Effects Of Prenatal Tetrandrine Treatment On The Expression Of Krüppel-like Factor 5 And Survivin In The Lung Of Congenital Diaphragmatic Hernia Rat

Objective: The aim of the study was to investigate the effect of prenatal administration of tetrandrine on the expression of Krüppell like factor 5(KLF5)and Survivin in the lung of congenital diaphragmatic hernia(CDH)rat model.Methods :This experiment took simple random group method,11 pregnant S-D rats were randomly divided into three groups: Normal+Control group(group NC,3);Model+Control group(group MC,4);Model+Tetrandrine group(group MT,4).The rats in group MC and MT were gave gavage administration with 125 mg nitrofen(Solubled in 2ml olive oil)to induce CDH and the rats in group MT were administered with 30 mg /kg tetrandrine for three consecutive days(gestational age of18.5d,19.5d and 20.5d).The rats in group NC and MC were gave the equivalent normal saline.All fetuses were delivered by caesarian section at the gestational age of21.5d,observing the formation situation of diaphragmatic hernia in 3 groups and recording the ratio of Lung weight and Body weight(Lw/Bw).The maturity situation of fetal lung tissue was evaluated by HE staining.The expression of KLF5 and Survivin were detected by immunohistochemical staining and quantitatively analyzed by Western blot.Single factor analysis was taken among the 3 groups to compare means of variables and the chi-square test was used to compare the difference of birth defect rate.Results: There was significant difference on CDH incidence between MC(56.06%(37/66))and MT group(50%(28/56))(p<0.001,x2 =122).The Lw/Bw of the three groups indicated no significant difference(F=0.985,p=0.376).The results of HE staining showed that pulmonary vascular-related and pulmonary alveolar-related developmental indexes in MC group were lower than those in the group of NC,and the MT group was during group NC and MC.In addition,immunohistochemical staining showed that the average optical density(AOD)of KLF5 in NC,MC,MT groups were0.1948±0.0074,0.2121±0.0049 and 0.1927±0.0019,respectively,between which there were significant differences(F=14.531,P=0.002);and Survivin was 0.1852±0.0087,0.2092±0.0036 and 0.1928±0.0075(F=12.308,P=0.003).The expression level of Survivin was positively correlated with KLF5(r=0.993,p=0.039).Western blot revealed that the KLF5 and Survivin protein expression in 3 groups were significant different(F=4.29?10.13,P <0.05,respectively).Conclousions: The rat CDH animal model which is induction by Nitrofen can simulate the pathogenesis of human diaphragmatic hernia.The management that prenatal gavage of tetrandrine to CDH rats promoted the development of fetal lung parenchyma and vascellum,and reduce the expression level of KLF5 and Survivin protein in fetal lung,which may be the mechanism of tetrandrine in improving lung hypoplasia of CDH.