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Background Mitochondrial dynamin-related protein 1 (DRP1) regulates mitochondrial division and plays an important role in maintaining hepatocyte function. However, the role of DRP1 in cadmium exposure-induced maternal liver damage in pregnant mice remains unclear. Objective To investigate the role and mechanism of DRP1 in maternal liver damage induced by cadmium exposure during pregnancy. Methods This study consisted of animal experiments and cell experiments. (1) Animal experiments. Mice at 14 days of gestation were randomly divided into three groups: a control group, a low-dose cadmium group (LCd group: 2.5 mg·kg−1), and a high-dose cadmium group (HCd group: 5 mg·kg−1). The pregnant mice were intraperitoneally injected with cadmium chloride (CdCl2) for 6 and 24 h in the next morning. The weights of pregnant mice, uterus, maternal liver, and fetal mice were recorded after sacrifice. Serum and liver of pregnant mice were collected, the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were detected, and liver tissues were stained with HE to observe changes in liver function and liver tissue structure. The expressions of oxidative phosphorylation-related proteins, hypoxia inducible factor-1α (HIF-1α) and DRP1 proteins in liver of pregnant mice were detected by Western blotting. (2) Cell experiments. AML12 cells were treated with CdCl2 (10 μmol·L−1) for 0, 2, 6, 12, and 24 h. The expressions of oxidative phosphorylation-related proteins, DRP1, and hypoxia inducible factor-1α (HIF-1α) proteins were detected. AML12 cells were pretreated with DRP1 inhibitor Mdivi-1 for 1 h and then CdCl2 (10 μmol·L−1) for 12 h to detect the expression of oxidative phosphorylation-related proteins and DRP1 protein. AML12 cells were treated with Hif-1α siRNA for 48 h and CdCl2 (10 μmol·L−1) for 6 h to detect the expression of HIF-1α and DRP1 proteins. Results The results of animal experiments showed that cadmium exposure in pregnant mice had no effects on maternal liver weight and liver coefficient. However, the histomorphological changes and necrosis in hepatocytes were observed. Compared with the control group, the serum ALT and AST levels of pregnant mice in the LCd group were significantly increased after 6 h (P＜0.05), and the levels in the HCd group were significantly increased after 6 and 24 h (P＜0.05). Cadmium exposure during pregnancy significantly up-regulated HIF-1α and DRP1 expressions and down-regulated the expressions of oxidative phosphorylation-related proteins in maternal livers. In vitro cell experiments showed that the expressions of oxidative phosphorylation-related proteins was significantly decreased and HIF-1α and DRP1 protein expressions were significantly increased in the AML12 cells treated with CdCl2 for 6 h. Mdivi-1 pretreatment significantly antagonized the inhibitory effect of cadmium on the expressions of oxidative phosphorylation-related proteins in AML12 cells, while Hif-1α siRNA pretreatment significantly antagonized the up-regulative effect of cadmium on DRP1 expression in AML12 cells. Conclusion Cadmium exposure in pregnant mice may up-regulate DRP1 expression by activating HIF-1α signaling, then inhibit oxidative phosphorylation level of hepatic cells, and ultimately lead to maternal liver damage.
