| objectivePreeclampsia(PE)Preeclampsia(PE)is a serious complication of pregnancy.Its main clinical symptoms are hypertension and proteinuria,which is one of the main causes of maternal and perinatal death.Its pathogenesis has not been clearly reported,and it is widely recognized as follows: 1.placental trophoblast infiltration and apoptosis 2.Oxidative stress disorder and inflammation.Long non coding RNA(LncRNA)can participate in the physiological and pathological processes of organisms by regulating cell growth,proliferation and apoptosis.Research shows that,LncRNA-GHET1 can participate in the regulation of PE by affecting the invasion and migration of trophoblast cells J.Because LncRNA-GHET1 shows a potential correlation with PE,three models were used to study whether LncRNA-GHET1 has a positive effect on PE,and the mechanism of its effect was discussed.Method(1)Clinical samples were collected and placental tissues of healthy pregnant women and PE patients were frozen immediately and stored at-80 ° C.The m RNA expressions of LncRNA-GHET1 were detected by q RT PCR,the key factors related to inflammation,oxidative stress and placental growth were detected by ELISA,and the protein levels of COX-2,PGE2 and β-Catenin were detected by Western Blot.(2)The hypoxia / reoxygenation(H / R)model of HTR-8 / Svneo cells was established.LncRNA-GHET1 was overexpressed and silenced by cell transfection.The m RNA expression of LncRNA-GHET1 was detected by q RT-PCR,the cell proliferation level was detected by CCK8,the apoptosis level was detected by TUNEL,Western The protein expression of apoptosis and COX-2,PGE2 and β-Catenin in each group was measured by blot,the expression level of key indicators of inflammation and oxidative stress in each group was measured by ELISA,Cell migration and invasion potential were detected by cell scratch and cell invasion experiments.(3)Endotoxin was used to induce the construction of rat PE model and the overexpression and silencing of LncRNA-GHET1.The expression of LncRNA-GHET1 was detected by q RT-PCR,the expression of inflammation related factors and oxidative stress key indexes in serum was detected by ELISA,the apoptosis of placental tissue was detected by TUNEL,the proliferation and differentiation of placental tissue was detected by HE staining,immunohistochemistry and Western Blot The expression level of apoptosis and COX-2,PGE2 and β-Catenin was determined by blot.Result(1)In the placenta of PE patients,the m RNA level of LncRNA-GHET1 decreased significantly,the levels of TNF-α,IL-1 β and IL-6 increased significantly,the activities of CAT and SOD decreased,ROS and MDA increased significantly.The results of ELISA showed that the expression of s Eng and s Flt-1 was up-regulated,while the expression of VEGF and PLGF was down regulated.Western Blot showed that the protein levels of COX-2 and PGE2 in placenta of PE patients increased significantly,while the protein levels of β-Catenin decreased significantly.The above results showed that PE patients had obvious inflammatory reaction and oxidative stress disorder,accompanied by abnormal expression of various genes.(2)In H / R-induced HTR-8 / Svneo cells,the cell proliferation rate of LncRNA-GHET1 overexpression group was significantly higher than that of LncRNA-GHET1 gene silencing group.TUNEL results showed that the degree of apoptosis of LncRNA-GHET1 overexpression group decreased.Western Blot showed that the protein levels of Bcl-2 and Bax and caspase-9 in LncRNA-GHET1 overexpression group increased.The results of ELISA showed that the levels of TNF-α,IL-1 β,IL-6,ROS and MDA in the over expression group of LncRNA-ghet1 decreased significantly,and the activities of CAT and SOD increased significantly.The results of cell scratch and cell invasion showed that the overexpression of LncRNA-GHET1 could enhance HTR-8 / Svneo cell migration and induce cell infiltration potential.Western Blot showed that the overexpression of LncRNA-GHET1 significantly down regulated the protein level of COX-2 and PGE2,and up regulated the protein level of β-Catenin.The above results indicate that LncRNA-GHET1 can promote the proliferation of H /R-induced HTR-8 /Svneo cells,inhibit their apoptosis,enhance their migration and invasion potential,alleviate their inflammation and oxidative stress response.A series of regulatory effects may be related to COX-2 / PGE2 / β-Catenin pathway.(3)In the placenta of PE rats,the m RNA expression of LncRNA-GHET1 was down regulated.The results of ELISA showed that the levels of TNF-α,IL-1 β,IL-6,ROS and MDA in serum of the rats in the LncRNA-GHET1 overexpression group decreased significantly,and the activities of CAT and SOD increased significantly.TUNEL results showed that the degree of apoptosis in placenta decreased in LncRNA-GHET1 overexpression group.Western Blot showed that the protein levels of Bcl-2 and Bax and caspase-9 in LncRNA-ghet1 overexpression group increased.He staining showed that overexpression of LncRNA-GHET1 could reduce the pathological changes of placenta in PE rats.The results of immunohistochemistry and Western Blot showed that the overexpression of LncRNA-GHET1 significantly down regulated the protein level of COX-2 and PGE2,and up regulated the protein level of β-Catenin in PE rats.The above results in vivo verify the previous results in vitro.conclusionThrough the above three models,this paper studies the intervention of LncRNA-GHET1 on PE process,and discusses its potential mechanism.The results showed that LncRNA-GHET1 might affect COX-2 / PGE2 / β-catenin pathway,and then affect oxidative stress,inflammatory response,and participate in the growth,migration,invasion and apoptosis regulation of placental trophoblast cells.In conclusion,our study suggests that the intervention of LncRNA-GHET1 on PE may be related to COX-2 / PGE2 / β-Catenin pathway. |
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