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Objective:To investigate the effect of heme oxygenase-1 (HO-1) modified rat bone marrow mesenchymal stem cells (BMSCs) on fibrotic rats.Methods:110 male SD rats aged 6-8 weeks were selected randomly divided into model group, BMSCs group and HO-1/BMSCs group with 11 rats in each group after intraperitoneal injection of CCl 4, PBS, BMSCs and HO-1/BMSCs were injected respectively. Another 11 rats were selected as control group. After 4 weeks of intervention, tracer experiment was used to detect the location of BMSCs. Rats in each group were executed, and liver function were detected by biochemical analyzer, liver fibrosis indexes were detected by ELISA, liver histopathology were detected by HE and Sirius red staining. Immunohistochemistry, Western blot and RT-PCR were used to detect the protein and mRNA expression of E-cadherin and Vimentin. Results:The rat fibrosis model was successfully established. Tracer experiment showed that BMSCs were implanted in rat liver after transplantation. Compared with the model group, the liver function and liver fibrosis indexes of BMSCs group and HO-1/BMSCs group were improved, and Ishak score and stage were significantly decreased, and HO-1/BMSCs group was superior to BMSCs group. The expression of E-cadherin in HO-1/BMSCs group (0.92±0.21), (0.84±0.03) were higher than those in BMSCs group [(0.54±0.16), (0.53±0.04)] and model group [(0.49±0.06), (0.11±0.06)] both at protein and mRNA level, while protein and mRNA level of Vimentin (1.21±0.23), (3.82±0.80) were lower than that in BMSCs group [(1.32±0.17), (6.39±0.75)] and model group [(1.41±0.18), (16.94±1.30)]. The difference was statistically significant ( P<0.05). Conclusion:HO-1/BMSCs can improve liver function and liver fibrosis in fibrotic rats more effectively than BMSCs alone. The mechanism was possibly through inhibiting liver epithelial mesenchymal transition.
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Objective:To investigate the effect and mechanism of bone marrow mesenchymal stem cells (BMMSCs) on liver transplantation with 50% reduced-size in rat models.Methods:For 40 normal male Brown Norway(BN) and Lewis rats weighing 210-250 g were included respectively to generate the acute rejection models following 50% reduced-size liver transplantation in rats. The recipients were divided into BMMSCs group ( n=20) and normal saline group ( n=20). Healthy male BN rats were used to prepare BMMSCs. Transplanted liver tissues were collected at 0h, 1d, 3d, 7d post the transplantation for further analysis. Pathological changes and the extent of rejection were evaluated under the light microscope. The levels of microtubule-associated protein 1 light 3 (LC3) and autophagy regulator Beclin-1 proteins were detected by immunohistochemical analysis and Western blotting. Results:Rejection activity indices of the normal saline group at 0d, 3d, 7d after the surgery were (2.33±0.58), (4.00±0.00), (6.33±0.58). The BMMSCs group were (2.10±0.58), (3.73±0.58), (5.67±1.15), which was decreased comparing with normal saline group, difference was statistically significant ( P<0.05). On the 1d, 3d, 7d after the transplantation, compared with normal saline group, expression of autophagy-related proteins LC3 and Beclin-1 in BMMSCs group was increased ( P<0.05). Conclusions:It showed that autophagy has an effect on the protection of BMMSCs liver graft of rats.
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Objective:To investigate the effects of heme oxygenase-1 (HO-1)-modified rat bone marrow mesenchymal stem cells (BMMSCs) on T lymphocyte subsets in cirrhotic rats.Methods:A rat model of liver cirrhosis was established by intraperitoneal injection of carbon tetrachloride (CCL 4) in 110 rats. These rats were randomly divided into three groups, model group, BMMSCs group and HO-1/BMMSCs group, and injected with PBS, BMMSCs and HO-1/BMMSCs through dorsal penile vein, respectively. Another 10 rats were selected as control group. All rats were executed four weeks after intervention. Pathological changes in liver tissues were observed with HE and Sirius red staining. Serum albumin (ALB) and alanine aminotransferase (ALT) were detected by biochemical analyzer. Serum hyaluronidase (HA) and collagen type Ⅳ (Ⅳ-C) were detected by ELISA. T lymphocyte subsets in peripheral blood and spleen were detected by flow cytometry. Results:The rat model of cirrhosis was successfully established. Compared with the model group and BMMSCs group, the HO-1/BMMSCs group had significantly lower Ishak score and disease stage ( P<0.05), increased serum ALB level and CD4 + T/CD8 + T cell ratio ( P<0.05), and decreased serum ALT, HA and Ⅳ-C levels and Th17/Treg ratio ( P<0.05). Conclusions:HO-1/BMMSCs could improve liver function and liver fibrosis in cirrhotic rats more effectively than BMMSCs alone. The mechanism was possibly through regulating the immunomodulatory function of T lymphocyte subsets in cirrhotic rats.
