BACKGROUND Chronic biliary obstruction results in ischemia and hypoxia of hepatocytes,and leads to apoptosis.Apoptosis is very important in regulating the homeostasis of the hepatobiliary system.Endoplasmic reticulum(...BACKGROUND Chronic biliary obstruction results in ischemia and hypoxia of hepatocytes,and leads to apoptosis.Apoptosis is very important in regulating the homeostasis of the hepatobiliary system.Endoplasmic reticulum(ER)stress is one of the signaling pathways that induce apoptosis.Moreover,the protein kinase RNA-like endoplasmic reticulum kinase(PERK)-induced apoptotic pathway is the main way;but its role in liver injury remains unclear.Yinchenhao decoction(YCHD)is a traditional Chinese medicine formula that alleviates liver injury and apoptosis,yet its mechanism is unknown.We undertook this study to investigate the effects of YCHD on the expression of ER stress proteins and hepatocyte apoptosis in rats with obstructive jaundice(OJ).AIM To investigate whether YCHD can attenuate OJ-induced liver injury and hepatocyte apoptosis by inhibiting the PERK-CCAAT/enhancer-binding protein homologous protein(CHOP)-growth arrest and DNA damage-inducible protein 34(GADD34)pathway and B cell lymphoma/leukemia-2 related X protein(Bax)/B cell lymphoma/leukemia-2(Bcl-2)ratio.METHODS For in vivo experiments,30 rats were divided into three groups:control group,OJ model group,and YCHD-treated group.Blood was collected to detect the indicators of liver function,and liver tissues were used for histological analysis.For in vitro experiments,30 rats were divided into three groups:G1,G2,and G3.The rats in group G1 had their bile duct exposed without ligation,the rats in group G2 underwent total bile duct ligation,and the rats in group G3 were given a gavage of YCHD.According to the serum pharmacology,serum was extracted and centrifuged from the rat blood to cultivate the BRL-3A cells.Terminal deoxynucleotidyl transferase mediated dUTP nick end-labelling(TUNEL)assay was used to detect BRL-3A hepatocyte apoptosis.Alanine aminotransferase(ALT)and aspartate transaminase(AST)levels in the medium were detected.Western blot and quantitative real-time polymerase chain reaction(qRT-PCR)analyses were used to detect protein and gene expression leve展开更多
Objective: To test the hypothesis that the inhibition of endoplasmic reticulum(ER) stress-induced apoptosis in oxidized low-density lipoproteins(ox-LDL)-induced human aortic-vascular smooth muscle cells(HA-VSMCs) was ...Objective: To test the hypothesis that the inhibition of endoplasmic reticulum(ER) stress-induced apoptosis in oxidized low-density lipoproteins(ox-LDL)-induced human aortic-vascular smooth muscle cells(HA-VSMCs) was associated with suppression of the protein kinase RNA-like ER kinase(PERK)-eukaryotic translation initiation factor 2α(e IF2α)-activating transcription factor 4(ATF4)-CCAAT/enhancer binding protein homologous protein(CHOP) signaling pathway by Pollen Typhae total flavone(PTF). Methods: Primary HA-VSMCs were cultured and identified. The cultured HA-VSMCs were randomized into 5 groups, including a normal control group, an ox-LDL group(70 μg/m L high ox-LDL), an HPTF group(70 μg/m L high ox-LDL+500 μg/m L PTF), an MPTF group(70 μg/m L high ox-LDL+250 μg/m L PTF), and a LPTF group(70 μg/m L high ox-LDL+100 μg/m L PTF) in