Gangula 2007
Gangula 2007
Gangula 2007
doi:10.1152/ajpgi.00406.2006.
Gangula PRR, Maner WL, Micci M-A, Garfield RE, Pasricha has been reported in diabetic patients (28). Although the
PJ. Diabetes induces sex-dependent changes in neuronal nitric oxide pathogenesis of diabetic gastropathy is not completely under-
synthase dimerization and function in the rat gastric antrum. Am J stood, it appears to be much more common (up to four times)
Physiol Gastrointest Liver Physiol 292: G725–G733, 2007; in women (46, 22). Such a sex bias is also seen in the idiopathic
doi:10.1152/ajpgi.00406.2006.—Diabetic gastroparesis is a disorder variety of gastric dysfunction, reinforcing the concept of
that predominantly affects women. However, the biological basis of
http://www.ajpgi.org 0193-1857/07 $8.00 Copyright © 2007 the American Physiological Society G725
G726 DIABETES IMPAIRED GASTRIC NNOS DIMER FUNCTION
PDZ/GLGF domain interacts with various proteins and deter- fasting male and female diabetic as well as control rats at the time the
mines nNOS␣ subcellular localization and function (9, 47). In experiment was performed (12 wk after STZ). Diabetes was con-
contrast, the NH2-terminally truncated nNOS and nNOS␥ firmed if blood glucose levels ranged from 300 – 400 mg/dl in both
lack the PDZ/GLGF motif for protein-protein interaction and male and female rats. No differences in blood glucose levels were
possible membrane association. nNOS⫺/⫺ mice lacking exon 2 noticed between male and female rats 12 wk after the induction of
diabetes with STZ. Age-matched control male and female rats showed
and subsequent membrane-associated full-length nNOS␣ ex-
blood glucose levels between 80 –95 mg/dl. Both control and diabetic
hibit delayed gastric emptying of solids and liquids (19, 32). rats were selected during the diestrous stage of the estrous cycle. As
The catalytic activity of NOS depends on dimerization of two reported earlier, we noticed that 60 –70% of diabetic rats show a
NOS polypeptides (25). Dimerization results in the creation of persistent diestrous stage of the estrous cycle (data not shown) (8, 24).
high-affinity binding sites for (6R)-tetrahydrobiopterin (BH4) Solid gastric emptying studies. Solid gastric emptying studies were
and arginine in the oxygenase domain and enables electron performed in healthy male and female rats. Similarly, gastric empty-
transfer between flavin and heme groups (7, 17). Enzymatic ing studies were performed in both male and female ras 8 wk after
uncoupling of NOS due to lack of BH4 may in part account for diabetes induction. The gastric emptying rate of a solid meal was
reduced NO production and increased oxidative stress factors measured as reported previously (31). Groups of healthy and diabetic
such as superoxide (26, 51). The importance of dimerization rats from both sexes were fasted for 20 –24 h. A known amount of
for the catalytic activity of endothelial NOS and its disruption solid meal with free access to water was supplied for a 3-h period.
in diabetes has been reported by several independent investi- Food and water were then removed, and the gastric emptying rate of
RESULTS
levels (C) in diabetic male and diabetic females after normal- diabetic gastric dysmotility are common to both men and
ization to their own controls. As can be seen, diabetes signif- women but the latter are disproportionately symptomatic be-
icantly (P ⬍ 0.05) reduced gastric emptying, nitrergic relax- cause the motility of their stomachs is slower to begin with. On
ation, and nNOS␣ protein dimer levels in females compared the other hand, it is also possible that there are fundamental
with male rats (Fig. 7). The decline in gastric antrum nNOS␣ sex-specific differences between how the gastric neuromuscu-
dimer levels correlated with changes in gastric antrum nitrergic lar apparatus is affected by diabetes. The results of our study
relaxation and solid gastric emptying in diabetic female rats suggest that the latter explanation may be more likely.
(Fig. 7). In this study, we have demonstrated sex differences in solid
gastric emptying, antral nNOS expression, nitrergic relaxation,
DISCUSSION
IGP, and nNOS␣ protein dimerization in both diabetic and
Abnormal gastrointestinal motility in diabetes mellitus is nondiabetic rats. Compared with control male rats, healthy
likely multifactorial in origin, reflecting disturbances in enteric females during the diestrous stage of the estrous cycle showed
and vagal neural activity as well as interstitial cells of Cajal and delayed gastric emptying (Fig. 2). However, this was not
smooth muscle function (20, 28, 33, 38). Of these, enteric statistically significant. Previous studies have demonstrated
neuropathy may be particularly important (5, 6, 48). Several that estradiol-17 administration delays gastric emptying for
studies of animal models of diabetes have convincingly shown liquids in both male and female rats (16). Thus, more studies
disturbances in enteric nerves, particularly involving nitrergic are warranted to address whether solid gastric emptying rate is
nerves (14, 49, 52). Since these studies have used male rats different during various stages of the estrous cycle when
exclusively, it has been difficult to correlate these findings with estrogen levels are higher than in the diestrous phase. How-
the clinical finding that 80% of all gastroparetic patients are ever, nitrergic relaxation of the antrum is clearly greater in
female. It is possible that the pathogenetic mechanisms of healthy diestrous females (Fig. 3) compared with males, and
AJP-Gastrointest Liver Physiol • VOL 292 • MARCH 2007 • www.ajpgi.org
G730 DIABETES IMPAIRED GASTRIC NNOS DIMER FUNCTION
unconscious. Both of these alternatives may lead to spurious vary depending upon the type of method employed, i.e., liquids
data. By using a telemetric device in conscious, unrestrained vs. solids; type, duration, and severity of diabetes. Human
animals, we obtained the most accurate measurement for in- studies have demonstrated that diabetic women have more
traluminal pressure. In our study, L-NAME infusion increased delayed gastric emptying compared with diabetic men (22, 46).
IGP significantly in female but not in male rats, suggesting that In the present study, we have demonstrated that solid gastric
nonnitrergic mechanisms may be more important in the latter emptying rate is significantly slower in both male and female
(Fig. 4, C and D). Thus it appears that females rely on nitrergic rats after diabetic induction compared with their respective
control of gastric motility to a greater extent than males and controls. Furthermore, these studies suggested that diabetic
hence may be more vulnerable to alterations of this system females showed significant delayed gastric emptying compared
induced by diabetes. with diabetic males. These studies together with clinical find-
We next turned our attention to changes in these parameters ings suggested that gastric emptying is slower in diabetic
after induction of diabetes. The rate of gastric emptying may females than in diabetic males.
AJP-Gastrointest Liver Physiol • VOL 292 • MARCH 2007 • www.ajpgi.org
G732 DIABETES IMPAIRED GASTRIC NNOS DIMER FUNCTION
In diabetic gastric dysfunction, antral motility and the coor- tion for the greater vulnerability of females to develop diabetic
dination of pressures between the antrum and duodenum are gastric dysmotility problems.
diminished (28, 46, 53). Antral hypomotility has been recorded
with intraluminal pressure transducers in patients with diabetes ACKNOWLEDGMENTS
mellitus. In this study, we have shown that nitrergic relaxation The authors thank Xiemin Cao for technical assistance and John Winston
(Fig. 3) is significantly reduced with concomitant increases in and Raj Kumar for thoughtful discussions. Also, we thank Dr. Shao-Qing Shi
IGP in female diabetic rats (Fig. 4D). Furthermore, nNOS␣ for help with the surgeries for telemetric studies.
dimerization (Fig. 6) but not total (Fig. 5) nNOS expression is
significantly reduced in these animals. After normalization REFERENCES
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