Embryonic stem cells (ESC) are suggested to be immune-privileged, but they carry the risk of uncontrolled expansion and malignancy. Upon differentiation they lose their tumor-forming capacity, but they become immuno...Embryonic stem cells (ESC) are suggested to be immune-privileged, but they carry the risk of uncontrolled expansion and malignancy. Upon differentiation they lose their tumor-forming capacity, but they become immunogenic by the expression of a normal set of MHC molecules. This immunogenicity might trigger rejection after application in regenerative therapies. In this study MHC expression of and immune responses to endothelial derivatives of rat embryonic stem cell-like cells (RESC) under inflammatory conditions were determined in comparison to primary rat aortic endothelial cells (ECs). Cellular as well as humoral allo-recognition was analyzed in vitro. In addition, im- mune reactions in vivo were assessed by allo-antibody production and determination of interferon-y (IFNy)-secreting alio-reactive T cells. RESC derivatives expressed low but significant levels of MHC class I, and no MHC class II. In response to IFNy MHC class I expression was enhanced, while class II transactivator induction failed completely in these cells; MHC class II expression remained consistently absent. Functionally, the RESC derivatives showed a reduced allo-stimulatory capacity, protection against humoral allo-recognition in vitro and a slightly diminished sus- ceptibility to cytotoxic T cell lysis. Furthermore, in vivo experiments demonstrated that these ceils do not trigger host immune reactions, characterized by no allo-antibody production and no induction of allo-reactive memory T cells. Our results show that endothelial derivatives of RESC have a distinctive reduced immunogenic potency even under inflammatory conditions.展开更多
AIM:To explore the role of high-mobility group box 1 (HMGB1) protein during liver fibrogenesis and investigate the functional effects of HMGB1 gene silencing in hepatic stellate cells (HSCs) using siRNA.METHODS:Hepati...AIM:To explore the role of high-mobility group box 1 (HMGB1) protein during liver fibrogenesis and investigate the functional effects of HMGB1 gene silencing in hepatic stellate cells (HSCs) using siRNA.METHODS:Hepatic fibrosis in rats was induced through serial subcutaneous injections of dimethylnitrosamine,and expression of HMGB1 was detected by immunohistochemistry.HMGB1 siRNAs were developed and transiently transfected into HSC-T6 cells using Lipofectamine 2000.HMGB1 expression was evaluated by real-time polymerase chain reaction (PCR) and Western blotting analysis.Expression of α-smooth muscle actin (α-SMA) and collagen typesⅠand Ⅲ was evaluated by real-time PCR.Cell proliferation and the cell cycle were determined using the methyl thiazolyl tetrazolium method.Finally,collagen content in HSC supernatant was evaluated by an enzyme-linked immunosorbent assay.RESULTS:The results showed that HMGB1 was upregulated during liver fibrosis and that its expression was closely correlated with the deposition of collagen.siRNA molecules were successfully transfected into HSCs and induced inhibition of HMGB1 expression in a time-dependent manner.Moreover,HMGB1 siRNA treatment inhibited synthesis of α-SMA and collagen types Ⅰ and Ⅲ in transfected HSCs.CONCLUSION:This study suggests a significant functional role for HMGB1 in the development of liver fibrosis.It also demonstrates that downregulation of HMGB1 expression might be a potential strategy to treat liver fibrosis.展开更多
Objective Radiation-induced lung injury (RILl) is the most common, dose-limiting complication in thoracic malignancy radiotherapy. Considering its negative impact on patients and restrictions to efficacy, the mechan...Objective Radiation-induced lung injury (RILl) is the most common, dose-limiting complication in thoracic malignancy radiotherapy. Considering its negative impact on patients and restrictions to efficacy, the mechanism of RILl was studied. Methods Wistar rats were locally irradiated with a single dose of 0, 16, and 20 Gy to the right half of the lung to establish a lung injury model. Two and six months after irradiation, the right half of the rat lung tissue was removed, and the concentrations of TGF-[31, angiotensin II, and aldosterone were determined via enzyme-linked immunosorbent assay. Results Statistical differences were observed in the expression levels of angiotensin II and aldosterone between the non-irradiation and irradiation groups. Moreover, the expression level of the angiotensin II-aldosterone system increased with increasing doses, and the difference was still observed as time progressed. Conclusions Angiotensin II-aldosterone system has an important pathophysiological function in the progression of RILI.展开更多
