Al-Mg-Si alloys with high strength and good corrosion resistance are regarded as desirable materials for all-aluminum vehicles.However,the traditional trial-and-error experimental methods are insufficient to address t...Al-Mg-Si alloys with high strength and good corrosion resistance are regarded as desirable materials for all-aluminum vehicles.However,the traditional trial-and-error experimental methods are insufficient to address the trade-offbetween strength and corrosion resistance.In this work,a non-dominated sorting genetic machine-learning algorithm(NSGA-II)was employed to optimize the chemical composition,so as to simultaneously improve the strength and corrosion resistance.Three high-performance Al-Mg-Si alloys with low Mg,Si,and Cu contents were successfully developed,where the yield strength(YS),ultimate tensile strength(UTS),and the elongation(δ)reached 375-380 MPa,410-416 MPa,and 13.7%-15.2%,re-spectively.Compared with higher-Cu-content 6013 alloy,the YS and UTS of the present alloys increase by about 60 MPa,and the intergranular corrosion resistance is also significantly improved.Microstruc-ture characterization demonstrated thatβ’’and QP phases introduced a significant synergistic precipita-tion strengthening effect;the dispersoids formed by trace Mn,Cr,Fe,Zr,and Ti contributed dispersion strengthening effect;and the good pitting corrosion resistance is attributed to lower Mg and Si contents.展开更多
Epimedii Folium(EF)combined with Psoraleae Fructus(PF)is a common modern preparation,but liver injury caused by Chinese patent medicine preparations containing EF and PF has been frequently reported in recent years.Zh...Epimedii Folium(EF)combined with Psoraleae Fructus(PF)is a common modern preparation,but liver injury caused by Chinese patent medicine preparations containing EF and PF has been frequently reported in recent years.Zhuangguguanjiewan pills(ZGW),which contain EF and PF,could induce immune idiosyncratic liver injury according to clinical case reports and a nonhepatotoxic dose of lipopolysaccharide(LPS)model.This present study evaluated the liver injury induced by EF or PF alone or in combination and investigated the related mechanism by using the LPS model.Liver function indexes and pathological results showed that either EF or PF alone or in combination led to liver injury in normal rats;however,EF or PF alone could lead to liver injury in LPS-treated rats.Moreover,EF combined with PF could induce a greater degree of injury than that caused by EF or PF alone in LPS-treated rats.Furthermore,EF or PF alone or in combination enhanced the LPS-stimulated inflammatory cytokine production,implying that IL-1β,which is processed and released by activating the NLRP3 inflammasome,is a specific indicator of EF-induced immune idiosyncratic hepatotoxicity.Thus,EF may induce liver injury through enhancing the LPS-mediated proinflammatory cytokine production and activating the NLRP3 inflammasome.In addition,the metabolomics analysis results showed that PF affected more metabolites in glycerophospholipid and sphingolipid metabolic pathways compared with EF in LPS model,suggesting that PF increased the responsiveness of the liver to LPS or other inflammatory mediators via modulation of multiple metabolic pathways.Therefore,EF and PF combination indicates traditional Chinese medicine incompatibility,considering that it induces idiosyncratic hepatotoxicity under immunological stress conditions.展开更多
Idiosyncratic drus-induced liver injury(IDILI)is an intrequent but potentially serious disease that develops the main reason for post-marketing safety warnings and withdrawals of drugs.Epimedii Folium(EF),the widely u...Idiosyncratic drus-induced liver injury(IDILI)is an intrequent but potentially serious disease that develops the main reason for post-marketing safety warnings and withdrawals of drugs.Epimedii Folium(EF),the widely used herbal medicine,has shown to cause idiosyncratic liver injury,but the underlying mechanisms are poorly understood.Increasing evidence has indicated that most cases of IDILI are immune mediated.Here,we report that icarisideⅡ(ICSⅡ),the major active and metabolic constituent of EF,causes idiosyncratic liver injury by promoting NLRP3 inflammasome activation.ICSⅡexacerbates NLRP3 inflammasome