Background:Green tea,mulberry leaf and corn silk are traditional herbs used in the prevention and treatment of diabetes in China for a long time,but their synergistic hypoglycemic effects and mechanisms remain unclear...Background:Green tea,mulberry leaf and corn silk are traditional herbs used in the prevention and treatment of diabetes in China for a long time,but their synergistic hypoglycemic effects and mechanisms remain unclear.Methods:The effective components of green tea,mulberry leaf and corn silk were extracted and enriched.Mixture design of experiments was used to study the influences of different combinations on the cell viability and glucose uptake level of L6 myoblasts,so as to determine the optimal synergistic hypoglycemic combination.The possible hypoglycemic mechanism of the optimal synergistic combination was explored by cytotoxicity assay,glucose uptake assay,and western blot.Results:Three polyphenol enrichment fractions of the herbs,30%ethanol elution fraction of green tea(GT),50%ethanol elution fraction of mulberry leaf(ML)and 60%ethanol elution fraction of corn silk(CS)were obtained.The antioxidant activities of GT-30%,ML-50%and CS-60%were superior to those of crude extracts,and showed strong potential inα-amylase andα-glucosidase inhibition activities.The optimal synergistic combination of crude extracts G7(crude extract of green tea:crude extract of mulberry leaf:crude extract of corn silk=1:5:3),polyphenol enrichment fractions R3(GT-30%:ML-50%:CS-60%=1:7:1)and monomers X2(epigallocatechin gallate:morusin:formononetin=3:1:2)were selected,respectively.G7,R3,and X2 showed promoting effects on the cell viability and glucose uptake of L6 myoblasts within the detected concentration range.In addition,G7,R3,and X2 could increase the expression levels of p-PI3K/PI3K and p-Akt/Akt in L6 myoblasts,and promote the translocation of Glut4,but G7 and R3 showed more significant effects.Conclusion:The synergistic hypoglycemic effects of green tea,mulberry leaf and corn silk had the characteristics of multiple-components and multiple-targets with p-PI3K/PI3K,p-Akt/Akt and the translocation of Glut4 signal pathways involved.The three traditional herbs might have the potential to be combined used for the prevention and treatment of diabetes based on the synergistic hypoglycemic effects.展开更多
[Objective]This paper was to investigate the action targets and pathways of tea polyphenols in alleviating heat stress-induced injury by using network pharmacological analysis and an H9C2 cell model.[Method]First,the ...[Objective]This paper was to investigate the action targets and pathways of tea polyphenols in alleviating heat stress-induced injury by using network pharmacological analysis and an H9C2 cell model.[Method]First,the corresponding targets of tea polyphenols were obtained from the PubChem database.Then,the core targets were screened based on topological parameters.The relevant metabolism pathways of tea polyphenols related to diseases were identified through GO functional annotation and KECG signaling pathway enrichment.Moreover,common targets for thermal injury and targets of tea polyphenols were obtained.Then,GO functional annotation was performed to explore the pathway of tea polyphenols in alleviating heat stress damage.H9C2 cells were cultured at 42℃ to construct the heat stress model,and the cells were treated with 10μg/mL tea polyphenols.The key genes were confirmed using RT-PCR technology.[Result]The study yielded 364 targets corresponding to tea polyphenols,including 68 core targets.These targets are related to various biological processes such as involve oxidative stress,cancer,lipopolysaccharide-mediated signaling pathways,antiviral responses,regulation of cellular response to heat,apoptosis,and cellular lipid metabolic metabolism.Tea polyphe nols alleviate thermal damage by targeting BCL2,HSP90AA1,HSPA1A,JUN,MAPK1,NFKB1,NFKBIA,NOS3,and TP53.Moreover,10 mg/L tea polyphenols were found to upregulate the transcription levels of Hsp70,HO-1,NQ-O1,Nrf2,and MAPKI,and the transcription levels of Bax/Bcl2,p38,and JNK were downregulated to alleviate the heat stress-induced injury.[Conclusion]Tea polyphenols may enhance the antioxidant ability of H9C2 cells and inhibit cell apoptosis,thereby reducing heat stress injury.展开更多
基金the grant from National Key Research and Development Program of China(Grant No.2021YFE0110000)the grant from Tianjin Municipal Science and Technology Foundation(Grant No.22JCYBJC00160).
