In regenerating liver of mice, marked increase of the activity of phosphotyrosyl protein phosphatase (PTPP) in cytosol was observed. The PTPP activity varied with time and reached the highest level between 24 to 48 ho...In regenerating liver of mice, marked increase of the activity of phosphotyrosyl protein phosphatase (PTPP) in cytosol was observed. The PTPP activity varied with time and reached the highest level between 24 to 48 hours after partial hepatectomy. In H22a cells the PTPP activity found in every subcellular fraction was lower than that of the normal liver. The PTPP activity was mostly concentrated in lysosomes of normal liver, but mainly distributed in nucleus, cytosol and microsome of regenerating liver. In H22a cells PTPP activity seemed distribute evenly. Five similar major PTPP peaks (I-V) were obtained on DEAE cellulose chromatography of cytosols from all three of liver cells studied. However, two additional PTPP peaks, a and b, were also obtained from cytosol of liver.展开更多
目的通过检测NOD样受体蛋白3(NLRP3)/半胱氨酸天冬氨酸蛋白水解酶-1(Caspase-1)/消皮素D(GSDMD)通路,探讨阿霉素纳米凝胶载药(Nanogel drug loading,NG)对H22肝癌荷瘤小鼠的作用机制。方法将H22肝癌细胞珠每只1.25×10^(6)个/100μ...目的通过检测NOD样受体蛋白3(NLRP3)/半胱氨酸天冬氨酸蛋白水解酶-1(Caspase-1)/消皮素D(GSDMD)通路,探讨阿霉素纳米凝胶载药(Nanogel drug loading,NG)对H22肝癌荷瘤小鼠的作用机制。方法将H22肝癌细胞珠每只1.25×10^(6)个/100μl接种于Balb/c小鼠右前肢腋窝皮下,待肿瘤体积至50~60mm^(3)时,将40只Balb/c小鼠随机分为5组(n=8),分别为肿瘤对照组、阿霉素3mg组(DOX-3.0)、阿霉素6mg组(DOX-6.0)、阿霉素纳米凝胶载药3mg组(NG/DOX-3.0)和阿霉素纳米凝胶载药6mg组(NG/DOX-6.0)。各组均采用尾静脉注射给药治疗,分别给予生理盐水(5ml/kg)、DOX(3mg/kg)、DOX(6mg/kg)、NG/DOX(3mg/kg)和NG/DOX(6mg/kg),1次/3d,给药4次,于给药结束第3d处死动物取材。通过苏木精-伊红(hematoxylin-eosin,HE)染色,观察肿瘤生长情况和坏死面积;采用免疫组织化学染色检测NLRP3/Caspase-1/GSDMD凋亡基因的表达情况。结果HE染色与肿瘤对照组比较,各治疗组肿瘤生长状态较差,且DOX-3.0、NG/DOX-3.0、DOX-6.0、NG/DOX-6.0肿瘤坏死面积百分率依次增大,组间差异均有统计学意义(P<0.05);与肿瘤对照组比较,DOX-3.0、NG/DOX-3.0、DOX-6.0、NG/DOX-6.0组肿瘤组织中NLRP3、Caspase-1、GSDMD的蛋白阳性表达水平依次减弱,组间差异均有统计学意义(P<0.05),纳米凝胶载药组作用优于单纯阿霉素给药组。结论纳米凝胶载药组治疗H22肝癌的作用优于阿霉素单独用药,它能通过下调NLRP3/Caspase-1/GSDMD通路,有效抑制肿瘤生长、降低药物使用剂量及提高抗癌效果,具有良好发展前景,其作用机制有待于进一步探讨。展开更多
BACKGROUND Jianpi-Huatan-Huoxue-Anshen formula[Tzu-Chi cancer-antagonizing&lifeprotecting II decoction(TCCL)]is a Chinese medical formula that has been clinically shown to reduce the gastrointestinal side effects ...BACKGROUND Jianpi-Huatan-Huoxue-Anshen formula[Tzu-Chi cancer-antagonizing&lifeprotecting II decoction(TCCL)]is a Chinese medical formula that has been clinically shown to reduce the gastrointestinal side effects of chemotherapy in cancer patients and improve their quality of life.However,its effect and mechanism on the intestinal microecology after chemotherapy are not yet clear.AIM To discover the potential mechanisms of TCCL on gastrointestinal inflammation and microecological imbalance in chemotherapy-treated mice transplanted with hepatocellular carcinoma(HCC).METHODS Ninety-six mice were inoculated subcutaneously with HCC cells.One week later,the mice