黄芩苷对大鼠胰腺纤维化的保护作用
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摘要
目的：本实验采用二乙基二硫代氨基甲酸盐(DDC)致大鼠胰腺纤维化模型,比较加入黄芩苷后胰腺组织TGF-β1、CTGF、Fn蛋白表达量的变化,并观察大鼠胰腺组织病理情况,从胰腺组织TGF-β1、CTGF、Fn的蛋白表达量及病理学改变两方面探讨黄芩苷干预实验研究大鼠胰腺纤维化形成的机制与作用。方法：47只SPF级雄性SD大鼠随机分为5组(A组：空白组、B组：模型组、C组：小剂量黄芩苷组、D组中剂量黄芩苷组、E组大剂量黄芩苷组),B、C、D、E组用二乙基二硫代氨基甲酸盐(DDC)每周(周一、周四)腹腔注射两次,每次剂量为750mg/kg,共注射8次。A组每周两次注射等剂量的生理盐水,共注射8次。造模后第3周C、D、E组每日用不同剂量(25、50、100 mg/kg)的黄芩苷腹腔注射,B组注射生理盐水作为对照,造模及给药期间各组均予常规饲料。于造模开始后第6周实验结束前一天将各组大鼠禁食12个小时,心脏采血后处死,打开腹腔收集新鲜胰腺组织经生理盐水漂洗,肉眼观察胰腺组织改变后用4%的多聚甲醛液固定,常规石蜡包埋切片。放射免疫法和酶动力法检测血清透明质酸(HA)和淀粉酶,HE染色观察大鼠胰腺组织的病理学变化,免疫组化分析大鼠胰腺组织中的转化生长因子TGF-β1、CTGF、Fn含量,光镜下观察照相。
结果：1、黄芩苷对大鼠一般情况的影响
空白组大鼠一般情况良好,饮食、活动、精神均正常,3周后体重明显增加。模型组大鼠造模2周后大鼠多饮、少食、多尿、体重有所减轻,精神倦怠,反应较迟钝,鼻腔内有出血,皮毛疏松无光泽,大便糖稀,少数腹腔积液明显,2只死于腹部积液量过多。各黄芩苷组早期进食均减少,精神不佳,鼻腔内均有出血,大便溏稀。给药3周后进食无明显减少,精神、活动较正常,大便性状无明显改变。其中小、中剂量组分别有一只大鼠死于腹部注射部位撕咬,大剂量组一只大鼠造模后腹水过多经抽腹水抗感染治疗无效后死亡。
2、胰腺组织病理形态学改变：HE染色镜下改变。
空白组：胰腺腺泡呈嗜碱性,腺细胞为锥体形,泡浆丰满,细胞核呈圆形位于基底部,细胞大小一致,排列整齐,胰腺小叶轮廓清晰,小叶间结缔组织中可见胰导管及血管等,胰岛圆润饱满。
模型组：胰管扩张,胰腺腺泡变形,水肿。小叶间纤维组织增生,大量炎细胞浸润,小叶结构消失,有的形成炎性包裹,部分腺体萎缩坏死代之以纤维瘢痕。
小剂量黄芩苷组：可见胰腺腺泡水肿,但见散在坏死腺泡及炎性浸润,小叶内、小叶间腺泡组织局限性纤维化或无改变。
中剂量组：可见水肿的腺泡,小叶间少量炎细胞浸润,小叶内、小叶间腺泡组织少有纤维化改变。
大剂量组：可见水肿的腺泡,浸润的炎性细胞,部分小叶结构消失,小叶内、小叶间腺泡组织局限性纤维化或无改变。
3、血清生化：淀粉酶各给药组与模型组比较P<0.05,各组与空白组比较P<0.05,各给药组见两两比较无差异。透明质酸各组与空白组比较P<0.01,各给药组与模型组比较P<0.05,各给药组之间相互比较无差异。
4、各组TGF-β1、CTGF、Fn蛋白表达量变化：TGF-β1、CTGF免疫组化标记后,光镜下可见模型组TGF-β1主要表达于间质的纤维组织、成纤维细胞、巨噬细胞及血管内皮细胞胞浆和细胞膜,呈棕黄或棕褐色。而空白组大鼠不表达或仅微弱地表达于间质细胞内,呈淡黄色。CTGF在胰腺组织中主要分布于邻近间质区的部分腺上皮细胞的细胞质和间质的成纤维细胞的细胞质中,以细胞浆染成棕黄或棕褐色为阳性表达。而空白组大鼠无或仅有极少量的CTGF在腺上皮细胞的细胞质中表达。采用CMIA系列-多功能彩色病理图像分析TGF-β1、CTGF表达程度,黄芩苷处理各组TGF-β1、CTGF蛋白表达量与模型组相比差异均有统计学意义(P<0.05),黄芩苷处理组间相互比较无差异。Fn免疫组化标记后,光镜下可见模型组Fn主要表达在小叶间及腺泡周围纤维组织中,呈树枝状分布呈网格状包绕腺泡,在小血管周围和导管周围也有少量表达,呈棕黄或棕褐色,并随间质病变程度的增加而增加。空白组大鼠较弱地表达于间质中,呈淡黄色。半定量分析其表达程度,黄芩苷处理各组Fn蛋白表达量与模型组相比差异均有统计学意义(P<0.05),黄芩苷各处理组间相互比较无差异。结论：1、腹腔内反复注射DDC后通过氧化刺激使胰腺组织内氧自由基增高而激活胰腺星状细胞(PSCs)制造大鼠慢性胰腺炎胰腺纤维化模型。
2、造模后胰腺外分泌功能下降,而黄芩苷处理组淀粉酶和模型组比较明显升高表明黄芩苷有改善胰腺外分泌功能的作用。造模后透明质酸含量明显升高提示胰腺组织损伤并有纤维增生改变,且黄芩苷处理后透明质酸含量下降,显示黄芩苷有改善组织纤维化程度的作用。
3、黄芩苷可能通过下调TGF-β1水平,抑制胰腺星状细胞活化增值,降低CTGF蛋白表达,减少纤维连接蛋白(Fn)而具有抗大鼠胰腺纤维化作用。
Objective:To establish the rats model of chronic fibrosing pancreatitis with intraperitoneal injection of DDC and To study the efect and potential mechanism of baicalin on pancreatic fibrotic rats