activation triggered by adenosine triphosphate(ATP)and nigericin,but not silicon dioxide(SiO2),monosodium urate(MSU)crystal or cytosolic lipopolysaccharide(LPS).Additionally,the activation of NLRC4 and AIM2 inflammasomes is not affected by ICSⅡ.Mechanistically,synergistic induction of mitochondrial reactive oxygen species(mtROS)is a crucial contributor to the enhancing effect of ICSⅡon ATP-or nigericin-induced NLRP3 inflammasome activation.Importantly,in vivo data show that a combination of non-hepatotoxic doses of LPS and ICSⅡcauses the increase of aminotransferase activity,hepatic inflammation and pyroptosis,which is attenuated by Nlrp3 deficiency or pretreatment with MCC950(a specific NLRP3 inflammasome inhibitor).In conclusion,these findings demonstrate that ICSⅡcauses idiosyncratic liver injury through enhancing NLRP3 inflammasome activation and suggest that ICSⅡmay be a risk factor and responsible for EF-induced liver injury.展开更多
Aberrant activation of NLRP3 inflammasome has been implicated in the pathogenesis of diverse inflammation-related diseases, and pharmacological molecules targeting NLRP3 inflammasome are of considerable value to ident...Aberrant activation of NLRP3 inflammasome has been implicated in the pathogenesis of diverse inflammation-related diseases, and pharmacological molecules targeting NLRP3 inflammasome are of considerable value to identifying potential therapeutic interventions. Cardamonin(CDN), the major active ingredient of the traditional Chinese medicinal herb Alpinia katsumadai, has exerted an excellent anti-inflammatory activity, but the mechanism underlying this role is not fully understood. Here, we show that CDN blocks canonical and noncanonical NLRP3 inflammasome activation triggered by multiple stimuli. Moreover, the suppression of CDN on inflammasome activation is specific to NLRP3, not to NLRC4 or AIM2 inflammasome. Besides, the inhibitory effect is not dependent on the expression of NF-κB-mediated inflammasome precursor proteins. We also demonstrate that CDN suppresses the NLRP3 inflammasome through blocking ASC oligomerization and speckle formation in a dose-dependent manner.Importantly, CDN improves the survival of mice suffering from lethal septic shock and attenuates IL-1βproduction induced by LPS in vivo, which is shown to be NLRP3 dependent. In conclusion, our results identify CDN as a broad-spectrum and specific inhibitor of NLRP3 inflammasome and a candidate therapeutic drug for treating NLRP3 inflammasome-driven diseases.展开更多
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 ...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-ip 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-ip 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.展开更多
To predict the fatigue life for oblique hyperbola-and bilinear-mode S-N curves of metallic materials with various strengths,a machine-learning approach for direct analysis was employed.Additionally,to determine the fa...To predict the fatigue life for oblique hyperbola-and bilinear-mode S-N curves of metallic materials with various strengths,a machine-learning approach for direct analysis was employed.Additionally,to determine the fatigue limit of the utilized materials(AISI 316,AISI 4140 and CA6 NM series)with different S-N curve modes using finite-fatigue life data,a Bayesian optimization-based inverse analysis was performed.The results indicated that predictions of the fatigue life for the utilized datasets via the random forest(RF)algo rithm for AISI 4140 and CA6 NM,and artificial neural network(ANN)for AISI 316,distribute within 2 factor error lines for most data.In the Bayesian optimization-based inverse analysis,the specific explanatory variables corresponding to the optimized maximum fatigue life were treated as the fatigue limits.The predicted fatigue limits either approximated to or slightly underestimated the experimental results,except for several cases with large errors.Using the inverse analysis to predict the fatigue limit for both S-N curve modes is applicable for current employed data-set.However,the explored maximum fatigue lives via BO corresponding to the predicted fatigue limit were underestimated for AISI 4140 and CA6 NM,and was overestimated for AISI 316 because of effect of shape of S-N curves.By combining the ANN or RF direct and BO inverse algorithms,whole S-N curves(including the fatigue limit)were evaluated for the S-N curve shapes of the oblique hyperbola and bilinear modes.展开更多
