ObjectiveThe study aims to investigate the intervention effect of modified Xiangsha Liujunzitang （M-XSLJZ） on intestinal-derived lipopolysaccharide （LPS）-activated Kupffer cell inflammation in rats with hyperlipidemia spleen deficiency syndrome. MethodsSeventy male SD rats were randomly divided into seven groups （n=10）： blank control （CON）， high-fat diet without spleen deficiency （HFD）， high-fat diet with spleen deficiency （SD-HFD）， M-XSLJZ low-， medium-， and high-dose groups （XS-L， XS-M， XS-H）， and western medicine control （R）. Spleen deficiency was induced in SD-HFD， XS-L， XS-M， XS-H， and R groups via irregular diet combined with exhaustive swimming for 15 days. The CON group received a standard diet， while other groups were fed a high-fat diet for 10 weeks to establish the hyperlipidemia model. After successful modeling， rats were treated for 8 weeks： M-XSLJZ was administered at 3.51， 7.02， 14.04 g·kg^-1 in XS-L， XS-M， and XS-H groups， respectively. The R group received 9×10^-4 g·kg^-1 of a reference drug. D-xylose excretion rate was measured by the phloroglucinol method. Blood lipids were assessed using an automated biochemical analyzer. Hematoxylin-eosin （HE） staining was used to evaluate the pathological conditions of the liver， and oil red O staining was used to observe the lipid deposition in the liver. The levels of LPS， portal vein serum LPS， LPS-binding protein （LBP）， serum interleukin-6 （IL-6）， IL-1β， and tumor necrosis factor-α （TNF-α） were detected by enzyme-linked immunosorbent assay （ELISA）. Immunofluorescence was used to evaluate CD86 expression and CD68/TLR4 co-localization in the liver. Protein levels of TLR4， MyD88， NF-κB p65， and p-NF-κB p65 in Kupffer cells were analyzed via Western blot automated protein analysis. Hepatic IL-6， TNF-α， and IL-1β mRNA and protein levels were measured using Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. ResultsCompared with the CON group， the SD-HFD group showed a decrease in D-xylose excretion （P<0.01）. TC， TG， HDL-C， and LDL-C increased （P<0.05， P<0.01）. A large number of hepatic lipid vacuoles and orange-red lipid droplet deposition appeared in the liver. Ileal LPS， portal LPS， and LBP increased （P<0.05， P<0.01）. The levels of serum IL-6， TNF-α， and IL-1β increased （P<0.01）. The expression of CD86 was upregulated （P<0.01）， and the co-expression of CD68 and TLR4 was enhanced. The protein levels of TLR4， MyD88， and p-p65 in Kupffer cells increased （P<0.01）. The mRNA and protein levels of IL-6， TNF-α， and IL-1β increased （P<0.05， P<0.01）. Compared with the HFD group， the SD-HFD group exhibited decreased D-xylose excretion （P<0.01）， higher HDL-C， LDL-C （P<0.05）， increased portal LBP and LPS （P<0.05）， increased serum IL-6 and TNF-α （P<0.01）， upregulated CD86 （P<0.01）， enhanced CD68/TLR4 co-expression， and higher TNF-α mRNA/protein （P<0.05）. Compared with the SD-HFD group， all M-XSLJZ treatment groups showed reduced TC， TG， and LDL-C （P<0.05， P<0.01）. XS-H and R groups displayed improved hepatic lipid deposition. XS-H and R groups had lower ileal LPS， portal LPS， and LBP levels （P<0.05， P<0.01）. All M-XSLJZ treatment groups exhibited reduced serum IL-6， IL-1β， and TNF-α （P<0.01）. The XS-H group showed downregulated CD86 （P<0.01） and weakened CD68/TLR4 co-expression. The XS-H group had reduced TLR4， MyD88， and p-NF-κB p65 in Kupffer cells （P<0.01）. XS-H and R groups showed lower IL-6， TNF-α， and IL-1β mRNA/protein （P<0.05， P<0.01）. ConclusionM-XSLJZ may exert its lipid-lowering effects by inhibiting intestinal-derived LPS and alleviating Kupffer cell inflammation in the liver.

