AIM To investigate the underlying mechanisms of the protective role of remote ischemic perconditioning(RIPerC) in rat liver transplantation. METHODS Sprague-Dawley rats were subjected to sham, orthotopic liver transpl...AIM To investigate the underlying mechanisms of the protective role of remote ischemic perconditioning(RIPerC) in rat liver transplantation. METHODS Sprague-Dawley rats were subjected to sham, orthotopic liver transplantation(OLT), ischemic postconditioning(IPostC) or RIPerC. After 3 h reperfusion, blood samples were taken for measurement of alanine aminotransferase, aspartate aminotransferase, creatinine(Cr) and creatinine kinase-myocardial band(CK-MB). The liver lobes were harvested for the following measurements: reactive oxygen species(ROS), H2O2, mitochondrial membrane potential(ΔΨm) and total nitric oxide(NO). These measurements were determined using an ROS/H2O2, JC1 and Total NOx Assay Kit, respectively. Endothelial NO synthase(e NOS) was analyzed by reverse transcription-polymerase chain reaction(RTPCR) and western blotting, and peroxynitrite was semiquantified by western blotting of 3-nitrotyrosine. RESULTS Compared with the OLT group, the grafts subjected to RIPerC showed significantly improved liver and remote organ functions(P < 0.05). ROS(P < 0.001) including H2O2(P < 0.05) were largely elevated in the OLT group as compared with the sham group, and RIPerC(P < 0.05) reversed this trend. The collapse of ΔΨm induced by OLT ischemia/reperfusion(I/R) injury was significantly attenuated in the RIPerC group(P < 0.001). A marked increase of NO content and phosphoserine eN OS, both in protein and mR NA levels, was observed in liver graft of the RIPer C group as compared with the OLT group(P < 0.05). I/R-induced 3-nitrotyrosine content was significantly reduced in the RIPerC group as compared with the OLT group(P < 0.05). There were no significant differences between the RIPerC and IPostC groups for all the results except Cr. The Cr level was lower in the RIPerC group than in the IPostC group(P < 0.01).CONCLUSION Liver graft protection by RIPerC is similar to or better than that of IPostC, and involves inhibition of oxidative stress and up-regulation of the PI3K/Akt/e NOS/NO pathway.展开更多
To the Editor:Liver transplantation (LT) is an effective therapeutic method for end-stage liver diseases. Although rejection is commonly mild or moderate in LT compared with other solid organs transplantation, it is s...To the Editor:Liver transplantation (LT) is an effective therapeutic method for end-stage liver diseases. Although rejection is commonly mild or moderate in LT compared with other solid organs transplantation, it is still requiring a life-long immunosuppressive therapy. Meanwhile, LT is less sensitive to rejection than other organs transplantation due to liver immune privilege which even allows the success of LT with positive across mismatches of blood group and complete major histocompatibility complex.[1, 2] During the early period of transplantation,clinical operational tolerance (COT) could be achieved on account of the genetic identity between recipient and donor.[3] Over the last decades, there is solid evident supporting that approximately 20% of LT recipients could develop COT, defined as maintaining normal graft function beyond one year completely without administering any immunosuppressive drugs.[4]展开更多
