Taxol production of Taxus chinensis (Pilger) Rehd. var. mairei (Lemee et Lévl.) Cheng et L. K. Fu induced by oligosaccharide from Fusarium oxysporum f. vasinfectum (Atkinson) Snyder et Hansen was ...Taxol production of Taxus chinensis (Pilger) Rehd. var. mairei (Lemee et Lévl.) Cheng et L. K. Fu induced by oligosaccharide from Fusarium oxysporum f. vasinfectum (Atkinson) Snyder et Hansen was studied in suspension cultures, and it was found that oligosaccharide triggered cell apoptosis. Under transmission electron microscope the following morphological changes were observed: cell shrinkage, condensation of cytoplasm, nuclear fragmentation, and the increase of high electron density bodies in vacuole in great quantity. In oligosaccharide_treated cells, agarose gel electrophoresis revealed that DNA was digested into oligonucleosomal fragments that were times of 200 bp appearing as DNA ladders. Control cells were in normal physiological state, they were intact, abundant in organelle and with integral nucleus DNA, and the rate of taxol biosynthesis in these cells was very low. After the oligosaccharide to the culture system, the defense system of cells was elicited and the secondary metabolism was strengthened, i.e. phenolics were accumulated in the medium, the activity of polyphenol oxidase (PPO) was increased quickly and secondary wall of cells was thickened. The activity of L _phenylalanine ammonia lyase (PAL), the critical enzyme of the phenylpropanoid pathway, was increased promptly 1 h after elicitation. The rate of taxol production was improved sharply and the maximal taxol concentration at 72 h was six times that of control. Appearance of cell apoptosis was accompanied with the highest concentration of taxol in suspension cultures.展开更多
OBJECTIVE: To investigate the effects of Tangnaikang(TNK), a mixture of five herbal plant extracts,on inflammation-mediated insulin resistance andβ-cell apoptosis in SHR.Cg-Leprcp/NDmcr(SHR-cp)rats.METHODS: Seven-wee...OBJECTIVE: To investigate the effects of Tangnaikang(TNK), a mixture of five herbal plant extracts,on inflammation-mediated insulin resistance andβ-cell apoptosis in SHR.Cg-Leprcp/NDmcr(SHR-cp)rats.METHODS: Seven-week-old SHR-cp rats were randomly divided into a control(CON) group and a TNK(3.24 g/kg) group. Wistar-Kyoto rats at the same age were used as the normal control group.After 7 weeks of continuous intragastric administration of TNK, the glucose metabolic status and insulin sensitivity of the rats were evaluated by assessing fasting serum glucose(FBG), the oral glucose tolerance test(OGTT), fasting serum insulin(FINS),and the insulin sensitivity index(ISI). Serum tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), C-reactive protein(CRP) and adiponectin were measured via enzyme-linked immunosorbent assays.Macrophage infiltration and apoptosis in adipose tissues were detected through F4/80 immunohistochemistry and the terminal deoxynucleotidyl transferase d UTP nick end labeling(TUNEL) assay. Islet morphology and β-cell apoptosis were investigated using double immunofluorescence staining and the TUNEL assay. The expression of cytokine genes in adipose tissue, the liver, and the pancreas was detected in real-time polymerase chain reaction assays. The expression and phosphorylation levels of insulin signaling-, inflammation-, and β-cell survival-related proteins in the liver and pancreas of SHR-cp rats were detected by western blotting.RESULTS: TNK(3.24 g/kg) treatment significantly decreased body weight, FBG and FINS; improved impaired glucose tolerance; increased the ISI; reduced serum levels of TNF-α, CRP and IL-6; and increased serum adiponectin. The m RNA expression of inflammatory markers was markedly reduced in the liver, pancreas, and adipose tissue. F4/80-and TUNEL-positive cells in adipose tissues were decreased, as was the number of TUNEL-positiveβ-cells. The phosphorylation of c-Jun N-terminal kinase and that of insulin receptor substrate-1 at serines 307 and 1101 was significantly decreased.In the pancreas, the expression of nuclear factor kappa-light chain-enhancer of activated B cells-p65 was significantly decreased, and phosphoinositide3-kinase and IRS-2 were significantly increased.CONCLUSION: TNK was able to improve insulin resistance and β-cell apoptosis in SHR-cp rats, which might be associated with its ability to relieve the overall and local metabolic inflammatory responses observed in SHR-cp rats.展开更多
