探討 DOW 發酵蛹蟲草燕麥對 TAA 誘發小鼠肝纖維化之影響

第四章研究結果

第二節探討 DOW 發酵蛹蟲草燕麥對 TAA 誘發小鼠肝纖維化之影響

一、不同水分發酵之蛹蟲草對肝纖維化小鼠肝重、體重與肝重/體重比值之變化 過去研究顯示，經 TAA 誘發肝纖維化後，負控制組之體重皆比正常組有顯著降低，而負控制組之肝重相較於正常組則呈現顯著上升，且肝重與體重之比值均較正常組來的高 (Abdulaziz Bardi et al., 2014; Chen et al., 2012)。本研究結果與過去相符，如表 4-2 結果顯示，雄性 BALB/c 小鼠經腹腔注射 TAA 8 周後，

其體重均顯著比 NOR 組低 (p<0.001)，而肝重及肝重/體重之比值皆顯著高於 NOR 組 (p<0.001)，在餵食試驗物質後，相較於 TAA 組，各組肝重及肝重/體 重比值均有降低，且體重也有回升。

二、不同水分發酵之蛹蟲草對肝纖維化小鼠血清中肝功能指標 AST、ALT、ALP 及 TBIL 含量之變化

血清肝功能生化參數不僅能夠評估肝臟損傷程度，還能依據各項參數之變化判斷為何種肝損傷類型，血清中 AST 與 ALT 是一個重要且被廣泛使用於檢測肝功能的指標，而 ALP 與 TBIL 則被用於檢測肝膽功能，當肝細胞受損或肝細胞膜通透性改變時，AST 與 ALT 會藉由周邊血管釋放到血液中，另外，ALP 位於膽小管與上皮細胞，當膽管阻塞、膽汁淤積時，血液中 ALP 含量便會提高，

但 ALP 如何釋放到血液中的機制目前尚未清楚 (Ramaiah, 2007)。

血清肝功能檢測結果如表 4-3 所示，在 AST 與 ALT 方面，TAA 組活性

圖 4-6 不同之氯鹽對蛹蟲草發酵燕麥中蟲草素 (A) 與腺苷 (B) 含量之影響 Fig. 4-6 Effect of various of chloride salt on the cordycepin (A) and adenosine (B) production of C. militaris-fermentedoats in solid culture. The 30X concentration of Mg²⁺, Na⁺, K⁺, Ca²⁺, Fe²⁺ in MgCl₂, NaCl, KCl, CaCl₂, FeCl₂ were equal to that in 30X-DOW. Data are presented as mean ± SD (n=3). ^*, ^**, ^*** indicated the significant difference (p<0.05, p<0.01, p<0.001, respectively) as compared with UPW.

表 4-2、不同水分發酵之蛹蟲草燕麥對肝纖維化小鼠肝重、體重與肝重/體重比值之影響

Table 4-2. The effects of C. militaris-fermented oats cultured with various water on the body weight gain, liver weight, and liver weight/body weight ratio of the TAA-induced fibrosis mice

Liver weight gain (g) Body weight (g) Liver weight/Body weight ratio (%)

