目的观察艾灸对结肠炎相关性结肠癌(CAC)大鼠的干预作用,从嘌呤受体P2X7R与Wnt/b-catenin信号通路探讨可能的效应机制。方法将SD大鼠随机分为正常组、CAC组、隔药灸组、隔姜灸组。CAC组、隔药灸组、隔姜灸组均采用腹腔注射AOM联合DSS法...目的观察艾灸对结肠炎相关性结肠癌(CAC)大鼠的干预作用,从嘌呤受体P2X7R与Wnt/b-catenin信号通路探讨可能的效应机制。方法将SD大鼠随机分为正常组、CAC组、隔药灸组、隔姜灸组。CAC组、隔药灸组、隔姜灸组均采用腹腔注射AOM联合DSS法制备CAC大鼠模型,隔药灸与隔姜灸组均取天枢(双)、气海穴进行治疗。记录各组大鼠体质量、疾病活动指数(DAI)和成瘤率;通过HE染色观察艾灸对CAC大鼠结肠损伤的干预效应;通过RT-q PCR和Western Blot技术,检测艾灸对CAC大鼠结肠组织C-myc、Wnt1、b-catenin、GSK-3bm RNA和P2X7R蛋白表达的调节作用。结果与正常组相比,CAC组大鼠体质量显著降低、DAI增高、成瘤率明显增加(P<0.05),结肠组织可见腺管共壁背靠背和筛样结构,高级别腺癌形成。CAC组大鼠结肠组织P2X7R蛋白表达显著下调(P<0.05),C-myc、b-catenin、GSK-3b、Wnt1 m RNA的表达均显著上调(P<0.05)。与CAC组相比,隔药灸组和隔姜灸组大鼠体质量增加,DAI降低(P<0.05),结肠组织P2X7R蛋白表达显著上调,C-myc m RNA下调(P<0.05);隔姜灸组成瘤率明显降低(P<0.05),且Wnt1、b-catenin、GSK-3bm RNA表达显著下调(P<0.05)。结论隔药灸、隔姜灸均能调节CAC大鼠结肠组织P2X7R及C-myc的异常表达,且隔姜灸还能下调CAC大鼠结肠组织Wnt1、b-catenin、GSK-3bm RNA的表达。展开更多
Objective:To observe the anti-inflammatory effect,as well as the effect on the expression of microtubule-associated protein light chain 3B(LC3B)and Beclin-1 of herbal cake-partitioned moxibustion in rats with experime...Objective:To observe the anti-inflammatory effect,as well as the effect on the expression of microtubule-associated protein light chain 3B(LC3B)and Beclin-1 of herbal cake-partitioned moxibustion in rats with experimental autoimmune thyroiditis(EAT).Methods:Female Sprague-Dawley rats were randomly divided into a normal group and a modeling group.The EAT rat model was prepared by a combination of antigen immunization plus iodine agent induction.After the model was prepared,rats in the modeling group were randomly and equally divided into a model group and a herbal cake-partitioned moxibustion group.In the herbal cake-partitioned moxibustion group,moxibustion was alternately applied to two groups of points[Dazhui(GV14)-Mingmen(GV4)and Tiantu(CV22)-Guanyuan(CV4)],and the treatment continued for 30 d.Rats in the normal and model groups were only fixed identically without intervention.Histopathological manifestations of thyroid glands were observed by hematoxylin-eosin staining;the concentrations of thyroid peroxidase antibodies(TPOAb),thyroglobulin antibodies(TGAb),interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay;real-time fluorescence quantitative polymerase chain reaction and immunohistochemistry were used to detect the mRNA and protein expression of autophagy-related factors LC3B and Beclin-1 in thyroid tissue.Results:There were massive follicular destruction,lymphocytic infiltration,and interstitial fibrous tissue hyperplasia of the thyroid glands in the model group.Some follicles of the thyroid glands were destroyed with few lymphocyte infiltrations and fibrous tissue hyperplasia in the moxibustion group.Compared with the normal group,the concentrations of serum TPOAb,TGAb,IL-1β,IL-6,and TNF-αwere increased in the model rats(P<0.05);the mRNA and protein expression levels of LC3B and Beclin-1 in thyroid tissue were reduced in the model group(P<0.05).Compared with the model group,the concentrations of serum TPOAb,TGAb,IL-1β,IL-6,and TNF-αwere reduced in the herbal cake-partitioned moxibustion group(P<0.05);the mRNA and protein expression levels of LC3B and Beclin-1 in thyroid tissue were increased in the herbal cake-partitioned moxibustion group(P<0.05).The mRNA and protein expression of LC3B and Beclin-1 in thyroid tissue was negatively correlated with the serum levels of TPOAb and TGAb.Conclusion:Herbal cake-partitioned moxibustion reduces the inflammatory response in the thyroid glands of EAT rats and lowers the levels of serum TPOAb and TGAb.This may be related to the regulation of mRNA and protein expression of the autophagy-associated factors LC3B and Beclin-1 in rat thyroid tissue.展开更多