the first part;and a normal control group, an ox-LDL group(70 μg/mL high ox-LDL), an MPTF group(70 μg/m L high ox-LDL+250 μg/m L PTF), a sh RNA group(transducted with PERK shRNA lentiviral particles), a scramble shRNA group(transducted with control shRNA lentiviral particles), an MPTF+ox-LDL+shRNA group(250 μg/mL PTF+70 μg/mL high ox-LDL+PERK shRNA lentiviral particles) and an ox-LDL+shRNA group(70 μg/mL high ox-LDL+PERK shRNA lentiviral particles) in the second part. The protein expression levels of ER-associated apoptosis proteins were detected by Western blot, and their m RNA expression levels were detected by quantitative real-time reverse transcription-polymerase chain reaction. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay was applied to test cel viability, and the level of apoptosis was monitored by flow cytometry. Results: The MTT assay and flow cytometry showed that the ox-LDL group had a significant increase in apoptosis, which was attenuated in PTF treatment groups and sh RNA groups. Moreover, the ox-LDL group had increased protein and m RNA levels of binding immunoglobulin protein and ER-associated apoptosis proteins, such as PERK,展开更多
In the process of tumor proliferation and metastasis,tumor cells encounter hypoxia,low glucose,acidosis,and other stressful environments.These conditions prompt tumor cells to generate endoplasmic reticulum stress(ERS...In the process of tumor proliferation and metastasis,tumor cells encounter hypoxia,low glucose,acidosis,and other stressful environments.These conditions prompt tumor cells to generate endoplasmic reticulum stress(ERS).As a signal mechanism that mitigates ERS in eukaryotic cells,the unfolded protein response(UPR)pathway can activate cells and tissues,regulating pathological activities in various cells,and maintaining ER homeostasis.It forms the most crucial adaptive and defensive mechanism for cells.However,under the continuous influence of chemotherapy drugs,the quantity of unfolded proteins and erroneous proteins produced by tumor cells significantly increases,surpassing the normal regulatory range of UPR.Consequently,ERS fails to function properly,fostering tumor cell proliferation and the development of drug resistance.This review delves into the study of three UPR pathways(PERK,IRE1,and ATF6),elucidating the mechanisms of drug resistance and research progress in the signal transduction pathway of UPR related to cancers.It provides a profound understanding of the role and relationship between UPR and anti-tumor drugs,offering a new direction for effective clinical treatment.展开更多
【目的】探讨四逆汤对阿霉素诱导的慢性心力衰竭(CHF)大鼠内质网应激的影响。【方法】从90只大鼠中随机选取15只作为正常组,其余大鼠腹腔注射阿霉素构建CHF模型,同时,正常组给予腹腔注射生理盐水。造模结束后,正常组大鼠全部存活,造模...【目的】探讨四逆汤对阿霉素诱导的慢性心力衰竭(CHF)大鼠内质网应激的影响。【方法】从90只大鼠中随机选取15只作为正常组,其余大鼠腹腔注射阿霉素构建CHF模型,同时,正常组给予腹腔注射生理盐水。造模结束后,正常组大鼠全部存活,造模大鼠存活60只。再将60只CHF大鼠随机分为5组,即模型组,四逆汤低、中、高剂量组,曲美他嗪组,每组12只。四逆汤低、中、高剂量组分别给予1.4、4.2、12.6(生药)g·kg^(-1)·d^(-1)灌胃,曲美他嗪组给予曲美他嗪10 mg·kg^(-1)·d^(-1)灌胃,正常组和模型组给予相同体积的生理盐水灌胃。连续给药4周。采用小动物超声测量左心室舒张末期直径(LVEDD)、左心室射血分数(LVEF)和左心室缩短分数(LVFS)变化;采用酶联免疫吸附分析(ELISA)检测血浆脑钠肽(BNP)和血管紧张素Ⅱ(AngⅡ)水平;采用实时荧光定量聚合酶链反应(PCR)技术检测心肌组织葡萄糖调节蛋白78(GRP78)、RNA依赖的蛋白激酶样内质网激酶(PERK)和活化转录因子4(ATF4)mRNA表达。【结果】与正常组比较,模型组大鼠LVEDD显著增大,LVEF、LVFS均显著降低,血浆BNP和AngⅡ含量升高,心肌组织GRP78、PERK及ATF4 m RNA表达量升高(P <0.01);与模型组比较,四逆汤低、中、高剂量组及曲美他嗪组LVEDD降低,LVEF、LVFS均显著增加,血浆BNP和AngⅡ含量减少,心肌组织GRP78、PERK及ATF4 m RNA表达量降低,且呈剂量依赖性,其中,四逆汤高剂量组及曲美他嗪组差异均有统计学意义(P <0.05或P <0.01)。【结论】四逆汤可改善慢性心力衰竭大鼠模型心脏功能,其机制可能与抑制PERK/ATF4信号通路从而减轻内质网应激有关。展开更多
