Summary： In order to investigate whether Yinchenhao decoction （YCHD） attenuates hepatic fibro- genesis in the bile duct ligation （BDL） model via recovering and restoring the self-regulation and bal- ance of the r...Summary： In order to investigate whether Yinchenhao decoction （YCHD） attenuates hepatic fibro- genesis in the bile duct ligation （BDL） model via recovering and restoring the self-regulation and bal- ance of the renin-angiotensin system （RAS）, 33 specific-pathogen-free （SPF） male Sprague-Dawley rats with common BDL and scission were randomly divided into five groups as follows： G1, the sham group （n=4）; G2, BDL 7-day group （n=5）; G3, BDL＋YCHD 430 mg/mL （n=8）; G4, BDL＋losartan 0.65 mg/mL （ARB group, n=8）; G5, model group （BDL without any treatment, n=8）. YCHD and losartan （10 mL.kgl.day-1） were given by gastric gavage for 16 days following BDL in G3 and G4 groups, respec- tively. The effect of YCHD on liver fibrosis and the detailed molecular mechanisms were assessed by liver function including total bilirubin （TBIL）, direct bilirubin （DBIL）, indirect bilirubin （IDBIL）, alanine aminotransferase （ALT）, and aspartate aminotransferase （AST）. Histological changes were ob-. served by transmission electron microscopy （TEM） and Masson trichrome staining. Western blotting was used to detect the protein expression level of the renin-angiotensin system （RAS） components in- cluding angiotensin converting enzyme （ACE）, angiotensin II type 1 receptor （AT1R）, ACE2, angio- tensin II （Ang II） as well as transforming growth factor 131 （TGF131）. The experimental data were ana- lyzed by principle component analytical method of pattern recognition. The results showed that bio- chemically, serum TBIL, DBIL, IDBIL, ALT and AST levels were markedly increased following BDL as compared with the sham group （P〈0.05）. Serum TBIL, IDBIL and DBIL levels in G3 group were dramatically decreased as compared with G5 and G4 groups （P〈0.05）. Serum AST level in G3 was sig- nificantly lowered than in G5 group （P〈0.05）, but there was no significant difference in ALT among G3, G4 and G5 groups （P〉0.05）. Histologically, livers in G3 group showed less hepatocytes necrosis, less bile duct hyperplasia and less collagen formation than in G4 and G5 groups. The protein expression lev- els of ACE2, ACE, Ang II, AT1R and TGF131 in G2, G3 and G4 groups were significantly higher than in sham group （P〈0.05）, and lower than in G5 group （P〈0.05）. However, the differences among G2, G3 and G4 groups were not significant （P〉0.05）. ACE2 protein expression in G3 group was significantly higher than in G2 group （P〈0.05） and there was no significant difference in comparison with G4 group （P〉0.05）. Moreover, the protein expression of TGF131 in G3 group was significantly lower than in G5 and G4 groups （P〈0.05）. Our findings suggest that the antifibrotic effects of YCHD may be associated with the decreased classical RAS pathway components and TGFI31 downexpression so as to recover and rebuild self-regulation of the RAS by elevating the protein expression of ACE2.展开更多
Here we developed a saccharic colorimetric method based on the combination of chemoselective ligation and enzyme-specific catalysis using aminooxy/ hydrazine-functionalized gold nanoparticles （AO/AuNPs or H/AuNPs）. ...Here we developed a saccharic colorimetric method based on the combination of chemoselective ligation and enzyme-specific catalysis using aminooxy/ hydrazine-functionalized gold nanoparticles （AO/AuNPs or H/AuNPs）. In the detection of galactose （Gal）, galactohexodialdose （GHDA）, the galactose oxidase （GalOx）-catalyzed product, has an aldehyde group, which allows it to chemoselectively react with an aminooxy or hydrazine group at the outer layer of AO/AuNPs or H/AuNPs by oxime/hydrazone click chemistry to form oxime or hydrozone. Consequent134 through the specific recognition of 