文摘目的观察艾灸对结肠炎相关性结肠癌(CAC)大鼠的干预作用,从嘌呤受体P2X7R与Wnt/b-catenin信号通路探讨可能的效应机制。方法将SD大鼠随机分为正常组、CAC组、隔药灸组、隔姜灸组。CAC组、隔药灸组、隔姜灸组均采用腹腔注射AOM联合DSS法制备CAC大鼠模型,隔药灸与隔姜灸组均取天枢(双)、气海穴进行治疗。记录各组大鼠体质量、疾病活动指数(DAI)和成瘤率;通过HE染色观察艾灸对CAC大鼠结肠损伤的干预效应;通过RT-q PCR和Western Blot技术,检测艾灸对CAC大鼠结肠组织C-myc、Wnt1、b-catenin、GSK-3bm RNA和P2X7R蛋白表达的调节作用。结果与正常组相比,CAC组大鼠体质量显著降低、DAI增高、成瘤率明显增加(P<0.05),结肠组织可见腺管共壁背靠背和筛样结构,高级别腺癌形成。CAC组大鼠结肠组织P2X7R蛋白表达显著下调(P<0.05),C-myc、b-catenin、GSK-3b、Wnt1 m RNA的表达均显著上调(P<0.05)。与CAC组相比,隔药灸组和隔姜灸组大鼠体质量增加,DAI降低(P<0.05),结肠组织P2X7R蛋白表达显著上调,C-myc m RNA下调(P<0.05);隔姜灸组成瘤率明显降低(P<0.05),且Wnt1、b-catenin、GSK-3bm RNA表达显著下调(P<0.05)。结论隔药灸、隔姜灸均能调节CAC大鼠结肠组织P2X7R及C-myc的异常表达,且隔姜灸还能下调CAC大鼠结肠组织Wnt1、b-catenin、GSK-3bm RNA的表达。
基金This work was supported by the National TCM Leading Talents Support Program-Qihuang Scholars(国家中医药领军人才支持计划-岐黄学者)National Program on Key Basic Research Project“973 Program”(国家重点基础研究发展计划“973计划项目”,No.2015CB554501)+1 种基金Projects of National Natural Science Foundation of China(国家自然科学基金项目,No.82074551,No.81704176)Youth Project from Department of Science and Education,Shanghai Health Commission(上海市卫健委科教处青年基金,No.20194Y0013).
文摘Objective:To observe the anti-inflammatory effect,as well as the effect on the expression of microtubule-associated protein light chain 3B(LC3B)and Beclin-1 of herbal cake-partitioned moxibustion in rats with experimental autoimmune thyroiditis(EAT).Methods:Female Sprague-Dawley rats were randomly divided into a normal group and a modeling group.The EAT rat model was prepared by a combination of antigen immunization plus iodine agent induction.After the model was prepared,rats in the modeling group were randomly and equally divided into a model group and a herbal cake-partitioned moxibustion group.In the herbal cake-partitioned moxibustion group,moxibustion was alternately applied to two groups of points[Dazhui(GV14)-Mingmen(GV4)and Tiantu(CV22)-Guanyuan(CV4)],and the treatment continued for 30 d.Rats in the normal and model groups were only fixed identically without intervention.Histopathological manifestations of thyroid glands were observed by hematoxylin-eosin staining;the concentrations of thyroid peroxidase antibodies(TPOAb),thyroglobulin antibodies(TGAb),interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay;real-time fluorescence quantitative polymerase chain reaction and immunohistochemistry were used to detect the mRNA and protein expression of autophagy-related factors LC3B and Beclin-1 in thyroid tissue.Results:There were massive follicular destruction,lymphocytic infiltration,and interstitial fibrous tissue hyperplasia of the thyroid glands in the model group.Some follicles of the thyroid glands were destroyed with few lymphocyte infiltrations and fibrous tissue hyperplasia in the moxibustion group.Compared with the normal group,the concentrations of serum TPOAb,TGAb,IL-1β,IL-6,and TNF-αwere increased in the model rats(P<0.05);the mRNA and protein expression levels of LC3B and Beclin-1 in thyroid tissue were reduced in the model group(P<0.05).Compared with the model group,the concentrations of serum TPOAb,TGAb,IL-1β,IL-6,and TNF-αwere reduced in the herbal cake-partitioned moxibustion group(P<0.05);the mRNA and protein expression levels of LC3B and Beclin-1 in thyroid tissue were increased in the herbal cake-partitioned moxibustion group(P<0.05).The mRNA and protein expression of LC3B and Beclin-1 in thyroid tissue was negatively correlated with the serum levels of TPOAb and TGAb.Conclusion:Herbal cake-partitioned moxibustion reduces the inflammatory response in the thyroid glands of EAT rats and lowers the levels of serum TPOAb and TGAb.This may be related to the regulation of mRNA and protein expression of the autophagy-associated factors LC3B and Beclin-1 in rat thyroid tissue.