Objective Based on apolipoprotein a-Ⅰ(Apoa-Ⅰ)gene knockout mice,the role and mechanism of phosphotidylcholine(PC)in improving cholesterol reverse transport were explored.Methods Thirty Apoa-Ⅰ-/-mice were randomly divided into an Apoa-Ⅰ-/-group,Apoa-Ⅰ-/-+HFD group,and Apoa-Ⅰ-/-+HFD+PC group using the random number table method;30 C57BL/6J mice were randomly divided into a WT group,WT+HFD group,and WT+HFD+PC control groups,with 10 mice in each group.The Apoa-Ⅰ-/-group and WT groups were fed basic feed,while the other groups were fed high-fat feed for 8 weeks to establish a hyperlipidemia model.From the 9th week,the WT+HFD+PC group and Apoa-Ⅰ-/-+HFD+PC group were given PC 2.5 g/(kg·d),while the remaining mice were given physiological saline by gavage for a total of 4 weeks of intervention.The serum lipid levels of the mice were detected using a fully automated analyzer.Hematoxylin and eosin and Oil red O staining were used to observe pathological and morphological changes,and the COD-PAP method was used to detect cholesterol levels in mouse liver tissue.The ELISA method was used to detect LCTA levels in mouse serum,and RT-qPCR and Western Blot method were used to detect the mRNA and protein expression of cholesterol ATP binding cassette transporter A1(ABCA1),ATP binding cassette transporter G1(ABC A1),lecithin cholesterol acyltransferase(LCAT),hepatic lipase(HL),scavenger receptor class B type Ⅰ(SR-B1),and low-density lipoprotein receptor(LDL-R)in liver tissue.Results Compared with the WT group,the serum lipid levelsof WT+HFD group mice were significantly increased(P＜0.01),LCAT levels were significantly reduced(P＜0.05),hepatic fat vacuoles were obvious,hepatic lipid deposition was significant,and liver tissue TC levels were significantly increased(P＜0.01).The mRNA and protein expression of ABCA1,ABCG1,LCAT,SR-B1,HL,and LDL-R were significantly reduced(P＜0.05,P＜0.01).Compared with the WT+HFD group,serum lipid levels in the WT+HFD+PC group were significantly reduced(P＜0.05,P＜0.01),LCAT levels were significantly increased(P＜0.05),hepatic fat vacuoles were significantly reduced,hepatic lipid deposition was alleviated,and liver tissue TC levels were significantly reduced(P＜0.05);mRNA and protein expression of ABCA1,LCAT,SR-B1,HL and LDL-R were significantly increased(P＜0.05,P＜0.01).The serum levels of TC,TG,and LDL-C were significantly increased,while the levels of LCAT、HDL-C were significantly reduced(P＜0.05,P＜0.01)in the Apoa-Ⅰ-/-+HFD group mice.Hepatocytes underwent balloon-like transformation,liver lipid deposition was significantly aggravated,and liver tissue TC levels were significantly increased(P＜0.05).The mRNA and protein expression of ABCA1,LCAT and HL were significantly reduced(P＜0.05,P＜0.01).Compared with the WT+HFD+PC group mice,the Apoa-Ⅰ-/-+HFD+PC group mice showed a significant increase in serum lipid levels(P＜0.05,P＜0.01),LCAT levels were significantly reduced(P＜0.05),significant hepatic lipid vacuoles,significant hepatic lipid deposition,and a significant increase in TC levels in liver tissue(P＜0.05).Their mRNA and protein expression of ABCA1,ABCG1,LCAT,SR-B1,and HL were also significantly reduced(P＜0.05,P＜0.01).Conclusions Phosphatidylcholine can improve dyslipidemia by interfering with Apoa-Ⅰ and thus regulating cholesterol reverse transport.

