| The study was carried out to compare about the effect of baicalin and quercetin On protection of mouse deciduas cells indurced by LPS.We tried to expore the effection and the possible mechanisms through tree parts of experiments.First: Use the decidua cell culture in vitro to observate the effection intuitively.Firstly,we improved the method of decidua cell culture. We confirmed that the explant culture is better than the traditional enzyme digest culture which laid the foundation for the next trials; The different doses of LPS were added to the plate(0ng/ml,10ng/ml,100ng/ml, 500ng/ml,1000ng/ml) and observed under a light microscope.The 100ng/ml dose of LPS were determinded to use the follow experiment due to the dose induced 50% of death; To determined the effct of baicalin and quercetin,the attached cells were devived into blank group, LPS group,LPS+baicalin(4μg/ml) group,and LPS+quercetin(25μmol/ml) group,incubated for 48h. The result showed that the treatment of decidual cells with quercetin and bacilin can significantly increase the live cells.Second:The mice were injected LPS and gived baicalin and quercetin by oral gavage method,which were used for the pharmacodynamics studies of the fetus protection action in vivo. Six days pregnant females were divided into eight groups of 5 animals each.Control and LPS groups(groupA and groupB) of mice were injected i.v with PBS and LPS(5μg/ml,0.1ml/10g) on day 6 of pregnancy.The baicalin(group C, D, E) and quercetin(group F, G, H) treatment groups were injected LPS with the same dose of LPS on day 6 of gestation.The decidual treatment groups were administrated with the different dose of quercetin (0.25mg/ml,1.25mg/ml,2.5mg/ml),when the excess groups were received the doses of baicalin (0.25mg/ml,1.25mg/ml,2.5mg/ml) on 7 and 8 day of pregnancy.The control group received PBS instead.LPS induced embryo resorption and the effect of drugs were studied on day 9 of gestation. Blood samples(about 0.5 mL) were collected to analyze the TNF-αand Progestrone. The uterus were immediately weighted and the percentage of resorptions was calculated.The tissues were stained with hematoxillin and eosin and examined under a light microscope. It can be seen that LPS increased the resorption to 27% and baicalin and quercetin can increase the weight of uterine compared to LPS group and protect the decidua cells.We found that both baicalin and quercetin could significantly down-regulate TNF-αexpression(P<0.01). In relation to progesterone,no significant difference was detected between groups.Third: Dectected the apoptotic protein Caspase-3 by immunohistochemistry.We attempted to elucidate the relationship between spontaneous abortion and abnormal cells apoptosis from gene level,which provide a new understanding for the mechanism of spontaneous and the fetus protection effect of traditional chinense medicine.We had deceted the Caspase-3 expression on the decidua tissues of 6 groups(control group, LPS group, middle and high group of baicalin and quercetin).The positive cells were calculated for statiscal analysis.The result has shown that LPS can significantly increase the expression of Caspase-3(P<0.01);Bacilin and quercetin can decrease the number significantly(P<0.01).Quercetin can dual-regulate the Caspase-3, high-dose of drug increase the expression Caspase-3 .Conclusion: Baicalin, quercetin can effectively protect the decidual cell damaged by LPS ; effectively reduce the abortion rate, reducing the embryo resorption rate;reduce the expression of pregnancy, maternal TNF-α; LPS can increase the expression of Caspase-3 in decidual cells (P<0.01); The protect action of baicalinand quercetin may relate to that they can decrease the expression of Caspase-3. |
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