Objective: To investigate the effect of moxibustion at Shenque (CV8) on the immune system in rats with different levels of exhaustive exercise. Methods: Fifty-six male Sprague-Dawley (SD) rats were randomly divided in...Objective: To investigate the effect of moxibustion at Shenque (CV8) on the immune system in rats with different levels of exhaustive exercise. Methods: Fifty-six male Sprague-Dawley (SD) rats were randomly divided into a blank group (n=8), an exhaustive group (n=24), and a moxibustion group (n=24). The exhaustive group was randomly divided into a 1-time exhaustive group, a 4-time exhaustive group and a 7-time exhaustive group, with 8 rats in each group. According to the treatment time, the moxibustion group was randomly divided into a 1-time moxibustion group, a 4-time moxibustion group and a 7-time moxibustion group, with 8 rats in each group. Rats in the exhaustive groups and the moxibustion groups were subjected to replicating the exhaustive swimming models. Rats in each moxibustion group received mild moxibustion for 15 min immediately after the exhaustive modeling, once every other day. Twenty-four hours after the corresponding exhaustive exercise, the rats in each group were tested for the levels of serum immunoglobulin (Ig) G, IgA, IgM and acid phosphatase (ACP), and the morphological changes of spleen tissues were observed. The level of IgA was detected by immunoturbidimetric assay, and the levels of IgG, IgM and ACP were detected by en zyme-linked imm uno sorb ent assay (ELISA). Results: Compared with the 1-time exhaustive group, swimming time of rats in the 4-time exhaustive group was significantly proIonged (P<0.01), and swimming time of rats in the 7-time exhaustive group was significantly shortened (P<0.01). Compared with the 7-time exhaustive group, exhaustive swimming time of rats in the 7-time moxibustion group was significantly proIonged (P<0.01). Compared with the blank group, the IgG level in the 1-time exhaustive group was significantly decreased (PvO.Ol), and the levels of IgG, IgA and IgM in the 4-time exhaustive group and the 7-time exhaustive group were all sign ifica ntly decreased (P<0.05 or PvO.Ol), while the ACP level was in creased sign ifica ntly (both P<0.01). Microscopically, the number of splenic corpuscles in the 1-time exhaustive group was reduced;the center of some splenic corpuscles in the 4-time exhaustive group was damaged;the number of splenic corpuscles in the 7-time exhaustive group was reduced, and there was no obvious germinal center. Compared with the 4-time exhaustive group, the IgA level in the 4-time moxibusti on group was sign ifica ntly in creased (P<0.01), and the ACP level was sign ifica ntly decreased (P<0.01). Compared with the 7-time exhaustive group, the levels of IgG, IgA and IgM in the 7-time moxibustion group were sign ifica ntly in creased (all PvO.Ol), and the ACP level was sign ifica ntly decreased (P<0.01). Microscopically, the nu mber of splenic corpuscles in the 1-time moxibustion group was reduced;the center of some splenic corpuscles in the 4-time moxibustion group was damaged together with hyperplasia of some splenic corpuscles;blast cells were proliferated in the center of some splenic corpuscles in the 7-time moxibustion group. Conclusion: Moxibustion at Shenque (CV 8) can improve the levels of IgG, IgA and IgM, reduce the ACP level, repair damaged spleen tissues, and enhance the immunity of the body to some extent in the Iong-term fatigue rats.展开更多
