Objective: The study aimed to explore the protective mechanism of Ganoderic acid A (GAA) in renal fibrosisand to verify that GAA can ameliorate renal fibrosis by regulating the Niemann-pick C1-like 1 (NPC1L1) gene. Meth...Objective: The study aimed to explore the protective mechanism of Ganoderic acid A (GAA) in renal fibrosisand to verify that GAA can ameliorate renal fibrosis by regulating the Niemann-pick C1-like 1 (NPC1L1) gene. Methods:Transforming growth factor beta1 (TGF-β1) was used to treat Human Kidney-2 (HK-2) cells to establish a renal fibrosismodel. The differentially expressed genes in the control (CTRL) group, TGF-β1 group, and TGF-β1 + GAA group werescreened via transcriptome sequencing technology and verified by qPCR and Western blot experiments. The NPC1L1gene overexpression plasmid was constructed. The expression levels of N-cad, E-cad, and Slug-related proteins inCTRL, TGF-β1, TGF-β1+GAA (25 μg/mL), and TGF-β1+GAA (25 μg/mL) + NPC1L1 Overexpression (OE) groupswere detected by qPCR and Western blot analysis. Western blot analysis was used to identify the extracellular matrixassociated proteins Tenascin-C, α-SMA, and fibrosis-related protein Collagen I. Fibrosis marker protein Fibronectinwas detected and quantified by immunofluorescence. Results: Transcriptomic sequencing revealed that TGF-β1stimulation led to 267 differentially regulated genes, with 118 up-regulated and 149 down-regulated, while furthermodulation of 213 genes, comprising 112 up-regulated and 101 down-regulated genes, was observed in the GAAintervention group. The target gene in these processes was found to be NPC1L1 by investigations using GeneOntology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG). qPCR and Western blot resultsconfirmed that TGF-β1 increased NPC1L1 expression, which was attenuated by GAA. Additionally, TGF-β1upregulated N-cad and Slug. However, GAA reversed this effect and NPC1L1 overexpression partially rescued theGAA effect. TGF-β1 also decreased E-cad expression, reversed by GAA, and NPC1L1 overexpression antagonized thisreversal. Furthermore, TGF-β1 promoted Collagen I, α-SMA, and Tenascin-C expression, and GAA reduced theselevels, effects that were reversed by NPC1L1 overexpression. Immunofluorescence results showed that TGF-β1increased fibronectin expression, which was decreased by GAA, and increased by NPC1L1 overexpression.Conclusion: GAA ameliorates renal fibrosis by antagonizing NPC1L1 gene expression inhibiting epithelialmesenchymal transition and reducing extracellular matrix formation.展开更多
Niemann-Pick C1 Like 1(NPC1L1)是肠道胆固醇吸收的关键蛋白,也是胆固醇吸收抑制剂依泽替米贝的分子作用靶点。NPC1L1的表达受一些核因子受体调控。敲除apoE-/-小鼠NPC1L1基因能显著抑制动脉粥样硬化的发生和发展,为动脉粥样硬化和冠...Niemann-Pick C1 Like 1(NPC1L1)是肠道胆固醇吸收的关键蛋白,也是胆固醇吸收抑制剂依泽替米贝的分子作用靶点。NPC1L1的表达受一些核因子受体调控。敲除apoE-/-小鼠NPC1L1基因能显著抑制动脉粥样硬化的发生和发展,为动脉粥样硬化和冠状动脉性心脏病提供了新的治疗靶点。展开更多
研究黑米对高脂高胆固醇膳食饲喂雌性小鼠血脂水平及小肠胆固醇代谢相关基因调控的影响。将48只雌性C57BL/6J小鼠随机分成高脂对照组和三个实验组(高脂饲料中添加白米和黑米代替玉米淀粉),测定血清中总胆固醇(TC)、甘油三酯(TG)及高密...研究黑米对高脂高胆固醇膳食饲喂雌性小鼠血脂水平及小肠胆固醇代谢相关基因调控的影响。将48只雌性C57BL/6J小鼠随机分成高脂对照组和三个实验组(高脂饲料中添加白米和黑米代替玉米淀粉),测定血清中总胆固醇(TC)、甘油三酯(TG)及高密度脂蛋白胆固醇(HDL-C)水平,气相色谱法检测肝脏中胆固醇含量,Real-time PCR法分析调控小肠胆固醇代谢相关基因HMG-Co A-R、MTP、ABCG5/8、ABCA1、NPC1L1等的m RNA表达水平。与高脂组相比,黑米组血清中TC和TG含量较对照组极显著降低,但白米组血清中TC、TG、HDL-C含量较对照组无统计学差异。与高脂组相比,白米组小鼠肝脏中胆固醇含量无显著变化,黑米低剂量组显著降低(P<0.05),黑米高剂量组极显著降低(P<0.01)。Real-time PCR结果显示,与高脂组相比,黑米添加组ABCG5、ABCA1 m RNA表达水平均显著上调(P<0.01),MTP m RNA表达水平均显著下调(P<0.05);黑米低剂量组ABCG8 m RNA表达水平显著升高(P<0.05),高剂量组上调极显著(P<0.01);黑米低剂量组NPC1L1 m RNA表达水平降低(P<0.05),高剂量组下调极显著(P<0.01);黑米高剂量组HMG-Co A-Rm RNA表达水平显著降低(P<0.05)。结果表明,黑米对高脂高胆固醇膳食饲喂小鼠胆固醇代谢平衡的调节,可能是通过增加小肠中胆固醇排泄并抑制固醇的吸收实现的。展开更多
