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Correlation of heat shock protein expression to gender difference in development of stress-induced gastric mucosal injury in rats.

Takahashi T, Otaka M, Odashima M, Tamaki K, Takada M, Izumi Y, Shibuya T, Sakamoto N, Itoh H, Watanabe S - J Clin Biochem Nutr (2010)

Bottom Line: The gastric ulcer index was significantly higher in male rats compared to female rats observed after 12 h water immersion stress exposure.At this time point, the expression levels of HSP60 and HSP90 in the gastric mucosa were significantly higher in females than those in males.Inversely, an ovariectomy dramatically reduced the expression of HSP60, HSP70 and HSP90 in the gastric mucosa.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology, Juntendo University School of Medicine, Bunkyo-Ku, Tokyo 113-8421, Japan.

ABSTRACT
Recent studies have indicated that heat shock proteins (HSPs), which function as molecular chaperones, play important roles in cellular responses to stress-related events. However, the gender difference in the expression of HSP in the gastric mucosa remains unclear. In order to understand the mechanism of gender difference in the prevalence or severity of gastric mucosal lesions, the expression level of HSP and the correlation of estrogen to HSP induction in the gastric mucosa were evaluated in this study. The basal expression levels of HSP60 and HSP90 in the gastric mucosa were significantly higher in females than those in males. The gastric ulcer index was significantly higher in male rats compared to female rats observed after 12 h water immersion stress exposure. At this time point, the expression levels of HSP60 and HSP90 in the gastric mucosa were significantly higher in females than those in males. An estrogen-treatment significantly induced the expression of HSP60, HSP70 and HSP90 in the gastric mucosa. Inversely, an ovariectomy dramatically reduced the expression of HSP60, HSP70 and HSP90 in the gastric mucosa. Our results suggested that estrogen might play an important role in gastric mucosal protection with the induction of gastric mucosal HSPs.

No MeSH data available.


Related in: MedlinePlus

In order to study the gender difference in the expression levels of the HSPs, we designed the experimental protocol to study the effect of estradiol on the male gastric mucosa. Each rat was euthanized before and 1 h, 3 h, or 6 h after the intra-peritoneal injection of 10 mg/kg estradiol (n = 4 at each time point). In the control group, the rats were treated with the vehicle (n = 4 at each time point). a: The expression level of HSP60 was significantly higher in the estradiol-administered group (estrogen group) compared to the control group after the 1 h (Bonferroni correction, p = 0.032) and 6 h (Bonferroni correction, p = 0.034). b: The expression level of HSP70 was significantly higher in the estradiol-administered group (estrogen group) compared to the control group after the 1 h (Bonferroni correction, p = 0.04), 3 h (Bonferroni correction, p = 0.013) and 6 h (Bonferroni correction, p = 0.016). c: The expression level of HSP90 was significantly higher in the estradiol-administered group (estrogen group) compared to the control group after the 1 h (Bonferroni correction, p<0.001), 3 h (Bonferroni correction, p = 0.003). E2: estradiol; 1 h: 1 h after intra-peritoneal injection of vehicle (DMSO/PBS, 0.5 ml) or estradiol (10 mg/kg); 3 h: 3 h after intra-peritoneal injection of vehicle (DMSO/PBS, 0.5 ml) or estradiol (10 mg/kg); 6 h: 6 h after intra-peritoneal injection of vehicle (DMSO/PBS, 0.5 ml) or estradiol (10 mg/kg); Ratio of the density (%) = density(E2 ip) / density(control) × 100, *p<0.05 was considered to be statistically significant.
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Figure 2: In order to study the gender difference in the expression levels of the HSPs, we designed the experimental protocol to study the effect of estradiol on the male gastric mucosa. Each rat was euthanized before and 1 h, 3 h, or 6 h after the intra-peritoneal injection of 10 mg/kg estradiol (n = 4 at each time point). In the control group, the rats were treated with the vehicle (n = 4 at each time point). a: The expression level of HSP60 was significantly higher in the estradiol-administered group (estrogen group) compared to the control group after the 1 h (Bonferroni correction, p = 0.032) and 6 h (Bonferroni correction, p = 0.034). b: The expression level of HSP70 was significantly higher in the estradiol-administered group (estrogen group) compared to the control group after the 1 h (Bonferroni correction, p = 0.04), 3 h (Bonferroni correction, p = 0.013) and 6 h (Bonferroni correction, p = 0.016). c: The expression level of HSP90 was significantly higher in the estradiol-administered group (estrogen group) compared to the control group after the 1 h (Bonferroni correction, p<0.001), 3 h (Bonferroni correction, p = 0.003). E2: estradiol; 1 h: 1 h after intra-peritoneal injection of vehicle (DMSO/PBS, 0.5 ml) or estradiol (10 mg/kg); 3 h: 3 h after intra-peritoneal injection of vehicle (DMSO/PBS, 0.5 ml) or estradiol (10 mg/kg); 6 h: 6 h after intra-peritoneal injection of vehicle (DMSO/PBS, 0.5 ml) or estradiol (10 mg/kg); Ratio of the density (%) = density(E2 ip) / density(control) × 100, *p<0.05 was considered to be statistically significant.

