Objective: he effect of M3 receptor antagonist 4-DAMP (4-diphenylacetoxy-N-methylpiperidine-methiodide) on the proliferation of melanoma were explored, and the development prospect and significance of M3 receptor anta...Objective: he effect of M3 receptor antagonist 4-DAMP (4-diphenylacetoxy-N-methylpiperidine-methiodide) on the proliferation of melanoma were explored, and the development prospect and significance of M3 receptor antagonist in the field of anti-tumor were discussed. Methods: The viability of tumor cells was detected by MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) colorimetry. The optimal concentration of 4-DAMP was screened and the effect of the drug was confirmed by EdU (5-ethynyl-2'-deoxyuridine) fluorescence staining. Adult male BALB/c nude mice were divided into control group, 5-Fu (5-Fluorouracil, 5-Fluorouracil, 20 mg/kg/d) group and 4-DAMP (2 mg/kg/d) group to establishtumor-bearing model in vivo. The tumor volume curve was calculated and plotted. Mice were killed after 21 days of continuous administration. Strip the tumor, take pictures and meatured the weighs, and calculate the inhibition rate. [1]The spleen was weighed and the spleen index was culculated. H & E staining was used to observe the pathological changes of tumor sections. The gene expression of M3R、MIA (Melanoma inhibitory activity, MIA)、SP-1 (Transcription factor Sp1, SP-1) and Lnc-SPRY4-IT1 (SPRY4 Intronic Transcript 1,Lnc-SPRY4-IT1) in A375 cell line was detected by real-time quantitative PCR (quantitative reverse-transcription polymerase chain reaction, qRT-PCR). The protein level of ERK1/2 (extracellular signal-regulated kinase 1/2,ERK1/2) was detected by Western Blot. Results: 4-DAMP could significantly inhibit the activity of A375 cell line (P < 0.01), and EdU staining demonstrated that 4-DAMP inhibited the growth of A375 cell line. The results of animal experiments showed that compared with the control group, the volume and mass of tumor in 5-Fu group and 4-DAMP group were significantly reduced (P < 0.05) and H & E staining showed that the intercellular space became larger, connective tissue increased, and tumor growth was obstructed. Compared with the control group, the tumor inhibition rate of 4-DAMP group was 72.01% (P < 0.01). There was no significant difference in body mass and spleen index. Compared with the control group, the expression of MIA, SP-1 and LncRNA-SPRY4-IT1 was decreased by 4-DAMP or si-M3R transfection. Compared with the control group, the expression of ERK1/2 was decreased by 4-DAMP or si-M3R transfection. Conclusions: These results suggest that M3 receptor antagonist 4-DAMP can inhibit the proliferation of melanoma and reduce the expression of mRNA of MIA, SP-1 and LncRNA-SPRY4-IT1 and the protein level of ERK1/2 in A375 tumor cells.展开更多
目的:探讨参苓白术散对脾虚湿困型溃疡性结肠炎(ulcerative colitis,UC)大鼠结肠组织中水通道蛋白4(aquaporin 4,AQP4)蛋白及mRNA表达的影响.方法:Wistar大鼠40只,随机分为正常对照组、模型对照组、柳氮磺吡啶组、参苓白术散组,每组10只...目的:探讨参苓白术散对脾虚湿困型溃疡性结肠炎(ulcerative colitis,UC)大鼠结肠组织中水通道蛋白4(aquaporin 4,AQP4)蛋白及mRNA表达的影响.方法:Wistar大鼠40只,随机分为正常对照组、模型对照组、柳氮磺吡啶组、参苓白术散组,每组10只,脾虚湿困型UC大鼠模型的复制采用环境与饮食干预结合三硝基苯磺酸(trinitrobenzene sulfonic acid,TNBS)/乙醇灌肠法,运用免疫组织化学法、荧光定量PCR法检测并比较各组大鼠结肠组织AQP4蛋白及mRNA表达的变化.结果:模型对照组AQP4蛋白及mRNA表达量低于正常对照组(0.764±0.051 vs 0.358±0.033,28.288±1.578 vs 28.296±1.142,P<0.05),参苓白术散组和柳氮磺吡啶组大鼠结肠组织AQP4表达平均光密度及AQP4mRNA表达较模型对照组显著升高,其间差异有统计学意义(0.752±0.059 vs 0.358±0.033,28.103±1.395 vs 28.288±1.578,0.487±0.046vs 0.358±0.033,27.711±1.638 vs 28.288±1.578,P<0.05),参苓白术散组大鼠结肠组织A Q P4表达平均光密度与柳氮磺吡啶组比较差异亦有统计学意义(0.752±0.059 vs 0.487±0.046,28.103±1.395 vs 27.711±1.638,P<0.05).结论:参苓白术散治疗可以明显改善UC大鼠结肠组织中AQP4的表达,促进肠黏膜的修复.展开更多
基金High-level Talents Fund of Natural Science Foundation of Hainan Province(No.2019RC376).
