Cryptotanshinone reverses ovarian insulin resistance in mice through activation of insulin signaling and the regulation of glucose transporters and hormone synthesizing enzymes  Yangang Huang, M.D., Wei Li, M.D., Chi Chiu Wang, M.D., Ph.D., Xiaoke Wu, M.D., Ph.D., Jianhua Zheng, M.D.  Fertility and Sterility  Volume 102, Issue 2, Pages 589-596.e4 (August 2014) DOI: 10.1016/j.fertnstert.2014.05.012 Copyright © 2014 Terms and Conditions

Figure 1 Insulin sensitivity after in vivo CRY treatment. 3H-deoxy-glucose uptakes in different tissues were determined by hyperinsulinemic clamp. CON = control group; DEX = DEX treatment group; DEX+CRY = combined DEX and CRY treatment group. The data are presented as means ± SE (n = 6). aP<.01, CON vs. DEX; bP<.05, CON vs. DEX; cP<.01, DEX vs. DEX+CRY; dP<.05, DEX vs. DEX+CRY. Fertility and Sterility 2014 102, 589-596.e4DOI: (10.1016/j.fertnstert.2014.05.012) Copyright © 2014 Terms and Conditions

Figure 2 Ovarian insulin signaling transduction after in vitro CRY treatment. Relative in vitro ovarian expression of ERK1/2, P-ERK1/2, AKT2, P-AKT2, GSK3β, and P-GSK3β protein were determined by Western blot. The data are presented as means ± SE (n = 5). aP<.01, CON vs. DEX; bP<.05, CON vs. DEX; cP<.01, DEX vs. DEX+CRY; dP<.05, DEX vs. DEX+CRY. Fertility and Sterility 2014 102, 589-596.e4DOI: (10.1016/j.fertnstert.2014.05.012) Copyright © 2014 Terms and Conditions

Figure 3 Ovarian hormone synthesizing enzymes after in vitro CRY treatment. Semiquantitative reverse transcription–polymerase chain reaction of ovarian mRNA expression of hormone synthesis enzymes, including STAR, CYP11, CYP17, and CYP19, in in vitro cultured ovaries. The results from two independent experiments are shown after 2% agarose gel electrophoresis. aP<.01, CON vs. DEX; bP<.05, CON vs. DEX; cP<.01, DEX vs. DEX+CRY; dP<.05, DEX vs. DEX+CRY. Fertility and Sterility 2014 102, 589-596.e4DOI: (10.1016/j.fertnstert.2014.05.012) Copyright © 2014 Terms and Conditions

Supplemental Figure 1 Ovarian insulin sensitivity after in vitro CRY treatment. 3H-deoxy-glucose uptakes in ovaries were determined in in vitro culture. The data are presented as means ± SE (n = 6). aP<.01, CON vs. DEX; dP<.05, DEX vs. DEX+CRY. CPM = counts per minute. Fertility and Sterility 2014 102, 589-596.e4DOI: (10.1016/j.fertnstert.2014.05.012) Copyright © 2014 Terms and Conditions

Supplemental Figure 2 Ovarian hormonal secretion after in vitro CRY treatment. Levels of T, P, and E2 were measured in the culture medium. The white bar is the basal hormone level, and the black bar is the hormone level after forskolin stimulation. The data are presented as means ± SE (n = 8). aP<.01, CON vs. DEX; bP<.05, CON vs. DEX; cP<.01, DEX vs. DEX+CRY; dP<.05, DEX vs. DEX+CRY. Fertility and Sterility 2014 102, 589-596.e4DOI: (10.1016/j.fertnstert.2014.05.012) Copyright © 2014 Terms and Conditions

Supplemental Figure 3 Ovarian glucose transporter expression after in vitro CRY treatment. Relative in vitro ovarian mRNA expression of GLUT1 and GLUT4 were determined by semiquantitative reverse transcription–polymerase chain reaction. The results from two independent experiments are shown after 2% agarose gel electrophoresis. bP<.05, CON vs. DEX; cP<.01, DEX vs. DEX+CRY. Fertility and Sterility 2014 102, 589-596.e4DOI: (10.1016/j.fertnstert.2014.05.012) Copyright © 2014 Terms and Conditions