Mentions: The expression of each HSP was evaluated by a previously reported method [14]. Briefly, the samples (25 µg of protein/lane) were electrophoresed on 9% SDS-polyacrylamide gels, electrophoretically transferred to a PVDF membrane (0.45 µm pore size; Amersham Pharmacia Biotech, Buckinghamshire, England), and processed as described by Towbin et al. [15]. The membrane was incubated with the rabbit anti-HSP60 antibody [16], rabbit anti-HSP70 antibody [17], or rabbit anti-HSP90 antibody [18] and treated with horseradish peroxidase-conjugated anti-rabbit IgG (1:1000). The stained bands were visualized with 0.02% 3,3-diaminobenzidine tetrahydrochloride containing 0.005% H2O2. The intensity of the visualized band was quantified using Quantify One.4.0 software (Bio-Rad, Richmond, California). The ratio of the density of the stained band was calculated using the following formula as previously reported [19]. In Fig. 1d–g, Ratio of the density (%) = density (female HSP) / density (male HSP) × 100; in Fig. 2a–c, Ratio of the density (%) = density (esreadiol (E2) i.p.) / density (control) × 100; in Fig. 3, Ratio of the density (%) = density (ovariectomy) / density (sham operation) × 100.


Correlation of heat shock protein expression to gender difference in development of stress-induced gastric mucosal injury in rats.

Takahashi T, Otaka M, Odashima M, Tamaki K, Takada M, Izumi Y, Shibuya T, Sakamoto N, Itoh H, Watanabe S - J Clin Biochem Nutr (2010)

