Fig 1.
Effects of a chronic administration of E2, P4 and combined treatment on the uteri and vagina of mice.
(A) Uterine weights, epithelial proliferation and luminal epithelial height after the different treatments. Representative sections from uterus stained with Ki-67 antigen. (B) Vagina weights, epithelial proliferation and luminal epithelial height after the different treatments. Representative sections from vagina stained with Ki-67 antigen are shown. (C) Vaginal lubrication (mg). Data are presented as mean ± SD. To test the respective roles of each treatment, a one-way ANOVA was performed and a Bonferroni’s multiple comparison test. * t test vs OVX. $ t test vs E2.
Fig 2.
Increased bleeding time in E2, P4 and E2+P4-treated mice.
The tail-bleeding times of untreated (OVX, n = 12), E2 (n = 9)-, P4 (n = 10)- and E2+P4 (n = 12)- treated mice were measured as described in Methods. Each point represents 1 individual
Fig 3.
E2+P4 treatment protects mice against thromboembolism.
Thromboembolism was induced by injection of a collagen (0.4 mg/kg) and epinephrine (60 μg/kg) mixture into the jugular vein. (A) All ovariectomized mice (OVX) died within 5 minutes. E2- and E2+P4- treated mice were protected from thromboembolism. (B) Representative sections of hematoxylin-eosin–stained lungs from P4- and E2+P4 -treated mouse are shown. Arrows point to thrombi in the pulmonary vasculature of OVX and treated mice. Original magnification ×250. (C) Representative images of the heart of P4- and E2+P4 -treated mouse at the indicate times are shown. The left ventricular (LV) sphericity index was calculated by the ratio between the cross-sectional diameter of the LV at the point of insertion of the right ventricular wall (D1) and the LV diameter measured at right angles (D2) (D) Quantification of the left ventricular sphericity index measured before, at time of induction of embolism and 5 and 10 minutes after.
Fig 4.
Inferior vena cava stasis in OVX and E2-, P4- and E2+P4- treated mice.
(A) 24 hours after the induction of the stasis in the inferior vena cava by ligation, mice were killed and thrombi were harvested. The mean weight of the thrombi is presented ± SD. * t test vs OVX, $ t test vs E2. (B) Following ferric chloride injury of the carotid artery, the thrombus formed was visualized by high-frequency ultrasound after. Longitudinal views of a mouse without thrombus and with a stable thrombus are shown.
Fig 5.
(A) Blood cells counts in ovariectomized (OVX), E2, P4- and E2+P4-treated mice. OVX (n = 5), +E2 (n = 5), P4 (n = 5), E2+P4 (n = 5). (B) Platelet count recovery after immune thrombocytopenia. Thrombocytopenia was induced after 3 weeks of treatment by intraperitoneal injection of anti-mouse GPIbα antibody. Blood samples were collected before injection (D = 0) and 2, 4, 7, 9, 12 and 15 days after. OVX (n = 5), E2 (n = 5), E2+P4 (n = 5). (C) Total bone marrow cell numbers between OVX, E2-, P4- and E2+P4- treated mice. Bone marrow was flushed from 2 femurs and total bone marrow cell numbers was obtained using a Beckman Coulter Counter. OVX (n = 8), E2 (n = 5), P4 (n = 5), E2+P4 (n = 6). Mean ± SD. To test the respective roles of each treatment, a one-way ANOVA was performed and a Bonferroni’s multiple comparison test. * t test vs OVX. $ t test vs P4.
Table 1.
Coagulation tests (prothrombin time (PT) and activated partial thromboplastin time (APPT)), fibrinogen and coagulation factors levels in ovariectomized (OVX) and treated mice with E2, P4 and E2+P4. Means ± SD. ** p<0.001 indicates significantly different from OVX.