Liver transplantation is an established therapy for end-stage liver diseases. Graft rejection occurs unless the recipient receives immunosuppression after transplantation. This study aimed to explore the mechanism of ...Liver transplantation is an established therapy for end-stage liver diseases. Graft rejection occurs unless the recipient receives immunosuppression after transplantation. This study aimed to explore the mechanism of acute rejection of liver allografts in rats pre-treated with total body irradiation to eliminate passenger lymphocytes and to define the role of CD4+CD25+ regulatory T cells in the induction of immunotolerance in the recipient. Male Lewis rats were used as donors and male DA rats were re- cipients. Rats were randomly assigned to the following four groups: control group, homogeneity liver transplantation group, idio-immunotolerance group and acute rejection group. After transplantation, the survival time of each group, serum alanine aminotransferase, total bilirubin levels, number of Foxp3+CD4+CD25+ regulatory T cells, expression of glucocorticoid-induced tumor necrosis factor receptor on T cell subgroups, histopathology of the hepatic graft and spleen cytotoxic T lymphocyte lytic activity were measured. In the acute rejection group, where donors were preconditioned with total body in'adiation before liver transplantation, all recipients died between day 17 and day 21. On day 14, serum alanine aminotransferase increased signifi- cantly to (459.2±76.9) U L^- 1, total bilirubin increased to (124.1±33.7) μmol L-1 (P〈0.05) and the ratio of Foxp3+CD4+CD25+ regulatory T cells decreased significantly to 1.50%±0.50% (P〈0.05) compared with the other groups. Analysis of the T cell subpopulations in the acute rejection group varied from the other groups. Histological analysis showed typical changes of acute rejection in the acute rejection group only. Preconditioning of the donors with total body irradiation eliminated passenger lymphocytes of the liver graft, and thus affected the course of tolerance and induced acute rejection after liver transplantation.展开更多
Elucidation of the mechanisms of liver fibrogenesis is important to treat liver fibrosis.In this study,we established rat models of liver fibrosis with stages from 0–1,2,and 3–4 to 4 at 2,4,6,and 8 weeks,respectivel...Elucidation of the mechanisms of liver fibrogenesis is important to treat liver fibrosis.In this study,we established rat models of liver fibrosis with stages from 0–1,2,and 3–4 to 4 at 2,4,6,and 8 weeks,respectively,by injection of pig serum.Liver fibrogenesis was detected by Masson’s trichrome staining.Rat non-parenchymal cells(NPCs)were enriched 4-fold by Percoll density gradient centrifugation.Protein extracts from NPCs were prepared at 4 and 8 weeks,separated by two-dimensional electrophoresis,and then stained with Coomassie Blue G-250.At 4 weeks,we identified 18 non-redundant differentially expressed proteins of which protein disulfide-isomerase associated protein 3(PDIA3)and NDUV showed consistent expression at protein and mRNA levels from 4 to 8 weeks.PDIA3 was found to be down-regulated by Western blotting in the rat model and immunohistochemically in human liver.Our results revealed important aspects of the pathogenesis/progression of liver fibrosis and demonstrated important changes in protein expression levels of NPCs at various stages of liver fibrosis.展开更多
OBJECTⅣE: To investigate the protective role of Sijunzi decoction in neuromuscular junction(NMJ)and muscle cell mitochondria ultrastructure; as well as its effects on the amount of adenosine triphosphate(ATP) and the...OBJECTⅣE: To investigate the protective role of Sijunzi decoction in neuromuscular junction(NMJ)and muscle cell mitochondria ultrastructure; as well as its effects on the amount of adenosine triphosphate(ATP) and the activities of mitochondrial respiratory chain complexes I, Ⅱ, Ⅲ, and Ⅳ in autoimmune myasthenia gravis rats.METHODS: An experimental autoimmune myasthenia gravis(EAMG) rat model was established by inoculating rats with acetylcholine receptors extracted from Torpedo. Rats were divided into three groups: model, prednisone, and Sijunzi decoction, and were fed physiological saline, prednisone, or Sijunzi decoction, respectively. NMJ and muscle cell mitochondria ultrastructure were observed by transmission electron microscope. The amount of ATP was assessed by high performance liquid chromatography. The activities ofmitochondrial respiratory chain complexes I, Ⅱ, Ⅲ,and Ⅳ was determined using the Clark oxygen electrode method.RESULTS: In the model group, there were