BACKGROUND Inhibition of liver fibrosis plays a crucial role in curbing the advancement of chronic disease to cirrhosis and even liver cancer.However,modern medicine currently lacks direct anti-fibrotic drugs.He-He-Sh...BACKGROUND Inhibition of liver fibrosis plays a crucial role in curbing the advancement of chronic disease to cirrhosis and even liver cancer.However,modern medicine currently lacks direct anti-fibrotic drugs.He-He-Shu-Yang formula(HHSY)is a renowned Chinese medicine for the treatment of liver fibrosis.However,its mechanism of action has not been fully unraveled.AIM To explore the efficacy and mechanism of action of HHSY through in vitro and in vivo experiments.METHODS A liver fibrosis rat model(carbon tetrachloride-induced)was treated with low-or high-dose HHSY(10.42 g/kg or 20.84 g/kg)or with colchicine(1 mg/kg)for 9 weeks.In vitro,LX-2 human hepatic stellate cells(HSCs)were activated using transforming growth factor-β1 and subsequently treated with HHSY-containing serum or a nicotinamide adenine dinucleotide phosphate oxidase 4(NOX4)inhibitor.Through high-performance liquid chromatography,histopathology(hematoxylin and eosin,Masson),immunohistochemistry,western blot,and quantitative reverse transcription polymerase chain reaction analyses,we demonstrated that HHSY inhibited HSC activation and suppressed the NOX4/reactive oxygen species(ROS)/nucleotide binding oligomerization domain-like receptor family pyrin domain-containing 3(NLRP3)pathway.RESULTS In vivo,HHSY improved liver function and alleviated liver pathology,including reducing inflammatory cell infiltration,and liver fibrosis in carbon tetrachloride rats.with more significant effects at higher doses.Immunohistochemistry revealed that HHSY could decrease alpha-smooth muscle actin,NOX4,and NLRP3 expression,as well as serum ROS levels(O_(2)-and H_(2)O_(2),P<0.05).Western blot analysis confirmed HHSY also reduced NLRP3 protein levels(P<0.05).In vitro,HHSY at 1.25%or 2.5%reduced the levels of ACTA2 mRNA,NOX4 protein and NOX4 mRNA,ROS production,and NLRP3 and IL-1βmRNA in activated LX-2 cells(P<0.05).CONCLUSION HHSY effectively treats liver fibrosis,likely by inhibiting HSC activation through the NOX4/ROS/NLRP3 pathway.This underscores HHSY’s clinical relevance as a potential therapeutic option for liver fibrosis.展开更多
Objective:To identify the underlying mechanism by which quercetin(Que)alleviates sepsis-related acute respiratory distress syndrome(ARDS).Methods:In vivo,C57BL/6 mice were assigned to sham,cecal ligation and puncture(...Objective:To identify the underlying mechanism by which quercetin(Que)alleviates sepsis-related acute respiratory distress syndrome(ARDS).Methods:In vivo,C57BL/6 mice were assigned to sham,cecal ligation and puncture(CLP),and CLP+Que(50 mg/kg)groups(n=15 per group)by using a random number table.The sepsisrelated ARDS mouse model was established using the CLP method.In vitro,the murine alveolar macrophages(MH-S)cells were classified into control,lipopolysaccharide(LPS),LPS+Que(10μmol/L),and LPS+Que+acetylcysteine(NAC,5 mmol/L)groups.The effect of Que on oxidative stress,inflammation,and apoptosis in mice lungs and MH-S cells was determined,and the mechanism with reactive oxygen species(ROS)/p38 mitogen-activated protein kinase(MAPK)pathway was also explored both in vivo and in vitro.Results:Que alleviated lung injury in mice,as reflected by a reversal of pulmonary histopathologic changes as well as a reduction in lung wet/dry weight ratio and neutrophil infiltration(P<0.05or P<0.01).Additionally,Que improved the survival rate and relieved gas exchange impairment in mice(P<0.01).Que treatment also remarkedly reduced malondialdehyde formation,superoxide dismutase and catalase depletion,and cell apoptosis both in vivo and in vitro(P<0.05 or P<0.01).Moreover,Que treatment diminished the release of inflammatory factors interleukin(IL)-1β,tumor necrosis factor-α,and IL-6 both in vivo and in vitro(P<0.05 or P<0.01).Mechanistic investigation clarified that Que administration led to a decline in the phosphorylation of p38 MAPK in addition to the suppression of ROS expression(P<0.01).Furthermore,in LPS-induced MH-S cells,ROS inhibitor NAC further inhibited ROS/p38 MAPK pathway,as well as oxidative stress,inflammation,and cell apoptosis on the basis of Que treatment(P<0.05 or P<0.01).Conclusion:Que was found to exert anti-oxidative,anti-inflammatory,and anti-apoptotic effects by suppressing the ROS/p38 MAPK pathway,thereby conferring protection for mice against sepsis-related ARDS.展开更多