Objective:Although some studies have linked Asari Radix et Rhizoma(Asari Radix)administration to hepatocellular carcinoma(HCC),few studies have examined the association between the development of HCC and use of Asari ...Objective:Although some studies have linked Asari Radix et Rhizoma(Asari Radix)administration to hepatocellular carcinoma(HCC),few studies have examined the association between the development of HCC and use of Asari Radix among patients in China's Mainland.This study aimed to evaluate the realworld association between Asari Radix and HCC in patients to strengthen the understanding of Asari Radix safety.Methods:A retrospective cohort study among hepatitis B virus(HBV)-monoinfected patients and nonHBV-monoinfected patients were performed.Patients over 18 years of age were eligible for inclusion.Prescription records of inpatients and outpatients were inquired to distinguish Asari Radix users and nonusers.The risk of developing HCC among Asari Radix users and nonusers in the HBV cohort and the non-HBV cohort was analyzed.Results:There were 49500 HBV and 133148 non-HBV patients involved in the two cohorts.Among HBV patients(2901 users;46599 nonusers),the prevalence of HCC in Asari Radix users was lower than that in nonusers(145.70 vs.265.43 per 105).Among non-HBV patients(5042 users;128106 nonusers),the prevalence of HCC in Asari Radix users was lower than that in nonusers(81.62 vs.134.11 per 105).None of the hazard ratios(HRs)of Asari Radix exposure ranging from 1 g to 200 g in the two cohorts showed correlation between Asari Radix exposure and hepatocarcinogenesis.Conclusion:An obvious irrelevancy was found between the consumption of Asari Radix and HCC development both in patients with and in those without HBV infection.Use of Asari Radix under 200 g appears safe in terms of HCC risk in the Chinese population;further prospective studies are needed to confirm our results.展开更多
Background and aims:Berberine is one of the most promising clinically tested natural alkaloids,and immunotherapy using natural killer(NK)cells is a potentially effective treatment for hepatocellular carcinoma(HCC).Thi...Background and aims:Berberine is one of the most promising clinically tested natural alkaloids,and immunotherapy using natural killer(NK)cells is a potentially effective treatment for hepatocellular carcinoma(HCC).This study aims to elucidate the effect of berberine on the anti-HCC effect of NK92-MI cells.Materials and methods:Human HCC SMMC-7721 and Hep3B cells were co-incubated with NK92-MI cells,berberine(60 or 80 mmol/L),or their combination for 36 h.To evaluate the killing effect of NK92-MI cells on HCC cells,the release of lactate dehydrogenase(LDH)in HCC cells was measured.The anti-tumor effects of berberine,NK92-MI cells,and their combinations were evaluated by MTS,EdU,Tunel,and Western blot assays.A male BALB/c nude mouse subcutaneous tumor model was used to investigate the anti-HCC effect of berberine and NK92-MI cells in vivo.Results:The LDH assay showed that berberine enhanced the cytotoxicity of NK92-MI cells on HCC cells.The combination of berberine and NK92-MI cells demonstrated more obvious anti-HCC effect than did the berberine or NK92-MI single treatment in inhibiting cell proliferation and inducing apoptosis both in vitro and in vivo.Mechanistically,the expression of programmed cell death-ligand 1(PD-L1)in HCC cells was upregulated after co-culture with NK-92MI cells.PD-L1 expression was knocked down,thereby inhibiting the proliferation and promoting apoptosis of HCC cells,and inhibited by berberine that blocked the secretion of interferon gamma(IFN-g),thereby enhancing the anti-tumor effect of NK92-MI cells.Conclusions:Current data show that the immunomodulatory role of berberine is to enhance the cytotoxic effect of NK92-MI cells and inhibit tumor immune escape by reducing the expression of PD-L1.Our study provides a rationale for the clinical application of berberine in combination with NK cells for the treatment of HCC.展开更多
基金supported by the Key Scientific and Technologi-cal Project of Foshan City(No.1920001000409).
