目的通过RT-PCR检测IECs中TLR2与TLR4的表达,通过荧光定量PCR检测IECs中IL-1和TNF-α的表达,以对双歧杆菌预防与辅助治疗UC作用机制加以探讨。方法将36只Babl/c小鼠随即分为3组,即正常对照组(N)、DSS诱导组(D)、双歧杆菌喂养组(B)。N组...目的通过RT-PCR检测IECs中TLR2与TLR4的表达,通过荧光定量PCR检测IECs中IL-1和TNF-α的表达,以对双歧杆菌预防与辅助治疗UC作用机制加以探讨。方法将36只Babl/c小鼠随即分为3组,即正常对照组(N)、DSS诱导组(D)、双歧杆菌喂养组(B)。N组不施加作用,D组生理盐水灌肠2周,并在第2周给以2%的DSS自由饮用;双歧杆菌组双歧杆菌灌肠2周,并在第2周再给以2%的DSS自由饮用。2周后处死小鼠做组织切片和分离IECs并提取IECs中的总RNA,用RT-PCR检测TLR2、TLR4的表达,用Real time RT-PCR检测IL-1和TNF-α表达。结果双歧杆菌组IECs表达TLR2远远高于单纯的DSS诱导组,结果具有统计学意义,而双歧杆菌组IECs表达TLR4、IL-1和TNF-α则远远低于单纯的DSS诱导组,结果具有统计学意义。结论双歧杆菌通过与诱导IECs中TLR2的表达而抑制其TLR4、IL-1和TNF-α的表达,从而达到预防与辅助性治疗DSS诱导的UC作用。展开更多
[Objective] The paper was to investigate the protective effects of different concentrations of Bacillus subtilis peptidoglycan(PG) on β-conglycinin-induced inflammatory injury in intestinal epithelial cells of juveni...[Objective] The paper was to investigate the protective effects of different concentrations of Bacillus subtilis peptidoglycan(PG) on β-conglycinin-induced inflammatory injury in intestinal epithelial cells of juvenile carp(Cyprinus carpio).[Method] In 24-cell microplates, the intestinal epithelial cells(IECs) of juvenile carp were primarily cultured for 72 h at 26°C and 6% CO2, and then the IECs were randomly divided into6 groups with 4 replicates per group. One of the six groups was set as negative control group, and the other groups were all supplemented with 1.0 mg/mL β-conglycinin in culture medium to establish inflammatory injury. At 24 h post induction, the culture media were changed into B. subtilis PG culture media with the concentrations of 0(positive control group), 0.15, 0.30, 0.45 and 0.60 mg/mL, respectively. The samples were collected to measure the antioxidant and anti-inflammatory indices at 12, 24 and 36 h post culture.[Result]β-conglycinin exposure significantly decreased the activity of ASA, AHR, SOD, CAT, GPx, and increased the PC content and the mRNA expression of inflammatory cytokines(IL-1β, IL-8, TNF-α1,IL-10 and TGF-β). At 12, 24 and 36 h post PG treatment, the activities of ASA, AHR, SOD, CAT, GPx and the content of PC in cells decreased in a dose-dependent manner;the mRNA levels of IL-1β, IL-8 and TNF-α1 were down-regulated and those of IL-10 and TGF-β were up-regulated.[Conclusion] Different concentrations of B. subtilis PG could protect IECs oxidative damage induced by β-conglycinin and improve the antioxidant capacity of IECs. High concentration of PG could improve the anti-inflammatory ability of IECs by inhibiting inflammatory factors and promoting the gene expression of anti-inflammatory cytokines.展开更多
文摘目的通过RT-PCR检测IECs中TLR2与TLR4的表达,通过荧光定量PCR检测IECs中IL-1和TNF-α的表达,以对双歧杆菌预防与辅助治疗UC作用机制加以探讨。方法将36只Babl/c小鼠随即分为3组,即正常对照组(N)、DSS诱导组(D)、双歧杆菌喂养组(B)。N组不施加作用,D组生理盐水灌肠2周,并在第2周给以2%的DSS自由饮用;双歧杆菌组双歧杆菌灌肠2周,并在第2周再给以2%的DSS自由饮用。2周后处死小鼠做组织切片和分离IECs并提取IECs中的总RNA,用RT-PCR检测TLR2、TLR4的表达,用Real time RT-PCR检测IL-1和TNF-α表达。结果双歧杆菌组IECs表达TLR2远远高于单纯的DSS诱导组,结果具有统计学意义,而双歧杆菌组IECs表达TLR4、IL-1和TNF-α则远远低于单纯的DSS诱导组,结果具有统计学意义。结论双歧杆菌通过与诱导IECs中TLR2的表达而抑制其TLR4、IL-1和TNF-α的表达,从而达到预防与辅助性治疗DSS诱导的UC作用。
基金Supported by Science and Technology Research Project of Henan Province(172102110205)
文摘[Objective] The paper was to investigate the protective effects of different concentrations of Bacillus subtilis peptidoglycan(PG) on β-conglycinin-induced inflammatory injury in intestinal epithelial cells of juvenile carp(Cyprinus carpio).[Method] In 24-cell microplates, the intestinal epithelial cells(IECs) of juvenile carp were primarily cultured for 72 h at 26°C and 6% CO2, and then the IECs were randomly divided into6 groups with 4 replicates per group. One of the six groups was set as negative control group, and the other groups were all supplemented with 1.0 mg/mL β-conglycinin in culture medium to establish inflammatory injury. At 24 h post induction, the culture media were changed into B. subtilis PG culture media with the concentrations of 0(positive control group), 0.15, 0.30, 0.45 and 0.60 mg/mL, respectively. The samples were collected to measure the antioxidant and anti-inflammatory indices at 12, 24 and 36 h post culture.[Result]β-conglycinin exposure significantly decreased the activity of ASA, AHR, SOD, CAT, GPx, and increased the PC content and the mRNA expression of inflammatory cytokines(IL-1β, IL-8, TNF-α1,IL-10 and TGF-β). At 12, 24 and 36 h post PG treatment, the activities of ASA, AHR, SOD, CAT, GPx and the content of PC in cells decreased in a dose-dependent manner;the mRNA levels of IL-1β, IL-8 and TNF-α1 were down-regulated and those of IL-10 and TGF-β were up-regulated.[Conclusion] Different concentrations of B. subtilis PG could protect IECs oxidative damage induced by β-conglycinin and improve the antioxidant capacity of IECs. High concentration of PG could improve the anti-inflammatory ability of IECs by inhibiting inflammatory factors and promoting the gene expression of anti-inflammatory cytokines.
