OBJECTIVE The pathological characteristics of nonalcoholic steatohepatitis(NASH)include liver steatosis,inflammation,and fibrosis.Fibrosis is the most severe and significant pathological feature in NASH.Effective drug...OBJECTIVE The pathological characteristics of nonalcoholic steatohepatitis(NASH)include liver steatosis,inflammation,and fibrosis.Fibrosis is the most severe and significant pathological feature in NASH.Effective drug treatment could reverse early liver fibrosis and is of significance to prevent NASH from progressing into cirrhosis and liver cancer.Identification of drug targets for NASH treatment has been an active research area and is essential for the development of anti-NASH medications.Naringenin(NGN)is a flavonoid compound rich in citrus fruits.Our preliminary data demonstrated that NGN reduced diet-induced lipid accumulation and inflammation in the mouse liver,but whether NGN can attenuate liver fibrosis of NASH is not known.METHODS To study the effect of NGN on NASH fibrosis.WT mice were fed with high fat diet(HFD)and injected intraperitoneally 20%carbon tetrachloride at the same time for 8 weeks to induce NASH,and NGN was administrated by gavage in the meantime.In vitro,LO2 cells and LX2 cells were stimulated by oleic acid(OA)combined with lipopolysaccharide(LPS),respectively.RESULTS Treating the WT mice with NGN 100 mg·kg^(-1)·d-1 significantly attenuated hepatic lipid accumulation,hepatic fibrosis,plasma ALT and AST levels,inhibited protein expression of p-ERK,p-FoxO3a in the mouse livers.In vitro,on OA and LPS stimulated LO2 or LX2 cells,NGN significantly promoted apoptosis of activated hepatic stellate cells while inhibited apoptosis of hepatocytes.Mechanism study indicated that NGN inhibited MAPK pathway and promoted activation of FoxO3a,consequently promoted apoptosis of the activated LX2 cells and inhibited liver fibrosis.CONCLUSION NGN preventes NASH fibrosis via regulating MAPK/FoxO3a pathway,thus promoting apoptosis of the activated hepatic stellate cells.展开更多
Aim The present study developed a CYP3A4-expressed Caco-2 monolayer model at which effects of the efflux-metabolism alliance on the transport and uptake of clausenamide（CLA） enantiomers as CYP3A4 substrates were inv...Aim The present study developed a CYP3A4-expressed Caco-2 monolayer model at which effects of the efflux-metabolism alliance on the transport and uptake of clausenamide（CLA） enantiomers as CYP3A4 substrates were investigated. The apparent permeability coefficients （Papp） of （ - ） and （ ＋ ）CLA were higher in the ab- sorptive direction than those in the secretory direction with efflux ratios（ER） of 0. 709 ± 0.411 and 0. 867± 0. 250 （ Х10^-6 -1 cm · s ）, respectively. Their bidirectional transports were significantly reduced by （75.6 ± 87.5）% af- ter treatment with verapamil （ a P-glycoprotein inhibitor） that increased the rate of metabolism by CYP3 A4, whereas the CYP3A4 inhibitor ketoconazole treatment markedly enhanced the basolateral to apical flux of （ - ） and （ ＋ ） CLA with ERs being 2. 934 ± 1. 432 and 1. 877 ± 0. 148 （ Х 10^-6 cm/s） respectively. These changes could be blocked by the duel CYP3A4/P-glycoprotein inhibitor cyclosporine A, consequently, Papp values for CLA enanti- omers in both directions were significantly greater than those obtained by using verapamil or ketoconazole, and their ERs were similar to those following （ - ） or （ ＋ ）-isomer treatment alone. Furthermore, the uptake of （ - ）CLA was more than that of （ ＋ ）CLA in the transfected cells. Incubation with ketoeonazole decreased the intracellular concentrations of the two enantiomers. This effect disappeared in the presence of a CYP3A4 inducer dexametha- sone. These results indicated that CYP3A4 could influence P-gp efflux, transport and uptake of CLA enantiomers as CYP3A4 substrates and that a duel inhibition to CYP3A4/ P-glycoprotein could enhance their absorption and bioavailability, which provides new insight into the efflux-metabolism alliance and will benefit the clinical pharma- cology of （？） CLA as a candidate drug for treatment of Alzheimer＇ s disease.展开更多