目的探讨血清外泌体源性环状RNA Tousled样激酶1(circTLK1)与老年急性脑梗死病情严重程度及预后的关系。方法选取2021年10月至2022年11月恩施土家族苗族自治州中心医院脑病科收治的老年急性脑梗死患者128例(急性脑梗死组),采用美国国立...目的探讨血清外泌体源性环状RNA Tousled样激酶1(circTLK1)与老年急性脑梗死病情严重程度及预后的关系。方法选取2021年10月至2022年11月恩施土家族苗族自治州中心医院脑病科收治的老年急性脑梗死患者128例(急性脑梗死组),采用美国国立卫生研究院卒中量表(NIHSS)评分评估病情严重程度。急性脑梗死组90 d时又根据改良的Rankin量表(mRS)评分分为预后不良组46例和预后良好组82例。另选取健康体检者80例(对照组)。RT-PCR检测血清外泌体源性circTLK1表达。出院后对老年急性脑梗死患者进行随访,90 d时用mRS评分对预后进行评价。结果急性脑梗死组血清外泌体源性circTLK1表达明显高于对照组(1.86±0.46 vs 0.87±0.12,P<0.05)。轻度、中度和重度急性脑梗死患者血清外泌体源性circTLK1表达逐渐升高(1.34±0.28 vs 1.78±0.19 vs 2.35±0.33,P<0.05)。预后不良组入院时NIHSS评分和circTLK1表达明显高于预后良好组[(13.21±3.24)分vs(9.55±2.16)分,P=0.001;2.15±0.77 vs 1.47±0.58,P=0.001]。多因素logistic回归分析显示,入院时NIHSS评分和circTLK1表达是老年急性脑梗死患者不良预后的独立影响因素(OR=4.123,95%CI:1.109~12.520,P=0.001;OR=2.734,95%CI:1.067~6.135,P=0.001)。ROC曲线分析显示,血清外泌体源性circTLK1表达预测急性脑梗死患者预后的曲线下面积为0.831,敏感性和特异性分别为96.19%、85.20%。结论老年急性脑梗死患者血清外泌体源性circTLK1表达升高,其表达越高,病情可能越重,且预后不良。展开更多
基金Supported by the National Natural Science Foundation of China,No.81273952
文摘BACKGROUND Chronic biliary obstruction results in ischemia and hypoxia of hepatocytes,and leads to apoptosis.Apoptosis is very important in regulating the homeostasis of the hepatobiliary system.Endoplasmic reticulum(ER)stress is one of the signaling pathways that induce apoptosis.Moreover,the protein kinase RNA-like endoplasmic reticulum kinase(PERK)-induced apoptotic pathway is the main way;but its role in liver injury remains unclear.Yinchenhao decoction(YCHD)is a traditional Chinese medicine formula that alleviates liver injury and apoptosis,yet its mechanism is unknown.We undertook this study to investigate the effects of YCHD on the expression of ER stress proteins and hepatocyte apoptosis in rats with obstructive jaundice(OJ).AIM To investigate whether YCHD can attenuate OJ-induced liver injury and hepatocyte apoptosis by inhibiting the PERK-CCAAT/enhancer-binding protein homologous protein(CHOP)-growth arrest and DNA damage-inducible protein 34(GADD34)pathway and B cell lymphoma/leukemia-2 related X protein(Bax)/B cell lymphoma/leukemia-2(Bcl-2)ratio.METHODS For in vivo experiments,30 rats were divided into three groups:control group,OJ model group,and YCHD-treated group.Blood was collected to detect the indicators of liver function,and liver tissues were used for histological analysis.For in vitro experiments,30 rats were divided into three groups:G1,G2,and G3.The rats in group G1 had their bile duct exposed without ligation,the rats in group G2 underwent total bile duct ligation,and the rats in group G3 were given a gavage of YCHD.According to the serum pharmacology,serum was extracted and centrifuged from the rat blood to cultivate the BRL-3A cells.Terminal deoxynucleotidyl transferase mediated dUTP nick end-labelling(TUNEL)assay was used to detect BRL-3A hepatocyte apoptosis.Alanine aminotransferase(ALT)and aspartate transaminase(AST)levels in the medium were detected.Western blot and quantitative real-time polymerase chain reaction(qRT-PCR)analyses were used to detect protein and gene expression leve
基金Supported by the National Natural Science Foundation of China(No.81573922)the Traditional Chinese Medicine Scientific Research Project of Guangdong Province,China(No.20151076)the Sanming Project of Medicine in Shenzhen,China(No.SZSM201612033)