Objective: To explore the effect and mechanism of action of bufalin in triple-negative breast cancer(TNBC) drug-resistant cell lines. Methods: The normal human mammary epithelial cell line, TNBC cell line, TNBC adriam...Objective: To explore the effect and mechanism of action of bufalin in triple-negative breast cancer(TNBC) drug-resistant cell lines. Methods: The normal human mammary epithelial cell line, TNBC cell line, TNBC adriamycin-resistant cell line, and TNBC docetaxel-resistant cell line were treated with different doses of bufalin(0–1,000 nmol/L) at different time points(0–72 h). Propidium iodide staining, AV-FITC/PI double staining, Hoechst 33342/PI double staining and transmission electron microscopy(TEM) were used to evaluate the death patterns of the cell lines. Results: Bufalin killed the TNBC cell line and its drug-resistant cell lines in a dose/time-dependent manner(all P<0.01). After treatment with bufalin for 24 h, the adriamycinresistant cell line showed a co-existing pattern of necroptosis and apoptosis. However, at 48 h, necroptosis was the main manifestation. After treatment with bufalin, the expressions of tumor necrosis factor α, phospho-tumor necrosis factor receptor 1, phospho-receptor interacting protein 1 and c-caspase 3 increased(all P<0.01), the killing effect of bufalin could be mostly inhibited by NEC-1, and by z-VAD-fmk(both P <0.01). Besides, the intracellular reactive oxygen species(ROS) levels increased considerably(P<0.01), the antioxidant N-acetyl cysteine or Nec-1 could inhibit the increase of ROS level and the killing effect of bufalin(all P<0.01). The adriamycin-resistant cell line exhibited necroptosis characteristic after 48 h of bufalin treatment under TEM. Conclusions: Bufalin could induce necroptosis through RIP1/ROS-mediated pathway to kill the drug-resistant TNBC cell lines. This finding provides critical experimental data and theoretical basis for the clinical application of bufalin to overcome the difficulties in the treatment of TNBC.展开更多
基金Supported by the Guangdong Basic and Applied Basic Research Fund Project,No.2022A1515110825 and No.2023A1515011092the Incubation Program for the Science and Technology Development of Chinese Medicine Guangdong Laboratory,No.HQL2024PZ033+5 种基金the Chi Xiaoling National Famous Traditional Chinese Medicine Expert Inheritance Studio,Teaching Letter from the State Traditional Chinese Medicine Office[2022],No.75the Advantage Disease Project of Guangdong Provincial Hospital of Traditional Chinese Medicine[2020],No.37the Science and Technology Planning Project of Guangdong Province,No.2023B1212060063the Project of Guangdong Provincial Key Laboratory of Chinese Medicine for Prevention and Treatment of Refractory Chronic Diseases,No.YN2023MB04the Clinical Research Projects of Guangdong Provincial Hospital of Chinese Medicine,No.YN2022QN05the Guangzhou Basic Research Program for Young Doctors in Basic and Applied Basic Research,No.2024A04J3004.
文摘BACKGROUND Inhibition of liver fibrosis plays a crucial role in curbing the advancement of chronic disease to cirrhosis and even liver cancer.However,modern medicine currently lacks direct anti-fibrotic drugs.He-He-Shu-Yang formula(HHSY)is a renowned Chinese medicine for the treatment of liver fibrosis.However,its mechanism of action has not been fully unraveled.AIM To explore the efficacy and mechanism of action of HHSY through in vitro and in vivo experiments.METHODS A liver fibrosis rat model(carbon tetrachloride-induced)was treated with low-or high-dose HHSY(10.42 g/kg or 20.84 g/kg)or with colchicine(1 mg/kg)for 9 weeks.In vitro,LX-2 human hepatic stellate cells(HSCs)were activated using transforming growth factor-β1 and subsequently treated with HHSY-containing serum or a nicotinamide adenine dinucleotide phosphate oxidase 4(NOX4)inhibitor.Through high-performance liquid chromatography,histopathology(hematoxylin and eosin,Masson),immunohistochemistry,western blot,and quantitative reverse transcription polymerase chain reaction analyses,we demonstrated that HHSY inhibited HSC activation and suppressed the NOX4/reactive oxygen species(ROS)/nucleotide binding oligomerization domain-like receptor family pyrin domain-containing 3(NLRP3)pathway.RESULTS In vivo,HHSY improved liver function and alleviated liver pathology,including reducing inflammatory cell infiltration,and liver fibrosis in carbon tetrachloride rats.with more significant effects at higher doses.Immunohistochemistry revealed that HHSY could decrease alpha-smooth muscle actin,NOX4,and NLRP3 expression,as well as serum ROS levels(O_(2)-and H_(2)O_(2),P<0.05).Western blot analysis confirmed HHSY also reduced NLRP3 protein levels(P<0.05).In vitro,HHSY at 1.25%or 2.5%reduced the levels of ACTA2 mRNA,NOX4 protein and NOX4 mRNA,ROS production,and NLRP3 and IL-1βmRNA in activated LX-2 cells(P<0.05).CONCLUSION HHSY effectively treats liver fibrosis,likely by inhibiting HSC activation through the NOX4/ROS/NLRP3 pathway.This underscores HHSY’s clinical relevance as a potential therapeutic option for liver fibrosis.
