第五章 結論與建議
第二節 建議
在現代，糖尿病的病人愈來愈多，且外傷的照護愈受重視，而人的 生命價值及生活品質也愈受尊重。進行有關已是糖尿病人，又因外傷而 出血性休克所導致的傷害的研究，雖然變因增多了，但是相信更符合臨 床的可能，也會對臨床醫學與基礎醫學都有相當的助益。在動物模型的 研究結果完善之後，也可以對臨床病患的照護產生不同程度的啟發和思 考方向。
圖表
Control HS DM DM+HS DM+HS+R
RV
LV
RV RV RV RV
LV LV LV LV
LV= Left ventricle； RV= Right ventricle
Figure 1：以 H＆E 染色觀察心臟整體型態變化之情形
利用組織切片觀察所有組別其心臟橫切面。Control 組為一正常心臟橫切 面圖，其左右心室腔壁、孔徑大小正常；HS 其左右心室均有擴張的現象，
其心室壁厚度均有變薄、變細，已走向擴張性的肥大；DM、DM+HS 與 DM+HS+組其左右心室均有擴張的現象，其心室壁厚度均有變薄、變細，
特別是DM+HS 組更為明顯；已走向擴張性的肥大。
Control HS DM
DM+HS DM+HS+R
Figure 2：以 Trichrome Masson 染色觀察心肌細胞纖維化情形
與 Control 組相比，HS 心肌組織，有較多解不規則斷裂與些微藍色的膠 原蛋白纖維產生，表示有纖維化的情形出現，DM 組同樣有些微的不規則 與藍色膠原蛋白的表現，而DM+HS 與 HS、DM 組相比有明顯且嚴重的 不規則斷裂與藍色的膠原蛋白纖維產生，而 DM+HS+R 組不規則的斷裂 與藍色的膠原蛋白纖維產生的現象有減少的情形，表示纖維化的情況有 被改善的現象。
Figure 3：以 TUNEL assay 染色法觀察心肌細胞凋亡情形
將心臟組織以PBS 清洗乾淨，再用 10%中性福馬林固定，以 TUNEL assay 染色法觀察心肌細胞的凋亡情形，藍色螢光點是染DAPI stain，表示同一 平面中所有細胞的所在位置；而綠色螢光點是染TUNEL stain，若出現綠 螢光色點為凋亡的心肌細胞。由結果發現HS、DM、DM+HS 與 DM+HS+R 相較於控制組，心肌細胞凋亡的情形，都有明顯的增加。其中 DM +HS 與HS、DM 組相比，凋亡情形有較為嚴重；而 DM+HS+R 與 DM +HS 組 相比有明顯的減少。
C o n t r o l H S
D M D M + H S
D M + H S + R
IGFIIR 300 kDa
Control HS DM DM+HS DM+HS+R
β- actin 46 kDa
% of IGFIIR/actin
Control HS DM DM+HS DM+HS+R
DM: diabetes mellitus HS: Hemorrhagic shock
R: resuscitation
# Δ
Figure 4：以Western blotting方法分析IGFIIR蛋白表現量
從實驗結果發現HS、DM、DM+HS相較於Control，IGFIIR蛋白的表現有 上升的趨勢，DM+HS與HS、DM相比，蛋白的表現都有加乘性上升的趨 勢，特別是DM與DM+HS相比後，IGFIIR其蛋白的表現有明顯加乘性上 升的現象，並達到統計上顯著差異；而DM+HS+R與DM+HS相比後，
IGFIIR其蛋白的表現有明顯下降的現象，並達到統計上顯著差異。
(#表示 P <0.05，DM與DM+HS組比較，具顯著差異;Δ表示 P <0.05，與 DM+HS+R組比較，具顯著差異)
0
Control HS DM DM+HS DM+HS+R
p-PKC-α 82 kDa
β- actin 46 kDa
Control HS DM DM+HS DM+HS+R DM: diabetes mellitus HS: Hemorrhagic shock
R: resuscitation
**
**
DM: diabetes mellitus HS: Hemorrhagic shock
R: resuscitation
Δ Δ
# Δ
#
Figure 5：以Western blotting方法分析Gαq 、p-PKCα蛋白表現量
從實驗結果發現HS、DM、DM+HS組相較於控制組，Gαq、p-PKCα蛋白 的表現有明顯的上升，特別是p-PKCα蛋白表現達到統計上顯著差異，而 DM+HS組，與HS組相比，p-PKCα蛋白的表現有加乘性上升的趨勢，特 別是DM+HS+R組與DM+HS組相比，其Gαq 、p-PKCα蛋白的表現有明 顯被壓制的現象，並達到統計上顯著差異。 (**表示 P <0.05，與控制組 比較，具顯著差異；＃表示 P <0.05，與DM+HS組比較，具顯著差異; Δ 表示 P <0.05，ΔΔ表示 P <0.01，與DM+HS+R組比較，具顯著差異)
0
% of calcineurin/actin
Control HS DM DM+HS DM+HS+R
Calcineurin 61kDa
β- actin 46kDa
Control HS DM DM+HS DM+HS+R
DM: diabetes mellitus HS: Hemorrhagic shock