Methods:Fourty seven male Sprague-Dawley rats were randomly divided into 5 equal groups:normal control group(A group), model control group(B group),low-dose baicalin-treated group (C group), mediate-dose baicalin-treated group(D group), andhigh-dose baicalin-treated group (E group). The rats of the later 4 groups underwent intraperitoneal injectin of 750mg/kg body weight (BW) of DDC twice a week for 4 weeks into the pancreatic duct so as to establish models of pancreatic fibrosis. At the third week the baicalin-treated rats were intraperitoneal injectin with different doses of baicalin(25, 50, and 100 mg/kg body weight) every day, while the normal and model control groups received 0.9% sodium chloride solution instead. lasting 4 weeks, the day befor the last day, the rats were fasted 24 hours and then blood samples were colected from rats'heart. Immediately the rats were put to death and their pancreases were taken out. The levels of hyaluronic acid (HA) and diastase were determined by radioimmunoassay and enzyme dynamia. Pancreatic tissue was embedded by paraffin and cut. The histopathological alterations of pancreatic tissue with HE staining were studied by optical microscopy.immunohistochemistry was used to detect the protein expression of transforming growth factor-β1 (TGF-β1) and CTGF and Fn, and measure the expression of TGF-β1 and Fn and CTGF for the observation by tramission electron microscope.
Results:The rats'common conditions of group A、C、D、E、were obviously better than that of model group B. The levels of diastase of the low-dose, mediate-dose, and high-dose baicalin-treated groups were all higher than that of the model control group (P<0.05). The serum level of HA of the low-dose, mediate-dose, and high-dose baicalin-treated groups were all significantly lower than that of the model control group (P< 0.05). Comparison of HA and diastase with the each group used baicalin was nonsense. The degrees of fibrosis of the baicalin-treated groups were ameliorated in comparison with the model control group. The protein content of TGF-β1 and fibronectin (Fn) and CTGF of the low-dose, mediate-dose, and high-dose baicalin-treated groups were all lower than that of the model control group (P<0.05). Comparison with the each group used baicalin was nonsense. Comparison of TGF-β1 and CTGF and Fn with the each group used baicalin was nonsense.
Conclusions:The models of chronic pancreatitis rats can be induced with in 4 weeks by intraperitoneal injection of DDC twice a week. Baicalin can improve the common conditions of chronic pancreatitis rats; decrease the levels of HA; improve the level s of amylase; Anti-fibrosis effect of baicalin on pancreatic fibrosis maybe related to its elimination oxygen free radical effect and the content of TGF-β1 and CTGF protein.