In order to study the gender difference in the expression levels of the HSPs, we designed the experimental protocol to study the effect of estradiol on the male gastric mucosa. Each rat was euthanized before and 1 h, 3 h, or 6 h after the intra-peritoneal injection of 10 mg/kg estradiol (n = 4 at each time point). In the control group, the rats were treated with the vehicle (n = 4 at each time point). a: The expression level of HSP60 was significantly higher in the estradiol-administered group (estrogen group) compared to the control group after the 1 h (Bonferroni correction, p = 0.032) and 6 h (Bonferroni correction, p = 0.034). b: The expression level of HSP70 was significantly higher in the estradiol-administered group (estrogen group) compared to the control group after the 1 h (Bonferroni correction, p = 0.04), 3 h (Bonferroni correction, p = 0.013) and 6 h (Bonferroni correction, p = 0.016). c: The expression level of HSP90 was significantly higher in the estradiol-administered group (estrogen group) compared to the control group after the 1 h (Bonferroni correction, p<0.001), 3 h (Bonferroni correction, p = 0.003). E2: estradiol; 1 h: 1 h after intra-peritoneal injection of vehicle (DMSO/PBS, 0.5 ml) or estradiol (10 mg/kg); 3 h: 3 h after intra-peritoneal injection of vehicle (DMSO/PBS, 0.5 ml) or estradiol (10 mg/kg); 6 h: 6 h after intra-peritoneal injection of vehicle (DMSO/PBS, 0.5 ml) or estradiol (10 mg/kg); Ratio of the density (%) = density(E2 ip) / density(control) × 100, *p<0.05 was considered to be statistically significant.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 2: In order to study the gender difference in the expression levels of the HSPs, we designed the experimental protocol to study the effect of estradiol on the male gastric mucosa. Each rat was euthanized before and 1 h, 3 h, or 6 h after the intra-peritoneal injection of 10 mg/kg estradiol (n = 4 at each time point). In the control group, the rats were treated with the vehicle (n = 4 at each time point). a: The expression level of HSP60 was significantly higher in the estradiol-administered group (estrogen group) compared to the control group after the 1 h (Bonferroni correction, p = 0.032) and 6 h (Bonferroni correction, p = 0.034). b: The expression level of HSP70 was significantly higher in the estradiol-administered group (estrogen group) compared to the control group after the 1 h (Bonferroni correction, p = 0.04), 3 h (Bonferroni correction, p = 0.013) and 6 h (Bonferroni correction, p = 0.016). c: The expression level of HSP90 was significantly higher in the estradiol-administered group (estrogen group) compared to the control group after the 1 h (Bonferroni correction, p<0.001), 3 h (Bonferroni correction, p = 0.003). E2: estradiol; 1 h: 1 h after intra-peritoneal injection of vehicle (DMSO/PBS, 0.5 ml) or estradiol (10 mg/kg); 3 h: 3 h after intra-peritoneal injection of vehicle (DMSO/PBS, 0.5 ml) or estradiol (10 mg/kg); 6 h: 6 h after intra-peritoneal injection of vehicle (DMSO/PBS, 0.5 ml) or estradiol (10 mg/kg); Ratio of the density (%) = density(E2 ip) / density(control) × 100, *p<0.05 was considered to be statistically significant.
Mentions: The expression of each HSP was evaluated by a previously reported method [14]. Briefly, the samples (25 µg of protein/lane) were electrophoresed on 9% SDS-polyacrylamide gels, electrophoretically transferred to a PVDF membrane (0.45 µm pore size; Amersham Pharmacia Biotech, Buckinghamshire, England), and processed as described by Towbin et al. [15]. The membrane was incubated with the rabbit anti-HSP60 antibody [16], rabbit anti-HSP70 antibody [17], or rabbit anti-HSP90 antibody [18] and treated with horseradish peroxidase-conjugated anti-rabbit IgG (1:1000). The stained bands were visualized with 0.02% 3,3-diaminobenzidine tetrahydrochloride containing 0.005% H2O2. The intensity of the visualized band was quantified using Quantify One.4.0 software (Bio-Rad, Richmond, California). The ratio of the density of the stained band was calculated using the following formula as previously reported [19]. In Fig. 1d–g, Ratio of the density (%) = density (female HSP) / density (male HSP) × 100; in Fig. 2a–c, Ratio of the density (%) = density (esreadiol (E2) i.p.) / density (control) × 100; in Fig. 3, Ratio of the density (%) = density (ovariectomy) / density (sham operation) × 100.

Bottom Line: The gastric ulcer index was significantly higher in male rats compared to female rats observed after 12 h water immersion stress exposure.At this time point, the expression levels of HSP60 and HSP90 in the gastric mucosa were significantly higher in females than those in males.Inversely, an ovariectomy dramatically reduced the expression of HSP60, HSP70 and HSP90 in the gastric mucosa.

View Article: PubMed Central - PubMed

Affiliation: Department of Gastroenterology, Juntendo University School of Medicine, Bunkyo-Ku, Tokyo 113-8421, Japan.

ABSTRACT
Recent studies have indicated that heat shock proteins (HSPs), which function as molecular chaperones, play important roles in cellular responses to stress-related events. However, the gender difference in the expression of HSP in the gastric mucosa remains unclear. In order to understand the mechanism of gender difference in the prevalence or severity of gastric mucosal lesions, the expression level of HSP and the correlation of estrogen to HSP induction in the gastric mucosa were evaluated in this study. The basal expression levels of HSP60 and HSP90 in the gastric mucosa were significantly higher in females than those in males. The gastric ulcer index was significantly higher in male rats compared to female rats observed after 12 h water immersion stress exposure. At this time point, the expression levels of HSP60 and HSP90 in the gastric mucosa were significantly higher in females than those in males. An estrogen-treatment significantly induced the expression of HSP60, HSP70 and HSP90 in the gastric mucosa. Inversely, an ovariectomy dramatically reduced the expression of HSP60, HSP70 and HSP90 in the gastric mucosa. Our results suggested that estrogen might play an important role in gastric mucosal protection with the induction of gastric mucosal HSPs.

No MeSH data available.


Related in: MedlinePlus