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Objective To explore the role and mechanism of thymosin β4 (Tβ4) in the treatment of non-alcoholic fatty liver disease (NAFLD).Methods Forty male C57BL/J6 mice were divided into normal group,NAFLD group,low dose Tβ4 group and high dose Tβ4 group with 10 mice in each group.NAFLD mice model was established by feeding with high fat and high sugar diet for 16 weeks.The mice in low-dose Tβ4 group and high dose Tβ4 group were intraperitonealy injected with Tβ4 at 0.05 mg · kg-1 · d-1 and 0.20 mg · kg-1 · d-1,respectively,for eight weeks.The liver function indexes and serum tumor necrosis factor-α (TNF-α) level were detected;the pathological changes of liver tissue were observed under optical microscope and non-alcoholic fatty liver disease activity score (NAS) was evaluated.The protein expression levels of nuclear factor-κB p65 (NF-κB p65) and nuclear factor κB inhibit protein a (IκBa) at the protein level in liver tissue were measured by Western blotting method.The expression of TNF-α in liver tissue was detected by immunohistochemistry.Mean integral absorbance (MIA) was calculated.T test was performed for groups comparison.Results The levels of alanine aminotransferase (ALT),γ-glutamine transferase (GGT) and serum TNF-α levels of high dose Tβ4 group were all lower than those of NAFLD group ((28±17) U/L vs.(76±29) U/L,(61±39) U/L vs.(102±56) U/L,(144.1± 48.2) ng/L vs.(187.3±58.8) ng/L,respectively),and the differences were statistically significant (t=4.52,2.78 and 2.30,all P＜0.05).The NAS of low dose Tβ4 group and high dose Tβ4 group were both lower than that of NAFLD group (3.7±40.4,2.3±0.3 vs.4.6±0.3),and the differences were statistically significant (t=5.69 and 17.14,both P＜0.01).The relative expression level of Tβ4 protein of NAFLD group was lower than that of normal group (0.2±0.1 vs.1.4±0.6),and the difference was statistically significant (t=6.24,P＜0.01).The relative expression levels of Tβ4 and IκBa of high dose Tβ4 group were higher than those of NAFLD group (1.0±0.3,0.5±0.3 vs.0.2±0.1),and the differences were statistically significant (t=8.00 and 3.00,both P＜0.01).The relative expression level of NF-κB p65 in liver tissue of high dose Tβ4 group was lower than that of NAFLD group (0.6±0.3 vs.1.5±0.7),and the difference was statistically significant (t=3.74,P＜0.01).The MIA of high dose Tβ4 group was lower than that of NAFLD group (0.4±0.2 vs.0.7±0.3),and the difference was statistically significant (t=2.63,P＜ 0.01).Conclusion Tβ4 can effectively treat NAFLD probably through inhibiting the NF-κB pathway.
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Objective To investigate the expression of humanβ-defensin 2 (HBD-2) in gastric mucosa of Helico-bacter pylori (H. pylori) associated gastric mucosa-associated lymphoid tissue (MALT) lymphoma, and the role of HBD-2 in gastric MALT lymphoma. Methods Forty gastric mucosa specimens from patients with H. pylori associated gastric MALT lymphoma were collected. And 36 gastric mucosa specimens from chronic superficial gastritis without H. pylori infection were included as control group. The expression of HBD-2 was detected by immunohistochemistry staining. Results The ex-pression of HBD-2 was significantly higher in H. pylori associated gastric MALT lymphoma than that of control group. (P＜0.01). The expression of HBD-2 was significantly decreased after the eradication of H. pylori (P＜0.01). The expression of HBD-2 was significantly higher in H. pylori associated gastric MALT lymphoma than that of lymphoma cells (P＜0.01). There was no expression of HBD-2 in lymphoma cells. Conclusion HBD-2 is possibly involved in the pathogenesis of H. pylori associated gastric MALT lymphoma. But whether it has anti-tumor effect is not clear.