CDC25A, TSSK3 and P53 expressions in vitro in cultured sertoli cells after FSH treatment were studied in order to provide some data for further researches of spermatogenesis. Different concentrations of FSH（0, 0.01, ...CDC25A, TSSK3 and P53 expressions in vitro in cultured sertoli cells after FSH treatment were studied in order to provide some data for further researches of spermatogenesis. Different concentrations of FSH（0, 0.01, 0.02, 0.04, and 0.08 IU ·m L^-1） were used to treat sertoli cells cultured in vitro. The expression of CDC25 A, TSSK3 and P53 was determined by real-time-PCR at 6 h,12 h and 24 h after FSH treatment of sertoli cells. The results showed that FSH had no significant effect on expression of CDC25A（p〈0.05）, could significantly improve the expression of TSSK3 and P53（p0.05）, and had no significant effect on expression of CDC25 A in sertoli cells, but it could significantly improve the expression of TSSK3. CDC25 A was likely to play a role in other signaling pathways in sertoli cells. Within the range of certain concentration of FSH, TSSK3 in sertoli cells had the highest expression at about 24 h. TSSK3 protein produced in sertoli cells was likely to play an important role in substrate-level phosphorylationbe in meiosis and mitosis of spermatogenic cells. FSH could promote P53 expression and the highest expression was at about 12 h, and P53 might control the division of spermatogenic cells as well as sertoli cells.展开更多
OBJECTIVE Basic fibroblast growth factor(b FGF)and platelet-derived growth factor(PDGF)produced by hepatocellular carcinoma(HCC)cells are responsible for the cell growth.Accumulating evidence shows that insulin-like g...OBJECTIVE Basic fibroblast growth factor(b FGF)and platelet-derived growth factor(PDGF)produced by hepatocellular carcinoma(HCC)cells are responsible for the cell growth.Accumulating evidence shows that insulin-like growth factor-binding protein-3(IGFBP-3)suppresses HCC cell proliferation in both IGF-dependent and independent manners.The present study is to investigate whether treatment with exogenous IGFBP-3 inhibits bF GF and PDGF production and the cell proliferation of HCC cells.METHODS Cell Counting Kit 8 assay were designed to detect HCC cell proliferation,transcription factor early growth response-1(EGR1)involving in IGFBP-3 regulation of b FGF and PDGF were detected by RT-PCR and Western blot assays.Western blot assay was adopted to detect the IGFBP-3 regulating insulin-like growth factor 1 receptor(IGF-1R)signaling pathway.RESULTS The present study demonstrates that IGFBP-3 suppressed IGF-1-induced b FGF and PDGF expression while it does not affect their expression in the absence of IGF-1.To delineate the underlying mechanism,Western-blot and RT-PCR assays confirmed that the transcription factor early growth response protein 1(EGR1)is involved in IGFBP-3 regulation of b FGF and PDGF.IGFBP-3 inhibition of type 1 insulin-like growth factor receptor(IGF1R),ERK and AKT activation is IGF-1-dependent.Furthermore,transient transfection with constitutively activated AKT or MEK partially blocks the IGFBP-3 inhibition of EGR1,b FGF and PDGF expression.CONCLUSION In conclusion,these findings suggest that IGFBP-3suppresses transcription of EGR1 and its target genes b FGF and PDGF through inhibiting IGF-1-dependent ERK and AKT activation.It demonstrates the importance of IGFBP-3 in the regulation of HCC cell proliferation,suggesting that IGFBP-3 could be a target for the treatment of HCC.展开更多
