柴胡疏肝散通过抗氧化反应对二氯二丁基酯联合乙醇诱发小鼠胰腺纤维化的防治作用

 目的:通过观察柴胡疏肝散对二氯二丁基酯（DBTC）联合乙醇所致慢性胰腺炎小鼠胰腺丙二醛(MDA)和超氧化物歧化酶(SOD)的影响，探讨柴胡疏肝散防治胰腺纤维化的机制。方法:健康昆明小鼠120只随机分为空白组、模型组和柴胡疏肝散治疗组（n=40）。除空白组外，其余小鼠经尾静脉一次性注射DBTC（8 mg/kg）后，联合饮用10％乙醇诱发慢性胰腺炎。造模后3 d 治疗组小鼠给予柴胡疏肝散 (6 g·kg-1·d-1）灌胃，空白组和模型组给予生理盐水灌胃。于造模前及造模后1、2、4、8周分批处死小鼠，HE染色检测各组小鼠胰腺组织的形态学改变及纤维化程度； 化学比色法和ELISA法分别检测血清淀粉酶和透明质酸的变化；酶化学法检测胰腺组织匀浆SOD和MDA 的水平； Western blotting检测胰腺组织Ⅰ型胶原的表达。结果:DBTC联合饮用乙醇2周和4周血清淀粉酶及透明质酸处于较高水平，8周时淀粉酶明显降低，而透明质酸水平进一步升高，各指标与相应时点空白组相比有显著差异（P<0.05）。 造模后2、4、8周胰腺组织匀浆MDA 的水平持续升高，SOD活性持续降低，与相应时点空白组相比有显著差异（P<0.05）；造模后1周胰腺实质肿胀、坏死，2周可见小叶间纤维化。造模4周和8周显示胰腺腺泡明显萎缩，出现典型、广泛的不规则纤维化，胰腺Ⅰ型胶原的表达亦呈现较高水平。柴胡疏肝散治疗后，血清淀粉酶活性和透明质酸含量降低,胰腺组织匀浆MDA 的水平降低,SOD活性升高，各时点胰腺的纤维化损伤程度明显减轻。结论: 柴胡疏肝散通过抗氧化机制，发挥防治慢性胰腺炎胰腺纤维化的作用。

Effect of Chaihushugansan on pancreatic fibrosis in mice with chronic pancreatitis induced by DBTC plus ethanol and its anti-oxidation mecha-nism

AIM:To explore the role of superoxide dismutase (SOD) and malondialdehyde (MDA) in chronic pancreatitis (CP) induced by dibutyltin dichloride (DBTC) combined with ethanol, and the mechanisms for prevention and treatment of pancreatic fibrosis by Chaihushugansan. METHODS:The KM mice were randomly divided into control group, CP group (DBTC combined with ethanol) and Chaihushugansan group (CP+Chaihushugansan). Except for control group, the mice in other groups were intravenously injected in tail with DBTC (8 mg/kg) and drank 10% ethanol. The mice in Chaihushugansan group were administered intragastrically with Chaihushugansan (6 g·kg-1·d-1) at the following experimenal period. Before modeling and 1 week, 2 weeks, 4 weeks and 8 weeks after modeling, the mice were anesthetized and sacrificed. The activity of amylase and the content of hyaluronic acid in the serum were measured. The morphology and the degree of fibrosis in the pancreas were observed by HE staining. The activity of SOD and the level of MDA in the pancreas homogenate were analyzed. The protein of pancreas was extracted to detect the expression of type I collagen by Western blotting. RESULTS:DBTC combined with ethanol induced CP with increased serum amylase and hyaluronic acid levels， while the serum amylase and hyaluronic acid levels in Chaihushugansan group were significantly lowered (P<0.05). In 1 week, 2 weeks, 4 weeks and 8 weeks, the pancreas were obviously injured and appeared different degrees of fibrosis. The content of MDA and the expression of type I collagen in the increased significantly, but the SOD was decreased. In Chaihushugansan group, the pathological damage and the degree of fibrosis of the pancreas were improved. The level of MDA and type I collagen expression in the pancreas were significantly reduced, but the SOD was increased. CONCLUSION: The oxidative stress may take part in the development of CP. Inhibition of oxidative stress in the pancreas is one of the mechanisms that Chaihushugansan attenuates the development of CP.