文摘Objective: To test the hypothesis that the inhibition of endoplasmic reticulum(ER) stress-induced apoptosis in oxidized low-density lipoproteins(ox-LDL)-induced human aortic-vascular smooth muscle cells(HA-VSMCs) was associated with suppression of the protein kinase RNA-like ER kinase(PERK)-eukaryotic translation initiation factor 2α(e IF2α)-activating transcription factor 4(ATF4)-CCAAT/enhancer binding protein homologous protein(CHOP) signaling pathway by Pollen Typhae total flavone(PTF). Methods: Primary HA-VSMCs were cultured and identified. The cultured HA-VSMCs were randomized into 5 groups, including a normal control group, an ox-LDL group(70 μg/m L high ox-LDL), an HPTF group(70 μg/m L high ox-LDL+500 μg/m L PTF), an MPTF group(70 μg/m L high ox-LDL+250 μg/m L PTF), and a LPTF group(70 μg/m L high ox-LDL+100 μg/m L PTF) in the first part;and a normal control group, an ox-LDL group(70 μg/mL high ox-LDL), an MPTF group(70 μg/m L high ox-LDL+250 μg/m L PTF), a sh RNA group(transducted with PERK shRNA lentiviral particles), a scramble shRNA group(transducted with control shRNA lentiviral particles), an MPTF+ox-LDL+shRNA group(250 μg/mL PTF+70 μg/mL high ox-LDL+PERK shRNA lentiviral particles) and an ox-LDL+shRNA group(70 μg/mL high ox-LDL+PERK shRNA lentiviral particles) in the second part. The protein expression levels of ER-associated apoptosis proteins were detected by Western blot, and their m RNA expression levels were detected by quantitative real-time reverse transcription-polymerase chain reaction. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay was applied to test cel viability, and the level of apoptosis was monitored by flow cytometry. Results: The MTT assay and flow cytometry showed that the ox-LDL group had a significant increase in apoptosis, which was attenuated in PTF treatment groups and sh RNA groups. Moreover, the ox-LDL group had increased protein and m RNA levels of binding immunoglobulin protein and ER-associated apoptosis proteins, such as PERK,
文摘In the process of tumor proliferation and metastasis,tumor cells encounter hypoxia,low glucose,acidosis,and other stressful environments.These conditions prompt tumor cells to generate endoplasmic reticulum stress(ERS).As a signal mechanism that mitigates ERS in eukaryotic cells,the unfolded protein response(UPR)pathway can activate cells and tissues,regulating pathological activities in various cells,and maintaining ER homeostasis.It forms the most crucial adaptive and defensive mechanism for cells.However,under the continuous influence of chemotherapy drugs,the quantity of unfolded proteins and erroneous proteins produced by tumor cells significantly increases,surpassing the normal regulatory range of UPR.Consequently,ERS fails to function properly,fostering tumor cell proliferation and the development of drug resistance.This review delves into the study of three UPR pathways(PERK,IRE1,and ATF6),elucidating the mechanisms of drug resistance and research progress in the signal transduction pathway of UPR related to cancers.It provides a profound understanding of the role and relationship between UPR and anti-tumor drugs,offering a new direction for effective clinical treatment.
