The Study Of Pregnancy Status In Patients With Lymphangiomyomatosis And The Exploration Of Autophagy Changes In The Process Of Pulmonary Fibrosis

PART1 Fertility status and safety in patients with lymphangioleiomyomatosis Objective:Our objective was to determine outcomes and safety of pregnancy and parturition in patients with lymphangioleiomyomatosis(LAM).Methods:We collected baseline data of patients with LAM in the project of LAM-China registry and analyzed the general fertility status of them.We also selected patients who got pregnant after diagnosis of LAM from the registry and our patient database to explore the outcomes and safety of fertility.Results:1.Among 188 patients from LAM-China registry,154(81.9%)had history of pregnancy and 148(78.7%)had history of parturition.All of the 154 patients had pregnancy before diagnosis of LAM and 4 of them had another pregnancy after diagnosis.Of a total of 362 pregnancies,50.6%turned out parturition,3.0%miscarriage and 42.8%induced abortion;2.There were 28 patients with 31 pregnancies after diagnosis of LAM.Of 23 pregnancies terminated in the first trimester,17.4%(4/23)reported dyspnea;and of 8 pregnancies terminated in the second and third trimesters,75%(6/8)reported dyspnea.1 patient had bilateral pneumothorax and 1 had spontaneous bleeding of renal angiomyolipoma during gestation.1 patient had stillbirth;3.There were 16 patients with 17 pregnancies after initiation of sirolimus.29.5%(5/17)had miscarriage and 41.2%(7/17)had induced abortion in the first trimester.All of the 4 live births got healthy babies and 2 patients were still pregnant.Conclusions:The rates of pregnancy and parturition in LAM patients are lower than that in general population,especially in groups of patients after diagnosis of LAM.Outcomes of pregnancy before diagnosis of LAM are almost not impacted by the potential disease.Pregnant patients after diagnosis reported dyspnea during gestation commonly and most of them can recover after labor.Taking sirolimus before pregnancy would increase the risk of miscarriage in the first trimester but might not result in structural malformation of the baby.PART 2 Dynamic alteration of autophagic activity in lungs in the process of bleomycin-induced pulmonary fibrosisObjective:We did this research to explore the dynamic alteration of autophagic activity in lungs of bleomycin-induced pulmonary fibrotic mice.Methods:Wildtype C57BL/6J mice was randomized to untreated(blank control),saline-treated and bleomycin-treated groups.Bleomycin and saline was intratracheally administrated with a dose of 2.5mg/kg of mouse weight.We sacrificed the mice 1,3,7,14 and 21 days after administration.We counted total cells in bronchoalveolar lavage fluid(BALF),scored the fibrotic lungs,did western blot to examine autophagy-related proteins and observed autophagosomes in lungs.Results:Bleomycin-treated mice generally weighed less than the saline-treated mice(p<0.05).The total cell counts in BALF were in an uptrend from day 1 to day 7(p<0.05),and kept this high levels from day 7 to day 21.The Ashcroft scores for pulmonary fibrosis showed a significant elevation from day 7(p<0.05)and continues increasing in the groups of 14 and 21 days after bleomycin(p<0.05).Immunohistochemistry showed that a-smooth muscle actin was positively in a fusiform shape in lung tissues in groups of day 14 and 21.Significantly higher levels of LC3B-?/LC3B-? and Beclinl in whole lysates from lungs on day 3 and rapid decrease of them on day 7 was observed by western blot.No significant differences of autophagosomes in lungs were observed in all groups by transmission electron microscope.Conclusions:Wildtype mice could be induced mild pulmonary fibrosis 7 days after single-dose of intratracheal bleomycin and severe pulmonary fibrosis 14 days after that,while inflammation would persist from day 3 to day 21.During this process,the autophagic activity may rise to peak on day 3 and decline on day 7,and low levels of autophagy may continue to day 21.