Identification and enumeration of both dendritic Ia^+ epi-dermal cells (Ia^+DECs) and dendritic Thy-1^+ epidermalcells (Thy-1^+DECa) from various parts of the body wereexamined by using epidermal sheets of C3H/He inbr...Identification and enumeration of both dendritic Ia^+ epi-dermal cells (Ia^+DECs) and dendritic Thy-1^+ epidermalcells (Thy-1^+DECa) from various parts of the body wereexamined by using epidermal sheets of C3H/He inbred miceof different age groups and indirect immunofluorescent tech-nique. A significant decline of both Ia^+DECs and Thy-1^+DECs in the mice of the aged group was demonstrated anddifferent densities and different distribution patterns betweenIa^+DECs and Thy-1^+DECs were obserged. These findingsmay imply that the decline of both Ia^+DECs and Thy-1^+DECs in the aged group may reflect the alterations of im-mune response in aging.展开更多
Aim To investigate the effects of JAK inhibitor （SHR0302） on adjuvant-induced arthritis （AA） rats and the partial mechanisms focused on T, B lymphocyte subsets through JAK1-STAT3 pathway, including Thl7, Treg, tot...Aim To investigate the effects of JAK inhibitor （SHR0302） on adjuvant-induced arthritis （AA） rats and the partial mechanisms focused on T, B lymphocyte subsets through JAK1-STAT3 pathway, including Thl7, Treg, total B cells and memory B cells. Methods Animals were divided randomly into 6 groups including normal control, AA, SHR0302 （0.3, 1.0, 3.0 nag · kg^-1, ig） and MTX （0.5 nag · kg^-1 , ig） . The effects of SHR0302 on AA rats by evaluating arthritis index, arthritis global assessment and paw swelling degree, histopathology of joint and spleen, inflammatory cytokine and antibody production in serum. We examined the proliferation of T, B and FLS by CCK8 kit; Thl7, Treg, total B and memory B cell proportion was measured by flow cytometry; Cytokines TNF-αβ, IL-1β, IL-10, IL-17 and antibody IgG1, IgG2a levels in serum were measured by ELISA kits; The ex- pression of p-JAK1 and p-STAT3 was measured by Western blot analysis. Results SHR0302 suppressed the se- verity of AA rats by attenuating the arthritis index, arthritis global assessment and paw swelling degree, and allevia- ted histopathology of spleen and joint of AA rats. SHR0302 can inhibit the proliferation of T, B and FLS, and down-regulated cytokines TNF-α, IL-1β, IL-17 and antibody IgG1, IgG2a levels, and suppressed the proportion of Thl7 and total B, and inhibited JAK1-STAT3 phosphorylation; There was no significant effect on Treg function and memory B cell proportion. Conclusion SHR0302 may attenuate the severity of AA rats, partially through signifi- cantly reducing Thl7 function and total B cell proportion by inhibiting JAK1-STAT3 phosphorylation.展开更多
Direct carbon solid oxide fuel cells(DC-SOFCs)are promising,green,and efficient power-generating devices that are fueled by solid carbons and comprise all-solid-state structures.Developing suitable anode materials for...Direct carbon solid oxide fuel cells(DC-SOFCs)are promising,green,and efficient power-generating devices that are fueled by solid carbons and comprise all-solid-state structures.Developing suitable anode materials for DC-SOFCs is a substantial scientific challenge.Herein we investigated the use of La_(0.75)Sr_(0.25)Cr_(0.5)Mn_(0.5)O_(3)-δ−Ce_(0.8)Gd_(0.2)O_(1.9)(LSCM−GDC)composite electrodes as anodes for La_(0.9)Sr_(0.1)Ga_(0.8)Mg_(0.2)O_(3)-δelectrolyte-based DC-SOFCs,with Camellia oleifera shell char as the carbon fuel.The LSCM−GDC-anode DC-SOFC delivered a maximum power density of 221 mW/cm^(2) at 800℃ and it significantly improved to 425 mW/cm^(2) after Ni nanoparticles were introduced into the LSCM−GDC anode through wet impregnation.The microstructures of the prepared anodes were characterized,and the stability of the anode in a DC-SOFC and the influence of catalytic activity on open circuit voltage were studied.The above results indicate that LSCM–GDC anode is promising to be applied in DC-SOFCs.展开更多
