K Ashokan, M Kurane, M Pillai
and brunner’s gland, cysteamine, duodenal ulcer, estradiol-17β, ovariectomy, ulcer index
K Ashokan, M Kurane, M Pillai. Effect Of Ovariectomy And Of Estrogen Administration Upon Duodenal Ulceration Induced By Cysteamine. The Internet Journal of Gastroenterology. 2008 Volume 8 Number 1.
Duodenal ulcers were induced in ovariectomized, intact and old mice using cyteamine hydrochloride. Under these experimental conditions ovariectomy and old age strikingly increased sensitivity to ulcer induction while estrogen administration showed a decrease in sensitivity to ulcer induction. Nevertheless, the administration of estrogen in old showed no effects in either intact or overiectomized mice. This change in ulcer sensitivity reflected from histological, histochemistry and biochemical studies. The histological study was performed by using haematoxylin-eosin staining technique. The histochemistry of the duodenal region was studied by using periodic acid Schiff reaction (PAS) for glycoproteins. The biochemical study was performed to study various constituents of glycoproteins like hexose, fucose, sialic acid and the protein. The result showed that ulcer severity was more in overiectomized cyteamine treated mice and old mice treated with cyteamine. The histological studies showed the ovariectomy decreased or not shown any change in the ulcer sensitivity considering cryptus Lieburkuhn and bruner’ glands. The same result reflected in histochemistry study by differential intensity in the staining property of the bruners gland. The biochemical study showed that the glycoprotein contents were reduced many folds in overiectomized cyteamine treated mice and their reversal in estrogen administered ovariectomized cysteamine injected mice. These findings prove that estrogen protect the duodenal ulcer form cysteamine administration.
Duodenal ulcer is a mucosal erosion of duodenum, due to multiple causes, including bacteria (Marshall and Warren 1984; Lykoudes 1958), chewing gum tobacco smoking, not eating properly, blood group, spice (NADDIC), chronic stress (Kim et al
Materials and methods
There were 60 mice in the present study. The mice were divided into two groups- 2 months old females (40 numbers) and 3 years old females (20 numbers). Ten mice were used as control in both groups. The 2 months old female mice were treated with cysteamine-HCl divided into tow groups. The group one contain 10 female mice, were used as cysteamine –HCl treated one, the second group contain 20 female mice were used for ovariectomy.
Ovariectomy and administration of estrogen
The ovariectomy was done under mild ether anesthesia. The operated mice were maintained fro 15 days in separate age with optimum care of light, temperature, humidity, food and water. On the 16th day half the number of ovariectomized cysteamine treated mice were injected (i.p) consecutively for 3 days with 2 mg/body weight estradiol- 17β (Sigma, Batch no. E9505) in olive oil. The remaining ovariectomized mice were injected with olive oil only. On the 4th day of the 1st injection the ovariectomized, ovariectomized cyteamine injected and ovariectomized cysteamine injected and estrogen administered mice were used for duodenal ulcer induction (Szabo 1978). Ten 3 years old mice were used for administration of cysteamine, after three days it was subjected for ulcer induction.
The pyloro-duodenal junctions were fixed in 10% neutral buffered fromaline, washed and routinely processed for histological technique. The sections were stained with haematoxyline-eosine (Gurr1962). Histology of pyloric glands, duodenal villi, crypts of Lieberkuhn and Brunner’s glands was studied.
To study the changes in the duodenal mucosa glycoproteins of crypts of Lieberkuhn, goblet cells, pyloric gland cells and Brunner’s glands of cyteamine treated and control mice PAS techniques (McManus 1946) were used.
The glycoprotein from Brunner’s gland was isolated by the method of Satakopan and Kurup (1977).To study various constituents of glycoproteins biochemical estimations of fucose (Dische and Shettles 1977), hexose (Dubois et al. 1956), sialic acid (Warren 1959) and protein (Lowry et al. 1951) were used.
Statistical analyses were performed using the Statistical Package for Social Science (version 13.0, SPSS, Inc) software. Results were expressed as means ± SE (standard Error). All reported p-values were made on the basis of 2-sided tests and compared to a significance level of 5%, differences were considered statistically significant at p< 0.05.
The ovariectomized mice showed little ulceration (Ulcer index 2.6) or no ulceration. The overiectomozed + estradiol- 17β injected mice showed ulceration very low compared to the overiectomized mice (ulcer index 1.64) (Table No.1) The ovariectomized + Cysteamine administered mice showed higher ulceration (Ulcer index 5.1) (Table No.1).The administration of estradiol- 17β to cysteamine injected ovariectopmized mice showed little recovery of ulceration (Ulcer index 4.2) (Table No.1). The old mice administered with cysteamine showed ulceration as high as ovariectomized + cysteamine administrated mice (Ulcer index 4.8) (Table No.2).
* Values are mean ± Standard error, p< 0.05 is significant
Administration of estradiol- 17β to cysteamine treated old mice fails to recover the severity of ulceration (Table No.2). The histology of ovariectomized mice (Fig.1) showed that the pyloric glands were simple tubular and situated deep in the sub mucosa. The duodenal villi were tall, leaf like and uniformly arranged with desquamation intermittently. The crypts of Lieburkuhn and Brunner’s glands are unaffected. The cysteamine administration to the ovariectomized mice (Fig.2) cause pyloric glands with dilated lumen and picnotic nuclei and increased eosinophilia. The pyloric villi showed fissures and ramifications, and ulcer formation. The goblets cells are less in it. The Brunner’s gland acini showed reduced height, dilated lumen and nuclei with abnormal size and shape. The histochemistry showed strong PAS reaction in pyloric glands, crypts of Lieberkuhn, Brunner’s glands in ovariectomized mice but less in pyloric pit (Fig.3).
The duodenal villi showed PAS activity but other cells are PAS negative. The cyteamine treated overiectomized mice showed reduction in PAS reactivity in all cells of pyloric glands, pyloric pits, goblet cells and Brunner’s glands (Fig 4). The biochemical studies showed that the hexose contents reduce 3 folds in cyteamine injected mice comparing to the normal (Table 3). In ovariectomized it was less than that of normal but more than that of cyteamine treated mice. But in ovariectomized cysteamine treated mice it was 6 times less than that of normal mice. The fucose contents, sialic acid contents and protein contents (Table 3) also showed the same trend as hexose.
Peptic duodenal and gastric ulcers raise serious health problems and significant economic cost worldwide. There are approximately 500,000 new cases and 4.5 million people suffering from these diseases each year in USA (Valle et al