Objective Based on apolipoprotein a-Ⅰ(Apoa-Ⅰ)gene knockout mice,the role and mechanism of phosphotidylcholine(PC)in improving cholesterol reverse transport were explored.Methods Thirty Apoa-Ⅰ-/-mice were randomly divided into an Apoa-Ⅰ-/-group,Apoa-Ⅰ-/-+HFD group,and Apoa-Ⅰ-/-+HFD+PC group using the random number table method;30 C57BL/6J mice were randomly divided into a WT group,WT+HFD group,and WT+HFD+PC control groups,with 10 mice in each group.The Apoa-Ⅰ-/-group and WT groups were fed basic feed,while the other groups were fed high-fat feed for 8 weeks to establish a hyperlipidemia model.From the 9th week,the WT+HFD+PC group and Apoa-Ⅰ-/-+HFD+PC group were given PC 2.5 g/(kg·d),while the remaining mice were given physiological saline by gavage for a total of 4 weeks of intervention.The serum lipid levels of the mice were detected using a fully automated analyzer.Hematoxylin and eosin and Oil red O staining were used to observe pathological and morphological changes,and the COD-PAP method was used to detect cholesterol levels in mouse liver tissue.The ELISA method was used to detect LCTA levels in mouse serum,and RT-qPCR and Western Blot method were used to detect the mRNA and protein expression of cholesterol ATP binding cassette transporter A1(ABCA1),ATP binding cassette transporter G1(ABC A1),lecithin cholesterol acyltransferase(LCAT),hepatic lipase(HL),scavenger receptor class B type Ⅰ(SR-B1),and low-density lipoprotein receptor(LDL-R)in liver tissue.Results Compared with the WT group,the serum lipid levelsof WT+HFD group mice were significantly increased(P＜0.01),LCAT levels were significantly reduced(P＜0.05),hepatic fat vacuoles were obvious,hepatic lipid deposition was significant,and liver tissue TC levels were significantly increased(P＜0.01).The mRNA and protein expression of ABCA1,ABCG1,LCAT,SR-B1,HL,and LDL-R were significantly reduced(P＜0.05,P＜0.01).Compared with the WT+HFD group,serum lipid levels in the WT+HFD+PC group were significantly reduced(P＜0.05,P＜0.01),LCAT levels were significantly increased(P＜0.05),hepatic fat vacuoles were significantly reduced,hepatic lipid deposition was alleviated,and liver tissue TC levels were significantly reduced(P＜0.05);mRNA and protein expression of ABCA1,LCAT,SR-B1,HL and LDL-R were significantly increased(P＜0.05,P＜0.01).The serum levels of TC,TG,and LDL-C were significantly increased,while the levels of LCAT、HDL-C were significantly reduced(P＜0.05,P＜0.01)in the Apoa-Ⅰ-/-+HFD group mice.Hepatocytes underwent balloon-like transformation,liver lipid deposition was significantly aggravated,and liver tissue TC levels were significantly increased(P＜0.05).The mRNA and protein expression of ABCA1,LCAT and HL were significantly reduced(P＜0.05,P＜0.01).Compared with the WT+HFD+PC group mice,the Apoa-Ⅰ-/-+HFD+PC group mice showed a significant increase in serum lipid levels(P＜0.05,P＜0.01),LCAT levels were significantly reduced(P＜0.05),significant hepatic lipid vacuoles,significant hepatic lipid deposition,and a significant increase in TC levels in liver tissue(P＜0.05).Their mRNA and protein expression of ABCA1,ABCG1,LCAT,SR-B1,and HL were also significantly reduced(P＜0.05,P＜0.01).Conclusions Phosphatidylcholine can improve dyslipidemia by interfering with Apoa-Ⅰ and thus regulating cholesterol reverse transport.