P2X7 receptor （P2X7r） is important in inflammation and fibrosis. The aim of the present study was to investigate the effect of P2X7r inhibition, using a specific inhibitor （A438079） to prevent the development of l...P2X7 receptor （P2X7r） is important in inflammation and fibrosis. The aim of the present study was to investigate the effect of P2X7r inhibition, using a specific inhibitor （A438079） to prevent the development of liver fibrosis on human hepatic stellate cells, LX-2. The supernatant from lipopolysaccharide （LPS）-stimulated RAW 264.7 mouse macrophages was supplemented to LX-2 cells for 24 h. LX-2 cells were primed with LPS for 4 h and subsequently stimulated for 30 rain with 3 mmol · L^-1 of adenosine 5＇-triphosphate （ATP）. A438079 （ 10 μmol · L^-1） was supplemented to LX-2 cells 10 rain prior to ATP. Directly treated with LPS on LX-2 cells, mRNA ex- pressions of IL-1β, IL-18 and IL-6 were increased, as well as P2X7r. And caspase-1, ASC and NLRP3 mRNA ex- pressions were increased with LPS stimulation. LPS stimulation also increased oL-SMA and collagen I mRNA expres- sions. Interestingly treatment of LX-2 cells with mediums from LPS-primed RAW 264.7 mouse macrophages exhibi- ted greater increase of mRNA expressions of above genes than those in LX-2 directly treated with LPS. Pretreatment of directly or indirectly LPS-stimulated LX-2 cells with A438079 both suppressed IL-1β mRNA expression. In addi- tion treatment of LPS-primed LX-2 cells with 3 mM ATP induced the significant increase of IL-1β, IL-6, caspase- 1, pannexin-1, α-SMA and collagen I mRNA expression, the increasing of oL-SMA protein expression and cleavage of IL-1β. These events were significantly suppressed by pretreatment with P2X7r antagonist A438079. P2XTr blockade also significantly reduced the protein expression of oL-SMA. Our results suggest that the involvement of the P2X7r-NLRP3 inflammasome pathway in the secretion of IL-1β from extracellular ATP/LPS-stimulated human he- patic stellate cells. This study demonstrated that repression of the P2XTr represents a novel potential therapeutic ap- proach to control liver fibrosis.展开更多
文摘OBJECTIVE The pathological characteristics of nonalcoholic steatohepatitis(NASH)include liver steatosis,inflammation,and fibrosis.Fibrosis is the most severe and significant pathological feature in NASH.Effective drug treatment could reverse early liver fibrosis and is of significance to prevent NASH from progressing into cirrhosis and liver cancer.Identification of drug targets for NASH treatment has been an active research area and is essential for the development of anti-NASH medications.Naringenin(NGN)is a flavonoid compound rich in citrus fruits.Our preliminary data demonstrated that NGN reduced diet-induced lipid accumulation and inflammation in the mouse liver,but whether NGN can attenuate liver fibrosis of NASH is not known.METHODS To study the effect of NGN on NASH fibrosis.WT mice were fed with high fat diet(HFD)and injected intraperitoneally 20%carbon tetrachloride at the same time for 8 weeks to induce NASH,and NGN was administrated by gavage in the meantime.In vitro,LO2 cells and LX2 cells were stimulated by oleic acid(OA)combined with lipopolysaccharide(LPS),respectively.RESULTS Treating the WT mice with NGN 100 mg·kg^(-1)·d-1 significantly attenuated hepatic lipid accumulation,hepatic fibrosis,plasma ALT and AST levels,inhibited protein expression of p-ERK,p-FoxO3a in the mouse livers.In vitro,on OA and LPS stimulated LO2 or LX2 cells,NGN significantly promoted apoptosis of activated hepatic stellate cells while inhibited apoptosis of hepatocytes.Mechanism study indicated that NGN inhibited MAPK pathway and promoted activation of FoxO3a,consequently promoted apoptosis of the activated LX2 cells and inhibited liver fibrosis.CONCLUSION NGN preventes NASH fibrosis via regulating MAPK/FoxO3a pathway,thus promoting apoptosis of the activated hepatic stellate cells.