sparse muscle fibers, with decreased mitochondria, and sparse, diffluent, or absent NMJ folds. After intervention with Sijunzi decoction, the above pathology changes were improved: muscle fiber structure was clear and complete; the mitochondria count was higher; and the NMJ structure was close to normal. Gastrocnemius muscle mitochondria in the model group produced significantly less ATP than those in the prednisone group(P<0.01). Conversely, the ATP of Sijunzi decoction group was significantly higher than prednisone group(P<0.01). The activities of gastrocnemius muscle mitochondrial respiratory chain complexes I, Ⅱ, Ⅲ, and Ⅳ in both the prednisone and Sijunzi decoction groups was dramatically higher compared with the model group(P<0.05). The activities of complexes I and Ⅲ in the Sijunzi decoction group were significantly higher than those in the prednisone group(P<0.05), but there was no obvious difference in complex Ⅱ or Ⅳ activities between the two groups(P>0.05).CONCLUSION: Sijunzi decoction improved pathological changes in muscle mitochondria and NMJ,enhanced the amount of ATP in gastrocnemius muscle mitochondria, and improved the activities of respiratory chain complexes I, Ⅱ, Ⅲ, and Ⅳ(especially I and Ⅲ) of the EAMG rats.展开更多
Objective:To observe the anti-inflammatory effect,as well as the effect on the expression of microtubule-associated protein light chain 3B(LC3B)and Beclin-1 of herbal cake-partitioned moxibustion in rats with experime...Objective:To observe the anti-inflammatory effect,as well as the effect on the expression of microtubule-associated protein light chain 3B(LC3B)and Beclin-1 of herbal cake-partitioned moxibustion in rats with experimental autoimmune thyroiditis(EAT).Methods:Female Sprague-Dawley rats were randomly divided into a normal group and a modeling group.The EAT rat model was prepared by a combination of antigen immunization plus iodine agent induction.After the model was prepared,rats in the modeling group were randomly and equally divided into a model group and a herbal cake-partitioned moxibustion group.In the herbal cake-partitioned moxibustion group,moxibustion was alternately applied to two groups of points[Dazhui(GV14)-Mingmen(GV4)and Tiantu(CV22)-Guanyuan(CV4)],and the treatment continued for 30 d.Rats in the normal and model groups were only fixed identically without intervention.Histopathological manifestations of thyroid glands were observed by hematoxylin-eosin staining;the concentrations of thyroid peroxidase antibodies(TPOAb),thyroglobulin antibodies(TGAb),interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay;real-time fluorescence quantitative polymerase chain reaction and immunohistochemistry were used to detect the mRNA and protein expression of autophagy-related factors LC3B and Beclin-1 in thyroid tissue.Results:There were massive follicular destruction,lymphocytic infiltration,and interstitial fibrous tissue hyperplasia of the thyroid glands in the model group.Some follicles of the thyroid glands were destroyed with few lymphocyte infiltrations and fibrous tissue hyperplasia in the moxibustion group.Compared with the normal group,the concentrations of serum TPOAb,TGAb,IL-1β,IL-6,and TNF-αwere increased in the model rats(P<0.05);the mRNA and protein expression levels of LC3B and Beclin-1 in thyroid tissue were reduced in the model group(P<0.05).Compared with the model group,the concentrations of serum TPOAb,TGAb,IL-1β,IL-6,and TNF-αwere reduced in the herbal cake-partitioned moxibustion group(P<0.05);the mRNA and protein expression levels of LC3B and Beclin-1 in thyroid tissue were increased in the herbal cake-partitioned moxibustion group(P<0.05).The mRNA and protein expression of LC3B and Beclin-1 in thyroid tissue was negatively correlated with the serum levels of TPOAb and TGAb.Conclusion:Herbal cake-partitioned moxibustion reduces the inflammatory response in the thyroid glands of EAT rats and lowers the levels of serum TPOAb and TGAb.This may be related to the regulation of mRNA and protein expression of the autophagy-associated factors LC3B and Beclin-1 in rat thyroid tissue.展开更多