De novo non-alcoholic fatty liver disease(NAFLD) is a common late complication for long-term survivors after liver transplantation. Genomic studies confirmed that PNPLA3 I148 M and TM6SF2 E167 K polymorphisms affected...De novo non-alcoholic fatty liver disease(NAFLD) is a common late complication for long-term survivors after liver transplantation. Genomic studies confirmed that PNPLA3 I148 M and TM6SF2 E167 K polymorphisms affected NAFLD susceptibility in the general population. However,this association was not validated in survivors after liver transplantation(LT). We performed a crosssectional survey to investigate this relationship. A comprehensive survey,including anthropometric measurements,fasting venous blood sampling,ultrasound,and questionnaires was performed in the shortterm. The clinical indications and patient's steatosis status before LT were collected from inpatient medical records. Sixty-five long-term recipients with a survival exceeding 10 years were enrolled in the final analysis. De novo NAFLD was more frequent in PNPLA3 GG carriers(0.33 vs 0.10 for GG vs CC + CG carriers,P = 0.018),while the genetic impact on NAFLD susceptibility was insignificant when categorized by the TM6SF2 polymorphism(0.19 in CC vs 0.14 in CT + TT carriers,P = 0.883). Multi-covariate analysis revealed that PNPLA3 exerted a significant genetic effect on de novoNAFLD following a recessive model(GG vs CC + CG,OR = 14.2,95%CI:1.78-113,P = 0.012). Compared to recipients with only the PNPLA3 GG allele or obesity(defined as body mass index > 25 kg/m2),steatosis was highly prevalent(71.4%) in PNPLA3 GG carriers Table 1 Current status of long-term recipients surviving more than 10 years Univariate Multivariate NAFLD(n = 12)Control(n = 53)P value OR P value Age(yr) 56.5 ± 8.4 53.6 ± 10.1 0.356 1.04(0.92-1.18)0.528Gender(M/F) 10/2 47/6 0.611 1.40(0.14-14.2)0.427 Indication for LT Hepatitis/cirrhosis/ cancer/others1/8/2/17/35/9/20.889 Survival time(yr) 11.2 ± 0.9 11.5 ± 1.4 0.541 BMI(kg/m2) 25.1 ± 3.0 22.5 ± 2.6 0.003 1.47(1.03-2.08)0.032 TG(mmol/L) 1.6 ± 1.1 1.1 ± 0.6 0.038 1.34(0.38-4.71)0.652 HDL-C(mmol/L) 1.2(1.0-1.4) 1.3(1.0-1.7) 0.267 FBG(mmol/L) 7.6 ± 3.4 5.7 ± 1.9 0.013 1.49(0.93-2.37)0.095 Hypertension(Yes/no)3/9 21/32 0.343 SUA(μmol/L) 381.6 ± 75.6 342.6 ± 76.4 0.116 Met S(Yes/no) 4/8 9/44 0.201 ALT(U/L) 36.7 ± 7.0 38.8 ± 6.7 0.882 Alcohol intake(g/wk)11.6 ± 7.3 21.0 ± 8.2 0.766 Smoking(cigar/d) 4.2 ± 3.4 4.3 ± 1.2 0.969 Exercise(min/d) 18.5 ± 6.0 22.9 ± 2.9 0.513 Immunosuppression Tacrolimus/ cyclosporine/ MMF/sirolimus/ none11/1/0/0/036/12/1/2/20.575 PNPLA3(CC/CG/GG)1/3/8 16/21/16 0.018 14.2(1.78-113)0.012 TM6SF2(CC/CT/TT)11/1/0 47/5/1 0.883 2.68(0.25-28.5)0.413 Continuous variables with equal variance are presented as the mean ± SD; Continuous variables with unequal variance are presented as the median(interquartile range); Categorical variables are presented as the number of subjects. One-way ANOVA was used for the comparison between continuous variables with equal variance,Mann-Whitney U test was used for the comparison between continuous variables with unequal variance,chi-square test was used for the comparison between categorical variables in univariate analysis,and logistic regression analysis was used in the multivariate analysis. The effect of the PNPLA3 I148 M polymorphism was evaluated by a recessive genetic model(GG vs CG + CC); the effect of the TM6SF2 E167 K polymorphism was evaluated by a dominant genetic model(CC + CT vs TT) for decreased prevalence of TT carrier. ALT:Alanine aminotransferase; BMI:Body mass index; F:Female; FBG:Fasting blood glucose; HBs Ag:Hepatitis B surface antigen; HDL-C:Highdensity lipoprotein cholesterol; LT:Liver transplantation; M:Male; Met S:Metabolic syndrome; MMF:Mycophenolatemofetil; SUA:Serum uric acid; TG:Triglyceride. De novo NAFLD Controlwith obesity. In conclusion,PNPLA3 I148 M,but not TM6SF2 E167 K,affects de novo NAFLD occurrence with a prominent interaction with obesity. Weight control might be a meaningful method to reduce the genetic susceptibility to NAFLD exerted by PNPLA3 variants.展开更多