NOR 1.24 ± 0.06 29.36 ± 1.17 4.51 ± 0.14

TAA 2.07 ± 0.09^### 23.34 ± 1.05^### 7.88 ± 0.19^###

SL 1.95 ± 0.07^*** 27.35 ± 1.03^*** 7.15 ± 0.24^***

UCM 1.91 ± 0.08^*** 27.31 ± 0.46^*** 7.20 ± 0.24^***

DCM-L 1.90 ± 0.06^*** 27.88 ± 0.59^*** 6.98 ± 0.36^***

DCM-H 1.93 ± 0.04^*** 27.26 ± 1.15^*** 7.09 ± 0.31^***

SCM 1.81 ± 0.04^*** 25.79 ± 0.55^*** 7.10 ± 0.18^***

DOW 1.89 ± 0.06^*** 26.96 ± 0.82^*** 7.17 ± 0.15^***

AS 1.69 ± 0.08^*** 25.76 ± 0.93^*** 7.10 ± 0.22^***

CC 1.75 ± 0.05^*** 25.01 ± 0.87^*** 6.95 ± 0.24^***

NOR: normal group, TAA: TAA-induced liver fibrosis mice (TAA 100 mg/kg/ip, three times per week), SL: TAA-induced liver fibrosis mice fed 75 mg/kg/day of silymarin, UCM: TAA-induced liver fibrosis mices fed 1.809 g/kg/day of DCM, DCM-L: TAA-induced liver fibrosis mices fed 0.603 g/kg/day of DCM, DCM-H: TAA-induced liver fibrosis mices fed 0.603 g/kg/day of DCM, SCM: TAA-induced liver fibrosis mices fed 0.603 g/kg/day of SCM, DOW: TAA-induced liver fibrosis mices fed with DOW equal to that in DCM-L group (0.603 mL/kg/day), AS:

TAA-induced liver fibrosis mices fed with AS equal to that in DCM-L group (0.329 mg/kg/day), CC: TAA-induced liver fibrosis mices fed with CC equal to that in DCM-L group (0.615 mg/kg/day). Data are presented as mean ± SD (n=8).^### indicated the significant difference (p<0.001) as compared with NOR. *** indicated the significant difference (p<0.001) as compared with TAA.

表 4-3、不同水發發酵之蛹蟲草對肝纖維化小鼠血清肝功能指標 AST、ALT 與 ALP 活性及 TBIL 含量之影響

Table 4-3. The effects of C. militaris-fermented oats cultured with various water on serum AST, ALT, ALP activities and TBIL content in the TAA-induced fibrosis mice

AST (U/L) ALT (U/L) ALP(U/L) TBIL (mg/dL)

NOR 80.63 ± 3.89 43.25 ± 2.55 97.63 ± 4.93 0.043 ± 0.007

TAA 124.50 ± 11.12^### 138.75 ± 10.12^### 202.63 ± 7.48^### 0.109 ± 0.010^###

SL 94.38 ± 4.84^*** 114.75 ± 5.68^* 175.00 ± 16.29^*** 0.091 ± 0.010^***

UCM 92.25 ± 3.45^*** 126.63 ± 12.23 170.63 ± 8.90^*** 0.094 ± 0.005^**

DCM-L 89.25 ± 5.57^*** 128.88 ± 10.79 174.63 ± 13.71^*** 0.086 ± 0.009^***

DCM-H 90.75 ± 6.92^*** 119.63 ± 7.44^* 171.63 ± 14.09^*** 0.086 ± 0.009^***

SCM 97.63 ± 9.02^*** 129.88 ± 14.51 176.13 ± 5.30^*** 0.086 ± 0.012^***

D 96.25 ± 8.97^*** 128.63 ± 13.90 177.13 ± 9.43^*** 0.095 ± 0.009^**

AS 99.00 ± 4.78^*** 125.38 ± 17.73^* 161.75 ± 11.52^*** 0.071 ± 0.011^***

CC 97.13 ± 10.59^*** 106.75 ± 10.50^* 176.38 ± 10.01^*** 0.080 ± 0.011^***

NOR: normal group, TAA: TAA-induced liver fibrosis mices (TAA 100 mg/kg/ip, three times per week), SL: TAA-induced liver fibrosis mices fed 75 mg/kg/day of silymarin, UCM: TAA-induced liver fibrosis mices fed 0.603 g/kg/day of UCM, DCM-L: TAA-induced liver fibrosis mices fed 0.603 g/kg/day of DCM, DCM-H: TAA-induced liver fibrosis mices fed 1.809 g/kg/day of DCM, SCM: TAA-induced liver fibrosis mices fed 0.603 g/kg/day of SCM, DOW: TAA-induced liver fibrosis mices fed with DOW equal to that in DCM-L group (0.603 mL/kg/day), AS:

64 圖 4-7 不同水分發酵之蛹蟲草對肝纖維化小鼠肝臟外觀之影響

Fig. 4-7 The effects of C. militaris fermented oats, which were cultured with various of water on appearance of liver on TAA-induced liver fibrosis in mice. (A) NOR normal group, (B) TAA TAA-induced liver fibrosis mices (TAA 100 mg/kg/ip, three times per week), (C) SL TAA-induced liver fibrosis mices fed 75 mg/kg/day of silymarin, (D) UCM TAA-induced liver fibrosis mices fed 1.809g/kg/day of DCM, (E) DCM-L TAA-induced liver fibrosis mices fed 0.603 g/kg/day of DCM, (F) DCM-H TAA-induced liver fibrosis mices fed 0.603g/kg/day of DCM, (G) SCM TAA-induced liver fibrosis mices fed 0.603 g/kg/day of SCM, (H) DOW TAA-induced liver fibrosis mices fed with DOW equal to that in DCM-L group (0.603 mL/kg/day), (I) AS TAA-induced liver fibrosis mices fed with A equal to that in DCM-L group (0.329 mg/kg/day), (J) CC TAA-induced liver fibrosis mices fed with CC equal to that in DCM-L group (0.615 mg/kg/day)

65 圖 4-8 不同水分發酵之蛹蟲草對肝纖維化小鼠肝組織病理變化之影響

Fig. 4-8 The effects of C. militaris-fermented-oats, which were cultured with various of water on pathological changes on TAA-induced liver fibrosis in mice. (A) NOR normal group, (B) TAA TAA-induced liver fibrosis mices (TAA 100 mg/kg/ip, three times per week), (C) SL TAA-induced liver fibrosis mices fed 75 mg/kg/day of silymarin, (D) UCM TAA-induced liver fibrosis mices fed 1.809g/kg/day of DCM, (E) DCM-L TAA-induced liver fibrosis mices fed 0.603 g/kg/day of DCM, (F) DCM-H TAA-induced liver fibrosis mices fed 0.603g/kg/day of DCM, (G) SCM TAA-induced liver fibrosis mices fed 0.603 g/kg/day of SCM, (H) DOW TAA-induced liver fibrosis mices fed with DOW equal to that in DCM-L group (0.603 mL/kg/day), (I) AS TAA-induced liver fibrosis mices fed with A equal to that in DCM-L group (0.329 mg/kg/day), (J) CC TAA-induced liver fibrosis mices fed with CC equal to that in DCM-L group (0.615 mg/kg/day)

66 圖 4-9 不同水分發酵之蛹蟲草對肝纖維化小鼠肝組織膠原堆積之影響

Fig. 4-9 The effects of C. militaris-fermented-oats, which were cultured with various of water on collagen accmulation on TAA-induced liver fibrosis in mice. (A) NOR normal group, (B) TAA TAA-induced liver fibrosis mices (TAA 100 mg/kg/ip, three times per week), (C) SL TAA-induced liver fibrosis mices fed 75 mg/kg/day of silymarin, (D) UCM TAA-induced liver fibrosis mices fed 1.809g/kg/day of DCM, (E) DCM-L TAA-induced liver fibrosis mices fed 0.603 g/kg/day of DCM, (F) DCM-H TAA-induced liver fibrosis mices fed 0.603g/kg/day of DCM, (G) SCM TAA-induced liver fibrosis mices fed 0.603 g/kg/day of SCM, (H) DOW TAA-induced liver fibrosis mices fed with DOW equal to that in DCM-L group (0.603 mL/kg/day), (I) AS TAA-induced liver fibrosis mices fed with A equal to that in DCM-L group (0.329 mg/kg/day), (J) CC TAA-induced liver fibrosis mices fed with CC equal to that in DCM-L group (0.615 mg/kg/day)

圖 4-10 不同水分發酵之蛹蟲草燕麥對 TAA 誘發肝纖維化小鼠肝組織 IL-1β 蛋白表現之影響

Fig. 4-10 The effect of C. militaris-fermented oats, which were cultured with various of water on haptatic IL-1β expression on TAA-induced liver fibrosis in mice. Target protein expressions were visualized using immunoblotting and quantified using gel analysis software.

