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1.
Article in Chinese | WPRIM | ID: wpr-1018673

ABSTRACT

Objective To investigate the preventive effect of schisandrin(SCH)on fetal neural tube defects(NTDs)of mice and its mechanism.Methods C57BL/6 mice were mated with female and male at a ratio of 2:1.Pregnant female mice with vaginal plug after mating were randomly divided into control group,model group,SCH group,and folic acid group,with 9 mice in each group.The NTDs fetal mice model was induced by intraperitoneal injection of all-trans retinoic acid(atRA)(7.5 mg/kg)on embryonic day 7.5(E 7.5 d).During E 0.5 d-E 11.5 d,pregnant rats in folic acid group were given folic acid[61.0 μg/(kg·d)]by gavage once a day,and pregnant rats in SCH group were given SCH[8.0 mg/(kg·d)]by gavage once a day.Fetal mice were removed by cesarean section on E 11.5 d.PC12 cells were divided into control group,model group and SCH group.PC12 cells were treated with atRA(20 μmol/L)for 12 hours to establish cell damage model in model group,and treated with SCH(2.5 μmol/L)for 24 hours in SCH group.Fetuses were identified NTDs by stereoscopic microscopy.HE staining was used to observe the closure of the neural tube.The expression levels of p-PI3K,Akt and p-Akt molecules in PI3K/Akt signaling pathway were detected by Western Blotting.Results Compared with control group,the incidence of NTDs was significantly increased in mice of model group(P<0.01);compared with model group,the incidence of NTDs was decreased in folic acid group and SCH group(P<0.01);compared with folic acid group,SCH group had a lower incidence of NTDs(P<0.01).Western Blotting results showed that compared with control group,the expression of p-PI3K and p-Akt protein in fetal tissues of model group was significantly decreased(P<0.01,P<0.05);compared with model group,there was no significant difference in expression of p-PI3K and p-Akt in fetal tissues of folic acid group(P>0.05),while the expression of p-PI3K and p-Akt protein in SCH group was significantly higher(P<0.05).Compared with control group,PC12 cells in model group showed lower expression levels of p-PI3K and p-Akt(P<0.05);compared with model group,PC12 cells in SCH group showed higher expression levels of p-PI3K and p-Akt(P<0.05).Conclusions SCH can reduce the incidence of atRA-induced NTDs in fetal mice,and its preventive effect is better than folic acid,which may be related to the activation of the PI3K/Akt signaling pathway.

2.
Article in Chinese | WPRIM | ID: wpr-802261

ABSTRACT

Objective:To investigate the regulatory effect of Sinisan(SNS) on the polarization of RAW264.7 macrophages induced by lipopolysaccharide (LPS). Method:RAW264.7 cells stimulated by LPS were used as the in vitro model. The cells were intervened with the different concentrations of SNS in advance. The effects of different concentrations of SNS on the proliferation of RAW264.7 cells were detected by methyl thiazolyl tetrazolium (MTT) colorimetry. The degree of cell differentiation was detected by enzyme-linked immuno sorbent assay(ELISA) method. The contents of M1 polarization factors tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1β (IL-1β) and M2 polarization factors interleukin-10 (IL-10) in cell culture supernatant were detected by ELISA method, mRNA levels of M1 polarization factors TNF-α, IL-6 and M2 polarization factors IL-10, arginase-1 (Arg-1) were detected by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) method. Result:SNS had no effect on the cell viability of RAW264.7 cells, inhibited LPS-induced cell proliferation, decreased LPS-stimulated cell differentiation, down-regulated M1 polarizing factors TNF-α, IL-6, IL-1β release and TNF-α, IL-6 mRNA levels, and increased the release of IL-10 and mRNA levels of IL-10 and Arg-1. Conclusion:SNS inhibits the inflammation of RAW264.7 cells induced by LPS, and its mechanism may be related to the regulation of polarization balance of M1/M2 macrophages.

3.
Article in Chinese | WPRIM | ID: wpr-701203

ABSTRACT

AIM:To investigate the effects of kaempferol-3-O-rutinoside(KR)on the proliferation,migration of vascular smooth muscle cells(VSMC)and the activation of transforming growth factor βreceptor 1(TGFBR1)signaling pathway in the cells.METHODS: The viability of VSMC was detected by MTT assay.The proliferation of VSMC was measured by EdU staining.The migration ability of VSMC was examined by Transwell assay.The protein levels of the mi-gration-associated proteins matrix metalloproteinase 2(MMP2)and matrix metalloproteinase 9(MMP9)were detected by Western blot.Molecular docking study was conducted to explore the interaction between KR and TGFBR 1.The protein le-vels of the phosphorylated TGFBR1,Smad2 and Smad3 were determined by Western blot.RESULTS: KR inhibited the viability of VSMC in a dose-and time-dependent manner.KR reduced the ratio of EdU-positive cells in a dose-dependent manner.KR dose-dependently suppressed the migration ability of VSMC and decreased the protein levels of MMP 2 and MMP9(P<0.05).KR docked into TGFBR1 with the binding energy of -9.804 kcal/mol by forming hydrogen bonds with SER-280,ARG-215,ASP-290 and LYS-335 of TGBFR1.KR dose-dependently suppressed the activation of TGFBR 1 and its downstream proteins Smad2 and Smad3(P<0.05).CONCLUSION: KR inhibits the proliferation and migration of VSMC,possibly via blocking the TGFBR1 signaling pathway.

4.
Article in Chinese | WPRIM | ID: wpr-676448

ABSTRACT

Objective To observe the effects of genistein and 17?-estradiol on microstructure of cancellous bone in ovariectomized (OVX) rats.Methods Ninty 7-month-old SD rats were randomly divided into baseline group,ovariectomized (OVX),sham-operated (SHAM),17?-estradiol treated (10?g?kg~(-1).day~(-1),EST) and genistein treated (5 mg?kg~(-1)?day~(-1),GEN) groups,and were killed at the beginning of the experiment,the 3rd and 15th week after operation.MicroCT scanning was performed on the left tibia in vitro.The regions involving 0.5 mm slice thickness and 1.6 mm distal to the tibial growth plate were selected as the regions of interest.Results At the 3rd week after operation,the tissue bone mineral density (tBMD) and trabecular thickness (sTh.Th) in group GEN were significantly higher than those in OVX and EST groups (all P

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