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Objective To study the effects of different doses of T-2 toxin on organs and bones of BALB/c pregnant mice and their offspring mice.Methods Sixty female SPF BALB/c mice at the age of 6-weeks were mated with 30 6-week-old male SPF BALB/c mice.Female mice were divided into control,low dose,medium dose and high dose groups according to body weight via the random digital table method,with 15 mice in each group.After mating with male mice,the pregnant mice in each group were 14,10,9 and 15,respectively.Nutritional interventions (feed:the doses of T-2 toxin were 0,600,1 200 and 2 400 ng/g,respectively) were initiated from the gestation day 0 until the first generation mice were grown up (6-weeks-old).The growth status and organ coefficient (heart,liver,kidney,thymus,spleen and brain) of the two generations in each period were recorded.Skeletal X-ray photographs of the two generations were taken by digital radiography.The histopathological changes in the organs (liver,thymus,spleen and epiphyseal cartilage) of the two generations were observed under light microscope.Results Among the pregnant mice,there were no significant differences in organ coefficients (P ＞ 0.05).No abnormalities were observed in each group of skeletal X-ray photographs.In the female generation mice,there were no significant differences in the coefficients of heart,liver and kidney (F =0.233,2.196,0.430,P ＞ 0.05),while there were significant differences in the coefficients of thymus,spleen and brain (F=3.683,3.148,4.498,P ＜ 0.05),and the thymus coefficient of medium dose group was higher than that of control group;the thymus coefficient of high dose group was lower than that of other three groups;the spleen coefficients of the three dose groups were higher than that of control group;the brain coefficients of the three dose groups were lower than that of control group (P ＜ 0.05).In the male generation mice,there were no significant differences in the coefficients of thymus and brain (F =2.447,1.620,P ＞ 0.05),while there were significant differences in the coefficients of heart,liver,kidney and spleen (F =5.339,2.738,11.435,2.872,P ＜ 0.05),and the heart coefficient of high dose group was lower than that of control group and low dose group;the coefficients of liver and kidney in medium dose and high dose groups were lower than those in control and low dose groups;the spleen coefficients of the three dose groups were higher than that of control group (P ＜ 0.05).The pathological changes of liver,thymus,spleen and epiphyseal cartilage were found in dose exposure groups;epiphyseal hyperplasia was found in the skeletal X-ray photographs of medium and high dose groups.Conclusion T-2 toxin has no significant effects on pregnant mice,but it could cause damage to the organs and epiphyseal plate cartilage of the first generation,and the location of the injury is related to gender.
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T-2 toxin is a secondary fungal metabolite that belongs to the trichothecene mycotoxin family,it is the most toxic in class A trichothecene mycotoxin.T-2 toxin can induce apoptosis in cells bearing high proliferating activity.The mycotoxin readily passes the placenta and is distributed to embryo tissues,which results in fetal brain damage,bone malformation,thymic atrophy and even death.This article reviewed the effects of T-2 toxin on immune system,blood system and cell toxicities in pregnant mice and generation mice.
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Objective:To explore the abortion effect and mechanism induced by Chinese herbal medicine rhubarb extract.Methods:The pregnant mice at day 3 of gestation were Drenched by Aqueous extract of rhubarb at different doses (7,5,2.5 g/kg)once per day for five days,and mices in control group were oral administration of same volume of saline.All mices at 12 day of gestation were sacrificed.estradiol and progesterone levels in serum were detected by Radioimmunoassay;Content of IFN-γ, IL-2 and TNF-αin uterine lysates were measured by ELISA, macrophages in endometrium were determined by immunohistochemical method, mast cells in endometrium were tested by Toluidine blue staining, respectively.Results: showed as follows:With increasing dose of rhubarb extract,abortion rate and resorption rate were gradually increased and was 100%abortion in 7g/kg treatment groups.Estrogen and progesterone content showed a downward trend,IFN-γ,IL-2 and TNF-αin uterine lysates and the numbers of macrophages and mast cells were significantly higher in mices Drenched by Aqueous extract of rhubarb than in the control.Conclusion: These results show that rhubarb extract not only interfere stability of pregnancy state in pregnant mices because of its diarrhea,but also can directly affect endometrial environment in early embryonic mice,Resulting in abortion.
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Objective Uterine epithelial cells were isolated from pregnant mice and cultured in vitro , and exam-ined the effect of CD82 on the expression of integrin αV,β3, E-cadherin and β-catenin in the cells.Methods The uter-ine epithelial cells were primarily isolated from pregnant mouse uterus .The recombinant adenovirus containing mouse CD 82 gene which had been constructed in our lab infected the uterine epithelial cells .Immunocytochemistry was used to detect the protein expressions of integrin αV,β3, E-cadherin and β-catenin in the uterine epithelial cells , which were infected with CD82 adenovirus or not .Results 1.The purity of primary cultured cells was (93.2 ±0.6)%.2.The transfection efficiency of CD82 recombinant adenovirus was ( 92 ±4.5 )%.The adenoviral particles carrying CD 82 gene indeed ex-pressed CD82 gene and protein in the primary uterine epithelial cells after 24 hours or 48 hours.3.The uterine epithelial cells of pregnant mice on d4 expressed integrin αV, β3, E-cadherin and β-catenin.4.In contrast to the control group, when CD82 adenovirus infected cells , the uterine epithelial cells of pregnant mice on d 4 increased the expression of integrinαV,β3 and β-catenin protein, had no significant changes of E-cadherin.Conclusions CD82 may have effect on the ex-pression of integrin αV,β3 and β-catenin in mouse uterine epithelial cells before implantation .