(A) NOR normal group, (B) TAA TAA-induced liver fibrosis mices (TAA 100 mg/kg/ip, three times per week), (C) SL TAA-induced liver fibrosis mices fed 75 mg/kg/day of silymarin, (D) UCM TAA-induced liver fibrosis mices fed 1.809g/kg/day of DCM, (E) DCM-L TAA-induced liver fibrosis mices fed 0.603 g/kg/day of DCM, (F) DCM-H TAA-induced liver fibrosis mices fed 0.603g/kg/day of DCM, (G) SCM TAA-induced liver fibrosis mices fed 0.603 g/kg/day of SCM, (H) DOW TAA-induced liver fibrosis mices fed with DOW equal to that in DCM-L group (0.603 mL/kg/day), (I) AS TAA-induced liver fibrosis mices fed with A equal to that in DCM-L group (0.329 mg/kg/day), (J) CC TAA-induced liver fibrosis mices fed with CC equal to that in DCM-L group (0.615 mg/kg/day)

圖 4-11 不同水分發酵之蛹蟲草燕麥對 TAA 誘發肝纖維化小鼠肝組織 IL-6 蛋白表現之影響

Fig. 4-11 The effect of C. militaris-fermented oats, which were cultured with various of water on haptatic IL-6 expression on TAA-induced liver fibrosis in mice. Target protein expressions were visualized using immunoblotting and quantified using gel analysis software. ^**, ^*** indicated the significant difference (p<0.01, p<0.001, respectively) as compared with TAA. ^##, ^### indicated the significant difference (p<0.01, p<0.001, respectively) as compared with TAA. (A) NOR normal group, (B) TAA TAA-induced liver fibrosis mices (TAA 100 mg/kg/ip, three times per week), (C) SL TAA-induced liver fibrosis mices fed 75 mg/kg/day of silymarin, (D) UCM TAA-induced liver fibrosis mices fed 1.809g/kg/day of DCM, (E) DCM-L TAA-induced liver fibrosis mices fed 0.603 g/kg/day of DCM, (F) DCM-H TAA-induced liver fibrosis mices fed 0.603g/kg/day of DCM, (G) SCM TAA-induced liver fibrosis mices fed 0.603 g/kg/day of SCM, (H) DOW TAA-induced liver fibrosis mices fed with DOW equal to that in DCM-L group (0.603 mL/kg/day), (I) AS TAA-induced liver fibrosis mices fed with A equal to that in DCM-L group (0.329 mg/kg/day), (J) CC TAA-induced liver fibrosis mices fed with CC equal to that in DCM-L group (0.615 mg/kg/day)

組比起 NOR 組 IL-6 之表現量提升 341 % (p<0.001)，而餵食試驗物質後，相 較於 TAA 組，DCM-L、DCM-H、SCM、D、AS 與 CC 分別降低 37% (p<0.001)、

45% (p<0.001)、33% (p<0.001)、31% (p<0.001)、51% (p<0.001) 與 57% (p<0.001)，

而 SL 則降低 20% (p<0.01)，表示 DCM-L、DCM-H、SCM、D、AS 與 CC 均 1264% (p<0.001)，經試驗物質作用後，DCM-L、DCM-H、SCM、D、AS 與 CC 分別降低 52% (p<0.001)、51% (p<0.001)、36% (p<0.001)、29% (p<0.001)、42%

(p<0.001) 與 46% (p<0.001)，而 SL 使其下降 17% (p<0.01)，UCM 則不影響 and Benyon, 2004)。另外，TGF-β1 還能藉由 trans-modulating PDGF receptor 來調控 HSCs 的增生 (Olaso and Friedman, 1998)。因此，TGF-β1 的合成或是其訊息傳遞路經已被認為是抗發炎和潛在的抗纖維化指標 (Shek and Benyon, 2004)。

TGF-β1 主要透過 TGF-β1/Smad 訊息傳遞路徑活化 HSCs，從而造成肝纖維化

圖 4-12 不同水分發酵之蛹蟲草燕麥對 TAA 誘發肝纖維化小鼠肝組織 TNF-α 蛋白表現之影響

Fig. 4-12 The effect of C. militaris-fermented oats, which were cultured with various of water on haptatic TNF-α expression on TAA-induced liver fibrosis in mice. Target protein expressions were visualized using immunoblotting and quantified using gel analysis software. ^**,

*** indicated the significant difference (p<0.01, p<0.001, respectively) as compared with TAA. ^##, ^### indicated the significant difference (p<0.01, p<0.001, respectively) as compared with TAA.