基金Supported by National Natural Science Foundation of China,No.81421062the Science and Technology Bureau of Zhejiang Province,China,No.2016C33145+1 种基金the National Natural Science Foundation of China,No.81470891the 863 National High Technology Research and Development Program of China for young scientist No.2015AA020923
文摘AIM To investigate the underlying mechanisms of the protective role of remote ischemic perconditioning(RIPerC) in rat liver transplantation. METHODS Sprague-Dawley rats were subjected to sham, orthotopic liver transplantation(OLT), ischemic postconditioning(IPostC) or RIPerC. After 3 h reperfusion, blood samples were taken for measurement of alanine aminotransferase, aspartate aminotransferase, creatinine(Cr) and creatinine kinase-myocardial band(CK-MB). The liver lobes were harvested for the following measurements: reactive oxygen species(ROS), H2O2, mitochondrial membrane potential(ΔΨm) and total nitric oxide(NO). These measurements were determined using an ROS/H2O2, JC1 and Total NOx Assay Kit, respectively. Endothelial NO synthase(e NOS) was analyzed by reverse transcription-polymerase chain reaction(RTPCR) and western blotting, and peroxynitrite was semiquantified by western blotting of 3-nitrotyrosine. RESULTS Compared with the OLT group, the grafts subjected to RIPerC showed significantly improved liver and remote organ functions(P < 0.05). ROS(P < 0.001) including H2O2(P < 0.05) were largely elevated in the OLT group as compared with the sham group, and RIPerC(P < 0.05) reversed this trend. The collapse of ΔΨm induced by OLT ischemia/reperfusion(I/R) injury was significantly attenuated in the RIPerC group(P < 0.001). A marked increase of NO content and phosphoserine eN OS, both in protein and mR NA levels, was observed in liver graft of the RIPer C group as compared with the OLT group(P < 0.05). I/R-induced 3-nitrotyrosine content was significantly reduced in the RIPerC group as compared with the OLT group(P < 0.05). There were no significant differences between the RIPerC and IPostC groups for all the results except Cr. The Cr level was lower in the RIPerC group than in the IPostC group(P < 0.01).CONCLUSION Liver graft protection by RIPerC is similar to or better than that of IPostC, and involves inhibition of oxidative stress and up-regulation of the PI3K/Akt/e NOS/NO pathway.
基金supported by grants from National Natural Science Foundation of China(81373160)Major Program of National Natural Science Foundation of China(91542205)Zhejiang Provincial Natural Science Foundation of China(LQ15H030003)
文摘To the Editor:Liver transplantation (LT) is an effective therapeutic method for end-stage liver diseases. Although rejection is commonly mild or moderate in LT compared with other solid organs transplantation, it is still requiring a life-long immunosuppressive therapy. Meanwhile, LT is less sensitive to rejection than other organs transplantation due to liver immune privilege which even allows the success of LT with positive across mismatches of blood group and complete major histocompatibility complex.[1, 2] During the early period of transplantation,clinical operational tolerance (COT) could be achieved on account of the genetic identity between recipient and donor.[3] Over the last decades, there is solid evident supporting that approximately 20% of LT recipients could develop COT, defined as maintaining normal graft function beyond one year completely without administering any immunosuppressive drugs.[4]
基金Supported by National S and T Major Project,No.2012ZX 10002017Foundation for Innovative Research Groups of the National Natural Science Foundation of China,Grant No.81421062China Postdoctoral Science Foundation Project,No.2015M570518