received a large dose of 5-fluorouracil by intraperitoneal injection to establish a HCC chemotherapy model.Thirty-six mice were randomly selected before administration,and feces,ileal tissue,and ileal contents were collected from each mouse.The remaining mice were randomized into normal saline,continuous chemotherapy,Yangzheng Xiaoji capsulestreated,and three TCCL-treated groups.After treatment,feces,tumors,liver,spleen,thymus,stomach,jejunum,ileum,and colon tissues,and ileal contents were collected.Morphological changes,serum levels of IL-1β,IL-6,IL-8,IL-10,IL-22,TNF-α,and TGF-β,intestinal SIgA,and protein and mRNA expression of ZO-1,NF-κB,Occludin,MUC-2,Claudin-1,and IκB-αin colon tissues were documented.The effect of TCCL on the abundance and diversity of intestinal flora was analyzed using 16S rDNA sequencing.RESULTS TCCL treatment improved thymus and spleen weight,thymus and spleen indexes,and body weight,decreased tumor volumes and tumor tissue cell density,and alleviated injury to gastric,ileal,and colonic mucosal tissues.Among proteins and genes associated with inflammation,IL-10,TGF-β,SIgA,ZO-1,MUC-2,and Occludin were upregulated,whereas NF-κB,IL-1β,IL-6,TNF-α,IL-22,IL-8,and IκB-αwere downregulated.Additionally,TCCL increased the proportions of fecal Actinobacteria,AF12,Adlercreutzia,Clostridium,Coriobacteriaceae,and Paraprevotella in the intermediate stage of treatment,decreased the proportions of Mucipirillum,Odoribacter,RF32,YS2,and Rikenellaceae but increased the proportions of p_Deferribacteres and Lactobacillus at the end of treatment.Studies on ileal mucosal microbiota showed similar findings.Moreover,TCCL improved community richness,evenness,and the diversity of fecal and ileal mucosal flora.CONCLUSION TCCL relieves pathological changes in tumor tissue and chemotherapy-induced gastrointestinal injury,potentially by reducing the release of pro-inflammatory factors to repair the gastrointestinal mucosa,enhancing intestinal barrier function,and maintaining gastrointestinal microecological balance.Hence,TCCL is a very effective adjuvant to chemotherapy.展开更多
文摘In regenerating liver of mice, marked increase of the activity of phosphotyrosyl protein phosphatase (PTPP) in cytosol was observed. The PTPP activity varied with time and reached the highest level between 24 to 48 hours after partial hepatectomy. In H22a cells the PTPP activity found in every subcellular fraction was lower than that of the normal liver. The PTPP activity was mostly concentrated in lysosomes of normal liver, but mainly distributed in nucleus, cytosol and microsome of regenerating liver. In H22a cells PTPP activity seemed distribute evenly. Five similar major PTPP peaks (I-V) were obtained on DEAE cellulose chromatography of cytosols from all three of liver cells studied. However, two additional PTPP peaks, a and b, were also obtained from cytosol of liver.