结果：1、黄芩苷对大鼠一般情况的影响
空白组大鼠一般情况良好,饮食、活动、精神均正常,3周后体重明显增加。模型组大鼠造模2周后大鼠多饮、少食、多尿、体重有所减轻,精神倦怠,反应较迟钝,鼻腔内有出血,皮毛疏松无光泽,大便糖稀,少数腹腔积液明显,2只死于腹部积液量过多。各黄芩苷组早期进食均减少,精神不佳,鼻腔内均有出血,大便溏稀。给药3周后进食无明显减少,精神、活动较正常,大便性状无明显改变。其中小、中剂量组分别有一只大鼠死于腹部注射部位撕咬,大剂量组一只大鼠造模后腹水过多经抽腹水抗感染治疗无效后死亡。
2、胰腺组织病理形态学改变：HE染色镜下改变。
空白组：胰腺腺泡呈嗜碱性,腺细胞为锥体形,泡浆丰满,细胞核呈圆形位于基底部,细胞大小一致,排列整齐,胰腺小叶轮廓清晰,小叶间结缔组织中可见胰导管及血管等,胰岛圆润饱满。
模型组：胰管扩张,胰腺腺泡变形,水肿。小叶间纤维组织增生,大量炎细胞浸润,小叶结构消失,有的形成炎性包裹,部分腺体萎缩坏死代之以纤维瘢痕。
小剂量黄芩苷组：可见胰腺腺泡水肿,但见散在坏死腺泡及炎性浸润,小叶内、小叶间腺泡组织局限性纤维化或无改变。
中剂量组：可见水肿的腺泡,小叶间少量炎细胞浸润,小叶内、小叶间腺泡组织少有纤维化改变。
大剂量组：可见水肿的腺泡,浸润的炎性细胞,部分小叶结构消失,小叶内、小叶间腺泡组织局限性纤维化或无改变。
3、血清生化：淀粉酶各给药组与模型组比较P<0.05,各组与空白组比较P<0.05,各给药组见两两比较无差异。透明质酸各组与空白组比较P<0.01,各给药组与模型组比较P<0.05,各给药组之间相互比较无差异。
4、各组TGF-β1、CTGF、Fn蛋白表达量变化：TGF-β1、CTGF免疫组化标记后,光镜下可见模型组TGF-β1主要表达于间质的纤维组织、成纤维细胞、巨噬细胞及血管内皮细胞胞浆和细胞膜,呈棕黄或棕褐色。而空白组大鼠不表达或仅微弱地表达于间质细胞内,呈淡黄色。CTGF在胰腺组织中主要分布于邻近间质区的部分腺上皮细胞的细胞质和间质的成纤维细胞的细胞质中,以细胞浆染成棕黄或棕褐色为阳性表达。而空白组大鼠无或仅有极少量的CTGF在腺上皮细胞的细胞质中表达。采用CMIA系列-多功能彩色病理图像分析TGF-β1、CTGF表达程度,黄芩苷处理各组TGF-β1、CTGF蛋白表达量与模型组相比差异均有统计学意义(P<0.05),黄芩苷处理组间相互比较无差异。Fn免疫组化标记后,光镜下可见模型组Fn主要表达在小叶间及腺泡周围纤维组织中,呈树枝状分布呈网格状包绕腺泡,在小血管周围和导管周围也有少量表达,呈棕黄或棕褐色,并随间质病变程度的增加而增加。空白组大鼠较弱地表达于间质中,呈淡黄色。半定量分析其表达程度,黄芩苷处理各组Fn蛋白表达量与模型组相比差异均有统计学意义(P<0.05),黄芩苷各处理组间相互比较无差异。结论：1、腹腔内反复注射DDC后通过氧化刺激使胰腺组织内氧自由基增高而激活胰腺星状细胞(PSCs)制造大鼠慢性胰腺炎胰腺纤维化模型。
2、造模后胰腺外分泌功能下降,而黄芩苷处理组淀粉酶和模型组比较明显升高表明黄芩苷有改善胰腺外分泌功能的作用。造模后透明质酸含量明显升高提示胰腺组织损伤并有纤维增生改变,且黄芩苷处理后透明质酸含量下降,显示黄芩苷有改善组织纤维化程度的作用。
3、黄芩苷可能通过下调TGF-β1水平,抑制胰腺星状细胞活化增值,降低CTGF蛋白表达,减少纤维连接蛋白(Fn)而具有抗大鼠胰腺纤维化作用。
Objective:To establish the rats model of chronic fibrosing pancreatitis with intraperitoneal injection of DDC and To study the efect and potential mechanism of baicalin on pancreatic fibrotic rats
Methods:Fourty seven male Sprague-Dawley rats were randomly divided into 5 equal groups:normal control group(A group), model control group(B group),low-dose baicalin-treated group (C group), mediate-dose baicalin-treated group(D group), andhigh-dose baicalin-treated group (E group). The rats of the later 4 groups underwent intraperitoneal injectin of 750mg/kg body weight (BW) of DDC twice a week for 4 weeks into the pancreatic duct so as to establish models of pancreatic fibrosis. At the third week the baicalin-treated rats were intraperitoneal injectin with different doses of baicalin(25, 50, and 100 mg/kg body weight) every day, while the normal and model control groups received 0.9% sodium chloride solution instead. lasting 4 weeks, the day befor the last day, the rats were fasted 24 hours and then blood samples were colected from rats'heart. Immediately the rats were put to death and their pancreases were taken out. The levels of hyaluronic acid (HA) and diastase were determined by radioimmunoassay and enzyme dynamia. Pancreatic tissue was embedded by paraffin and cut. The histopathological alterations of pancreatic tissue with HE staining were studied by optical microscopy.immunohistochemistry was used to detect the protein expression of transforming growth factor-β1 (TGF-β1) and CTGF and Fn, and measure the expression of TGF-β1 and Fn and CTGF for the observation by tramission electron microscope.
