肝纖維化是諸多病因導致慢性肝病的共同病理過程，目前尚未有很好的
抗肝纖維化藥物，草藥在抗肝纖維化應用深具潛力。本研究探討草藥鯽魚
膽(Pluchea indica)的根水萃物能否降低硫代乙醯胺(thioacetamide; TAA)所
誘發小鼠(BALB/c)肝纖維化。實驗組小鼠每週腹腔注射三次TAA(200
mg/Kg)；治療組則除TAA注射外，每週腹腔注射一次鯽魚膽的根水萃物
(PIAE)，水萃物濃度分成低中高三種不同劑量(0.5, 1.0, 和1.5mg/ml)；控制
組每週腹腔注射三次磷酸鹽緩衝液(PBS, 2 ml/Kg)。小鼠連續處理四週或八
週後，檢測血清glutamyl pyruvic transaminase (GPT)活性，結果發現注射TAA
四週及八週的GPT活性都高於控制組，中劑量及高劑量治療組GPT活性明
顯降低。肝組織切片以Hematoxylin & Eosin、Reticular fiber和Sirius red染色
結果發現TAA誘發纖維化，而治療組纖維化程度減輕。Hydroxyproline分析
指出TAA會使肝臟collagen含量增加，治療組在四週與八週collagen含量會
降低。膠原蛋白α1型和α-肌動蛋白基因表現量則在PIAE治療四週與八週顯
著降低。CD163(ED2)、α-肌動蛋白、p53、和磷酸化p53之蛋白質表現量，
PIAE治療明顯低於TAA處理。本研究結果證實鯽魚膽的根水萃物可抑制肝
臟膠原蛋白的合成，降低肝纖維化形成。

Typically chronic injury leads to hepatic fibrosis. No effective antifibrotic
drugs have been approved, but herbal drugs have potential on the therapy of
hepatic fibrosis. The objective of this study used TAA-induced liver fibrosis
mouse as a model to elucidate whether aqueous extract of the root of Pluchea
indica (PIAE) can reduce liver fibrosis triggered by TAA. Mice were
intraperitoneally injected with TAA (200 mg/Kg) three times per week as the
TAA group, and those of injected with PIAE once per week as the treatment
group. Three PIAE dosages of low- (0.5 mg/ml), medium- (1.0 mg/ml), and
high- (1.5 mg/ml) doses were applied. Control mice were intraperitoneally
injected with phosphate-buffered saline (2 ml/Kg) three times per week. Mice
were sacrificed after 4 or 8 week treatment. Mice serum glutamyl pyruvic
transaminases (GPT) were increased in the TAA group while the treatment
group effects were declined after 4 or 8 weeks. H&E, Reticular fiber, and Sirius
red staining revealed that TAA induced liver fibrosis and fibrotic lesions were
reduced by PIAE treatment. Hydroxyproline assay showed that TAA increased
collagen contents and PIAE significantly decreased collagen contents after 4 or
8 weeks. Collagen α1 and α-SMA mRNA levels were decreased after 4- or 8-
week PIAE treatments. The protein levels of ED2, α-SMA, p53, and
phospho-p53 were all significantly declined on 4 or 8 weeks after PIAE
treatment. In conclusion, these results demonstrated that the aqueous extract of
P. indica shows anti-fibrotic effects on fibrogenesis of mouse liver.

Contents
Abstract in Chinese ---------------------------------------------------------------------I
Abstract in English ---------------------------------------------------------------------II
Acknowledgements --------------------------------------------------------------------III
Abbreviation ----------------------------------------------------------------------------V
Introduction ------------------------------------------------------------------------------1
Mechanism of liver fibrosis --------------------------------------------------------1
Treatment of liver fibrosis ----------------------------------------------------------2
Pluchea indica (L.) Less. -----------------------------------------------------------3
Apply of Pluchea indica (L.) Less. ------------------------------------------------5
Research of Pluchea indica (L.) Less. --------------------------------------------6
Materials and Methods ----------------------------------------------------------------8
Plant material -------------------------------------------------------------------------8
Animal grouping and treatment ----------------------------------------------------8
Specimen collection and Biochemistry assay ------------------------------------9
Histological examination -----------------------------------------------------------9
Hydroxyproline assay --------------------------------------------------------------13
Quantitative real-time PCR analysis ---------------------------------------------13
Western blot analysis --------------------------------------------------------------15
VII
Statistical analysis ------------------------------------------------------------------18
Results -----------------------------------------------------------------------------------19
Serum GPT activity ----------------------------------------------------------------19
Histological changes of the liver -------------------------------------------------19
Liver hydroxyproline contents ----------------------------------------------------20
mRNA expression of collagen α1 and α-SMA ---------------------------------20
Protein expression of ED2, α-SMA, p53, and Phospho-p53 -----------------21
Discussion -------------------------------------------------------------------------------23
References -------------------------------------------------------------------------------26
Table -------------------------------------------------------------------------------------36
Figure ------------------------------------------------------------------------------------44
Appendix --------------------------------------------------------------------------------58

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