文摘Aim The present study developed a CYP3A4-expressed Caco-2 monolayer model at which effects of the efflux-metabolism alliance on the transport and uptake of clausenamide（CLA） enantiomers as CYP3A4 substrates were investigated. The apparent permeability coefficients （Papp） of （ - ） and （ ＋ ）CLA were higher in the ab- sorptive direction than those in the secretory direction with efflux ratios（ER） of 0. 709 ± 0.411 and 0. 867± 0. 250 （ Х10^-6 -1 cm · s ）, respectively. Their bidirectional transports were significantly reduced by （75.6 ± 87.5）% af- ter treatment with verapamil （ a P-glycoprotein inhibitor） that increased the rate of metabolism by CYP3 A4, whereas the CYP3A4 inhibitor ketoconazole treatment markedly enhanced the basolateral to apical flux of （ - ） and （ ＋ ） CLA with ERs being 2. 934 ± 1. 432 and 1. 877 ± 0. 148 （ Х 10^-6 cm/s） respectively. These changes could be blocked by the duel CYP3A4/P-glycoprotein inhibitor cyclosporine A, consequently, Papp values for CLA enanti- omers in both directions were significantly greater than those obtained by using verapamil or ketoconazole, and their ERs were similar to those following （ - ） or （ ＋ ）-isomer treatment alone. Furthermore, the uptake of （ - ）CLA was more than that of （ ＋ ）CLA in the transfected cells. Incubation with ketoeonazole decreased the intracellular concentrations of the two enantiomers. This effect disappeared in the presence of a CYP3A4 inducer dexametha- sone. These results indicated that CYP3A4 could influence P-gp efflux, transport and uptake of CLA enantiomers as CYP3A4 substrates and that a duel inhibition to CYP3A4/ P-glycoprotein could enhance their absorption and bioavailability, which provides new insight into the efflux-metabolism alliance and will benefit the clinical pharma- cology of （？） CLA as a candidate drug for treatment of Alzheimer＇ s disease.
基金Supported by the National International Scientific and Technological Cooperation Project(2011DFA30760-2-1)Fund of Key Laboratory of Northeast Agricultural University,China(GXZDSYS-2012-07)
文摘CDC25A, TSSK3 and P53 expressions in vitro in cultured sertoli cells after FSH treatment were studied in order to provide some data for further researches of spermatogenesis. Different concentrations of FSH（0, 0.01, 0.02, 0.04, and 0.08 IU ·m L^-1） were used to treat sertoli cells cultured in vitro. The expression of CDC25 A, TSSK3 and P53 was determined by real-time-PCR at 6 h,12 h and 24 h after FSH treatment of sertoli cells. The results showed that FSH had no significant effect on expression of CDC25A（p〈0.05）, could significantly improve the expression of TSSK3 and P53（p0.05）, and had no significant effect on expression of CDC25 A in sertoli cells, but it could significantly improve the expression of TSSK3. CDC25 A was likely to play a role in other signaling pathways in sertoli cells. Within the range of certain concentration of FSH, TSSK3 in sertoli cells had the highest expression at about 24 h. TSSK3 protein produced in sertoli cells was likely to play an important role in substrate-level phosphorylationbe in meiosis and mitosis of spermatogenic cells. FSH could promote P53 expression and the highest expression was at about 12 h, and P53 might control the division of spermatogenic cells as well as sertoli cells.