文摘Background:Green tea,mulberry leaf and corn silk are traditional herbs used in the prevention and treatment of diabetes in China for a long time,but their synergistic hypoglycemic effects and mechanisms remain unclear.Methods:The effective components of green tea,mulberry leaf and corn silk were extracted and enriched.Mixture design of experiments was used to study the influences of different combinations on the cell viability and glucose uptake level of L6 myoblasts,so as to determine the optimal synergistic hypoglycemic combination.The possible hypoglycemic mechanism of the optimal synergistic combination was explored by cytotoxicity assay,glucose uptake assay,and western blot.Results:Three polyphenol enrichment fractions of the herbs,30%ethanol elution fraction of green tea(GT),50%ethanol elution fraction of mulberry leaf(ML)and 60%ethanol elution fraction of corn silk(CS)were obtained.The antioxidant activities of GT-30%,ML-50%and CS-60%were superior to those of crude extracts,and showed strong potential inα-amylase andα-glucosidase inhibition activities.The optimal synergistic combination of crude extracts G7(crude extract of green tea:crude extract of mulberry leaf:crude extract of corn silk=1:5:3),polyphenol enrichment fractions R3(GT-30%:ML-50%:CS-60%=1:7:1)and monomers X2(epigallocatechin gallate:morusin:formononetin=3:1:2)were selected,respectively.G7,R3,and X2 showed promoting effects on the cell viability and glucose uptake of L6 myoblasts within the detected concentration range.In addition,G7,R3,and X2 could increase the expression levels of p-PI3K/PI3K and p-Akt/Akt in L6 myoblasts,and promote the translocation of Glut4,but G7 and R3 showed more significant effects.Conclusion:The synergistic hypoglycemic effects of green tea,mulberry leaf and corn silk had the characteristics of multiple-components and multiple-targets with p-PI3K/PI3K,p-Akt/Akt and the translocation of Glut4 signal pathways involved.The three traditional herbs might have the potential to be combined used for the prevention and treatment of diabetes based on the synergistic hypoglycemic effects.
基金Supported by National Natural Science Foundation of China(32302919,32302918)Taishan Industrial Experts Program(tscx202306046)+1 种基金Key R&D Program Rural Revitalization Project of Shandong Province(2023TZXD083)Science and Technology Cooperation Project of Shandong and Chongqing(2022LYXZ030)。
文摘[Objective]This paper was to investigate the action targets and pathways of tea polyphenols in alleviating heat stress-induced injury by using network pharmacological analysis and an H9C2 cell model.[Method]First,the corresponding targets of tea polyphenols were obtained from the PubChem database.Then,the core targets were screened based on topological parameters.The relevant metabolism pathways of tea polyphenols related to diseases were identified through GO functional annotation and KECG signaling pathway enrichment.Moreover,common targets for thermal injury and targets of tea polyphenols were obtained.Then,GO functional annotation was performed to explore the pathway of tea polyphenols in alleviating heat stress damage.H9C2 cells were cultured at 42℃ to construct the heat stress model,and the cells were treated with 10μg/mL tea polyphenols.The key genes were confirmed using RT-PCR technology.[Result]The study yielded 364 targets corresponding to tea polyphenols,including 68 core targets.These targets are related to various biological processes such as involve oxidative stress,cancer,lipopolysaccharide-mediated signaling pathways,antiviral responses,regulation of cellular response to heat,apoptosis,and cellular lipid metabolic metabolism.Tea polyphe nols alleviate thermal damage by targeting BCL2,HSP90AA1,HSPA1A,JUN,MAPK1,NFKB1,NFKBIA,NOS3,and TP53.Moreover,10 mg/L tea polyphenols were found to upregulate the transcription levels of Hsp70,HO-1,NQ-O1,Nrf2,and MAPKI,and the transcription levels of Bax/Bcl2,p38,and JNK were downregulated to alleviate the heat stress-induced injury.[Conclusion]Tea polyphenols may enhance the antioxidant ability of H9C2 cells and inhibit cell apoptosis,thereby reducing heat stress injury.