1,4-phenylenediboronic acid （PDBA） on cis-diols, GHDA, which contains two pairs of hydroxyls in the cis form, can bind not only with AO/AuNPs or H/AuNPs, but also with PDBA to form boronate diester, thereby triggering the aggregation of AuNPs and causing the corresponding color change. As GalOx catalyzed specific substrates, the amount of Gal correlated with the production of GHDA and the extent of AuNPs aggregation, thus allowing a simple and easily operatable colorimetric method for Gal detection to be developed. Under the optimized experimental conditions, the ratios of absorbance at a wavelength of 617 nm to that at 536 nm vary linearly with the logarithmic values of Gal concentrations within a wide range of 500 nM to 5 mM. Moreover, this colorimetric method shows anti-interference capability and high sensitivity with a detection limit of 21 nM. Thus, a universal platform for accurate and specific colorimetric analysis can be established through the integration of chemoselective ligation with enzyme specific catalysis.展开更多
Artificial synthesis and site-specific modification of peptides and proteins has evolved into an indispensable tool for protein engineers and chemical biologists. Chemical and enzymatic approaches to peptide ligation ...Artificial synthesis and site-specific modification of peptides and proteins has evolved into an indispensable tool for protein engineers and chemical biologists. Chemical and enzymatic approaches to peptide ligation are important alternatives of recombinant DNA technology for protein synthesis and modification. Although as old as that of chemical procedures, enzyme-mediated peptide ligation is far less developed than that of chemical counterpart due to the difficult availability of peptide ligase.Fortunately, this situation has been changed slowly with the fast development of biological techniques. In the past decades, several natural peptide ligases have been discovered. Protein engineering to improve the ligation efficiencies of the natural peptide ligase and to reverse the functionality of protease provide more powerful peptide ligases. In this review, the advances of enzyme-mediated peptide ligation and their application in protein synthesis and modification will be discussed.展开更多
目的观察拳参正丁醇提取物对结扎大鼠左冠状动脉前降支损伤的保护作用。方法取40只雄性W istar大鼠,随机分为假手术组、结扎冠状动脉组、拳参正丁醇提取物高剂量组(120 mg/kg)、拳参正丁醇提取物低剂量组(60 mg/kg)。结扎冠状动脉左前...目的观察拳参正丁醇提取物对结扎大鼠左冠状动脉前降支损伤的保护作用。方法取40只雄性W istar大鼠,随机分为假手术组、结扎冠状动脉组、拳参正丁醇提取物高剂量组(120 mg/kg)、拳参正丁醇提取物低剂量组(60 mg/kg)。结扎冠状动脉左前降支使心肌缺血并持续120 m in,复制结扎冠状动脉损伤模型。记录缺血前、缺血15m in、给药30,60,90,120 m inⅡ导联ECG,统计各标测点Ⅱ导联ECG的ST段偏移∑-ST。治疗组于结扎冠状动脉左前降支前20 m in经十二指肠注入拳参正丁醇提取物。结扎冠状动脉组及假手术组经十二指肠注入等体积生理盐水。实验结束后心内取血测定血清乳酸脱氢酶(LDH)、磷酸肌酸激酶(CK)活性;测定心肌组织丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性。另用24只W ister大鼠同上述分组及动物模型制作方法,取其心肌作TTC染色测心肌梗死面积。结果治疗组与模型组比较,心肌梗死面积明显缩小,血清LDH、CK活性降低,血清MDA含量减小,SOD活性升高,∑-ST值减少,且呈一定剂量依赖性。结论拳参正丁醇提取物治疗组(高、低剂量)对大鼠结扎冠状动脉的损伤均有保护作用。展开更多
AIM:To determine the molecular mechanisms involved in experimental hepatic fibrosis prevention by caffeine(CFA).METHODS:Liver fibrosis was induced in Wistar rats by intraperitoneal thioacetamide or bile duct ligation ...AIM:To determine the molecular mechanisms involved in experimental hepatic fibrosis prevention by caffeine(CFA).METHODS:Liver fibrosis was induced in Wistar rats by intraperitoneal thioacetamide or bile duct ligation and they were concomitantly treated with CFA(15 mg/kg per day).Fibrosis and inflammatory cell infiltrate were evaluated and classified by Knodell index.Inflammatory infiltrate was quantified by immunohistochemistry(anti-CD11b).Gene expression was analyzed by quantitative reverse transcription-polymerase chain reaction for collagenⅠ?