β-Sitosterol is a type of phytosterol that occurs naturally in plants.Previous studies have shown that it has anti-oxidant,anti-hyperlipidemic,anti-inflammatory,immunomodulatory,and anti-tumor effects,but it is unkno...β-Sitosterol is a type of phytosterol that occurs naturally in plants.Previous studies have shown that it has anti-oxidant,anti-hyperlipidemic,anti-inflammatory,immunomodulatory,and anti-tumor effects,but it is unknown whetherβ-sitosterol treatment reduces the effects of ischemic stroke.Here we found that,in a mouse model of ischemic stroke induced by middle cerebral artery occlusion,β-sitosterol reduced the volume of cerebral infarction and brain edema,reduced neuronal apoptosis in brain tissue,and alleviated neurological dysfunction;moreover,β-sitosterol increased the activity of oxygen-and glucose-deprived cerebral cortex neurons and reduced apoptosis.Further investigation showed that the neuroprotective effects ofβ-sitosterol may be related to inhibition of endoplasmic reticulum stress caused by intracellular cholesterol accumulation after ischemic stroke.In addition,β-sitosterol showed high affinity for NPC1L1,a key transporter of cholesterol,and antagonized its activity.In conclusion,β-sitosterol may help treat ischemic stroke by inhibiting neuronal intracellular cholesterol overload/endoplasmic reticulum stress/apoptosis signaling pathways.展开更多
A new isochromanone,cladosporinisochromanone(1),accompanied by 15 known compounds(2–16)were obtained from secondary metabolites produced by marine-derived fungus Cladosporium sp.DLT-5.NMR and HRESIMS spectra elucidat...A new isochromanone,cladosporinisochromanone(1),accompanied by 15 known compounds(2–16)were obtained from secondary metabolites produced by marine-derived fungus Cladosporium sp.DLT-5.NMR and HRESIMS spectra elucidation determined the planar structure of 1.Subsequent electronic circular dichroism(ECD)experiment assigned the absolute configuration of 1.Compounds 1,2,4–6,and 10 displayed different degrees of neuroprotective activities on human neuroblastoma cells SH-SY5Y.Five compounds(1,3–5,and 13)emerged resistance to protein tyrosine phosphatase 1B(PTP1B),further kinetic analysis and molecular docking study indicated that the most potent compound 13(IC50value of 10.74±0.61μmol/L)was found as a noncompetitive inhibitor for PTP1B.Surface plasmon resonance(SPR)and molecular docking studies also demonstrated the interaction between compound 12 and Niemann-Pick C1 Like 1(NPC1L1),which has been identified as significant therapeutic target for hypercholesteremia.In addition,compounds 3,6,and 14 showed attractive inhibitory activity against the phytopathogenic fungi:Colletotrichum capsici.Therefore,library of Cladosporium metabolites is enriched and new active uses of known compounds are explored.展开更多
基金sponsored by KeyResearch and Development Project of Science andTechnology Department of Tibet (No. XZ202201ZY0033G).
