| PurposeThis study aims to further verify the relationship between the expression of Th17 and Treg cells and the Notch signaling pathway in patients with preeclampsia,and to observe the relationship between the three after giving Notch signaling pathway inhibitors to further clarify the onset of preeclampsia Reasons,and as far as possible to provide new targets and experimental basis for the treatment of preeclampsia.MethodSevere pre-eclampsia group(SP group): all the selected patients in this group were pregnant women diagnosed as severe pre-eclampsia in Gansu Provincial People’s Hospital from November 2018 to June 2019;a total of 30 cases;normal NC group(NC group): The selected patients in this group were all healthy pregnant women with normal prenatal examination from November 2018 to June 2019,a total of 30 cases.There was no significant difference in general data such as pregnancy time and age between SP group and NC group(P> 0.05).Compared with SP group and NC group,the gestational age of delivery in SP group was significantly lower than that in NC group(P <0.05).The peripheral blood T lymphocytes of SP group and NC group were treated with different concentrations of DAPT and the same volume of DMSO,and flow cytometry was used to detect Th17 cells,Treg cells,Th17 / Treg ratio and other indicators in each group.The combined immunoassay method detected the expression of IL-17 and IL-10 in each group,and analyzed and compared the expression characteristics and differences between the groups.Result1.The results of flow cytometry analysis showed that the Th17 / CD4 + T,Treg / CD4 +T,Th17 / Treg ratios in the serum of NC group were 1.63 ± 0.32%,3.73 ± 0.32%,0.44± 0.09,SP Serum Th17 / CD4 + T,Treg / CD4 + T,Th17 / Treg ratios were 3.90 ± 0.28%,2.24 ± 0.45%,1.83 ± 0.48 in SP group,compared with SP group and NC group.Th17 /CD4 + T expression level was higher than NC group(t =-29.209,P <0.001),SP group Treg /CD4 + T expression level was lower than NC group(t = 14.842,P <0.001),SP group Theexpression level of Th17 / Treg ratio in patients’ serum was higher than that in NC group(t =-15.505,P <0.001).2.The results of enzyme-linked immunoassay showed that the expression levels of IL-17 and IL-10 inflammatory factors in the serum of the NC group were 35.52 ± 3.52 and37.86 ± 3.98 ng / L,and the expression levels of IL-17 and IL-10 in the serum of the SP group were 68.54,respectively.± 9.86,27.58 ± 4.58 ng / L,compared with SP group and NC group,the expression of IL-17 in the serum of SP group was higher than that of NC group(t =-17.275,P <0.001).10 The expression level was lower than that in the NC group(t= 9.271,P <0.001).3.In order to verify whether DAPT can inhibit the Notch1 signaling pathway gene and clarify the role of Notch1 signaling pathway gene in Th17/Treg imbalance in patients with severe preeclampsia,Real-time PCR test results showed that SP patients were in control and10 and 20,respectively.At 30umol/L DAPT concentration,Notch1 mRNA was 1.24±0.65,1.19 ± 0.59,1.11 ± 0.43,1.06 ± 0.49,and the NC group patients had a Notch1 mRNA expression level of 0.67± at the control and 10,20,and 30umol/L DAPT concentrations,respectively.0.21,0.64± 0.16,0.55± 0.12,0.52 ±0.09,when DAPT was not added,the content of Notch1 mRNA in serum of SP group was significantly higher than that of NC group(t=5.533,P<0.05),indicating that Notch1 itself participated in the changes of the disease;With the increase of DAPT concentration,Notch1 mRNA expression level decreased,indicating that Notch1 mRNA content and DAPT concentration gradient have a certain relationship,but the change difference is small(P>0.05).4.DAPT inhibits the Notch signaling pathway,which significantly reduces the expression of Th17 cells and the ratio of Th17 / Treg in the serum of PE patients.The test results showed that the SP group patients had Th17 / CD4 + T expression levels of 3.90 ±0.28%,1.96 ± 0.14%,1.67 ± 0.12%,1.44 ± 0.11% at the control and 10,20,30 umol /L DAPT concentrations,respectively.Treg / CD4 + T expression levels were 2.24 ± 0.45%,2.19 ± 0.47%,2.33 ± 0.47%,2.28 ± 0.46%,Th17 / Treg ratios were 1.83 ± 0.48,1.01 ± 0.41,0.75 ± 0.20,0.66 ± 0.17.Under DMSO conditions,the content of Th17 cells in the serum of SP group was significantly higher than that of NC group(t = 29.240,P<0.05),the content of Treg cells was lower than that of NC group(t = 14.780,P <0.05),andthe ratio of Th17 / Treg was significantly higher In the NC group(t = 15.590,P <0.05).With the increase of DAPT concentration,the content of Th17 cells in SP group decreased significantly,the expression of Treg cells increased to a certain extent,but not significantly,and the Th17 / Treg ratio decreased,indicating that Th17 cells,Th17 / Treg ratio and DAPT concentration were significantly correlated,Th17 cells under 30 umol / LDAPT and Th17 /Treg ratio were significantly different from those under 10 umol / L DAPT(all P <0.05).After DAPT treatment in NC patients,the expression levels of Th17 cells,Treg cells and Th17 /Treg ratio changed little(all P> 0.05).5.DAPT inhibits the Notch signaling pathway and can significantly affect the expression of Th17 and Treg-related cytokines in PE patients.The test results showed that the SP group patients had IL-17 expression levels of 68.54 ± 9.86,61.73 ± 9.28,48.43 ±7.27,36.40 ± 4.37 ng / L,IL-10 at the control and 10,20,30 umol / L DAPT concentrations,respectively.The expression levels were 27.58 ± 4.58,29.63 ± 4.56,28.49 ± 4.49,29.44 ± 4.75 ng / L.Under the DMSO condition,the serum IL-10 content of the SP group was significantly lower than that of the NC group(t = 9.280,P <0.05),and the IL-17 content was significantly higher than that of the NC group(t = 17.270,P <0.05).With the increase of DAPT concentration,the IL-17 expression level in SP patients decreased significantly,and the IL-10 expression level slightly increased,indicating that IL-17 expression level was negatively correlated with DAPT concentration,while IL-10 expression The correlation between the level and DAPT concentration was small.The expression level of IL-17 under30 umol / L DAPT was significantly different from that under 10 umol / L DAPT(both P<0.05).After DAPT treatment in the NC group,the expression levels of IL-17 and IL-10 differed significantly(all P> 0.05).ConclusionTh17,Treg cell content and Th17 / Treg values in patients with preeclampsia are different from healthy pregnant women.At the same time,the expression levels of IL-17,IL-10 and other inflammatory factors also appear abnormal.After adding DAPT,it was found that the expression of Th17 cells and related cytokines in patients with preeclampsia was reduced,which further improved the development of PE patients.The principle may be related to the inhibition of the expression of Notch protein and mRNA.The application ofDAPT may be a new target for the treatment of PE in the future. |
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