文摘Embryonic stem cells (ESC) are suggested to be immune-privileged, but they carry the risk of uncontrolled expansion and malignancy. Upon differentiation they lose their tumor-forming capacity, but they become immunogenic by the expression of a normal set of MHC molecules. This immunogenicity might trigger rejection after application in regenerative therapies. In this study MHC expression of and immune responses to endothelial derivatives of rat embryonic stem cell-like cells (RESC) under inflammatory conditions were determined in comparison to primary rat aortic endothelial cells (ECs). Cellular as well as humoral allo-recognition was analyzed in vitro. In addition, im- mune reactions in vivo were assessed by allo-antibody production and determination of interferon-y (IFNy)-secreting alio-reactive T cells. RESC derivatives expressed low but significant levels of MHC class I, and no MHC class II. In response to IFNy MHC class I expression was enhanced, while class II transactivator induction failed completely in these cells; MHC class II expression remained consistently absent. Functionally, the RESC derivatives showed a reduced allo-stimulatory capacity, protection against humoral allo-recognition in vitro and a slightly diminished sus- ceptibility to cytotoxic T cell lysis. Furthermore, in vivo experiments demonstrated that these ceils do not trigger host immune reactions, characterized by no allo-antibody production and no induction of allo-reactive memory T cells. Our results show that endothelial derivatives of RESC have a distinctive reduced immunogenic potency even under inflammatory conditions.
基金Supported by The Select and Train Outstanding Young Teach-ers Foundation of Shanghai,No.jdy08086WUJieping Experimental Diagnosis of Liver Disease Medical Foundation,No.LDWMF-SY-2011B009
文摘AIM:To explore the role of high-mobility group box 1 (HMGB1) protein during liver fibrogenesis and investigate the functional effects of HMGB1 gene silencing in hepatic stellate cells (HSCs) using siRNA.METHODS:Hepatic fibrosis in rats was induced through serial subcutaneous injections of dimethylnitrosamine,and expression of HMGB1 was detected by immunohistochemistry.HMGB1 siRNAs were developed and transiently transfected into HSC-T6 cells using Lipofectamine 2000.HMGB1 expression was evaluated by real-time polymerase chain reaction (PCR) and Western blotting analysis.Expression of α-smooth muscle actin (α-SMA) and collagen typesⅠand Ⅲ was evaluated by real-time PCR.Cell proliferation and the cell cycle were determined using the methyl thiazolyl tetrazolium method.Finally,collagen content in HSC supernatant was evaluated by an enzyme-linked immunosorbent assay.RESULTS:The results showed that HMGB1 was upregulated during liver fibrosis and that its expression was closely correlated with the deposition of collagen.siRNA molecules were successfully transfected into HSCs and induced inhibition of HMGB1 expression in a time-dependent manner.Moreover,HMGB1 siRNA treatment inhibited synthesis of α-SMA and collagen types Ⅰ and Ⅲ in transfected HSCs.CONCLUSION:This study suggests a significant functional role for HMGB1 in the development of liver fibrosis.It also demonstrates that downregulation of HMGB1 expression might be a potential strategy to treat liver fibrosis.
基金supported by grants from the National Natural Science Foundation of China(No.30900384)Education Bureau Foundation of Liaoning Province,China (No.2009a723)
文摘Objective Radiation-induced lung injury (RILl) is the most common, dose-limiting complication in thoracic malignancy radiotherapy. Considering its negative impact on patients and restrictions to efficacy, the mechanism of RILl was studied. Methods Wistar rats were locally irradiated with a single dose of 0, 16, and 20 Gy to the right half of the lung to establish a lung injury model. Two and six months after irradiation, the right half of the rat lung tissue was removed, and the concentrations of TGF-[31, angiotensin II, and aldosterone were determined via enzyme-linked immunosorbent assay. Results Statistical differences were observed in the expression levels of angiotensin II and aldosterone between the non-irradiation and irradiation groups. Moreover, the expression level of the angiotensin II-aldosterone system increased with increasing doses, and the difference was still observed as time progressed. Conclusions Angiotensin II-aldosterone system has an important pathophysiological function in the progression of RILI.