文摘Objective: The study aimed to explore the protective mechanism of Ganoderic acid A (GAA) in renal fibrosisand to verify that GAA can ameliorate renal fibrosis by regulating the Niemann-pick C1-like 1 (NPC1L1) gene. Methods:Transforming growth factor beta1 (TGF-β1) was used to treat Human Kidney-2 (HK-2) cells to establish a renal fibrosismodel. The differentially expressed genes in the control (CTRL) group, TGF-β1 group, and TGF-β1 + GAA group werescreened via transcriptome sequencing technology and verified by qPCR and Western blot experiments. The NPC1L1gene overexpression plasmid was constructed. The expression levels of N-cad, E-cad, and Slug-related proteins inCTRL, TGF-β1, TGF-β1+GAA (25 μg/mL), and TGF-β1+GAA (25 μg/mL) + NPC1L1 Overexpression (OE) groupswere detected by qPCR and Western blot analysis. Western blot analysis was used to identify the extracellular matrixassociated proteins Tenascin-C, α-SMA, and fibrosis-related protein Collagen I. Fibrosis marker protein Fibronectinwas detected and quantified by immunofluorescence. Results: Transcriptomic sequencing revealed that TGF-β1stimulation led to 267 differentially regulated genes, with 118 up-regulated and 149 down-regulated, while furthermodulation of 213 genes, comprising 112 up-regulated and 101 down-regulated genes, was observed in the GAAintervention group. The target gene in these processes was found to be NPC1L1 by investigations using GeneOntology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG). qPCR and Western blot resultsconfirmed that TGF-β1 increased NPC1L1 expression, which was attenuated by GAA. Additionally, TGF-β1upregulated N-cad and Slug. However, GAA reversed this effect and NPC1L1 overexpression partially rescued theGAA effect. TGF-β1 also decreased E-cad expression, reversed by GAA, and NPC1L1 overexpression antagonized thisreversal. Furthermore, TGF-β1 promoted Collagen I, α-SMA, and Tenascin-C expression, and GAA reduced theselevels, effects that were reversed by NPC1L1 overexpression. Immunofluorescence results showed that TGF-β1increased fibronectin expression, which was decreased by GAA, and increased by NPC1L1 overexpression.Conclusion: GAA ameliorates renal fibrosis by antagonizing NPC1L1 gene expression inhibiting epithelialmesenchymal transition and reducing extracellular matrix formation.