文摘Taxol production of Taxus chinensis (Pilger) Rehd. var. mairei (Lemee et Lévl.) Cheng et L. K. Fu induced by oligosaccharide from Fusarium oxysporum f. vasinfectum (Atkinson) Snyder et Hansen was studied in suspension cultures, and it was found that oligosaccharide triggered cell apoptosis. Under transmission electron microscope the following morphological changes were observed: cell shrinkage, condensation of cytoplasm, nuclear fragmentation, and the increase of high electron density bodies in vacuole in great quantity. In oligosaccharide_treated cells, agarose gel electrophoresis revealed that DNA was digested into oligonucleosomal fragments that were times of 200 bp appearing as DNA ladders. Control cells were in normal physiological state, they were intact, abundant in organelle and with integral nucleus DNA, and the rate of taxol biosynthesis in these cells was very low. After the oligosaccharide to the culture system, the defense system of cells was elicited and the secondary metabolism was strengthened, i.e. phenolics were accumulated in the medium, the activity of polyphenol oxidase (PPO) was increased quickly and secondary wall of cells was thickened. The activity of L _phenylalanine ammonia lyase (PAL), the critical enzyme of the phenylpropanoid pathway, was increased promptly 1 h after elicitation. The rate of taxol production was improved sharply and the maximal taxol concentration at 72 h was six times that of control. Appearance of cell apoptosis was accompanied with the highest concentration of taxol in suspension cultures.
基金Supported by Grants from the International Scientific and Technological Cooperation Program of China:Innovative Chinese Medicine for the prevention and treatment of insulin resistance(No.2010DFB33260)the Key Project of the Chinese Ministry of Education:the Effects and Mechanisms of Tangnaikang on Insulin Signaling Pathway(No.311011)the Beijing Talents Fund(No.2016000021469G193)
文摘OBJECTIVE: To investigate the effects of Tangnaikang(TNK), a mixture of five herbal plant extracts,on inflammation-mediated insulin resistance andβ-cell apoptosis in SHR.Cg-Leprcp/NDmcr(SHR-cp)rats.METHODS: Seven-week-old SHR-cp rats were randomly divided into a control(CON) group and a TNK(3.24 g/kg) group. Wistar-Kyoto rats at the same age were used as the normal control group.After 7 weeks of continuous intragastric administration of TNK, the glucose metabolic status and insulin sensitivity of the rats were evaluated by assessing fasting serum glucose(FBG), the oral glucose tolerance test(OGTT), fasting serum insulin(FINS),and the insulin sensitivity index(ISI). Serum tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), C-reactive protein(CRP) and adiponectin were measured via enzyme-linked immunosorbent assays.Macrophage infiltration and apoptosis in adipose tissues were detected through F4/80 immunohistochemistry and the terminal deoxynucleotidyl transferase d UTP nick end labeling(TUNEL) assay. Islet morphology and β-cell apoptosis were investigated using double immunofluorescence staining and the TUNEL assay. The expression of cytokine genes in adipose tissue, the liver, and the pancreas was detected in real-time polymerase chain reaction assays. The expression and phosphorylation levels of insulin signaling-, inflammation-, and β-cell survival-related proteins in the liver and pancreas of SHR-cp rats were detected by western blotting.RESULTS: TNK(3.24 g/kg) treatment significantly decreased body weight, FBG and FINS; improved impaired glucose tolerance; increased the ISI; reduced serum levels of TNF-α, CRP and IL-6; and increased serum adiponectin. The m RNA expression of inflammatory markers was markedly reduced in the liver, pancreas, and adipose tissue. F4/80-and TUNEL-positive cells in adipose tissues were decreased, as was the number of TUNEL-positiveβ-cells. The phosphorylation of c-Jun N-terminal kinase and that of insulin receptor substrate-1 at serines 307 and 1101 was significantly decreased.In the pancreas, the expression of nuclear factor kappa-light chain-enhancer of activated B cells-p65 was significantly decreased, and phosphoinositide3-kinase and IRS-2 were significantly increased.CONCLUSION: TNK was able to improve insulin resistance and β-cell apoptosis in SHR-cp rats, which might be associated with its ability to relieve the overall and local metabolic inflammatory responses observed in SHR-cp rats.
