本研究探讨竹节参总皂苷(saponins from Panax japonicus,SPJ)对自然衰老大鼠内质网应激相关蛋白的调节作用以及对神经细胞凋亡的干预作用。雄性SD大鼠随机分为6月龄组、24月龄组、SPJ低剂量组(10mg/kg)和高剂量组(30mg/kg)。Westernblo...本研究探讨竹节参总皂苷(saponins from Panax japonicus,SPJ)对自然衰老大鼠内质网应激相关蛋白的调节作用以及对神经细胞凋亡的干预作用。雄性SD大鼠随机分为6月龄组、24月龄组、SPJ低剂量组(10mg/kg)和高剂量组(30mg/kg)。Westernblot法检测大鼠皮层和海马内质网应激相关蛋白(GRP78、p-PERK、p-EIF2α、ATF4、ATF6、p-IRE1、XBP1)以及凋亡相关蛋白(p-JNK、CHOP、Bcl-2、Bax)表达变化。结果显示,与6月龄组相比,24月龄组皮层/海马组织中GRP78、p-PERK、p-EIF2α、ATF4、ATF6、p-IRE1、XBP1和Bcl-2/Bax蛋白表达分别下降了80%/59%、82%/78%、79%/77%、94%/74%、72%/66%、78%/81%、82%/71%和90%/92%,p-JNK/JNK、CHOP蛋白表达分别上升了1/1.72、3.33/1.33倍。给予不同剂量竹节参总皂苷干预后,均能明显上调内质网应激相关蛋白和Bcl-2/Bax的表达水平,下调促凋亡蛋白表达水平。研究结果表明,竹节参总皂苷能通过上调内质网应激相关蛋白以及抗凋亡蛋白的表达水平,下调促凋亡蛋白的表达,从而维持衰老大鼠脑组织的内质网稳态,减少神经细胞凋亡。展开更多
OBJECTIVE Gastric ulcers affect people of all ages and half of the world's population,which is being considered as the new"plague of the 21st century".As is well known,gastric mucosa is known as the firs...OBJECTIVE Gastric ulcers affect people of all ages and half of the world's population,which is being considered as the new"plague of the 21st century".As is well known,gastric mucosa is known as the first guard to protect the stomach from ulcer injury,while the aetiology of gastric ulcer is relative to imbalances between gastric mucosal protective and aggressive factors.Therefore,reducing or eliminating the aggressive factors,returning to the balance of between mucosal protective and aggressive factors,and then restoring the normal functional of gastric mucosal barrier could be crucial for treating the gastric ulcer.The fruits of Chaenomeles speciosa(TCS),also known as"mugua",might be processed into edible and health care derived products,and used as a commonly used traditional medicine in China for thousands of years.In China folk,there is a saying that"apricot one benefit,pear two benefits,mgua hundred benefits",so it has a"hundred-benefit"fruit reputation.Tujia nationality inhabitants in Southwestern China should have rheumatic diseases and peptic ulcers for living in the damp environments and bingeing on spicy and pungent foods.For the exis⁃tence of the fruit derived products of Chaenomeles speciosa as their complementary foods or snacks,the habit makes them rarely suffer from the two kinds of diseases.Enlightened by these,we had investigated the structure-activity rela⁃tionships,screened out CSTT with gastroprotective activity.Our previous studies demonstrated that TCS owned effec⁃tively therapeutic effects on gastric ulcer patients and animals,and further confirmed that TFF1 and apoptotic pathway were closely interrelated with its exerting gastroprotection.However,its underlying molecular mechanisms involved have not been fully elucidated.The current study was to further investigate its protective effect on indomethacin(IND)-damaged RGM-1 cells and rats and its underlying mechanisms through modulating TFF1-mediated EGF/EGFR and apoptotic pathways.METHODS The gastroprotection of TCS was evaluated with IND-induced gastric lesions model in RGM-1 cells and rats.In vitro,the proliferation,migration,mitochondrial viability and apoptosis were assessed,In vivo,ulcer index,ulcer inhibition rate,gastric juice acidity,gastric wall mucus(GWM),histopathology of gastric mucosa were detected.The gastroprotective effects of TCS through the TFF1-mediated EGFR/EGFR and apoptotic pathways were presented and measured by qRT-PCR and Western blotting assays.RESULTS TCS had gastroprotective function,which was related to the amelioration in promoting IND-damaged RGM-1 cell proliferation and migration,hoisting gastric juice acidity and GWM,improving ulcer index and ulcer inhibition rate,attenuating the hemorrhage,edema,epithelial cell loss and inflammatory cell infiltration of gastric mucosa,upregulating proliferation cell nuclear antigen,Bcl-2,Bcl-xl mRNA and TFF1,EGF,p-EGFR,p-Src,pro-caspase-3,pro-caspase-9 protein expressions,mitochondrial viability,mitochondrial cytochrome C concentration and p-EGFR/EGFR,p-Src/Src,Bcl-2/Bax,Bcl-xl/Bad ratioes,downregulating Bax,Bad,Apaf-1 mRNA and cleaved-caspase-3,cleaved-caspase-9,cleaved-PARP-1 protein expressions,cytosol cytochrome C concentration.CONCLUSION TCS′s gastroprotective effect was closely connected with boosting TFF1 expression,acti⁃vating TFF1-mediated EGF/EGFR pathway,thus restraining mitochondrial-dependent apoptosis,which provided new insights into interpreting its underlying mechanism and promised to act as a candidate drug to treat gastric mucosal injury.展开更多