文摘Niemann-Pick C1 Like 1(NPC1L1)是肠道胆固醇吸收的关键蛋白,也是胆固醇吸收抑制剂依泽替米贝的分子作用靶点。NPC1L1的表达受一些核因子受体调控。敲除apoE-/-小鼠NPC1L1基因能显著抑制动脉粥样硬化的发生和发展,为动脉粥样硬化和冠状动脉性心脏病提供了新的治疗靶点。
文摘研究黑米对高脂高胆固醇膳食饲喂雌性小鼠血脂水平及小肠胆固醇代谢相关基因调控的影响。将48只雌性C57BL/6J小鼠随机分成高脂对照组和三个实验组(高脂饲料中添加白米和黑米代替玉米淀粉),测定血清中总胆固醇(TC)、甘油三酯(TG)及高密度脂蛋白胆固醇(HDL-C)水平,气相色谱法检测肝脏中胆固醇含量,Real-time PCR法分析调控小肠胆固醇代谢相关基因HMG-Co A-R、MTP、ABCG5/8、ABCA1、NPC1L1等的m RNA表达水平。与高脂组相比,黑米组血清中TC和TG含量较对照组极显著降低,但白米组血清中TC、TG、HDL-C含量较对照组无统计学差异。与高脂组相比,白米组小鼠肝脏中胆固醇含量无显著变化,黑米低剂量组显著降低(P<0.05),黑米高剂量组极显著降低(P<0.01)。Real-time PCR结果显示,与高脂组相比,黑米添加组ABCG5、ABCA1 m RNA表达水平均显著上调(P<0.01),MTP m RNA表达水平均显著下调(P<0.05);黑米低剂量组ABCG8 m RNA表达水平显著升高(P<0.05),高剂量组上调极显著(P<0.01);黑米低剂量组NPC1L1 m RNA表达水平降低(P<0.05),高剂量组下调极显著(P<0.01);黑米高剂量组HMG-Co A-Rm RNA表达水平显著降低(P<0.05)。结果表明,黑米对高脂高胆固醇膳食饲喂小鼠胆固醇代谢平衡的调节,可能是通过增加小肠中胆固醇排泄并抑制固醇的吸收实现的。
基金supported by the National Natural Science Foundation of China,Nos.82104158(to XT),31800887(to LY),31972902(to LY),82001422(to YL)China Postdoctoral Science Foundation,No.2020M683750(to LY)partially by Young Talent Fund of University Association for Science and Technology in Shaanxi Province of China,No.20200307(to LY).
文摘β-Sitosterol is a type of phytosterol that occurs naturally in plants.Previous studies have shown that it has anti-oxidant,anti-hyperlipidemic,anti-inflammatory,immunomodulatory,and anti-tumor effects,but it is unknown whetherβ-sitosterol treatment reduces the effects of ischemic stroke.Here we found that,in a mouse model of ischemic stroke induced by middle cerebral artery occlusion,β-sitosterol reduced the volume of cerebral infarction and brain edema,reduced neuronal apoptosis in brain tissue,and alleviated neurological dysfunction;moreover,β-sitosterol increased the activity of oxygen-and glucose-deprived cerebral cortex neurons and reduced apoptosis.Further investigation showed that the neuroprotective effects ofβ-sitosterol may be related to inhibition of endoplasmic reticulum stress caused by intracellular cholesterol accumulation after ischemic stroke.In addition,β-sitosterol showed high affinity for NPC1L1,a key transporter of cholesterol,and antagonized its activity.In conclusion,β-sitosterol may help treat ischemic stroke by inhibiting neuronal intracellular cholesterol overload/endoplasmic reticulum stress/apoptosis signaling pathways.
基金Supported by the China Agriculture Research System of MOF and MARA(CARS-21)the Financial Fund of the Ministry of Agriculture and Rural Affairs,China(No.NFZX2021)the National Natural Science Foundation of China(No.81973568)。
文摘A new isochromanone,cladosporinisochromanone(1),accompanied by 15 known compounds(2–16)were obtained from secondary metabolites produced by marine-derived fungus Cladosporium sp.DLT-5.NMR and HRESIMS spectra elucidation determined the planar structure of 1.Subsequent electronic circular dichroism(ECD)experiment assigned the absolute configuration of 1.Compounds 1,2,4–6,and 10 displayed different degrees of neuroprotective activities on human neuroblastoma cells SH-SY5Y.Five compounds(1,3–5,and 13)emerged resistance to protein tyrosine phosphatase 1B(PTP1B),further kinetic analysis and molecular docking study indicated that the most potent compound 13(IC50value of 10.74±0.61μmol/L)was found as a noncompetitive inhibitor for PTP1B.Surface plasmon resonance(SPR)and molecular docking studies also demonstrated the interaction between compound 12 and Niemann-Pick C1 Like 1(NPC1L1),which has been identified as significant therapeutic target for hypercholesteremia.In addition,compounds 3,6,and 14 showed attractive inhibitory activity against the phytopathogenic fungi:Colletotrichum capsici.Therefore,library of Cladosporium metabolites is enriched and new active uses of known compounds are explored.
