2001 Fiscal Year Final Research Report Summary
Research on Apoptosis after Hemorrhagic Shock -Hypertonic Saline Resuscitation-
| Project/Area Number |
12671498
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Anesthesiology/Resuscitation studies
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| Research Institution | Nara Medical University |
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Principal Investigator |
MURAO Yoshinori Nara University, Associate Professor, 医学部, 助教授 (50211602)
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| Project Period (FY) |
2000 – 2001
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| Keywords | Hemorrhagic Shock / Hypertonic Saline / Apoptosis / TUNEL / DNA ladder / caspase-3 |
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| Research Abstract |
Male Sprague-Dawley rats were hemorrhaged until a mean arterial pressure (MAP) of 40 +/- 5 mmHg was reached. This level of hypotension was sustained for 1 h, and the animals were randomly assigned to the following groups by resuscitation methods. Control, Sham, HS + SB : (4 mg/Kg of 7.5 % NaCl solution: HS) and shed blood (SB) at the same time, 2LR + SB : hemorrhage shock for 1 h and resuscitated with Lactated Ringer's solution (2 times the volume of the shed blood) + SB, 3LR (3 times the volume of the shed blood). Lungs were harvested and total histology scores were examined at 48 h after hemorrhage and resuscitation. HS + SB group significantly prevented lung damage compared with other groups (p < O.05). The effect of hypertonic saline resuscitation on the time course of intestinal damage and apoptosis after hemorrhagic shock in mice was invesigated. After anesthesia, male BALB/c mice were hemorrhaged to the mean arterial pressure of 40 +/- 5 mmHg for 90 min.
Resuscitation was made by
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HS + SB or 2LR + SB. Intestinal injury was devised and based on the extent of the vacuolation at basal area of the intestinal villi. Apoptosis score of the small intestines was examined by TUNEL method and DNA ladder assay. Activation of caspase-3 expression was assessed, by Western blot. Hemorrhagic shock and resuscitation induced the apoptosis in 2LR + SB at 2 h and intestinal damage in 2LR + SB at 6h compared to other groups significantly (p < 0.01) Apoptosis index of HS + SB at 2h group was significantly reduced compared to 2LR+SB at 2h (p < 0.01) and also intestinal damage of HS + SB at 6h group was significantly reduced compared to 2LR + SB at 6h (p < 0.01).
Activation of caspase-3 of HS + SB at 2h group was reduced compared to 2LR + SB at 2h group. Conclusions: Hypertonic saline resuscitation reduced the apoptosis and intestinal damage after hemorrhagic shock and resuscitation. Thus, the effect of hypertonic saline resuscitation toward apoptosis may have important role in hemorrhagic shock and resuscitation. Less
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