結腸癌相關指標

在 aberrant crypt foci (ACF) 計數結果方面，結果顯示 DS0、DS0.5 以及 DS1 組發現皆有 1 至 6 個不等異常腺窩 (AC) 的 ACFs 的存在 (圖 4-5)。PS0 及 PS0.5 組則完全沒有 ACFs 發生。

依 ACFs 的大小分類比較，依據 Cassand 等人 (1997) 研究，將 1 至 3 個 AC 的 ACFs 定義為小的 ACFs，而大於 3 個 AC 的 ACFs 定義為 大的 ACFs。各組分別做比較，結果顯示 DS0 組 1 個及 2 個 AC 的 ACFs 數目顯著低於 DS0 組 (p < 0.05)。比較小的 ACFs 總數，DS0、DS0.5 及 DS1 三組間有負向的劑量差異 (p < 0.05)，也就是劑量越高，小的 ACFs 總數越少 (表 4-5)。而比較大的 ACFs 總數，DS1 組則顯著的低於 DS0 組 (p < 0.05) (表 4-6)。

依腸區域比較 ACFs 數目比較，各組分別做比較，結果顯示 DS1 組 在盲腸端 ACFs 總數顯著低於 DS0 組 (p < 0.05)。在中段結腸，DS1 組 ACFs 總數顯著低於 DS0 組和 DS0.5 組 (p < 0.05)。在肛門端，DS0.5 組 和 DS1 組 ACFs 總數顯著皆低於 DS0 組 (p < 0.05)。將結腸不分段，比 較總 ACFs 總數，DS0、DS0.5 及 DS1 三組間有負向的劑量差異 (p < 0.05) (表 4-7)。

在 COX-2 蛋白質表現量分析方面，在注射 DMH 的 DS0、DS0.5 及 DS1 三組有 COX-2 的表現，在注射 PBS 的組別則沒有 COX-2 的表現。

比較 DS0、DS0.5 及 DS1 三組蛋白質表現量 (圖 4-6A)，各組之間沒有 顯著差異。而依腸區域分開比較 DS0、DS0.5 及 DS1 三組，無論是在盲 腸端、中段或是肛門端 COX-2 蛋白質表現量，各組之間沒有顯著差異 (圖 4-6B~C)。在 iNOS 蛋白質表現量分析方面，本實驗各組皆沒有偵測 到其蛋白質表現。

在 PGE₂濃度分面，DS0 組為 16205 ± 2910 pg/mL；DS0.5 組為 6523

± 2823 pg/mL；DS1 組為 4507 ± 396 pg/mL；PS0 組為 2704 ± 415 pg/mL；

PS0.5 組為 3146 ± 537 pg/mL。在注射 DMH 的 DS0 組顯注高於注射 PBS 的 PS0 組 (p < 0.05)，DS0、DS0.5 及 DS1 三組之間則沒有顯著差異 (圖 4-7)。

在 β-glucuronidase 酵素相對活性方面，DS0 組為 346.9 ± 15.6 nmole/min/mg protein；DS0.5 組為 209.6 ± 17.1 nmole/min/mg protein；

DS1 組為 163.7 ± 5.2 nmole/min/mg protein；PS0 組為 158.4 ± 8.8 nmole/min/mg protein；PS0.5 組為 159.3 ± 6.3 nmole/min/mg protein。DS0 組顯著高於 PS0 組 (p < 0.05)，DS0.5 組及 DS1 組顯著低於 DS0 組 (p <

0.05) (圖 4-8)。

圖 4-5、以亞甲藍染色觀察老鼠結腸黏膜上 ACFs (10×40)^1,2

Figure 4-5. The pathological assessment by methylene blue stain in the ACFs of the colonic mucosa of rats^1,2

1ACFs：aberrant crypt foci

2PS0：received saline and fed AIN93G diet；PS0.5：received saline and fed AIN93G diet containing 0.5% saponins ； DSO ： received 1,2-dimethylhydrazine (DMH) and fed AIN93G diet；DS0.5：received DMH and fed AIN93G diet containing 0.5% saponins；

DS1：received DMH and fed AIN93G diet containing 1% saponins.

DS0-1 DS0-2 DS0-3 DS0-4

DS0.5-1 DS0.5-2 DS0.5-3 DS0.5-4

DS1-1 DS1-2 DS1-3 DS1-4

PS0-1 PS0-2 PS0.5-1 PS0.5-2

表 4-5、老鼠小型 ACFs 計數^1,2

Table 4-5. The counts of small ACFs in rats^1,2

The incidence of ACFs³

1AC 2AC 3AC small⁴

PS0 0/4 ND⁵ ND ND ND

PS0.5 0/4 ND ND ND ND

DS0 6/6 10.16 ± 1.22^a 9.00 ± 1.46^a 5.83 ± 0.70 25.00 ± 1.89^a DS0.5 6/6 5.67 ± 0.59^ab 6.00 ± 0.75^ab 3.83 ± 0.41 15.50 ±0.54^b DS1 6/6 2.83 ± 0.39^b 1.83 ± 0.50^b 3.00 ± 0.63 7.67 ± 1.32^c

1Data are expressed as Mean ± SEM (n=6)

2PS0：received saline and fed AIN93G diet；PS0.5：received saline and fed AIN93G diet containing 0.5% saponins ； DSO ： received 1,2-dimethylhydrazine (DMH) and fed AIN93G diet；DS0.5：received DMH and fed AIN93G diet containing 0.5% saponins；

DS1：received DMH and fed AIN93G diet containing 1% saponins.