基金supported by National Natural Science Foundation of China(81502123 and81330081)Natural Science Foundation of Anhui Province(1308085QH130)Anhui Province Nature Science Foundation in University(KJ2014A119)
文摘OBJECTIVE Basic fibroblast growth factor(b FGF)and platelet-derived growth factor(PDGF)produced by hepatocellular carcinoma(HCC)cells are responsible for the cell growth.Accumulating evidence shows that insulin-like growth factor-binding protein-3(IGFBP-3)suppresses HCC cell proliferation in both IGF-dependent and independent manners.The present study is to investigate whether treatment with exogenous IGFBP-3 inhibits bF GF and PDGF production and the cell proliferation of HCC cells.METHODS Cell Counting Kit 8 assay were designed to detect HCC cell proliferation,transcription factor early growth response-1(EGR1)involving in IGFBP-3 regulation of b FGF and PDGF were detected by RT-PCR and Western blot assays.Western blot assay was adopted to detect the IGFBP-3 regulating insulin-like growth factor 1 receptor(IGF-1R)signaling pathway.RESULTS The present study demonstrates that IGFBP-3 suppressed IGF-1-induced b FGF and PDGF expression while it does not affect their expression in the absence of IGF-1.To delineate the underlying mechanism,Western-blot and RT-PCR assays confirmed that the transcription factor early growth response protein 1(EGR1)is involved in IGFBP-3 regulation of b FGF and PDGF.IGFBP-3 inhibition of type 1 insulin-like growth factor receptor(IGF1R),ERK and AKT activation is IGF-1-dependent.Furthermore,transient transfection with constitutively activated AKT or MEK partially blocks the IGFBP-3 inhibition of EGR1,b FGF and PDGF expression.CONCLUSION In conclusion,these findings suggest that IGFBP-3suppresses transcription of EGR1 and its target genes b FGF and PDGF through inhibiting IGF-1-dependent ERK and AKT activation.It demonstrates the importance of IGFBP-3 in the regulation of HCC cell proliferation,suggesting that IGFBP-3 could be a target for the treatment of HCC.
文摘Identification and enumeration of both dendritic Ia^+ epi-dermal cells (Ia^+DECs) and dendritic Thy-1^+ epidermalcells (Thy-1^+DECa) from various parts of the body wereexamined by using epidermal sheets of C3H/He inbred miceof different age groups and indirect immunofluorescent tech-nique. A significant decline of both Ia^+DECs and Thy-1^+DECs in the mice of the aged group was demonstrated anddifferent densities and different distribution patterns betweenIa^+DECs and Thy-1^+DECs were obserged. These findingsmay imply that the decline of both Ia^+DECs and Thy-1^+DECs in the aged group may reflect the alterations of im-mune response in aging.
文摘Aim To investigate the effects of JAK inhibitor （SHR0302） on adjuvant-induced arthritis （AA） rats and the partial mechanisms focused on T, B lymphocyte subsets through JAK1-STAT3 pathway, including Thl7, Treg, total B cells and memory B cells. Methods Animals were divided randomly into 6 groups including normal control, AA, SHR0302 （0.3, 1.0, 3.0 nag · kg^-1, ig） and MTX （0.5 nag · kg^-1 , ig） . The effects of SHR0302 on AA rats by evaluating arthritis index, arthritis global assessment and paw swelling degree, histopathology of joint and spleen, inflammatory cytokine and antibody production in serum. We examined the proliferation of T, B and FLS by CCK8 kit; Thl7, Treg, total B and memory B cell proportion was measured by flow cytometry; Cytokines TNF-αβ, IL-1β, IL-10, IL-17 and antibody IgG1, IgG2a levels in serum were measured by ELISA kits; The ex- pression of p-JAK1 and p-STAT3 was measured by Western blot analysis. Results SHR0302 suppressed the se- verity of AA rats by attenuating the arthritis index, arthritis global assessment and paw swelling degree, and allevia- ted histopathology of spleen and joint of AA rats. SHR0302 can inhibit the proliferation of T, B and FLS, and down-regulated cytokines TNF-α, IL-1β, IL-17 and antibody IgG1, IgG2a levels, and suppressed the proportion of Thl7 and total B, and inhibited JAK1-STAT3 phosphorylation; There was no significant effect on Treg function and memory B cell proportion. Conclusion SHR0302 may attenuate the severity of AA rats, partially through signifi- cantly reducing Thl7 function and total B cell proportion by inhibiting JAK1-STAT3 phosphorylation.