文摘De novo non-alcoholic fatty liver disease(NAFLD) is a common late complication for long-term survivors after liver transplantation. Genomic studies confirmed that PNPLA3 I148 M and TM6SF2 E167 K polymorphisms affected NAFLD susceptibility in the general population. However,this association was not validated in survivors after liver transplantation(LT). We performed a crosssectional survey to investigate this relationship. A comprehensive survey,including anthropometric measurements,fasting venous blood sampling,ultrasound,and questionnaires was performed in the shortterm. The clinical indications and patient's steatosis status before LT were collected from inpatient medical records. Sixty-five long-term recipients with a survival exceeding 10 years were enrolled in the final analysis. De novo NAFLD was more frequent in PNPLA3 GG carriers(0.33 vs 0.10 for GG vs CC + CG carriers,P = 0.018),while the genetic impact on NAFLD susceptibility was insignificant when categorized by the TM6SF2 polymorphism(0.19 in CC vs 0.14 in CT + TT carriers,P = 0.883). Multi-covariate analysis revealed that PNPLA3 exerted a significant genetic effect on de novoNAFLD following a recessive model(GG vs CC + CG,OR = 14.2,95%CI:1.78-113,P = 0.012). Compared to recipients with only the PNPLA3 GG allele or obesity(defined as body mass index > 25 kg/m2),steatosis was highly prevalent(71.4%) in PNPLA3 GG carriers Table 1 Current status of long-term recipients surviving more than 10 years Univariate Multivariate NAFLD(n = 12)Control(n = 53)P value OR P value Age(yr) 56.5 ± 8.4 53.6 ± 10.1 0.356 1.04(0.92-1.18)0.528Gender(M/F) 10/2 47/6 0.611 1.40(0.14-14.2)0.427 Indication for LT Hepatitis/cirrhosis/ cancer/others1/8/2/17/35/9/20.889 Survival time(yr) 11.2 ± 0.9 11.5 ± 1.4 0.541 BMI(kg/m2) 25.1 ± 3.0 22.5 ± 2.6 0.003 1.47(1.03-2.08)0.032 TG(mmol/L) 1.6 ± 1.1 1.1 ± 0.6 0.038 1.34(0.38-4.71)0.652 HDL-C(mmol/L) 1.2(1.0-1.4) 1.3(1.0-1.7) 0.267 FBG(mmol/L) 7.6 ± 3.4 5.7 ± 1.9 0.013 1.49(0.93-2.37)0.095 Hypertension(Yes/no)3/9 21/32 0.343 SUA(μmol/L) 381.6 ± 75.6 342.6 ± 76.4 0.116 Met S(Yes/no) 4/8 9/44 0.201 ALT(U/L) 36.7 ± 7.0 38.8 ± 6.7 0.882 Alcohol intake(g/wk)11.6 ± 7.3 21.0 ± 8.2 0.766 Smoking(cigar/d) 4.2 ± 3.4 4.3 ± 1.2 0.969 Exercise(min/d) 18.5 ± 6.0 22.9 ± 2.9 0.513 Immunosuppression Tacrolimus/ cyclosporine/ MMF/sirolimus/ none11/1/0/0/036/12/1/2/20.575 PNPLA3(CC/CG/GG)1/3/8 16/21/16 0.018 14.2(1.78-113)0.012 TM6SF2(CC/CT/TT)11/1/0 47/5/1 0.883 2.68(0.25-28.5)0.413 Continuous variables with equal variance are presented as the mean ± SD; Continuous variables with unequal variance are presented as the median(interquartile range); Categorical variables are presented as the number of subjects. One-way ANOVA was used for the comparison between continuous variables with equal variance,Mann-Whitney U test was used for the comparison between continuous variables with unequal variance,chi-square test was used for the comparison between categorical variables in univariate analysis,and logistic regression analysis was used in the multivariate analysis. The effect of the PNPLA3 I148 M polymorphism was evaluated by a recessive genetic model(GG vs CG + CC); the effect of the TM6SF2 E167 K polymorphism was evaluated by a dominant genetic model(CC + CT vs TT) for decreased prevalence of TT carrier. ALT:Alanine aminotransferase; BMI:Body mass index; F:Female; FBG:Fasting blood glucose; HBs Ag:Hepatitis B surface antigen; HDL-C:Highdensity lipoprotein cholesterol; LT:Liver transplantation; M:Male; Met S:Metabolic syndrome; MMF:Mycophenolatemofetil; SUA:Serum uric acid; TG:Triglyceride. De novo NAFLD Controlwith obesity. In conclusion,PNPLA3 I148 M,but not TM6SF2 E167 K,affects de novo NAFLD occurrence with a prominent interaction with obesity. Weight control might be a meaningful method to reduce the genetic susceptibility to NAFLD exerted by PNPLA3 variants.