3The incidence of ACFs= (the number of ACF rats/the number of total rats)

4small ACFs：1~3AC/ACF

5ND：No detection

abcLetters in each column sharing the different superscripts are significantly different (p <

0.05)

表 4-6、老鼠大型的 ACFs 計數 ^1,2

Table 4-6. The counts of large ACFs in rats^1,2

The incidence of ACFs³

4AC 5AC 6AC large⁴

PS0 0/4 ND⁵ ND ND ND

PS0.5 0/4 ND ND ND ND

DS0 6/6 1.67 ± 0.27 0.17 ± 0.09 0.17 ± 0.09 2.00 ± 0.20^a

DS0.5 6/6 1.17 ± 0.17 0 0 1.17 ± 0.17^ab

DS1 6/6 0.17 ± 0.09 0.17 ± 0.09 0 0.33 ± 0.12^b

1Data are expressed as Mean ± SEM (n=6)

2PS0：received saline and fed AIN93G diet；PS0.5：received saline and fed AIN93G diet containing 0.5% saponins ； DSO ： received 1,2-dimethylhydrazine (DMH) and fed AIN93G diet；DS0.5：received DMH and fed AIN93G diet containing 0.5% saponins；

DS1：received DMH and fed AIN93G diet containing 1% saponins.

3The incidence of ACFs= (the number of ACF rats/the number of total rats)

4large ACFs：>3AC/ACF

5ND：No detection

abLetters in each column sharing the different superscripts are significantly different (p <

0.05)

表 4-7、老鼠各段結腸 ACFs 計數^1,2

Table 4-7. Distribution of ACFs according to different sections of colon^1,2

The incidence of ACFs³

proximal middle distal total

PS0 0/4 ND⁴ ND ND ND

PS0.5 0/4 ND ND ND ND

DS0 6/6 4.50 ± 0.67^a 7.17 ± 0.62^a 15.50 ± 1.35^a 27.00 ± 1.91^a DS0.5 6/6 2.00 ± 0.28^ab 4.83 ± 0.33^a 9.83 ± 0.43^b 16.67 ± 0.44^b DS1 6/6 1.00 ± 0.35^b 1.83 ± 0.50^b 5.17 ± 0.81^b 8.00 ± 1.41^c

1Data are expressed as Mean ± SEM (n=6)

2PS0：received saline and fed AIN93G diet；PS0.5：received saline and fed AIN93G diet containing 0.5% saponins ； DSO ： received 1,2-dimethylhydrazine (DMH) and fed AIN93G diet；DS0.5：received DMH and fed AIN93G diet containing 0.5% saponins；

DS1：received DMH and fed AIN93G diet containing 1% saponins.

3The incidence of ACFs= (the number of ACF rats/the number of total rats)

4ND：No detection

abcLetters in each column sharing the different superscripts are significantly different (p <

0.05)

0.10 0.20.3 0.40.5 0.60.7 0.80.9

PS0 PS0.5 DS0 DS0.5 DS1

COX-2 / β-actin ratio

0

PS0 PS0.5 DS0 DS0.5 DS1

COX-2 / β-actin ratio

A. B.

PS0 PS0.5 DS0 DS0.5 DS1

COX-2 / β-actin ratio

C. D.

PS0 PS0.5 DS0 DS0.5 DS1

COX-2 / β-actin ratio

圖 4-6、老鼠結腸黏膜 COX-2 蛋白質表現量^1,2

Figure 4-6. The expression of COX-2 protein in the colonic mucosa^1,2

A. The total expression of COX-2 protein in the colonic mucosa. B. The expression of COX-2 protein in proximal colonic mucosa. C. The expression of COX-2 protein in middle colonic mucosa. D. The expression of COX-2 protein in distal colonic mucosa.

1Data are expressed as Mean ± SEM (n=3~4)

2The image of western blot was representative.

abLetters at the top of each bar sharing the different superscripts are significantly different (p < 0.05)

0 5000 10000 15000 20000 25000

PS0 PS0.5 DS0 DS0.5 DS1

P G E 2 le v el (p g /m g p ro te in )

圖 4-7、結腸黏膜 PGE₂濃度^1,2

Figure 4-7. The PGE2level in the colonic mucosa^1,2

1Data are expressed as Mean ± SEM (n=3~4)

2PS0：received saline and fed AIN93G diet；PS0.5：received saline and fed AIN93G diet containing 0.5% saponins ； DSO ： received 1,2-dimethylhydrazine (DMH) and fed AIN93G diet；DS0.5：received DMH and fed AIN93G diet containing 0.5% saponins；

DS1：received DMH and fed AIN93G diet containing 1% saponins.

abLetters at the top of each bar sharing the different superscripts are significantly different (p < 0.05)

b

ab a a ab

0 100 200 300 400

PS0 PS0.5 DS0 DS0.5 DS1

E n zy m e a ct iv it y (n m o le /m in /m g p ro te in )

圖 4-8、結腸黏膜 β-glucuronidase 酵素活性^1,2,3

Figure 4-8. The activity of β-glucuronidase in the colonic mucosa^1,2,3

1Data are expressed as Mean ± SEM (n=3~4)

2PS0：received saline and fed AIN93G diet；PS0.5：received saline and fed AIN93G diet containing 0.5% saponins ； DSO ： received 1,2-dimethylhydrazine (DMH) and fed AIN93G diet；DS0.5：received DMH and fed AIN93G diet containing 0.5% saponins；

DS1：received DMH and fed AIN93G diet containing 1% saponins.

3The enzyme activities are derived from the changes of activity at 0 and 60 mins.

abcLetters at the top of each bar sharing the different superscripts are significantly different (p < 0.05)

b

ac

ac

a a