Objective:The effect of bone marrow soluble B cell maturation antigen (sBCMA) expression on the efficacy and side effects of chimeric antigen receptor (CAR) -modified T-cell-targeting B cell maturation antigen (BCMA) in patients with multiple myeloma (MM) .Methods:This study involved 29 patients with relapsed or refractory MM (RRMM) who received humanized anti-BCMA CAR-T cell clinical trials from January 2018 to December 2021. The expression of sBCMA in bone marrow before and after anti-BCMA CAR-T cell treatment was detected by flow cytometry and compared.Results:①Two months after BCMA CAR-T cell treatment, 20 patients (68.97%) achieved an overall response (OR), whereas nine patients had stable disease (SD) or miner emission (MR). ②The expression of sBCMA in the bone marrow of 20 patients with OR was higher before treatment than after [26 926 (18 215, 32 488) ng/L vs 9 968 (6 634, 11 459) ng/L; P<0.001]; no significant difference was observed in patients with MR and SD [41 187 (33 816, 47 046) ng/L vs. 33 954 (31 569, 36 256) ng/L; P=0.145]; sBCMA expression in patients with OR before CAR-T cell treatment was lower than in patients with MR and SD ( P=0.005). ③No significant linear correlation was found between the peak value of CAR-T cells and sBCMA expression in the bone marrow of all 29 patients with RRMM ( R2=0.035, P=0.330). ④No significant difference in sBCMA expression was found between grades 0-1 CRS group (13 patients) and grades 2-4 CRS group [16 patients; 32 045 (18 742, 40 801) ng/L vs 29 102 (24 679, 38 776) ng/L, P=0.879], nor between grade 0 ICANS group (22 patients) and grade 1-3 ICANS group [seven patients; 30 073 (19 375, 40 065) ng/L vs 33 816 (22 933, 43 459) ng/L, P=0.763]. Conclusion:sBCMA expression in the bone marrow is related to the efficacy of BCMA CAR-T cell therapy in patients with RRMM, but is not significantly correlated with the severity of adverse events. It may serve as a predictive biomarker for the efficacy of BCMA CAR-T cell therapy in these patients.

Objective To explore the mechanism of modified Xiangsha Liujunzi Decoction in regulating apolipoproteinA-Ⅰ (apoA-Ⅰ),improving endoplasmic reticulum stress,regulating glucose and lipid metabolism,and preventing and treating dyslipidemia in mice. Methods Wild-type (WT) C57BL/6J mice were randomly divided into the WT,WT+high-fat diet(HFD),and WT+HFD+Xiangsha Liujunzi Decoction(XSLJZ) groups according to the random number table method. ApoA-Ⅰ-/-mice were randomly divided into the apoA-Ⅰ-/-,apoA-Ⅰ-/-+HFD,and apoA-Ⅰ-/-+HFD+XSLJZ groups (n=10) according to the random number table method. D12492 was used for HFD feeding to establish a hyperlipidemic mouse model. Modified XSLJZ (23.66g/kg) was administered daily by gavage from the ninth week. Serum and liver tissue were collected for testing after 4 weeks. An automatic biochemical analyzer was used to detect blood lipid levels;an enzyme-linked immunosorbent assay was used to detect serum fasting blood glucose (FBG) and insulin (INS) levels,and the INS resistance index (HOMA-IR) was calculated. Hematoxylin and eosin staining was used to observe the pathological changes in the liver. Oil red O staining was used to observe the lipid deposition in the liver. TG levels in liver tissue were detected using the microplate method. Real-time PCR was used to detect apoA-Ⅰ,glucose-regulated proteins (GRP78),sterol regulatory element binding protein-1c (SREBP-1c),acetyl CoA carboxylase 1 (ACC1),and fatty acid synthase (FASN) mRNA expression levels in liver tissue. The WES fully automated protein expression analysis system was used to detect apoA-Ⅰ,GRP78,inositol-requiring enzyme 1 (IRE1),p-IRE1,c-Jun N-terminal kinase (JNK),p-JNK,insulin receptor substrate (IRS1),p-IRS1,protein kinase B (Akt),p-Akt,SREBP-1c,ACC1,and FASN protein expression levels in liver tissue. Results Compared to the WT group,the WT+HFD group showed a significant increase in serum lipids,FBG,INS levels,and the HOMA-IR index (P<0.05). The orange-red lipid droplets in liver tissue increased,fat vacuoles were apparent,and TG levels were significantly increased. ApoA-Ⅰ mRNA and protein expression levels were significantly reduced,whereas