti-inflammatory activity. However, the underlying mechanism of its anti-inflammatory activity remains poorly understood. Aberrant activation of the NLRP3 inflammasome is involved in diverse inflammation-related diseases, which should be controlled. The results showed that DCV specifically inhibited the activation of the NLRP3 inflammasome in association with reduced IL-1β secretion and caspase-1 activation, without effect on the NLRC4 and AIM2 inflammasomes. Furthermore, DCV disturbed the interaction between NEK7 and NLRP3, resulting in the inhibition of NLRP3 inflammasome activation. The C=C double bond of DCV was required for the NLRP3 inflammasome inhibition induced by DCV. Importantly, DCV ameliorated inflammation in vivo through inhibiting the NLRP3 inflammasome. Taken together, our study reveals a novel mechanism by which DCV suppresses inflammation, which indicates the potential role of DCV in NLRP3 inflammasome-driven inflammatory disorders.
Animals
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Mice
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Inflammasomes
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NLR Family, Pyrin Domain-Containing 3 Protein/genetics*
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Inflammation/drug therapy*
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Anti-Inflammatory Agents/pharmacology*
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Interleukin-1beta/genetics*
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Mice, Inbred C57BL
9.Effect of NLRP3 inflammasome on vascular diseases.
Zeng CAO ; Yapei LI ; Ruifang CHEN ; Pingyu ZENG
Journal of Central South University(Medical Sciences) 2016;41(11):1232-1236
The NLRP3 inflammasome, a protein complex belonging to the family of nucleotide-binding and oligomerization domain like receptors (NLRs), plays a vital role in the innate immune system. It promotes pro-caspase 1 cleavage into active caspase-1, which contributes to maturation and releases of IL-1β and IL-18 in response to the harmful signals and participates in the host immune response and sterile inflammation. Recently a large number of studies have shown that NLRP3 inflammasome closely relates to the pathogenesis of the vascular diseases. NLRP3 inflammasome, which involves in the sterile inflammation of the vascular wall, plays an important role in the pathogenesis of main, middle and small arteries.
Caspase 1
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immunology
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metabolism
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Gene Expression Regulation
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genetics
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immunology
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Humans
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Inflammasomes
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immunology
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Inflammation
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complications
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genetics
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Interleukin-18
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genetics
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immunology
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Interleukin-1beta
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genetics
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immunology
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NLR Family, Pyrin Domain-Containing 3 Protein
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immunology
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Signal Transduction
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genetics
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immunology
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Vascular Diseases
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etiology
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genetics
;
immunology
10.NLRC4 plays a regulatory role in F. nucleatum-induced pyroptosis in macrophages.
Wei Yi JIANG ; Zi Long DENG ; Wang Hong ZHAO
Journal of Southern Medical University 2022;42(10):1560-1565
OBJECTIVE:
To explore the mechanism of F.nucleatum-induced pyroptosis in macrophages and the regulatory role of inflammasomes.
METHODS:
Lactate dehydrogenase (LDH) cytotoxicity assay and Hoechst 33342/PI double fluorescence staining were used to analyze cytolysis in F.nucleatum-infected macrophage RAW264.7 cells.The expressions of pyroptosis-related proteins caspase-1, GSDMD and IL-1β were determined using Western blotting.Inflammasome activation in the cells was analyzed by detecting the mRNA expressions of NLRP3, NLRC4, AIM2, and NLRP1 with qRT-PCR.RNA interference technique was used to knock down the key molecules involved in pyroptosis regulation in the macrophages, and the pyroptosis and necrosis rates of the cells following F.nucleatum infection were examined.
RESULTS:
The results of LDH cytotoxicity assay and double-fluorescence staining showed that F.nucleatum infection caused swelling and lytic cell death in RAW264.7 cells.F.nucleatum infection resulted in the activation of caspase-1 and GSDMD and upregulated IL-1β expression in a multiplicity of infection (MOI)-and time-dependent manner (P < 0.05).qRT-PCR revealed significantly increased expression of NLRC4 mRNA in the macrophages after F.nucleatum infection (P < 0.05).NLRC4 silencing by siRNA strongly inhibited the activation of caspase-1/GSDMD pathway and reduced cell death (P < 0.05) and IL-1β expression in F.nucleatum-infected cells.
CONCLUSION
NLRC4 inflammasome drives caspase-1/GSDMD-mediated pyroptosis and inflammatory signaling in F.nucleatum-infected macrophages, suggesting the potential of NLRC4 inflammasome as a therapeutic target for F.nucleatum infections.
Pyroptosis/genetics*
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Inflammasomes/metabolism*
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Caspase 1/metabolism*
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Interleukin-1beta/metabolism*
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Macrophages/metabolism*
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RNA, Messenger/metabolism*
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NLR Family, Pyrin Domain-Containing 3 Protein/metabolism*