文摘目的通过检测NOD样受体蛋白3(NLRP3)/半胱氨酸天冬氨酸蛋白水解酶-1(Caspase-1)/消皮素D(GSDMD)通路,探讨阿霉素纳米凝胶载药(Nanogel drug loading,NG)对H22肝癌荷瘤小鼠的作用机制。方法将H22肝癌细胞珠每只1.25×10^(6)个/100μl接种于Balb/c小鼠右前肢腋窝皮下,待肿瘤体积至50~60mm^(3)时,将40只Balb/c小鼠随机分为5组(n=8),分别为肿瘤对照组、阿霉素3mg组(DOX-3.0)、阿霉素6mg组(DOX-6.0)、阿霉素纳米凝胶载药3mg组(NG/DOX-3.0)和阿霉素纳米凝胶载药6mg组(NG/DOX-6.0)。各组均采用尾静脉注射给药治疗,分别给予生理盐水(5ml/kg)、DOX(3mg/kg)、DOX(6mg/kg)、NG/DOX(3mg/kg)和NG/DOX(6mg/kg),1次/3d,给药4次,于给药结束第3d处死动物取材。通过苏木精-伊红(hematoxylin-eosin,HE)染色,观察肿瘤生长情况和坏死面积;采用免疫组织化学染色检测NLRP3/Caspase-1/GSDMD凋亡基因的表达情况。结果HE染色与肿瘤对照组比较,各治疗组肿瘤生长状态较差,且DOX-3.0、NG/DOX-3.0、DOX-6.0、NG/DOX-6.0肿瘤坏死面积百分率依次增大,组间差异均有统计学意义(P<0.05);与肿瘤对照组比较,DOX-3.0、NG/DOX-3.0、DOX-6.0、NG/DOX-6.0组肿瘤组织中NLRP3、Caspase-1、GSDMD的蛋白阳性表达水平依次减弱,组间差异均有统计学意义(P<0.05),纳米凝胶载药组作用优于单纯阿霉素给药组。结论纳米凝胶载药组治疗H22肝癌的作用优于阿霉素单独用药,它能通过下调NLRP3/Caspase-1/GSDMD通路,有效抑制肿瘤生长、降低药物使用剂量及提高抗癌效果,具有良好发展前景,其作用机制有待于进一步探讨。
基金Supported by Natural Science Foundation of Xiamen,China,No.3502Z20227171the Young Investigator Research Program of Xiang’an Hospital of Xiamen University,No.XAH23005+2 种基金the Traditional Chinese Medicine Foundation of Xiamen,No.XWZY-2023-0103Natural Science Foundation of Fujian,China,No.2018J01136National Natural Science Foundation of China,No.81202659.
文摘BACKGROUND Jianpi-Huatan-Huoxue-Anshen formula[Tzu-Chi cancer-antagonizing&lifeprotecting II decoction(TCCL)]is a Chinese medical formula that has been clinically shown to reduce the gastrointestinal side effects of chemotherapy in cancer patients and improve their quality of life.However,its effect and mechanism on the intestinal microecology after chemotherapy are not yet clear.AIM To discover the potential mechanisms of TCCL on gastrointestinal inflammation and microecological imbalance in chemotherapy-treated mice transplanted with hepatocellular carcinoma(HCC).METHODS Ninety-six mice were inoculated subcutaneously with HCC cells.One week later,the mice received a large dose of 5-fluorouracil by intraperitoneal injection to establish a HCC chemotherapy model.Thirty-six mice were randomly selected before administration,and feces,ileal tissue,and ileal contents were collected from each mouse.The remaining mice were randomized into normal saline,continuous chemotherapy,Yangzheng Xiaoji capsulestreated,and three TCCL-treated groups.After treatment,feces,tumors,liver,spleen,thymus,stomach,jejunum,ileum,and colon tissues,and ileal contents were collected.Morphological changes,serum levels of IL-1β,IL-6,IL-8,IL-10,IL-22,TNF-α,and TGF-β,intestinal SIgA,and protein and mRNA expression of ZO-1,NF-κB,Occludin,MUC-2,Claudin-1,and IκB-αin colon tissues were documented.The effect of TCCL on the abundance and diversity of intestinal flora was analyzed using 16S rDNA sequencing.RESULTS TCCL treatment improved thymus and spleen weight,thymus and spleen indexes,and body weight,decreased tumor volumes and tumor tissue cell density,and alleviated injury to gastric,ileal,and colonic mucosal tissues.Among proteins and genes associated with inflammation,IL-10,TGF-β,SIgA,ZO-1,MUC-2,and Occludin were upregulated,whereas NF-κB,IL-1β,IL-6,TNF-α,IL-22,IL-8,and IκB-αwere downregulated.Additionally,TCCL increased the proportions of fecal Actinobacteria,AF12,Adlercreutzia,Clostridium,Coriobacteriaceae,and Paraprevotella in the intermediate stage of treatment,decreased the proportions of Mucipirillum,Odoribacter,RF32,YS2,and Rikenellaceae but increased the proportions of p_Deferribacteres and Lactobacillus at the end of treatment.Studies on ileal mucosal microbiota showed similar findings.Moreover,TCCL improved community richness,evenness,and the diversity of fecal and ileal mucosal flora.CONCLUSION TCCL relieves pathological changes in tumor tissue and chemotherapy-induced gastrointestinal injury,potentially by reducing the release of pro-inflammatory factors to repair the gastrointestinal mucosa,enhancing intestinal barrier function,and maintaining gastrointestinal microecological balance.Hence,TCCL is a very effective adjuvant to chemotherapy.
