Maternal exposure to a high-fat diet showed unfavorable effects on the body weight, apoptosis and morphology of cardiac myocytes in offspring.

OBJECTIVE: The study intends to explore the functions of maternal high-fat diet exposure on progeny weight and heart. METHODS: Sprague-Dawley (SD) rats, fed on a high-fat diet, were used to establish a model of weight gain before and during pregnancy. The body and cardiac weight of neonatal, 1-month- and 3-month-old rats were measured. The morphology of myocardial cells was observed by hemotoxylin and eosin (H&E) staining. The expression of caspase-3, 8, 9 was measured by qRT-PCR and western blot. RESULTS: Normal pregnant rats, fed on a high-fat diet throughout pregnancy, had a significant increase in body and cardiac weight of their neonates, and more fat deposition in myocardial cells and an increased expression of caspase-3, 8, 9, compared with that of the normal pregnant rats + normal diet group. These phenomena were relieved through later diet control. Pregnant rats, which fed on a high-fat diet throughout pregnancy, showed more adverse effects on neonatal body and cardiac weight, myocardial cell fat deposition, and the expression of caspase-3, 8, 9, compared with pregnant rats exposed to high-fat diet + normal diet and pregnant rats exposed to high-fat diet + normal diet + exercise. These phenomena cannot be fully restored via controlling later diet. CONCLUSIONS: Our results stated that a proper diet before and during pregnancy was important for the cardiac health of offspring.

[Interaction among peroxisome proliferators-activated receptor alpha, cytochrome P450 oxysterol 7α-hydroxylase and estrogen receptor and its association with intrahepatic cholestasis in pregnant rats].

OBJECTIVE: To investigate the relationship between interaction of peroxisome proliferators-activated receptor alpha (PPARα), cytochrome P450 oxysterol 7α-hydroxylase (CYP7B1) and estrogen receptor (ER) and intrahepatic cholestasis in pregnant rats. METHODS: Eighty clean SD pregnant rats were selected and divided into four groups randomly with 20 in each. Since the 13th day of pregnancy, rats in the control group was injected subcutaneously with refined vegetable oil 2.0 ml×kg(-1)×d(-1), those in the low-dose, moderate-dose and high-dose groups received 17-α-ethynylestradiol (EE) 1.0 mg×kg(-1)×d(-1), 1.25 mg×kg(-1)×d(-1) and 1.5 mg×kg(-1)×d(-1), respectively. All rats were sacrificed at the 21(st) day of pregnancy and maternal hepatic tissues were collected. The serum levels of alanine aminotransferase (ALT), aspartate transaminase (AST), total bile acid (TBA) and bilirubin (BIL) were determined by enzyme linked immunosorbent assay (ELISA). The mRNA expressions of PPARα, CYP7B1, ERα and ERß in maternal rat livers were examined by real-time PCR. RESULTS: (1) Biochemical indicators: the serum levels of ALT, AST, TBA and BIL were significantly lower in the control group than in the rest 3 groups, respectively [control group: (41.1 ± 2.8) U/L, (44.4 ± 3.6) U/L, (26.4 ± 5.6) µmol/L and (2.8 ± 0.2) U/L; low-dose group: (48.2 ± 3.4) U/L, (47.9 ± 3.7) U/L, (36.4 ± 4.2) µmol/L and (4.2 ± 0.2) U/L; moderate-dose group: (70.4 ± 5.3) U/L, (68.4 ± 5.6) U/L, (64.3 ± 3.8) µmol/L and (6.2 ± 1.2) U/L; high-dose group: (72.4 ± 7.6) U/L, (70.2 ± 3.8) U/L, (72.4 ± 7.8) µmol/L and (8.2 ± 2.2) U/L, P < 0.05], and those in the moderate or high-dose groups were higher than in the low-dose group (P < 0.05). (2) mRNA expression of ERα and ERß: the mRNA expression of ERα in pregnant rat livers increased in a dose-dependent manner, which were all significantly higher than that in the control group, respectively (low-dose group: 0.76 ± 0.02); moderate-dose group: (0.99 ± 0.04; high-dose group: 1.21 ± 0.01; control group: 0.65 ± 0.01, P < 0.05), but no difference was found among the 4 groups in the mRNA expression of ERß (P > 0.05). (3) mRNA expression of CYP7B1 and PPARα: the mRNA expression of CYP7B1 in pregnant rat livers increased from the low-dose group to the high-dose group, and were all higher than that of the control group (low-dose group: 0.93 ± 0.01; moderate-dose group: 0.99 ± 0.06; high-dose group: 1.22 ± 0.04; control group: 0.75 ± 0.02, P < 0.05). However, the mRNA expression of PPARα decreased from the low-dose group to the high-dose group, and were all lower than that of the control group (low-dose group: 0.83 ± 0.05; moderate-dose group: 0.71 ± 0.02; high-dose group: 0.64 ± 0.03; control group: 1.35 ± 0.05; P < 0.05). CONCLUSIONS: The down regulated mRNA expression of PPARα, caused by higher dose of estrogen, may increase the expression of CYP7B1 due to the ineffectiveness of the inhibition of PPARα on CYP7B1, which may further stimulate the ERα activity and then induce intrahepatic cholestasis. Abnormal expression of PPARα, CYP7B1 and ER may play a role in the pathogenesis of estrogen-induced intrahepatic cholestasis.