文摘Al-Mg-Si alloys with high strength and good corrosion resistance are regarded as desirable materials for all-aluminum vehicles.However,the traditional trial-and-error experimental methods are insufficient to address the trade-offbetween strength and corrosion resistance.In this work,a non-dominated sorting genetic machine-learning algorithm(NSGA-II)was employed to optimize the chemical composition,so as to simultaneously improve the strength and corrosion resistance.Three high-performance Al-Mg-Si alloys with low Mg,Si,and Cu contents were successfully developed,where the yield strength(YS),ultimate tensile strength(UTS),and the elongation(δ)reached 375-380 MPa,410-416 MPa,and 13.7%-15.2%,re-spectively.Compared with higher-Cu-content 6013 alloy,the YS and UTS of the present alloys increase by about 60 MPa,and the intergranular corrosion resistance is also significantly improved.Microstruc-ture characterization demonstrated thatβ’’and QP phases introduced a significant synergistic precipita-tion strengthening effect;the dispersoids formed by trace Mn,Cr,Fe,Zr,and Ti contributed dispersion strengthening effect;and the good pitting corrosion resistance is attributed to lower Mg and Si contents.
基金supported by the National Natural Science Foundation of China(Nos.81874368 and 81630100)the Beijing Nova Program(No.Z181100006218001)+2 种基金the National Key Technology R&D Program(No.2015ZX 09501-004-001-008)the National Key R&D Program of China(No.2018YFC1707000)the National TCM Industry Science and Technology Program(No.201507004-4-2).
文摘Epimedii Folium(EF)combined with Psoraleae Fructus(PF)is a common modern preparation,but liver injury caused by Chinese patent medicine preparations containing EF and PF has been frequently reported in recent years.Zhuangguguanjiewan pills(ZGW),which contain EF and PF,could induce immune idiosyncratic liver injury according to clinical case reports and a nonhepatotoxic dose of lipopolysaccharide(LPS)model.This present study evaluated the liver injury induced by EF or PF alone or in combination and investigated the related mechanism by using the LPS model.Liver function indexes and pathological results showed that either EF or PF alone or in combination led to liver injury in normal rats;however,EF or PF alone could lead to liver injury in LPS-treated rats.Moreover,EF combined with PF could induce a greater degree of injury than that caused by EF or PF alone in LPS-treated rats.Furthermore,EF or PF alone or in combination enhanced the LPS-stimulated inflammatory cytokine production,implying that IL-1β,which is processed and released by activating the NLRP3 inflammasome,is a specific indicator of EF-induced immune idiosyncratic hepatotoxicity.Thus,EF may induce liver injury through enhancing the LPS-mediated proinflammatory cytokine production and activating the NLRP3 inflammasome.In addition,the metabolomics analysis results showed that PF affected more metabolites in glycerophospholipid and sphingolipid metabolic pathways compared with EF in LPS model,suggesting that PF increased the responsiveness of the liver to LPS or other inflammatory mediators via modulation of multiple metabolic pathways.Therefore,EF and PF combination indicates traditional Chinese medicine incompatibility,considering that it induces idiosyncratic hepatotoxicity under immunological stress conditions.
基金supported by National Natural Science Foundation of China(81874368,81630100,and 81903891)Beijing Nova Program(Z181100006218001,China)+1 种基金National Science&Technology Major Project“Key New Drug Creation and Manufacturing Program”(2017ZX09301022 and 2018ZX09101002-001-002,China)the Innovation Groups of the National Natural Science Foundation of China(81721002)
文摘Idiosyncratic drus-induced liver injury(IDILI)is an intrequent but potentially serious disease that develops the main reason for post-marketing safety warnings and withdrawals of drugs.Epimedii Folium(EF),the widely used herbal medicine,has shown to cause idiosyncratic liver injury,but the underlying mechanisms are poorly understood.Increasing evidence has indicated that most cases of IDILI are immune mediated.Here,we report that icarisideⅡ(ICSⅡ),the major active and metabolic constituent of EF,causes idiosyncratic liver injury by promoting NLRP3 inflammasome activation.ICSⅡexacerbates NLRP3 inflammasome activation triggered by adenosine triphosphate(ATP)and nigericin,but not silicon dioxide(SiO2),monosodium urate(MSU)crystal or cytosolic lipopolysaccharide(LPS).Additionally,the activation of NLRC4 and AIM2 inflammasomes is not affected by ICSⅡ.Mechanistically,synergistic induction of mitochondrial reactive oxygen species(mtROS)is a crucial contributor to the enhancing effect of ICSⅡon ATP-or nigericin-induced NLRP3 inflammasome activation.Importantly,in vivo data show that a combination of non-hepatotoxic doses of LPS and ICSⅡcauses the increase of aminotransferase activity,hepatic inflammation and pyroptosis,which is attenuated by Nlrp3 deficiency or pretreatment with MCC950(a specific NLRP3 inflammasome inhibitor).In conclusion,these findings demonstrate that ICSⅡcauses idiosyncratic liver injury through enhancing NLRP3 inflammasome activation and suggest that ICSⅡmay be a risk factor and responsible for EF-induced liver injury.