文摘Objective: To investigate the effect of moxibustion at Shenque (CV8) on the immune system in rats with different levels of exhaustive exercise. Methods: Fifty-six male Sprague-Dawley (SD) rats were randomly divided into a blank group (n=8), an exhaustive group (n=24), and a moxibustion group (n=24). The exhaustive group was randomly divided into a 1-time exhaustive group, a 4-time exhaustive group and a 7-time exhaustive group, with 8 rats in each group. According to the treatment time, the moxibustion group was randomly divided into a 1-time moxibustion group, a 4-time moxibustion group and a 7-time moxibustion group, with 8 rats in each group. Rats in the exhaustive groups and the moxibustion groups were subjected to replicating the exhaustive swimming models. Rats in each moxibustion group received mild moxibustion for 15 min immediately after the exhaustive modeling, once every other day. Twenty-four hours after the corresponding exhaustive exercise, the rats in each group were tested for the levels of serum immunoglobulin (Ig) G, IgA, IgM and acid phosphatase (ACP), and the morphological changes of spleen tissues were observed. The level of IgA was detected by immunoturbidimetric assay, and the levels of IgG, IgM and ACP were detected by en zyme-linked imm uno sorb ent assay (ELISA). Results: Compared with the 1-time exhaustive group, swimming time of rats in the 4-time exhaustive group was significantly proIonged (P<0.01), and swimming time of rats in the 7-time exhaustive group was significantly shortened (P<0.01). Compared with the 7-time exhaustive group, exhaustive swimming time of rats in the 7-time moxibustion group was significantly proIonged (P<0.01). Compared with the blank group, the IgG level in the 1-time exhaustive group was significantly decreased (PvO.Ol), and the levels of IgG, IgA and IgM in the 4-time exhaustive group and the 7-time exhaustive group were all sign ifica ntly decreased (P<0.05 or PvO.Ol), while the ACP level was in creased sign ifica ntly (both P<0.01). Microscopically, the number of splenic corpuscles in the 1-time exhaustive group was reduced;the center of some splenic corpuscles in the 4-time exhaustive group was damaged;the number of splenic corpuscles in the 7-time exhaustive group was reduced, and there was no obvious germinal center. Compared with the 4-time exhaustive group, the IgA level in the 4-time moxibusti on group was sign ifica ntly in creased (P<0.01), and the ACP level was sign ifica ntly decreased (P<0.01). Compared with the 7-time exhaustive group, the levels of IgG, IgA and IgM in the 7-time moxibustion group were sign ifica ntly in creased (all PvO.Ol), and the ACP level was sign ifica ntly decreased (P<0.01). Microscopically, the nu mber of splenic corpuscles in the 1-time moxibustion group was reduced;the center of some splenic corpuscles in the 4-time moxibustion group was damaged together with hyperplasia of some splenic corpuscles;blast cells were proliferated in the center of some splenic corpuscles in the 7-time moxibustion group. Conclusion: Moxibustion at Shenque (CV 8) can improve the levels of IgG, IgA and IgM, reduce the ACP level, repair damaged spleen tissues, and enhance the immunity of the body to some extent in the Iong-term fatigue rats.