GRP78,SREBP-1c,ACC1,and FASN mRNA expression levels were increased,GRP78,SREBP-1c,ACC1,and FASN protein levels and the IRE1,JNK,IRS1,and Akt phosphorylation degree were increased (P<0.05). The serum TG,HDL-C,LDL-C,FBG,and INS levels and the HOMA-IR index in the WT+HFD group were significantly reduced after administering modified XSLJZ (P<0.05). The orange-red lipid droplets in liver tissue were significantly reduced,fat vacuolization was alleviated,and TG levels were significantly reduced,ApoA-Ⅰ mRNA and protein expression levels were significantly increased,whereas GRP78,SREBP-1c,ACC1,and FASN mRNA expression levels were reduced,GRP78,SREBP-1c,ACC1,and FASN protein expression levels and the IRE1,JNK,IRS1,and Akt phosphorylation degree were reduced (P<0.05). Compared to the WT+HFD group,the TG,LDL-C,and FBG levels and HOMA-IR index in the serum of the apoA-Ⅰ-/-+HFD group were significantly increased,whereas the HDL-C levels were significantly decreased (P<0.05). Diffuse orange-red lipid droplets in liver tissue and a significant increase in fat vacuoles were observed. Furthermore,TG levels were significantly increased,SREBP-1c,ACC1,FASN mRNA,SREBP-1c,and ACC1 protein expression levels and IRE1,JNK,IRS1,and Akt phosphorylation levels were significantly increased (P<0.05). Compared to the WT+HFD+XSLJZ group,the apoA-Ⅰ-/-+HFD+XSLJZ group showed a significant increase in serum TG,LDL-C,FBG,and INS levels,and the HOMA-IR index,whereas HDL-C levels decreased significantly (P<0.05). The deposition of orange-red lipid droplets in liver tissue improved,and TG levels significantly decreased,GRP78,SREBP-1c,ACC1,and FASN mRNA expression levels,GRP78,SREBP-1c,and ACC1 protein levels,and IRE1,JNK,IRS1,and Akt phosphorylation levels increased (P<0.05). Conclusion Modified XSLJZ improves liver glucose and lipid metabolism disorder by regulating apoA-Ⅰ to alleviate endoplasmic reticulum stress.

Objective: To evaluate the application effect of smart classroom teaching mode in undergraduate teaching of endodontics. Methods: Through micro-lecture and massive open online course which were closely integrated with clinical practice and frontier advances, we build a new smart classroom teaching mode of endodontics relying on information technology such as the medical education cloud APP platform. The mode was applied to the undergraduate teaching of grade 2017 (110 students) and grade 2018 (107 students) in 2020 and 2021 respectively (experimental group). The theoretical examination was conducted for the grade 2016 (control group, 111 students applied traditional teaching methods) in 2019, and for two experimental grades in 2020 and 2021 respectively. A questionnaire survey was conducted for the 2018 undergraduates to investigate the experience of the smart classroom teaching mode, and the application effect of the smart classroom teaching mode was evaluated by comparing the offline theoretical test scores of grades 2016, 2017 and 2018. Results: The results of the questionnaire showed that students in grade 2018 recognized the overall form of smart classroom teaching mode, and 75.2% (79/105) of the students satisfied with the teaching process, considering that it could enhance learning interest and enthusiasm, improve self-learning ability, facilitate the understanding and memory of knowledge points, as well as increase the extension and expansion of professional knowledge. Thirty-seven point one percent (39/105) of the students thought that smart classroom teaching mode was not conducive to the interaction between teachers and students and couldn't improve learning efficiency. Comparing the final theoretical examination scores of students in three years, it was found that the average scores of 2021 (78.79±9.88) and 2020 (76.45±8.33) were significantly higher than that of 2019 (67.67±10.58) (t=6.77, P<0.001; t=8.51, P<0.001). The average score in 2021 was higher than that in 2020, although the difference was not significant (t=1.79, P=0.223). Conclusions: The application of smart classroom mode improved the teaching effect of endodontics, which is worthy of further promotion to provide a positive reference in improving the educating effects of oral medicine.
