Volume 141, Issue 3, Pages 854-863 (September 2011) Estrogen Regulation of Duodenal Bicarbonate Secretion and Sex-Specific Protection of Human Duodenum  Biguang Tuo, Guorong Wen, Jinqi Wei, Xuemei Liu, Xue Wang, Yalin Zhang, Huichao Wu, Xiao Dong, Jimmy Y.C. Chow, Volker Vallon, Hui Dong  Gastroenterology  Volume 141, Issue 3, Pages 854-863 (September 2011) DOI: 10.1053/j.gastro.2011.05.044 Copyright © 2011 AGA Institute Terms and Conditions

Figure 1 Duodenal tube with 5 channels and 3 balloons used to functionally isolate human proximal duodenum for the measurement of duodenal HCO3− secretion. The tube was positioned to the proximal duodenum using special placing-clips under gastroscopy. Gastroenterology 2011 141, 854-863DOI: (10.1053/j.gastro.2011.05.044) Copyright © 2011 AGA Institute Terms and Conditions

Figure 2 Epidemiologic study of duodenal ulcer prevalence. (A) Numbers of male and female subjects included in the study. There were no significant differences between the numbers of men and women in each age group (#P > .05). (B) H pylori infection of men and women. There were no significant differences between H pylori infection of men and women in each age group (#P > .05). (C) Prevalence of duodenal ulcer in men and women. There were significant differences between duodenal ulcer prevalence in men and women in each age group younger than 70 years, especially in age groups younger than 50 years (*P < .05, ***P < .001, and #P > .05). Male/female ratio of duodenal ulcer prevalence in different age groups is shown at the bottom. (D) Duodenal ulcer prevalences in H pylori–infected and non–H pylori–infected male and female patients younger than 50 years (***P < .001). Gastroenterology 2011 141, 854-863DOI: (10.1053/j.gastro.2011.05.044) Copyright © 2011 AGA Institute Terms and Conditions

Figure 3 Age and sex differences of basal and acid-stimulated DBS in human beings. (A and C) Time-courses of basal and acid-stimulated DBS in men and women at age (A) 20–29 years and (C) 60–69 years. After measuring basal DBS for 60 minutes, the duodenum was luminally exposed to 10 mL HCl (100 mmol/L) for 10 minutes as indicated, and then DBS was measured for an additional 60 minutes. Acid markedly stimulated DBS in all groups (*P < .05, **P < .01, or ***P < .001, comparison with basal value). Maximal responses were reached within 30 minutes after administration of HCl. (B and D) Basal and acid-stimulated net peak DBS in men and women at age (B) 20–29 years and (D) 60–69 years (*P < .05, **P < .01, or #P > .05, comparison between men and women). (E) Basal DBS of men and women at age 20–29 years and 60–69 years (*P < .05 or #P > .05, comparison between 2 age groups). (F) Acid-stimulated net peak DBS of men and women at age 20–29 years and 60–69 years (**P < .01 or #P > .05, comparison between 2 age groups). Gastroenterology 2011 141, 854-863DOI: (10.1053/j.gastro.2011.05.044) Copyright © 2011 AGA Institute Terms and Conditions

Figure 4 Basal and acid-stimulated DBS during the different stages of the menstrual cycle in premenopausal women. (A) Time-courses of basal and acid-stimulated DBS during the preovulatory and premenstrual phases in women at age 20–29 years. After measuring basal DBS for 60 minutes, the duodenum was luminally exposed to 10 mL HCl (100 mmol/L) for 10 minutes as indicated, and then DBS was measured for an additional 60 minutes. Acid markedly stimulated DBS in preovulatory and premenstrual women (*P < .05, **P < .01, or ***P < .001, comparison with basal value). (B) Basal and acid-stimulated net peak DBS during preovulatory and premenstrual phases (*P < .05, comparison between preovulatory and premenstrual). (C) Serum estradiol levels during preovulatory and premenstrual phases in the same women (***P < .001, comparison between preovulatory and premenstrual). Gastroenterology 2011 141, 854-863DOI: (10.1053/j.gastro.2011.05.044) Copyright © 2011 AGA Institute Terms and Conditions

Figure 5 Effect of 17β-estradiol applied to the duodenal lumen on DBS in men and women at different ages. (A and C) Time-courses of basal and 17β-estradiol–stimulated DBS in men and women at age (A) 20–29 years and (C) 60–69 years. After measuring basal DBS for 60 minutes, the duodenum was luminally exposed to 10 mL 17β-estradiol (0.1 mg/mL) for 10 minutes as indicated, and then DBS was measured for an additional 75 minutes (**P < .01, comparison with basal value). (B and D) Basal and 17β-estradiol–stimulated net peak DBS in men and women at age (B) 20–29 years and (D) 60–69 years (*P < .05, or #P > .05, comparison between men and women). Gastroenterology 2011 141, 854-863DOI: (10.1053/j.gastro.2011.05.044) Copyright © 2011 AGA Institute Terms and Conditions

Figure 6 Expression of ERα and Erβ proteins in human proximal duodenal mucosa. Duodenal mucosae from healthy men (M) and women (F) at the age of 20–29 years were subjected to Western blot analysis as described in Methods. ERα and ERβ were similarly expressed in duodenal mucosae of men and women. Representative blots are shown from 3 separate experiments with identical results. Gastroenterology 2011 141, 854-863DOI: (10.1053/j.gastro.2011.05.044) Copyright © 2011 AGA Institute Terms and Conditions

Figure 7 Expression of ERα and ERβ in duodenal epithelial cells by immunohistochemistry. Human proximal duodenal mucosae (from the same subjects shown in Figure 6) were subjected to immunohistochemistry analysis as described in the Materials and Methods section. Representative pictures are shown from 3 men and 3 women in separate experiments with identical results. Both ERα and ERβ were localized mainly to the plasma membrane and cytosol of villous and crypt cells in duodenal mucosae. (Ai and Aii) Expressions of ERα in (Ai) villous and (Aii) crypt cells. (Bi and Bii) Expressions of ERβ in (Bi) villous and (Bii) crypt cells. (Ci and Cii) Expressions of (Ci) ERα and (Cii) ERβ in breast carcinoma cells (positive control). (Di and Dii) Expressions of (Di) ERα and (Dii) ERβ in hepatic carcinoma cells, HepG2 (negative control). (A–D) Original magnifications, 400×. (E) Negative control in human proximal duodenal mucosae without primary antibody (original magnification, 100×). Gastroenterology 2011 141, 854-863DOI: (10.1053/j.gastro.2011.05.044) Copyright © 2011 AGA Institute Terms and Conditions