R: resuscitation
* *
*
# Δ
Figure 6：以Western blotting方法分析Calcineurin蛋白表現量
從實驗結果發現HS、DM+HS組相較於控制組，Calcineurin蛋白的表現有
Cytochrome c 11 kDa
Active form 35kDa
Caspase 9
β- actin 46 kDa
Control HS DM DM+HS DM+S+R
0
% of active caspase9/actin *
Control HS DM DM+HS DM+HS+R Control HS DM DM+HS DM+HS+R
% of Cytochromec/actin
# Δ
# Δ Δ Δ
##
###***
DM: diabetes mellitus HS: Hemorrhagic shock
R: resuscitation DM: diabetes mellitus
HS: Hemorrhagic shock R: resuscitation
Figure 7：以Western blotting方法分析Cytochrome c、Caspase 9蛋白表現 量
從實驗結果發現HS、DM、DM+HS相較於Control，Cytochrome c、Caspase 9 蛋白的表現有明顯的上升，而DM+HS達到統計上顯著差異，HS、DM 與DM+HS相比，蛋白的表現都有加乘上升的趨勢，特別是DM+HS與 DM+HS+R相比後，其蛋白的表現有明顯下降的現象，並達到統計上顯著 差異。 (*表示 P <0.05，與控制組比較，具顯著差異；#表示 P <0.05，
##表示 P <0.01，###表示 P <0.001，與DM+HS組比較，具顯著差異; Δ 表示 P <0.05，ΔΔΔ表示 P <0.001，與DM+HS+R組比較，具顯著差異)
0
Control HS DM DM+HS DM+HS+R
35kDa
20kDa
β- actin 46 kDa
% of active caspase3/actin
Control HS DM DM+HS DM+HS+R
***
* *
DM: diabetes mellitus HS: Hemorrhagic shock
R: resuscitation
## Δ Δ
###
Figure 8：以Western blotting方法分析Caspase 3蛋白表現量
從實驗結果發現HS、DM、DM+HS相較於Control， Caspase 3蛋白的表 現有明顯的上升，且達到統計上顯著差異，DM+HS與HS、DM相比，蛋
p-PI3k 110kDa 46kDa β-actin
Control HS DM DM+HS DM+HS+R
0
% of p-PI3k/actin
Control HS DM DM+HS DM+HS+R
DM: diabetes mellitus HS: hemorrhagic shock R: resuscitation
* * *
Figure 9：以Western blotting方法分析p-PI3K蛋白表現量
從實驗結果發現HS、DM與DM+HS組相較於控制組，p-PI3K蛋白的表現 有明顯的抑制，且達到統計上顯著差異，而DM+HS+R組有些微的回復，
與DM+HS相比。(*表示 P <0.05，與控制組比較，具顯著差異)
β-actin
% of p-Akt/actin
Control HS DM DM+HS DM+HS+R
*
**
DM: diabetes mellitus HS: Hemorrhagic shock
R: resuscitation
46kDa 60kDa
Figure 10：以Western blotting方法分析p-Akt蛋白表現量
從實驗結果發現HS、DM、DM+HS與DM+HS+R組相較於控制組，p-Akt 蛋白的表現有明顯的抑制，且達到統計上顯著差異，而DM+HS+R組些微 的回復，與DM+HS相比。(*表示 P <0.05，與控制組比較，具顯著差異)
p-Bad 25kDa
46kDa
Control HS DM DM+HS DM+HS+R
β-actin
14 0 DM: diabetes mellitus
HS: Hemorrhagic shock R: resuscitation
Control HS DM DM+HS DM+HS+R
% of p-Bad/actin * * * *
Figure 11：以Western blotting方法分析p-Bad蛋白表現量
從實驗結果發現HS、DM、DM+HS與 DM+HS+R組相較於控制組，p-Bad 蛋白的表現有明顯的下降，並達到統計上顯著差異，而DM+HS組，與HS 及DM 組 相 比 ， p-Bad 蛋 白 的 表 現 都 有 加 乘 性 下 降 的 趨 勢 ， 特 別 是 DM+HS+R與DM+HS組相較之下，其蛋白的表現有回復的現象。(*表示 P
<0.05，與控制組比較，具顯著差異)
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