Humans ; Learning ; Endodontics ; Students ; Dental Care ; Surveys and Questionnaires

OBJECTIVE: The present study investigated how mild moxibustion treatment affects the intestinal microbiome and expression of NLRP3-related immune factors in a rat model of intestinal mucositis (IM) induced with 5-fluorouracil (5-Fu). METHODS: Forty male Sprague-Dawley rats were randomly divided into control, chemotherapy, moxibustion and probiotics groups. The IM rat model was established by intraperitoneal injection of 5-Fu. Mild moxibustion treatment and intragastric probiotic administration were provided once daily for 15 days. Tissue morphology, serum levels of inflammatory factors and the expression levels of tight junction proteins, caspase-1, gasdermin D and NLRP3 were evaluated in colon tissue, through hematoxylin and eosin staining, electron microscopy, enzyme-linked immunosorbent assay, Western blotting, quantitative real-time reverse transcription polymerase chain reaction and immunofluorescence. Gut microbiome profiling was conducted through 16S rRNA amplicon sequencing. RESULTS: Moxibustion and probiotic treatments significantly increased the expression levels of tight junction proteins, reduced cell apoptosis and the expression levels of caspase-1, gasdermin D and NLRP3; they also decreased the serum levels of tumor necrosis factor-α, interleukin (IL)-6, IL-1β and IL-18, while increasing serum levels of IL-10. Moxibustion and probiotic treatments also corrected the reduction in α-diversity and β-diversity in IM rats, greatly increased the proportion of the dominant bacterial genus Lactobacillus and reduced the abundance of the genera Roseburia and Escherichia in chemotherapy-treated rats to levels observed in healthy animals. We also found that these dominant genera were firmly correlated with the regulation of pyroptosis-associated proteins and inflammatory factors. Finally, moxibustion and probiotic treatments elicited similar effects in regulating intestinal host-microbial homeostasis and the expression of NLRP3 inflammasome-related factors. CONCLUSION Moxibustion exerts its therapeutic effect on IM by ameliorating mucosal damage and reducing inflammation. Moreover, moxibustion modulates the gut microbiota, likely via decreasing the expression levels of the NLRP3 inflammasome.

The purpose of this experiment was to study the effects of different shading conditions on the growth,physiological characteristics and biomass allocation of Polygonatum cyrtonema,which offered a theoretical basis for its cultivation.Different light environments(100%,80%,60% and 35% light transmittance) were simulated with shading treatments.Growth and photosynthetic indexes of P.cyrtonema were measured and the variances were analyzed.The results show that shading decreased superoxide anion radical(O-·2)production rate and hydrogen peroxide(H_2O_2) accumulation,kept the activity of SOD,POD and CAT enzyme at a high level.Furthermore,The content of chlorophyll a and chlorophyll b,net photosynthetic rate(Pn),stomatal conductance(Gs),transpiration rate(Tr),maximal photochemical efficiency of photosystem Ⅱ(Fv/Fm),photochemical quenching index(q P) and effective quantum yield of photosystem II(ΦPSⅡ) of P.cyrtonema were increased while the intercellular CO2 concentration(Ci),Foand NPQ were decreased by shading.Shading is beneficial to P.cyrtonema growth,can increase the total biomass P.cyrtonema.The allocation proportion of biomass on the aerial portion of P.cyrtonema increased but underground parts decreased with increasing shading conditions.In this study,P.cyrtonema can grow well in shading conditions,shading is beneficial to the formation of the yield and quality of the rhizomes of P.cyrtonema,especially in 65% light transmittance.