基金supported by the National Natural Science Foundation of China (Grant No. 61141013)the Key Medical Talents of Jiangsu Province(Grant No. RC2011090)+1 种基金the Natural Science Foundation of Jiangsu Province (Grant No. SBK201120268)the 333 Program for High Level Talents of Jiangsu Province (Grant No. 2011III-2640)
文摘Liver transplantation is an established therapy for end-stage liver diseases. Graft rejection occurs unless the recipient receives immunosuppression after transplantation. This study aimed to explore the mechanism of acute rejection of liver allografts in rats pre-treated with total body irradiation to eliminate passenger lymphocytes and to define the role of CD4+CD25+ regulatory T cells in the induction of immunotolerance in the recipient. Male Lewis rats were used as donors and male DA rats were re- cipients. Rats were randomly assigned to the following four groups: control group, homogeneity liver transplantation group, idio-immunotolerance group and acute rejection group. After transplantation, the survival time of each group, serum alanine aminotransferase, total bilirubin levels, number of Foxp3+CD4+CD25+ regulatory T cells, expression of glucocorticoid-induced tumor necrosis factor receptor on T cell subgroups, histopathology of the hepatic graft and spleen cytotoxic T lymphocyte lytic activity were measured. In the acute rejection group, where donors were preconditioned with total body in'adiation before liver transplantation, all recipients died between day 17 and day 21. On day 14, serum alanine aminotransferase increased signifi- cantly to (459.2±76.9) U L^- 1, total bilirubin increased to (124.1±33.7) μmol L-1 (P〈0.05) and the ratio of Foxp3+CD4+CD25+ regulatory T cells decreased significantly to 1.50%±0.50% (P〈0.05) compared with the other groups. Analysis of the T cell subpopulations in the acute rejection group varied from the other groups. Histological analysis showed typical changes of acute rejection in the acute rejection group only. Preconditioning of the donors with total body irradiation eliminated passenger lymphocytes of the liver graft, and thus affected the course of tolerance and induced acute rejection after liver transplantation.
基金supported by the Major New Drug Discovery Science and Technology(2012ZX09303013)the National Basic Research Program of China(2011CB910700)+4 种基金National Natural Science Foundation of China(81271834)China Postdoctoral Science Foundation(20100471238)China Postdoctoral Science Special Foundation(201104516)National Key Technology R&D Program of China(2012ZX10001003)the Postdoctoral Science Foundation of Central South University,Science and Technology Commission of Shanghai Municipality(11DZ2292900)
文摘Elucidation of the mechanisms of liver fibrogenesis is important to treat liver fibrosis.In this study,we established rat models of liver fibrosis with stages from 0–1,2,and 3–4 to 4 at 2,4,6,and 8 weeks,respectively,by injection of pig serum.Liver fibrogenesis was detected by Masson’s trichrome staining.Rat non-parenchymal cells(NPCs)were enriched 4-fold by Percoll density gradient centrifugation.Protein extracts from NPCs were prepared at 4 and 8 weeks,separated by two-dimensional electrophoresis,and then stained with Coomassie Blue G-250.At 4 weeks,we identified 18 non-redundant differentially expressed proteins of which protein disulfide-isomerase associated protein 3(PDIA3)and NDUV showed consistent expression at protein and mRNA levels from 4 to 8 weeks.PDIA3 was found to be down-regulated by Western blotting in the rat model and immunohistochemically in human liver.Our results revealed important aspects of the pathogenesis/progression of liver fibrosis and demonstrated important changes in protein expression levels of NPCs at various stages of liver fibrosis.
基金Supported by the National Natural Science Foundation of China(No.30873244) in Guangzhou University of Traditional Chinese Medicine
文摘OBJECTⅣE: To investigate the protective role of Sijunzi decoction in neuromuscular junction(NMJ)and muscle cell mitochondria ultrastructure; as well as its effects on the amount of adenosine triphosphate(ATP) and the activities of mitochondrial respiratory chain complexes I, Ⅱ, Ⅲ, and Ⅳ in autoimmune myasthenia gravis rats.METHODS: An experimental autoimmune myasthenia gravis(EAMG) rat model was established by inoculating rats with acetylcholine receptors extracted from Torpedo. Rats were divided into three groups: model, prednisone, and Sijunzi decoction, and were fed physiological saline, prednisone, or Sijunzi decoction, respectively. NMJ and muscle cell mitochondria ultrastructure were observed by transmission electron microscope. The amount of ATP was assessed by high performance liquid chromatography. The activities ofmitochondrial respiratory chain complexes I, Ⅱ, Ⅲ,and Ⅳ was determined using the Clark oxygen electrode method.RESULTS: In the model group, there were sparse muscle fibers, with decreased mitochondria, and sparse, diffluent, or absent NMJ folds. After intervention with Sijunzi decoction, the above pathology changes were improved: muscle fiber structure was clear and complete; the mitochondria count was higher; and the NMJ structure was close to normal. Gastrocnemius muscle mitochondria in the model group produced significantly less ATP than those in the prednisone group(P<0.01). Conversely, the ATP of Sijunzi decoction group was significantly higher than prednisone group(P<0.01). The activities of gastrocnemius muscle mitochondrial respiratory chain complexes I, Ⅱ, Ⅲ, and Ⅳ in both the prednisone and Sijunzi decoction groups was dramatically higher compared with the model group(P<0.05). The activities of complexes I and Ⅲ in the Sijunzi decoction group were significantly higher than those in the prednisone group(P<0.05), but there was no obvious difference in complex Ⅱ or Ⅳ activities between the two groups(P>0.05).CONCLUSION: Sijunzi decoction improved pathological changes in muscle mitochondria and NMJ,enhanced the amount of ATP in gastrocnemius muscle mitochondria, and improved the activities of respiratory chain complexes I, Ⅱ, Ⅲ, and Ⅳ(especially I and Ⅲ) of the EAMG rats.