(Col-1),connective tissue growth factor(CTGF),transforming growth factorβ1(TGF-β1),tumor necrosis factor alpha(TNF-α),interleukin-1(IL-1),IL-6,superoxide dismutase(SOD)and catalase(CAT).Activation of Nrf2 and Snail-1 was analyzed by Westernblot.TNF-αexpression was proved by enzyme-linked immunosorbant assay,CAT activity was performed by zymography.RESULTS:CFA treatment diminished fibrosis index in treated animals.The Knodell index showed both lower fibrosis and necroinflammation.Expression of profibrogenic genes CTGF,Col-1 and TGF-β1 and proinflammatory genes TNF-α,IL-6 and IL-1 was substantially diminished with CFA treatment with less CD11b positive areas.Significantly lower values of transcriptional factor Snail-1 were detected in CFA treated rats compared with cirrhotic rats without treatment;in contrast Nrf2was increased in the presence of CFA.Expression of SOD and CAT was greater in animals treated with CFA showing a strong correlation between mRNA expression and enzyme activity.CONCLUSION:Our results suggest that CFA inhibits the transcriptional factor Snail-1,down-regulating profibrogenic genes,and activates Nrf2 inducing antioxidant enzymes system,preventing inflammation and fibrosis.展开更多
Recent years have seen an ever increasing number of enzyme mediated protein/peptide modification reactions, which contribute significantly to the elucidation of related biological functions. The many available enzymes...Recent years have seen an ever increasing number of enzyme mediated protein/peptide modification reactions, which contribute significantly to the elucidation of related biological functions. The many available enzymes have, however, caused difficulties for practitioners in choosing the most appropriate enzyme for a certain purpose. This review surveyed the widely used enzymes(i.e., sortases, butelase 1,subtiligase, formylglycine generating enzyme and farnesyltransferase) in the manipulation of proteins/peptides, and the application fields of these enzymes as well as the advantages and limitations of each enzyme are summarized.展开更多
基金supported by grants from the National Natural Science Foundation of China(No.81102692)the Natural Science Foundation of Hubei Province,China(No.JX6B09)the Fundamental Research Funds for the Central Universities,China(No.2015QN203)
文摘Summary： In order to investigate whether Yinchenhao decoction （YCHD） attenuates hepatic fibro- genesis in the bile duct ligation （BDL） model via recovering and restoring the self-regulation and bal- ance of the renin-angiotensin system （RAS）, 33 specific-pathogen-free （SPF） male Sprague-Dawley rats with common BDL and scission were randomly divided into five groups as follows： G1, the sham group （n=4）; G2, BDL 7-day group （n=5）; G3, BDL＋YCHD 430 mg/mL （n=8）; G4, BDL＋losartan 0.65 mg/mL （ARB group, n=8）; G5, model group （BDL without any treatment, n=8）. YCHD and losartan （10 mL.kgl.day-1） were given by gastric gavage for 16 days following BDL in G3 and G4 groups, respec- tively. The effect of YCHD on liver fibrosis and the detailed molecular mechanisms were assessed by liver function including total bilirubin （TBIL）, direct bilirubin （DBIL）, indirect bilirubin （IDBIL）, alanine aminotransferase （ALT）, and aspartate aminotransferase （AST）. Histological changes were ob-. served by transmission electron microscopy （TEM） and Masson trichrome staining. Western blotting was used to detect the protein expression level of the renin-angiotensin system （RAS） components in- cluding angiotensin converting enzyme （ACE）, angiotensin II type 1 receptor （AT1R）, ACE2, angio- tensin II （Ang II） as well as transforming growth factor 131 （TGF131）. The experimental data were ana- lyzed by principle component analytical method of pattern recognition. The results showed that bio- chemically, serum TBIL, DBIL, IDBIL, ALT and AST levels were markedly increased following BDL as compared with the sham group （P〈0.05）. Serum TBIL, IDBIL and DBIL levels in G3 group were dramatically decreased as compared with G5 and G4 groups （P〈0.05）. Serum