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Objective: To observe the effect of diosgenin on aplastic anemia (AA) mice and peroxisome proliferator activated receptor γ (PPARγ), CCAAT-enhancer binding protein α (C/EBPα), Adiponectin, Leptin, in order to discuss the potential mechanism of bone marrow mesenchymal stem cells in the process of adipemia. Method: BALB/c mice were randomly divided into control group and model group. The model group was established by 60Coy irradiation combined with tail vein infusion with lymphatic suspension cells of DBA/2 mice. After successful evaluation of the model, the mice were randomly divided into 6 groups:model group, low, medium and high-dose diosgenin groups (37.44,74.88,149.76 mg·kg-1·d-1), cyclosporine group (23.5 mg·kg-1·d-1), and tripterygium glycoside group (9.36 mg·kg-1·d-1), and given corresponding drugs by gavage for 14 days. After the intervention, the peripheral blood of mice in each group was detected, and bone marrow smears were collected to evaluate the proliferation of bone marrow. Bone marrow mesenchymal stem cells (BMMSCs) were isolated and cultured by adherent method and induced by adipogenesis. The mRNA and protein expressions of PPARγ, C/EBPα, Adiponectin, Leptin in BMMSCs were detected by quantitative real-time fluorescent quantitative polymerase chain reaction(Real-time PCR) and Western blot. Result: The white blood cell (WBC), hemoglobin (HGB) and blood platelet (PLT) in peripheral blood of model group were significantly lower than those of normal group (PPγ, C/EBPα, Adiponectin and Leptin in BMMSCs of the model group increased significantly (Pγ, C/EBPα, Adiponectin and Leptin in the middle-dose group diosgenin decreased obviously, which was better than those of Tripterygium glycoside group (P0.05). Conclusion: Diosgenin can promote the recovery of peripheral blood in aplastic anemia mice and improve the hematopoiesis of bone marrow. Diosgenin can reduce the expressions of PPARγ and C/EBPα, the formation of adipocytes and the secretion of Adiponectin and Leptin in adipocytes, and effectively inhibit the process of adipose tissue derived from bone BMMSCs.
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Objective To explore the effects of C/EBPα knockout in podocyte on diabetic nephropathy and its mechanisms.Methods C/EBPαloxp/loxp mice were crossed with podocin-cre mice to obtain F1 hybrids and then propagated until homozygous mice (C/EBPαf/f) were obtained.Diabetic nephropathy (DN) models were established by low-dose streptozotocin (STZ,100 mg/kg) administration after 25 weeks of normal diet or 45％ high-fat diet treatment,and biochemical indicators of blood and urea were measured.The morphological characteristics and the proteins regulating oxidative stress and mitochondrial function were detected.Results The type 2 DN models were successfully constructed based on transgenic mice.The kidneys of 8-month-old C/EBPαf/f mice did not show obvious morphological changes,but after constructing DN models,they showed obvious renal impairment,inflammation and oxidative stress.Compared with wild-type DN mice,the protein levels of nephrin and E-cadherin in DN C/EBPαf/f mice with DN were significantly decreased (P ＜ 0.01);fibronectin and Nrf2 protein levels were all increased (all P ＜ 0.05).Keap1,phospho-AMPK and mitochondrial function related genes Pgc-1α protein levels were all decreased (all P ＜ 0.05).Conclusion Podocyte C/EBPα knockout exacerbates diabetic nephropathy by promoting fibrosis and inhibiting Pgc-1α-mediated mitochondrial antioxidant function.
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Aim To explore the effect of water extract of Radix Isatidis(WERI)on proliferation and differentiation of 3T3-L1 preadipocytes.Methods 3T3-L1 preadipocytes were cultured to explore the effect of WERI on their proliferation measured by MTT and flow cytometry.Oil red O staining was applied to investigate the effect of different concentrations of WERI on the differentiation of 3T3-L1 preadipocytes.