基金This work was supported by the National TCM Leading Talents Support Program-Qihuang Scholars(国家中医药领军人才支持计划-岐黄学者)National Program on Key Basic Research Project“973 Program”(国家重点基础研究发展计划“973计划项目”,No.2015CB554501)+1 种基金Projects of National Natural Science Foundation of China(国家自然科学基金项目,No.82074551,No.81704176)Youth Project from Department of Science and Education,Shanghai Health Commission(上海市卫健委科教处青年基金,No.20194Y0013).
文摘Objective:To observe the anti-inflammatory effect,as well as the effect on the expression of microtubule-associated protein light chain 3B(LC3B)and Beclin-1 of herbal cake-partitioned moxibustion in rats with experimental autoimmune thyroiditis(EAT).Methods:Female Sprague-Dawley rats were randomly divided into a normal group and a modeling group.The EAT rat model was prepared by a combination of antigen immunization plus iodine agent induction.After the model was prepared,rats in the modeling group were randomly and equally divided into a model group and a herbal cake-partitioned moxibustion group.In the herbal cake-partitioned moxibustion group,moxibustion was alternately applied to two groups of points[Dazhui(GV14)-Mingmen(GV4)and Tiantu(CV22)-Guanyuan(CV4)],and the treatment continued for 30 d.Rats in the normal and model groups were only fixed identically without intervention.Histopathological manifestations of thyroid glands were observed by hematoxylin-eosin staining;the concentrations of thyroid peroxidase antibodies(TPOAb),thyroglobulin antibodies(TGAb),interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay;real-time fluorescence quantitative polymerase chain reaction and immunohistochemistry were used to detect the mRNA and protein expression of autophagy-related factors LC3B and Beclin-1 in thyroid tissue.Results:There were massive follicular destruction,lymphocytic infiltration,and interstitial fibrous tissue hyperplasia of the thyroid glands in the model group.Some follicles of the thyroid glands were destroyed with few lymphocyte infiltrations and fibrous tissue hyperplasia in the moxibustion group.Compared with the normal group,the concentrations of serum TPOAb,TGAb,IL-1β,IL-6,and TNF-αwere increased in the model rats(P<0.05);the mRNA and protein expression levels of LC3B and Beclin-1 in thyroid tissue were reduced in the model group(P<0.05).Compared with the model group,the concentrations of serum TPOAb,TGAb,IL-1β,IL-6,and TNF-αwere reduced in the herbal cake-partitioned moxibustion group(P<0.05);the mRNA and protein expression levels of LC3B and Beclin-1 in thyroid tissue were increased in the herbal cake-partitioned moxibustion group(P<0.05).The mRNA and protein expression of LC3B and Beclin-1 in thyroid tissue was negatively correlated with the serum levels of TPOAb and TGAb.Conclusion:Herbal cake-partitioned moxibustion reduces the inflammatory response in the thyroid glands of EAT rats and lowers the levels of serum TPOAb and TGAb.This may be related to the regulation of mRNA and protein expression of the autophagy-associated factors LC3B and Beclin-1 in rat thyroid tissue.