AST level in G3 was sig- nificantly lowered than in G5 group （P〈0.05）, but there was no significant difference in ALT among G3, G4 and G5 groups （P〉0.05）. Histologically, livers in G3 group showed less hepatocytes necrosis, less bile duct hyperplasia and less collagen formation than in G4 and G5 groups. The protein expression lev- els of ACE2, ACE, Ang II, AT1R and TGF131 in G2, G3 and G4 groups were significantly higher than in sham group （P〈0.05）, and lower than in G5 group （P〈0.05）. However, the differences among G2, G3 and G4 groups were not significant （P〉0.05）. ACE2 protein expression in G3 group was significantly higher than in G2 group （P〈0.05） and there was no significant difference in comparison with G4 group （P〉0.05）. Moreover, the protein expression of TGF131 in G3 group was significantly lower than in G5 and G4 groups （P〈0.05）. Our findings suggest that the antifibrotic effects of YCHD may be associated with the decreased classical RAS pathway components and TGFI31 downexpression so as to recover and rebuild self-regulation of the RAS by elevating the protein expression of ACE2.
文摘Here we developed a saccharic colorimetric method based on the combination of chemoselective ligation and enzyme-specific catalysis using aminooxy/ hydrazine-functionalized gold nanoparticles （AO/AuNPs or H/AuNPs）. In the detection of galactose （Gal）, galactohexodialdose （GHDA）, the galactose oxidase （GalOx）-catalyzed product, has an aldehyde group, which allows it to chemoselectively react with an aminooxy or hydrazine group at the outer layer of AO/AuNPs or H/AuNPs by oxime/hydrazone click chemistry to form oxime or hydrozone. Consequent134 through the specific recognition of 1,4-phenylenediboronic acid （PDBA） on cis-diols, GHDA, which contains two pairs of hydroxyls in the cis form, can bind not only with AO/AuNPs or H/AuNPs, but also with PDBA to form boronate diester, thereby triggering the aggregation of AuNPs and causing the corresponding color change. As GalOx catalyzed specific substrates, the amount of Gal correlated with the production of GHDA and the extent of AuNPs aggregation, thus allowing a simple and easily operatable colorimetric method for Gal detection to be developed. Under the optimized experimental conditions, the ratios of absorbance at a wavelength of 617 nm to that at 536 nm vary linearly with the logarithmic values of Gal concentrations within a wide range of 500 nM to 5 mM. Moreover, this colorimetric method shows anti-interference capability and high sensitivity with a detection limit of 21 nM. Thus, a universal platform for accurate and specific colorimetric analysis can be established through the integration of chemoselective ligation with enzyme specific catalysis.
基金The National Natural Science Foundation of China (Nos. 21462023, 21778025)the Education Department of Jiangxi Province (No.150297)
文摘Artificial synthesis and site-specific modification of peptides and proteins has evolved into an indispensable tool for protein engineers and chemical biologists. Chemical and enzymatic approaches to peptide ligation are important alternatives of recombinant DNA technology for protein synthesis and modification. Although as old as that of chemical procedures, enzyme-mediated peptide ligation is far less developed than that of chemical counterpart due to the difficult availability of peptide ligase.Fortunately, this situation has been changed slowly with the fast development of biological techniques. In the past decades, several natural peptide ligases have been discovered. Protein engineering to improve the ligation efficiencies of the natural peptide ligase and to reverse the functionality of protease provide more powerful peptide ligases. In this review, the advances of enzyme-mediated peptide ligation and their application in protein synthesis and modification will be discussed.