基金supported by the National Key Technology R&D Program(No.2017ZX09301022)the National Key Technology R&D Program(No.2015ZX09501-004-001-008)+1 种基金the BeijingNova program(No.Z181100006218001)the National Natural Science Foundation of China(No.81630100)
文摘Aberrant activation of NLRP3 inflammasome has been implicated in the pathogenesis of diverse inflammation-related diseases, and pharmacological molecules targeting NLRP3 inflammasome are of considerable value to identifying potential therapeutic interventions. Cardamonin(CDN), the major active ingredient of the traditional Chinese medicinal herb Alpinia katsumadai, has exerted an excellent anti-inflammatory activity, but the mechanism underlying this role is not fully understood. Here, we show that CDN blocks canonical and noncanonical NLRP3 inflammasome activation triggered by multiple stimuli. Moreover, the suppression of CDN on inflammasome activation is specific to NLRP3, not to NLRC4 or AIM2 inflammasome. Besides, the inhibitory effect is not dependent on the expression of NF-κB-mediated inflammasome precursor proteins. We also demonstrate that CDN suppresses the NLRP3 inflammasome through blocking ASC oligomerization and speckle formation in a dose-dependent manner.Importantly, CDN improves the survival of mice suffering from lethal septic shock and attenuates IL-1βproduction induced by LPS in vivo, which is shown to be NLRP3 dependent. In conclusion, our results identify CDN as a broad-spectrum and specific inhibitor of NLRP3 inflammasome and a candidate therapeutic drug for treating NLRP3 inflammasome-driven diseases.
基金This work has been supported by the Beijing Nova Program(No.Z181100006218001)the National Natural Science Foundation of China(Nos.81874368,81630100,81903891,and 81573676)+1 种基金the National Science and Technology Major Project “Key New Drug Creation and Manufacturing Program”(Nos.2017ZX09301022 and 2018ZX09101002-001-002)the Innovation Groups of the National Natural Science Foundation of China(No.81721002).
文摘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-ip 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-ip 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.
文摘To predict the fatigue life for oblique hyperbola-and bilinear-mode S-N curves of metallic materials with various strengths,a machine-learning approach for direct analysis was employed.Additionally,to determine the fatigue limit of the utilized materials(AISI 316,AISI 4140 and CA6 NM series)with different S-N curve modes using finite-fatigue life data,a Bayesian optimization-based inverse analysis was performed.The results indicated that predictions of the fatigue life for the utilized datasets via the random forest(RF)algo rithm for AISI 4140 and CA6 NM,and artificial neural network(ANN)for AISI 316,distribute within 2 factor error lines for most data.In the Bayesian optimization-based inverse analysis,the specific explanatory variables corresponding to the optimized maximum fatigue life were treated as the fatigue limits.The predicted fatigue limits either approximated to or slightly underestimated the experimental results,except for several cases with large errors.Using the inverse analysis to predict the fatigue limit for both S-N curve modes is applicable for current employed data-set.However,the explored maximum fatigue lives via BO corresponding to the predicted fatigue limit were underestimated for AISI 4140 and CA6 NM,and was overestimated for AISI 316 because of effect of shape of S-N curves.By combining the ANN or RF direct and BO inverse algorithms,whole S-N curves(including the fatigue limit)were evaluated for the S-N curve shapes of the oblique hyperbola and bilinear modes.