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Objective To explore the influences of proliferation and differentiation of preadipocytes on high fat diet-induced obesity and obesity prevention through exercises in rats.Methods Forty male Sprague-Dawley rats were divided into a normal diet control group (C,n=8),a normal diet with exercises group (E,n=8),a high fat diet control group (H,n=14) and a high fat diet with exercises group (HE,n=10).Group C and group H kept sedentary,while group E and group HE underwent treadmill exercises at about 75％VO2max level.After 12 weeks of intervention,the body weight,epididymal,perirenal and retroperitoneal fat mass were recorded,and the total fat mass was determinated.Stereology method was used to calculate the number and average diameter of fat cells.Western Blotting was conducted to measure the expression of PPARγ and C/EBPα protein in adipose tissues.Results (1) The body weight and total fat mass of group H were significantly higher than those of group C (P＜0.01).Perirenal,retroperitoneal fat and total fat mass of group E and HE were significantly lower than those of group C and H respectively (P＜0.01).(2) The total fat cell number of group H was significantly higher than that of group C.The average diameter of fat cells in perirenal and retroperitoneal fat pads of group E were significantly lower than that of group C (P＜0.05,P＜0.01),while that of group HE in epididymal,perirenal and retroperitoneal fat pads was significantly lower than that of group H (P＜0.01).(3) Compared with group C,the expression of PPARγ protein of group E and H increased significantly in epididymal,perirenal and retroperitoneal fat pads (P＜0.01).The expression of C/EBPα protein of group H was significantly higher than that of group C in epididymal and perirenal fat pads (P＜0.01),the expression of C/EBPα protein of group E was significantly higher than that of group C in perirenal and retroperitoneal fat pads (P＜0.01),while the expression of C/EBPα protein of group HE was significantly lower than that of group H in perirenal fat pads(P＜0.01).Conclusion The proliferation and differentiation of preadipocyte was enhanced in the development of high fat diet induced obesity and exercises to prevent obesity,but its role and mechanisms were different.The high fat diet increases the number of fat cells which is a compensatory mechanism for the body to adapt to fat accumulation,while exercises might promote cell renewal and decrease the average size of fat cells which is an adaptive mechanism to improve fat storage.
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Objective To investigate the expression of SOX4 and C/EBPα mRNA in chronic myeloid leukemia (CML) and their clinical significances. Methods Bone marrow samples from 68 cases of CML including 57 newly diagnosed patients and 11 patients treated with imatinib were collected, and peripheral blood mononuclear cells from 30 healthy people were collected as healthy control. The expression of SOX4 and C/EBPαmRNA and protein levels were detected by RT-PCR and Western blot, respectively. The relations between the expression of SOX4 and C/EBPα and the influences of imatinib on SOX4 and C/EBPα were analyzed. Results The expression level of SOX4 mRNA was increased in newly diagnosed CML patients compared with that of normal control group (6.545 5±1.495 2 vs. 0.059 6±0.018 8, t=3.139, P=0.002 3), but the expression level of C/EBPαmRNA was significantly decreased (0.238 8±0.033 8 vs. 0.810 5±0.056 2, t=9.240, P0.05). The expression level of SOX4 mRNA in 5 patients treated with imatinib was decreased (0.120 6 ±0.044 9 vs. 0.557 9±0.144 8, t=2.885, P=0.020 4), and the expression level of C/EBPαmRNA was increased (0.330 3±0.042 4 vs. 0.150 5±0.046 5, t=2.855, P=0.021 3). The expression level of SOX4 mRNA in 6 patients who developed blast phase during the treatment of imatinib was increased (0.469 9±0.123 0 vs. 0.050 2±0.036 6, t=2.370, P=0.039 3), and the expression level of C/EBPα mRNA was decreased (0.197 9 ±0.064 7 vs. 0.378 7±0.042 9, t=2.327, P=0.042 3). The expression of SOX4 mRNA was negatively correlated with C/EBPα mRNA (t=-0.554 6, P=0.002 8). Conclusions In newly diagnosed CML, the expression level of SOX4 is increased, C/EBPα is decreased compared with that of healthy control, and both have negative correlation. In the patients in blast phase after imatinib treatment, SOX4 gene is up-regulated, and C/EBPα is down-regulated. C/EBPα-SOX4 axis may play a role in the occurrence and development of CML. SOX4 may be a new molecular target for the treatment of CML.