文摘Objective: he effect of M3 receptor antagonist 4-DAMP (4-diphenylacetoxy-N-methylpiperidine-methiodide) on the proliferation of melanoma were explored, and the development prospect and significance of M3 receptor antagonist in the field of anti-tumor were discussed. Methods: The viability of tumor cells was detected by MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) colorimetry. The optimal concentration of 4-DAMP was screened and the effect of the drug was confirmed by EdU (5-ethynyl-2'-deoxyuridine) fluorescence staining. Adult male BALB/c nude mice were divided into control group, 5-Fu (5-Fluorouracil, 5-Fluorouracil, 20 mg/kg/d) group and 4-DAMP (2 mg/kg/d) group to establishtumor-bearing model in vivo. The tumor volume curve was calculated and plotted. Mice were killed after 21 days of continuous administration. Strip the tumor, take pictures and meatured the weighs, and calculate the inhibition rate. [1]The spleen was weighed and the spleen index was culculated. H & E staining was used to observe the pathological changes of tumor sections. The gene expression of M3R、MIA (Melanoma inhibitory activity, MIA)、SP-1 (Transcription factor Sp1, SP-1) and Lnc-SPRY4-IT1 (SPRY4 Intronic Transcript 1,Lnc-SPRY4-IT1) in A375 cell line was detected by real-time quantitative PCR (quantitative reverse-transcription polymerase chain reaction, qRT-PCR). The protein level of ERK1/2 (extracellular signal-regulated kinase 1/2,ERK1/2) was detected by Western Blot. Results: 4-DAMP could significantly inhibit the activity of A375 cell line (P < 0.01), and EdU staining demonstrated that 4-DAMP inhibited the growth of A375 cell line. The results of animal experiments showed that compared with the control group, the volume and mass of tumor in 5-Fu group and 4-DAMP group were significantly reduced (P < 0.05) and H & E staining showed that the intercellular space became larger, connective tissue increased, and tumor growth was obstructed. Compared with the control group, the tumor inhibition rate of 4-DAMP group was 72.01% (P < 0.01). There was no significant difference in body mass and spleen index. Compared with the control group, the expression of MIA, SP-1 and LncRNA-SPRY4-IT1 was decreased by 4-DAMP or si-M3R transfection. Compared with the control group, the expression of ERK1/2 was decreased by 4-DAMP or si-M3R transfection. Conclusions: These results suggest that M3 receptor antagonist 4-DAMP can inhibit the proliferation of melanoma and reduce the expression of mRNA of MIA, SP-1 and LncRNA-SPRY4-IT1 and the protein level of ERK1/2 in A375 tumor cells.
文摘目的:探讨参苓白术散对脾虚湿困型溃疡性结肠炎(ulcerative colitis,UC)大鼠结肠组织中水通道蛋白4(aquaporin 4,AQP4)蛋白及mRNA表达的影响.方法:Wistar大鼠40只,随机分为正常对照组、模型对照组、柳氮磺吡啶组、参苓白术散组,每组10只,脾虚湿困型UC大鼠模型的复制采用环境与饮食干预结合三硝基苯磺酸(trinitrobenzene sulfonic acid,TNBS)/乙醇灌肠法,运用免疫组织化学法、荧光定量PCR法检测并比较各组大鼠结肠组织AQP4蛋白及mRNA表达的变化.结果:模型对照组AQP4蛋白及mRNA表达量低于正常对照组(0.764±0.051 vs 0.358±0.033,28.288±1.578 vs 28.296±1.142,P<0.05),参苓白术散组和柳氮磺吡啶组大鼠结肠组织AQP4表达平均光密度及AQP4mRNA表达较模型对照组显著升高,其间差异有统计学意义(0.752±0.059 vs 0.358±0.033,28.103±1.395 vs 28.288±1.578,0.487±0.046vs 0.358±0.033,27.711±1.638 vs 28.288±1.578,P<0.05),参苓白术散组大鼠结肠组织A Q P4表达平均光密度与柳氮磺吡啶组比较差异亦有统计学意义(0.752±0.059 vs 0.487±0.046,28.103±1.395 vs 27.711±1.638,P<0.05).结论:参苓白术散治疗可以明显改善UC大鼠结肠组织中AQP4的表达,促进肠黏膜的修复.