基金Project(2019YFC1907405)supported by the National Key R&D Program of ChinaProject(GJJ200809)supported by the Education Department Project Fund of Jiangxi Province,ChinaProject(2020BAB214021)supported by the Natural Science Foundation of Jiangxi Province,China。
文摘Direct carbon solid oxide fuel cells(DC-SOFCs)are promising,green,and efficient power-generating devices that are fueled by solid carbons and comprise all-solid-state structures.Developing suitable anode materials for DC-SOFCs is a substantial scientific challenge.Herein we investigated the use of La_(0.75)Sr_(0.25)Cr_(0.5)Mn_(0.5)O_(3)-δ−Ce_(0.8)Gd_(0.2)O_(1.9)(LSCM−GDC)composite electrodes as anodes for La_(0.9)Sr_(0.1)Ga_(0.8)Mg_(0.2)O_(3)-δelectrolyte-based DC-SOFCs,with Camellia oleifera shell char as the carbon fuel.The LSCM−GDC-anode DC-SOFC delivered a maximum power density of 221 mW/cm^(2) at 800℃ and it significantly improved to 425 mW/cm^(2) after Ni nanoparticles were introduced into the LSCM−GDC anode through wet impregnation.The microstructures of the prepared anodes were characterized,and the stability of the anode in a DC-SOFC and the influence of catalytic activity on open circuit voltage were studied.The above results indicate that LSCM–GDC anode is promising to be applied in DC-SOFCs.
文摘P2X7 receptor （P2X7r） is important in inflammation and fibrosis. The aim of the present study was to investigate the effect of P2X7r inhibition, using a specific inhibitor （A438079） to prevent the development of liver fibrosis on human hepatic stellate cells, LX-2. The supernatant from lipopolysaccharide （LPS）-stimulated RAW 264.7 mouse macrophages was supplemented to LX-2 cells for 24 h. LX-2 cells were primed with LPS for 4 h and subsequently stimulated for 30 rain with 3 mmol · L^-1 of adenosine 5＇-triphosphate （ATP）. A438079 （ 10 μmol · L^-1） was supplemented to LX-2 cells 10 rain prior to ATP. Directly treated with LPS on LX-2 cells, mRNA ex- pressions of IL-1β, IL-18 and IL-6 were increased, as well as P2X7r. And caspase-1, ASC and NLRP3 mRNA ex- pressions were increased with LPS stimulation. LPS stimulation also increased oL-SMA and collagen I mRNA expres- sions. Interestingly treatment of LX-2 cells with mediums from LPS-primed RAW 264.7 mouse macrophages exhibi- ted greater increase of mRNA expressions of above genes than those in LX-2 directly treated with LPS. Pretreatment of directly or indirectly LPS-stimulated LX-2 cells with A438079 both suppressed IL-1β mRNA expression. In addi- tion treatment of LPS-primed LX-2 cells with 3 mM ATP induced the significant increase of IL-1β, IL-6, caspase- 1, pannexin-1, α-SMA and collagen I mRNA expression, the increasing of oL-SMA protein expression and cleavage of IL-1β. These events were significantly suppressed by pretreatment with P2X7r antagonist A438079. P2XTr blockade also significantly reduced the protein expression of oL-SMA. Our results suggest that the involvement of the P2X7r-NLRP3 inflammasome pathway in the secretion of IL-1β from extracellular ATP/LPS-stimulated human he- patic stellate cells. This study demonstrated that repression of the P2XTr represents a novel potential therapeutic ap- proach to control liver fibrosis.