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Objective To investigate the alteration of C/EBP α,C/EBP β and endoplasmic reticulum stress (ERS) related molecules (IRE1α and sXBP1) expression in pancreatic tissues in rats with hypertriglyceridemia related acute pancreatitis.Methods Ninety six Sprague Dawley (SD) rats were divided into 4 groups:control group,hypertriglyceridemia (HTG) group (n =24,fed with high fat diet for 2 weeks),AP group (n =24),HTG + AP group (n =24),and AP was induced by peritoneal injection of cerulein.The rats were sacrificed at 3,6,9,24 h after AP induction,respectively.The pathological changes of the pancreatic tissues were observed and scored by HE staining.Plasma levels of IL-1β,IL-6 and TNF-α were measured by ELISA.The expression of IRE1α,sXBP1,C/EBPoα,C/EBPβ mRNA were analyzed by real time PCR.The expressions of IRE1α,sXBP1,NF-kB,C/EBPα and C/EBPβ protein were determined by Western Blot.The expressions of C/EBPα and C/EBPβ proteins were also determined by immunohitochemistry.Results After two weeks of high fat diet,serum levels of triglyceride(TG) and total cholesterol (TC) in HTG group,HTG + AP group were much higher than those of the control group,and the difference was statistically significant (P ＜ 0.05).The pancreatic tissue injury was more severe in HTG + AP group,particularly at 9 h (P ＜ 0.05).And plasma IL-1β,IL-6 and TNF-α levels were also much higher in HTG + AP group when compared with that of AP group,the differences were all significant at 9 h (P=0.011;P=0.034;P =0.027).After AP induction,IRE1,sXBP1,C/EBP and C/EBPβ mRNA began to be up-regulated at 3 h,and IRE1 mRNA reached the highest level at 24 h,sXBP1 mRNA at 9 h,while C/EBP and C/EBPβ mRNA reached the highest level at 6 h.Compared with AP group,IRE1,sXBP1,C/EBP and C/EBPβ mRNA levels were much higher in HTG + AP group.In addition,as to IRE1 and sXBP1 mRNA,the difference was significant at 3,6,9,24 h,and C/EBP mRNA at 6,9,24 h,C/EBPβ mRNA at 6 and 9 h (P ＜ 0.05).After AP induction,IRE1α,sXBP1 and NF-kB proteins in the pancreatic tissue began to be up-regulated at 3 h,and all reached the highest level at 9 h.IRE1α,sXBP1 and NF-kB proteins were up-regulated more obviously in HTG + AP group,and the up-regulation in HTG + AP group was higher than that in AP group,and the high expressions of C/EBPα and C/EBPβ proteins could only be detected at 6 and 9 h in the HTG + AP group,while there was no expression detected in AP group.Conclusions C/EBPα,C/EBPβ,IRE1α and sXBP1 may be involved in the pathogenesis of HTG related AP,and IRE1α/sXBP1 pathway and C/EBPoα,C/EBPβ may mediate the pathologic injury and inflammation process of HTG related AP.
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Aim To explore the effect and mechanism of resveratrol (Res) on proliferation and differentiation of murine 3T3-L1 preadipocytes.Methods 3T3-L1 preadipocytes were cultured and treated with resveratrol in different dosages.Cell proliferation was analyzed by WST-1 method. Oil red O staining method and spectrophotography were applied to analyze the degree of differentiation. Real-time PCR was applied to detect the mRNA expression of adiponectin and leptin. Western blot was applied to detect the expression of silent information regulator 1 (Sirt1),peroxisome proliferator activated receptor γ (PPARγ) and CCTTA enhancer binding proteinα (C/EBPα).Results Res inhibited proliferation of murine 3T3-L1 preadipocytes in a time-dose dependent manner.The expression levels of adiponectin and leptin mRNA were decreased, and Res also inhibited 3T3-L1 preadipocytes to differentiate into mature adipocytes. Res increased the expression levels of Sirt1 and decreased the expression levels of PPARγ and C/EBPα.Conclusions Resveratrol can inhibit the proliferation and differentiation of 3T3-L1 preadipocytes.The underlying mechanisms may include enhancing expression of Sirt1 and inhibiting expression of PPARγ,C/EBPα which are related to cell differentiation.