文摘本研究探讨竹节参总皂苷(saponins from Panax japonicus,SPJ)对自然衰老大鼠内质网应激相关蛋白的调节作用以及对神经细胞凋亡的干预作用。雄性SD大鼠随机分为6月龄组、24月龄组、SPJ低剂量组(10mg/kg)和高剂量组(30mg/kg)。Westernblot法检测大鼠皮层和海马内质网应激相关蛋白(GRP78、p-PERK、p-EIF2α、ATF4、ATF6、p-IRE1、XBP1)以及凋亡相关蛋白(p-JNK、CHOP、Bcl-2、Bax)表达变化。结果显示,与6月龄组相比,24月龄组皮层/海马组织中GRP78、p-PERK、p-EIF2α、ATF4、ATF6、p-IRE1、XBP1和Bcl-2/Bax蛋白表达分别下降了80%/59%、82%/78%、79%/77%、94%/74%、72%/66%、78%/81%、82%/71%和90%/92%,p-JNK/JNK、CHOP蛋白表达分别上升了1/1.72、3.33/1.33倍。给予不同剂量竹节参总皂苷干预后,均能明显上调内质网应激相关蛋白和Bcl-2/Bax的表达水平,下调促凋亡蛋白表达水平。研究结果表明,竹节参总皂苷能通过上调内质网应激相关蛋白以及抗凋亡蛋白的表达水平,下调促凋亡蛋白的表达,从而维持衰老大鼠脑组织的内质网稳态,减少神经细胞凋亡。
基金Research Foundation of Hubei Biological Enzyme Engineering Technology Research Center(JS2018-06)
文摘OBJECTIVE Gastric ulcers affect people of all ages and half of the world's population,which is being considered as the new"plague of the 21st century".As is well known,gastric mucosa is known as the first guard to protect the stomach from ulcer injury,while the aetiology of gastric ulcer is relative to imbalances between gastric mucosal protective and aggressive factors.Therefore,reducing or eliminating the aggressive factors,returning to the balance of between mucosal protective and aggressive factors,and then restoring the normal functional of gastric mucosal barrier could be crucial for treating the gastric ulcer.The fruits of Chaenomeles speciosa(TCS),also known as"mugua",might be processed into edible and health care derived products,and used as a commonly used traditional medicine in China for thousands of years.In China folk,there is a saying that"apricot one benefit,pear two benefits,mgua hundred benefits",so it has a"hundred-benefit"fruit reputation.Tujia nationality inhabitants in Southwestern China should have rheumatic diseases and peptic ulcers for living in the damp environments and bingeing on spicy and pungent foods.For the exis⁃tence of the fruit derived products of Chaenomeles speciosa as their complementary foods or snacks,the habit makes them rarely suffer from the two kinds of diseases.Enlightened by these,we had investigated the structure-activity rela⁃tionships,screened out CSTT with gastroprotective activity.Our previous studies demonstrated that TCS owned effec⁃tively therapeutic effects on gastric ulcer patients and animals,and further confirmed that TFF1 and apoptotic pathway were closely interrelated with its exerting gastroprotection.However,its underlying molecular mechanisms involved have not been fully elucidated.The current study was to further investigate its protective effect on indomethacin(IND)-damaged RGM-1 cells and rats and its underlying mechanisms through modulating TFF1-mediated EGF/EGFR and apoptotic pathways.METHODS The gastroprotection of TCS was evaluated with IND-induced gastric lesions model in RGM-1 cells and rats.In vitro,the proliferation,migration,mitochondrial viability and apoptosis were assessed,In vivo,ulcer index,ulcer inhibition rate,gastric juice acidity,gastric wall mucus(GWM),histopathology of gastric mucosa were detected.The gastroprotective effects of TCS through the TFF1-mediated EGFR/EGFR and apoptotic pathways were presented and measured by qRT-PCR and Western blotting assays.RESULTS TCS had gastroprotective function,which was related to the amelioration in promoting IND-damaged RGM-1 cell proliferation and migration,hoisting gastric juice acidity and GWM,improving ulcer index and ulcer inhibition rate,attenuating the hemorrhage,edema,epithelial cell loss and inflammatory cell infiltration of gastric mucosa,upregulating proliferation cell nuclear antigen,Bcl-2,Bcl-xl mRNA and TFF1,EGF,p-EGFR,p-Src,pro-caspase-3,pro-caspase-9 protein expressions,mitochondrial viability,mitochondrial cytochrome C concentration and p-EGFR/EGFR,p-Src/Src,Bcl-2/Bax,Bcl-xl/Bad ratioes,downregulating Bax,Bad,Apaf-1 mRNA and cleaved-caspase-3,cleaved-caspase-9,cleaved-PARP-1 protein expressions,cytosol cytochrome C concentration.CONCLUSION TCS′s gastroprotective effect was closely connected with boosting TFF1 expression,acti⁃vating TFF1-mediated EGF/EGFR pathway,thus restraining mitochondrial-dependent apoptosis,which provided new insights into interpreting its underlying mechanism and promised to act as a candidate drug to treat gastric mucosal injury.