[Expression of FXR mRNA, PPAR alpha mRNA and bile acid metabolism related genes in intrahepatic cholestasis of pregnant rats].

OBJECTIVE: To study the expressions of FXR, PPARa and Bile acid metabolism related genes in intrahepatic cholestasis of pregnant rats. METHODS: 60 clean SD pregnant rats were selected and divided randomly into three groups. Since the 13th day of pregnancy rats in control group were injected subcutaneously with refined vegetable oil 2.0 mg/kg/d Rats in no-treated group were injected subcutaneously with the 17-a-ethynylestradiol (EE) 1.25 mg/kg/d until the 17th day. Those rat ih treated group were injected subcutaneously with the 17-a-ethynylestradiol (EE) 1.25 mg/kg/d until the 17th day and then were treated with fenofibrate for another four days until the 21th day. All rats were killed at the 21th day and livers were collected for study. The levels of serum TBA were examined by ELISA. The mRNA expressions of PPARa, FXR, CYP7A1, CYP27A1 and CYP8B1 were examined by real-time PCR. (1) RESULTS: The levels of TBA were significantly higher in no-treated group (68.7+/-4.2)mumol/L and treated group (69.5+/-3.8)mumol/L compared with that of control group (26.6+/-2.3)mumol/L at the 17th day (P value is less than 0.05) and no difference found between treated and no-treated groups (P value is more than 0.05). The levels of TBA were higher in no-treated group (69.4+/-3.7)mumol/L and treated group (48.5+/-4.8)mumol/L as compared to control group (27.1+/-3.2)mumol/L at the 21th day (P value is less than 0.05). The lever of TBA was significantly lower in Treated group compared with No-treated group (P value is less than 0.05). (2) The mRNA expressions of CYP7A1, FXR, CYP27A1 and CYP8B1 increased in No-treated group (1.55+/-0.03, 1.75+/-0.02, 2.45+/-0.01, 2.15+/-0.01, respectively) and were all higher as compared to control group (0.75+/-0.02, 1.25+/-0.03, 0.65+/-0.03, 1.50+/-0.02, respectively) (P value is less than 0.05). However, the mRNA expression of PPARa decreased in No-treated group (0.85+/-0.02) compared with control group (1.45+/-0.02) (P value is less than 0.05). The mRNA expressions of CYP27A1, PPARa and CYP8B1 increased in treated group (1.25+/-0.01, 1.65+/-0.05, 1.65+/-0.02, respectively) and were all higher than that of control group (P value is less than 0.05). CONCLUSION: Abnormal expressions of CYP7A1, FXR, CYP27A1, CYP8B1 and PPARa may play a role in pathogenesis of estrogen-induced intrahepatic cholestasis. Activator of PPARa may be used as therapeutical drug for ICP.

[Study on the association between estrogen receptor alpha gene polymorphism and intrahepatic cholestasis of pregnancy].

OBJECTIVE: To investigate the relationship between estrogen receptor alpha (ERalpha) gene polymorphism and intrahepatic cholestasis of pregnancy (ICP). METHODS: The Xba I and Pvu II polymorphisms in intron 1 of ER alpha gene were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 100 pregnant women with ICP (ICP group) and 100 normal pregnant women (control group). RESULTS: (1) The frequencies of XX, Xx and xx genotypes of Xba I polymorphism were 6%, 33% and 61% respectively in control group, and were 2%, 33% and 65% respectively in ICP group. There was no significant difference in the distribution of Xba I genotypes between the two groups (P > 0.05). The frequencies of the two alleles X and x were 23% and 78% in control group, and were 19% and 82% in ICP group, respectively. There was also no significant difference in the frequency of Xba I alleles between the two groups (P > 0.05). (2) The frequencies of PP, Pp and pp genotypes of Pvu II polymorphism were 18%, 42% and 40% respectively in control group, and were 12%, 53% and 35% respectively in ICP group. There was no significant difference in the distribution of Pvu II genotypes between the two groups (P > 0.05). The frequencies of the two alleles P and p were 39% and 61% in control group, and were 39% and 62% in ICP group, respectively. There was also no significant difference in the frequency of Pvu II alleles between the two groups (P > 0.05). (3) There was also no significant difference in the distribution of Xba I and Pvu II combinative genotype between the two groups (P > 0.05). CONCLUSION: The ERalpha gene polymorphism is not associated with the risk of ICP.