Biomass ; Chlorophyll ; Chlorophyll A ; Photosynthesis ; Plant Leaves ; Plant Stomata ; Plant Transpiration ; Polygonatum ; growth & development ; physiology ; Sunlight

The study is aimed to explore the effect of combination use of nitrogen(N) and zinc(Zn) fertilizers on the growth, yield and the effective components of Agastache rugosa. A. rugosa was grown under two N application rate (120, 300 kg·hm⁻²) and five Zn levels (0, 20, 50, 100，150 kg·hm⁻²) under field condition. The effect of the treatments on the physiological indicators, distribution of nitrogen and zinc and volatile oil components of A. rugosa were studied. The results showed that the combination use of N and Zn could significantly affect the growth and development, yield and volatile oil components of A. rugosa. Under the test conditions, the highest yield of Agastaches Herba was obtained when 50 kg·hm⁻² of Zn fertilizer was applied with high N application rate of 300 kg·hm⁻². Under the same N application rate, the increase of Zn production was positively correlated with the amount of Zn application in a certain concentration range, but excessive Zn application led to the decrease of yield. With the increase of N application level, the content of Zn also significantly increased. The combination use of N and Zn increased the yield of Agastaches Herba. High level of N application was beneficial to the absorption and accumulation of N and Zn of A. rugosa. Zn fertilizer could also promote the absorption and accumulation of N of A. rugosa. The interaction between N and Zn had significant influence on the main chemical constituents of the volatile oil of A. rugosa. Among the volatile oil chemical constituents of A. rugosa the content of pulegone (34.56%-53.91%) and piperonyl methyl ether (18.86%-42.27%) were much higher. Under the same N application rate, different Zn application rates also had significant effects on the main chemical components of volatile oil.

The aim of the present study was to investigate the mechanism of the inhibitory effect of luteolin on the proliferation of breast cancer cells induced by epidermal growth factor (EGF) in vitro. MTT assay was used to detect the inhibitory effect of luteolin on the proliferation of MCF-7 and MDA-MB-231 cells as well as the effect on the proliferation of MCF-7 cells induced by EGF. Western blotting was used to detect the effects of luteolin on the expression of epidermal growth factor receptor (EGFR), phosphatidylinositol 3-kinase (PI3K)/Akt, mitogen-activated protein kinase (MAPK)/extracellular-signal-regulated kinases (Erk) 1/2 and signal transducers and activators of transcription-3 (STAT3) in MCF-7 cells induced by EGF. The results showed that luteolin could significantly inhibit the proliferation of MCF-7 and MDA-MB-231 cells, and the inhibitory effect on MCF-7 cells was more prominent. Moreover, luteolin could inhibit the proliferation of MCF-7 cells induced by EGF. Western blotting results showed that luteolin and AG1478 (an inhibitor of EGFR signaling) could inhibit the expression of p-EGFR and p-STAT3 in MCF-7 cells induced by EGF. Luteolin, LY294002 (an inhibitor of Akt signaling) and PD98059 (an inhibitor of Erk1/2 signaling) could inhibit the expression of p-Akt and p-Erk1/2 respectively in MCF-7 cells induced by EGF. Our data suggest that luteolin may inhibit EGF-induced activities of EGFR signaling pathway in human breast cancer cell lines, and PI3K/Akt, MAPK/Erk1/2, STAT3 signal pathways may be the major pathways that mediate the inhibitory effect of luteolin on EGFR signaling. Overall, our results may provide a theoretical foundation for the development of luteolin as anti-tumor drug.
Breast Neoplasms ; Cell Line, Tumor ; Cell Proliferation ; Chromones ; Epidermal Growth Factor ; Humans ; Luteolin ; MAP Kinase Signaling System ; Mitogen-Activated Protein Kinase 1 ; Mitogen-Activated Protein Kinase 3 ; Morpholines ; Phosphatidylinositol 3-Kinases ; Quinazolines ; Receptor, Epidermal Growth Factor ; Tyrphostins