圖 4-13 不同水分發酵之蛹蟲草燕麥對 TAA 誘發肝纖維化小鼠肝組織 NF-κB 蛋白表現之影響

Fig. 4-13 The effect of C. militaris-fermented oats, which were cultured with various of water on haptatic NF-κB expression on TAA-induced liver fibrosis in mice. Target protein expressions were visualized using immunoblotting and quantified using gel analysis software. ^**,

*** indicated the significant difference (p<0.01, p<0.001, respectively) as compared with TAA. ^##, ^### indicated the significant difference (p<0.01, p<0.001, respectively) as compared with TAA.

72 (activated form) 的指標 (Ramadori et al., 1990; Rockey et al., 1992)。試驗物質對肝組織中 α-SMA 蛋白表現之影響，如圖 4-17 所示，和 NOR 組相比，TAA 組 α-SMA 蛋白含量提高 425% (p<0.001)，經試驗物質作用後，相較於 TAA 組，

DCM-L、DCM-H、SCM、D、AS 與 CC 分別降低 41% (p<0.001)、45% (p<0.001)、

23% (p<0.001)、22% (p<0.001)、32% (p<0.001) 與 38% (p<0.001)，而 SL 降低

圖 4-14 不同水分發酵之蛹蟲草燕麥對 TAA 誘發肝纖維化小鼠肝組織 TGF-β1 蛋白表現之影響

Fig. 4-14 The effect of C. militaris-fermented oats, which were cultured with various of water on haptatic TGF-β1 expression on TAA-induced liver fibrosis in mice. Target protein expressions were visualized using immunoblotting and quantified using gel analysis software.

圖 4-15 不同水分發酵之蛹蟲草燕麥對 TAA 誘發肝纖維化小鼠肝組織 Smad2/3 蛋白表現之影響

Fig. 4-15 The effect of C. militaris-fermented oats, which were cultured with various of water on haptatic Smad2/3 expression on TAA-induced liver fibrosis in mice.

Target protein expressions were visualized using immunoblotting and quantified using gel analysis software.

圖 4-16 不同水分發酵之蛹蟲草燕麥對 TAA 誘發肝纖維化小鼠肝組織 p-Smad2/3 蛋白表現之影響

Fig. 4- 16 The effect of C. militaris-fermented oats, which were cultured with various of water on haptatic p-Smad2/3 expression on TAA-induced liver fibrosis in mice.

Target protein expressions were visualized using immunoblotting and quantified using gel analysis software.

圖 4-17 不同水分發酵之蛹蟲草燕麥對 TAA 誘發肝纖維化小鼠肝組織 α-SMA 蛋白表現之影響

Fig. 4-17 The effect of C. militaris-fermented oats, which were cultured with various of water on haptatic α-SMA expression on TAA-induced liver fibrosis in mice. Target protein expressions were visualized using immunoblotting and quantified using gel analysis software. (n=3) ^**, ^*** indicated the significant difference (p<0.01, p<0.001, respectively) as compared with TAA. ^### indicated the significant difference (p<0.001) as compared with TAA.

圖 4-18 不同水分發酵之蛹蟲草燕麥對 TAA 誘發肝纖維化小鼠肝組織 PPAR-γ 蛋白表現之影響

Fig. 4-18 The effect of C. militaris-fermented oats, which were cultured with various of water on haptatic PPAR-γ expression on TAA-induced liver fibrosis in mice. Target protein expressions were visualized using immunoblotting and quantified using gel analysis software.

(A) NOR normal group, (B) TAA TAA-induced liver fibrosis mices (TAA 100 mg/kg/ip, three times per week), (C) SL TAA-induced liver fibrosis mices fed 75

在文檔中 Liver Fibrosis of Cordyceps militaris-Fermented Oats (頁 72-93)

探討 DOW 發酵蛹蟲草燕麥對 TAA 誘發小鼠肝纖維化之影響

第四章 研究結果

第二節 探討 DOW 發酵蛹蟲草燕麥對 TAA 誘發小鼠肝纖維化之影響

第四章研究結果

第二節探討 DOW 發酵蛹蟲草燕麥對 TAA 誘發小鼠肝纖維化之影響