[Association between polymorphisms of CYP17 and CYP3A4 genes and intrahepatic cholestasis of pregnancy in Chengdu].

OBJECTIVE: investigate whether polymorphisms in estrogen-metabolizing genes, CYP17 and CYP3A4, are associated with intrahepatic cholestasis of pregnancy (ICP) in Chengdu China. METHODS: The -1931T/C polymorphism in the 5'-untranslated region of CYP17 gene and the -290A/G polymorphism in the 5'-regulatory region of CYP3A4 gene were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 100 pregnant women with ICP (ICP group) and 100 normal pregnant women (control group) in Chengdu. RESULTS: The T/C polymorphism of the CYP17 gene was shown clearly in ICP and control groups, but the genotype distributions (TT, TC and CC) and allele frequencies (T and C) of CYP17 polymorphism were of no significant difference between the two groups (P > 0.05). No CYP3A4-V (CTP3A4 * 1B) allele was found in the 200 individuals studied. The genotype of CYP3A4 was wt/wt in all the pregnant women. CONCLUSION: The T/C polymorphism of CYP17 gene is not associated with the risk of ICP in Chengdu. There are not -290A/G polymorphism of CYP3A4 gene among pregnunt women in Chengdu.

[Effects of ursodeoxycholic acid on the liver plasma membrane fluidity, hepatic glutathione concentration, hepatic estrogen receptors and progesterone receptors in pregnant rats with ethinylestradiol and progesterone induced intrahepatic cholestasis].

OBJECTIVE: To explore the effects of ursodeoxycholic acid (UDCA) on the fluidity of hepatic plasma membrane, glutathione concentration in liver, hepatic estrogen receptors and progesterone receptors in pregnant rats with ethinylestradiol and progesterone induced intrahepatic cholestasis. METHODS: sixty clean SD pregnant rats were selected and divided into three groups at random. Since the 13th day of pregnancy after taking blood, normal group was injected subcutaneously with refined vegetable oil 2.5 ml x kg(-1) x d(-1). Control group and treatment group were injected subcutaneously with the solution of progesterone 75 mg x kg(-1) x d(-1) and 17-alpha-ethynylestradio 1.25 mg x kg(-1) x d(-1) till the 17th day. Since the 17th day control group, normal group were fedwish 0.9% natriichloridi solution 5 ml x kg(-1) x d(-1); Treatment group was fedwish UDCA 50 mg x kg(-1) x d(-1) every day. On the 21th day, all rats were killed. Then the livers were collected for study. Membrane fluidity was measured by fluorescence polarization using 1,6-diphenyl-1,3,5-hexatriene (DPH) as a probe. Glutathione concentration was measured by 5,5'-dithionbis (2-nitrobenzoic acid) (DTNB). Estrogen receptors and progesterone receptors were measured by flow cytometry. RESULTS: (1) Hepatic plasma membrane fluidity and glutathione (GSH) concentration: significantly lower level of GSH concentration and higher fluorescence polarization (P) were detected in control group (GSH: 1.13 +/- 0.03, P: 0.149 +/- 0.008) in comparison with normal group (GSH: 2.11 +/- 0.07, P: 0.132 +/- 0.004, P < 0.05). However, Significantly higher level of GSH concentration and lower fluorescence polarization were detected in treatment group (GSH: 1.82 +/- 0.04, P: 0.141 +/- 0.006) in comparison with control group (P < 0.05). The level of GSH concentration and fluorescence polarization were no difference between treatment group and normal group. Hepatic estrogen receptors (ER) and progesterone receptors (PR): The expression of ER and PR in control group (ER: 89.4 +/- 8.4, PR: 112.3 +/- 11.6) were higher than that of other two groups (P < 0.05). The expression of ER and PR in treatment group (ER: 56.4 +/- 7.5, PR: 70.1 +/- 9.3) were lower than that of control group (P < 0.05). But there was no difference between treatment group and normal group (ER: 39.5 +/- 7.3, PR: 59.6 +/- 7.4; P > 0.05). CONCLUSION: Ursodeoxycholic acid may be effective drug in treatment intrahepatic cholestasis of pregnancy.