文摘目的观察拳参正丁醇提取物对结扎大鼠左冠状动脉前降支损伤的保护作用。方法取40只雄性W istar大鼠,随机分为假手术组、结扎冠状动脉组、拳参正丁醇提取物高剂量组(120 mg/kg)、拳参正丁醇提取物低剂量组(60 mg/kg)。结扎冠状动脉左前降支使心肌缺血并持续120 m in,复制结扎冠状动脉损伤模型。记录缺血前、缺血15m in、给药30,60,90,120 m inⅡ导联ECG,统计各标测点Ⅱ导联ECG的ST段偏移∑-ST。治疗组于结扎冠状动脉左前降支前20 m in经十二指肠注入拳参正丁醇提取物。结扎冠状动脉组及假手术组经十二指肠注入等体积生理盐水。实验结束后心内取血测定血清乳酸脱氢酶(LDH)、磷酸肌酸激酶(CK)活性;测定心肌组织丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性。另用24只W ister大鼠同上述分组及动物模型制作方法,取其心肌作TTC染色测心肌梗死面积。结果治疗组与模型组比较,心肌梗死面积明显缩小,血清LDH、CK活性降低,血清MDA含量减小,SOD活性升高,∑-ST值减少,且呈一定剂量依赖性。结论拳参正丁醇提取物治疗组(高、低剂量)对大鼠结扎冠状动脉的损伤均有保护作用。
基金Supported by Conacyt grant No.25474 to Juan ArmendárizBorunda
文摘AIM:To determine the molecular mechanisms involved in experimental hepatic fibrosis prevention by caffeine(CFA).METHODS:Liver fibrosis was induced in Wistar rats by intraperitoneal thioacetamide or bile duct ligation and they were concomitantly treated with CFA(15 mg/kg per day).Fibrosis and inflammatory cell infiltrate were evaluated and classified by Knodell index.Inflammatory infiltrate was quantified by immunohistochemistry(anti-CD11b).Gene expression was analyzed by quantitative reverse transcription-polymerase chain reaction for collagenⅠ?(Col-1),connective tissue growth factor(CTGF),transforming growth factorβ1(TGF-β1),tumor necrosis factor alpha(TNF-α),interleukin-1(IL-1),IL-6,superoxide dismutase(SOD)and catalase(CAT).Activation of Nrf2 and Snail-1 was analyzed by Westernblot.TNF-αexpression was proved by enzyme-linked immunosorbant assay,CAT activity was performed by zymography.RESULTS:CFA treatment diminished fibrosis index in treated animals.The Knodell index showed both lower fibrosis and necroinflammation.Expression of profibrogenic genes CTGF,Col-1 and TGF-β1 and proinflammatory genes TNF-α,IL-6 and IL-1 was substantially diminished with CFA treatment with less CD11b positive areas.Significantly lower values of transcriptional factor Snail-1 were detected in CFA treated rats compared with cirrhotic rats without treatment;in contrast Nrf2was increased in the presence of CFA.Expression of SOD and CAT was greater in animals treated with CFA showing a strong correlation between mRNA expression and enzyme activity.CONCLUSION:Our results suggest that CFA inhibits the transcriptional factor Snail-1,down-regulating profibrogenic genes,and activates Nrf2 inducing antioxidant enzymes system,preventing inflammation and fibrosis.
基金The financial support from the National Recruitment Program of Global Youth Experts(1000 Talents Plan)the National Natural Science Foundation of China (No. 81703406)
文摘Recent years have seen an ever increasing number of enzyme mediated protein/peptide modification reactions, which contribute significantly to the elucidation of related biological functions. The many available enzymes have, however, caused difficulties for practitioners in choosing the most appropriate enzyme for a certain purpose. This review surveyed the widely used enzymes(i.e., sortases, butelase 1,subtiligase, formylglycine generating enzyme and farnesyltransferase) in the manipulation of proteins/peptides, and the application fields of these enzymes as well as the advantages and limitations of each enzyme are summarized.