基金supported by grants from "the National Science and Technoloyg Major Project" Key New Drug Creation and Manufacturing Program (2018ZX09101002-001-002)Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine (No:ZYYCXTD-C-202005)
文摘Objective:Although some studies have linked Asari Radix et Rhizoma(Asari Radix)administration to hepatocellular carcinoma(HCC),few studies have examined the association between the development of HCC and use of Asari Radix among patients in China's Mainland.This study aimed to evaluate the realworld association between Asari Radix and HCC in patients to strengthen the understanding of Asari Radix safety.Methods:A retrospective cohort study among hepatitis B virus(HBV)-monoinfected patients and nonHBV-monoinfected patients were performed.Patients over 18 years of age were eligible for inclusion.Prescription records of inpatients and outpatients were inquired to distinguish Asari Radix users and nonusers.The risk of developing HCC among Asari Radix users and nonusers in the HBV cohort and the non-HBV cohort was analyzed.Results:There were 49500 HBV and 133148 non-HBV patients involved in the two cohorts.Among HBV patients(2901 users;46599 nonusers),the prevalence of HCC in Asari Radix users was lower than that in nonusers(145.70 vs.265.43 per 105).Among non-HBV patients(5042 users;128106 nonusers),the prevalence of HCC in Asari Radix users was lower than that in nonusers(81.62 vs.134.11 per 105).None of the hazard ratios(HRs)of Asari Radix exposure ranging from 1 g to 200 g in the two cohorts showed correlation between Asari Radix exposure and hepatocarcinogenesis.Conclusion:An obvious irrelevancy was found between the consumption of Asari Radix and HCC development both in patients with and in those without HBV infection.Use of Asari Radix under 200 g appears safe in terms of HCC risk in the Chinese population;further prospective studies are needed to confirm our results.
基金This study was supported by grants from the General Project of Guangdong Natural Science Foundation(2021A1515010795,2020A1515011255)to H.Yangthe National Natural Science Foundation of China(no.81372634)to H.Yang+1 种基金the Medical Science and Technology Research Foundation of Guangdong Province(B2020114)to C.Guthe Guangzhou Health Science and Technology Project(20201A010054)to C.Gu.
文摘Background and aims:Berberine is one of the most promising clinically tested natural alkaloids,and immunotherapy using natural killer(NK)cells is a potentially effective treatment for hepatocellular carcinoma(HCC).This study aims to elucidate the effect of berberine on the anti-HCC effect of NK92-MI cells.Materials and methods:Human HCC SMMC-7721 and Hep3B cells were co-incubated with NK92-MI cells,berberine(60 or 80 mmol/L),or their combination for 36 h.To evaluate the killing effect of NK92-MI cells on HCC cells,the release of lactate dehydrogenase(LDH)in HCC cells was measured.The anti-tumor effects of berberine,NK92-MI cells,and their combinations were evaluated by MTS,EdU,Tunel,and Western blot assays.A male BALB/c nude mouse subcutaneous tumor model was used to investigate the anti-HCC effect of berberine and NK92-MI cells in vivo.Results:The LDH assay showed that berberine enhanced the cytotoxicity of NK92-MI cells on HCC cells.The combination of berberine and NK92-MI cells demonstrated more obvious anti-HCC effect than did the berberine or NK92-MI single treatment in inhibiting cell proliferation and inducing apoptosis both in vitro and in vivo.Mechanistically,the expression of programmed cell death-ligand 1(PD-L1)in HCC cells was upregulated after co-culture with NK-92MI cells.PD-L1 expression was knocked down,thereby inhibiting the proliferation and promoting apoptosis of HCC cells,and inhibited by berberine that blocked the secretion of interferon gamma(IFN-g),thereby enhancing the anti-tumor effect of NK92-MI cells.Conclusions:Current data show that the immunomodulatory role of berberine is to enhance the cytotoxic effect of NK92-MI cells and inhibit tumor immune escape by reducing the expression of PD-L1.Our study provides a rationale for the clinical application of berberine in combination with NK cells for the treatment of HCC.