文摘【目的】探讨四逆汤对阿霉素诱导的慢性心力衰竭(CHF)大鼠内质网应激的影响。【方法】从90只大鼠中随机选取15只作为正常组,其余大鼠腹腔注射阿霉素构建CHF模型,同时,正常组给予腹腔注射生理盐水。造模结束后,正常组大鼠全部存活,造模大鼠存活60只。再将60只CHF大鼠随机分为5组,即模型组,四逆汤低、中、高剂量组,曲美他嗪组,每组12只。四逆汤低、中、高剂量组分别给予1.4、4.2、12.6(生药)g·kg^(-1)·d^(-1)灌胃,曲美他嗪组给予曲美他嗪10 mg·kg^(-1)·d^(-1)灌胃,正常组和模型组给予相同体积的生理盐水灌胃。连续给药4周。采用小动物超声测量左心室舒张末期直径(LVEDD)、左心室射血分数(LVEF)和左心室缩短分数(LVFS)变化;采用酶联免疫吸附分析(ELISA)检测血浆脑钠肽(BNP)和血管紧张素Ⅱ(AngⅡ)水平;采用实时荧光定量聚合酶链反应(PCR)技术检测心肌组织葡萄糖调节蛋白78(GRP78)、RNA依赖的蛋白激酶样内质网激酶(PERK)和活化转录因子4(ATF4)mRNA表达。【结果】与正常组比较,模型组大鼠LVEDD显著增大,LVEF、LVFS均显著降低,血浆BNP和AngⅡ含量升高,心肌组织GRP78、PERK及ATF4 m RNA表达量升高(P <0.01);与模型组比较,四逆汤低、中、高剂量组及曲美他嗪组LVEDD降低,LVEF、LVFS均显著增加,血浆BNP和AngⅡ含量减少,心肌组织GRP78、PERK及ATF4 m RNA表达量降低,且呈剂量依赖性,其中,四逆汤高剂量组及曲美他嗪组差异均有统计学意义(P <0.05或P <0.01)。【结论】四逆汤可改善慢性心力衰竭大鼠模型心脏功能,其机制可能与抑制PERK/ATF4信号通路从而减轻内质网应激有关。
文摘目的探讨血清外泌体源性环状RNA Tousled样激酶1(circTLK1)与老年急性脑梗死病情严重程度及预后的关系。方法选取2021年10月至2022年11月恩施土家族苗族自治州中心医院脑病科收治的老年急性脑梗死患者128例(急性脑梗死组),采用美国国立卫生研究院卒中量表(NIHSS)评分评估病情严重程度。急性脑梗死组90 d时又根据改良的Rankin量表(mRS)评分分为预后不良组46例和预后良好组82例。另选取健康体检者80例(对照组)。RT-PCR检测血清外泌体源性circTLK1表达。出院后对老年急性脑梗死患者进行随访,90 d时用mRS评分对预后进行评价。结果急性脑梗死组血清外泌体源性circTLK1表达明显高于对照组(1.86±0.46 vs 0.87±0.12,P<0.05)。轻度、中度和重度急性脑梗死患者血清外泌体源性circTLK1表达逐渐升高(1.34±0.28 vs 1.78±0.19 vs 2.35±0.33,P<0.05)。预后不良组入院时NIHSS评分和circTLK1表达明显高于预后良好组[(13.21±3.24)分vs(9.55±2.16)分,P=0.001;2.15±0.77 vs 1.47±0.58,P=0.001]。多因素logistic回归分析显示,入院时NIHSS评分和circTLK1表达是老年急性脑梗死患者不良预后的独立影响因素(OR=4.123,95%CI:1.109~12.520,P=0.001;OR=2.734,95%CI:1.067~6.135,P=0.001)。ROC曲线分析显示,血清外泌体源性circTLK1表达预测急性脑梗死患者预后的曲线下面积为0.831,敏感性和特异性分别为96.19%、85.20%。结论老年急性脑梗死患者血清外泌体源性circTLK1表达升高,其表达越高,病情可能越重,且预后不良。