基金Supported by the Natural Science Foundation of Jiangsu Province(No.BK20211136)Translational Medicine Foundation of Jiangsu Research Hospital Association(Nos.SYHKJXF-2025-22 and SYHKJ-XF-2025-24)Development Fund Project of Affiliated Hospital of Xuzhou Medical University(No.XYFY202402)。
文摘Objective:To identify the underlying mechanism by which quercetin(Que)alleviates sepsis-related acute respiratory distress syndrome(ARDS).Methods:In vivo,C57BL/6 mice were assigned to sham,cecal ligation and puncture(CLP),and CLP+Que(50 mg/kg)groups(n=15 per group)by using a random number table.The sepsisrelated ARDS mouse model was established using the CLP method.In vitro,the murine alveolar macrophages(MH-S)cells were classified into control,lipopolysaccharide(LPS),LPS+Que(10μmol/L),and LPS+Que+acetylcysteine(NAC,5 mmol/L)groups.The effect of Que on oxidative stress,inflammation,and apoptosis in mice lungs and MH-S cells was determined,and the mechanism with reactive oxygen species(ROS)/p38 mitogen-activated protein kinase(MAPK)pathway was also explored both in vivo and in vitro.Results:Que alleviated lung injury in mice,as reflected by a reversal of pulmonary histopathologic changes as well as a reduction in lung wet/dry weight ratio and neutrophil infiltration(P<0.05or P<0.01).Additionally,Que improved the survival rate and relieved gas exchange impairment in mice(P<0.01).Que treatment also remarkedly reduced malondialdehyde formation,superoxide dismutase and catalase depletion,and cell apoptosis both in vivo and in vitro(P<0.05 or P<0.01).Moreover,Que treatment diminished the release of inflammatory factors interleukin(IL)-1β,tumor necrosis factor-α,and IL-6 both in vivo and in vitro(P<0.05 or P<0.01).Mechanistic investigation clarified that Que administration led to a decline in the phosphorylation of p38 MAPK in addition to the suppression of ROS expression(P<0.01).Furthermore,in LPS-induced MH-S cells,ROS inhibitor NAC further inhibited ROS/p38 MAPK pathway,as well as oxidative stress,inflammation,and cell apoptosis on the basis of Que treatment(P<0.05 or P<0.01).Conclusion:Que was found to exert anti-oxidative,anti-inflammatory,and anti-apoptotic effects by suppressing the ROS/p38 MAPK pathway,thereby conferring protection for mice against sepsis-related ARDS.
基金Supported by the National Natural Science Foundation of China (No.81573654)。
文摘Objective: To explore the effect and mechanism of action of bufalin in triple-negative breast cancer(TNBC) drug-resistant cell lines. Methods: The normal human mammary epithelial cell line, TNBC cell line, TNBC adriamycin-resistant cell line, and TNBC docetaxel-resistant cell line were treated with different doses of bufalin(0–1,000 nmol/L) at different time points(0–72 h). Propidium iodide staining, AV-FITC/PI double staining, Hoechst 33342/PI double staining and transmission electron microscopy(TEM) were used to evaluate the death patterns of the cell lines. Results: Bufalin killed the TNBC cell line and its drug-resistant cell lines in a dose/time-dependent manner(all P<0.01). After treatment with bufalin for 24 h, the adriamycinresistant cell line showed a co-existing pattern of necroptosis and apoptosis. However, at 48 h, necroptosis was the main manifestation. After treatment with bufalin, the expressions of tumor necrosis factor α, phospho-tumor necrosis factor receptor 1, phospho-receptor interacting protein 1 and c-caspase 3 increased(all P<0.01), the killing effect of bufalin could be mostly inhibited by NEC-1, and by z-VAD-fmk(both P <0.01). Besides, the intracellular reactive oxygen species(ROS) levels increased considerably(P<0.01), the antioxidant N-acetyl cysteine or Nec-1 could inhibit the increase of ROS level and the killing effect of bufalin(all P<0.01). The adriamycin-resistant cell line exhibited necroptosis characteristic after 48 h of bufalin treatment under TEM. Conclusions: Bufalin could induce necroptosis through RIP1/ROS-mediated pathway to kill the drug-resistant TNBC cell lines. This finding provides critical experimental data and theoretical basis for the clinical application of bufalin to overcome the difficulties in the treatment of TNBC.