Results:The rats'common conditions of group A、C、D、E、were obviously better than that of model group B. The levels of diastase of the low-dose, mediate-dose, and high-dose baicalin-treated groups were all higher than that of the model control group (P<0.05). The serum level of HA of the low-dose, mediate-dose, and high-dose baicalin-treated groups were all significantly lower than that of the model control group (P< 0.05). Comparison of HA and diastase with the each group used baicalin was nonsense. The degrees of fibrosis of the baicalin-treated groups were ameliorated in comparison with the model control group. The protein content of TGF-β1 and fibronectin (Fn) and CTGF of the low-dose, mediate-dose, and high-dose baicalin-treated groups were all lower than that of the model control group (P<0.05). Comparison with the each group used baicalin was nonsense. Comparison of TGF-β1 and CTGF and Fn with the each group used baicalin was nonsense.
Conclusions:The models of chronic pancreatitis rats can be induced with in 4 weeks by intraperitoneal injection of DDC twice a week. Baicalin can improve the common conditions of chronic pancreatitis rats; decrease the levels of HA; improve the level s of amylase; Anti-fibrosis effect of baicalin on pancreatic fibrosis maybe related to its elimination oxygen free radical effect and the content of TGF-β1 and CTGF protein.
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24.王兴鹏,龚自华,吴凯.三硝基苯磺酸诱导大鼠慢性胰腺炎模型的建立[J].中华病理学杂志,2003,32(3)：267-269.
25.吴其夏,余应年,卢建,等.新编病理生理学.中国协和医科大学出版社,1999,174-190.
26.Heikkila RE,Cabbat FS,Cohen G. In vivo inhibition of superoxode dismutase in mice by diethyldithiocarbamate. J Biol Chem,1976,251:2182-2185.
27. Sasaki T. Studies on oxygen-derived free radicals in acute pancreatic damage in rats-pancreatic damage experimentally induced by diethyldithiocarbamate. Nippon Shokakibyo Gakkai Zasshi,1990,87:1436-1443.
28.张汝玲,吴恺,龚自华,等.三硝基苯磺酸诱导大鼠胰腺纤维化的病理学演变.胰腺病学,2002,2(3)：137.
29.张宇,李进京,孙红岩,等.黄芩叶对损伤胰腺的保护作用[J].中药材,2007,30(11)：1433-1434.
30. Masamune A,Shimosegawa T. Signal transduction in pancreatic stellate cells. J gastroenterol.2009,44(4):249-60.
31.杨鹤梅,李素婷,梅立新,等.黄芩茎叶总黄酮对纤维化大鼠肝脏星形细胞活化的影响[J].中国中医基础医学杂志,2006,12(1)：42-44.
32. Masamune A, Kikuta K, Watanabe T, et al. Fibrinogen induces cytokine and collagen production in pancreatic stellate cells.Gut,2009,58(4):550-9.
33. Aoki H, Ohnishi H, Hama K, et al Existence of autocrine loop between interleukin-6 and transforming growth factor-betal in activated rat pancreatic stellate cells [J].J cell Biochem,2006,99(1):221-228.
34.孙蕴伟,章永平,乔敏敏,等.转化生长因子β1对胰腺星状细胞中结缔组织生长因子基因的调控作用.中华医学杂志,2004,85(15)：1240-1242.
35. Vogelmann R,Rut D, Wagner M, et al.Effects of fibrogenic mediators on the development of pancreatic fibrosis in a TGF-betal transgenic mouse model. Am J Physiol Gastrointest Liver Physiol,2001,280:G164-172.
1.钱家鸣.慢性胰腺炎[M].内科学第六版,2004,6(44)：472-473.
2.巫协宁.慢性胰腺炎胰纤维化形成机制新解[M].胃肠病学,1999,4(3)：166-167.
3. Bachem MG, Scheider E, Gross H, et al. Identification, culture, and characterization of pancreatic stellate cells in rats and humans Gastroenterology 1998,115 (2):421-432
4.Apte MV, Haber PS, Darby SJ,et al. Pancreatic stellate cells are activated by proinflammatory cytokines:Implications for Pancreatic fibrogenesis. Gut,1999,44(4):534-541.
5. Haber PS, Keogh GW, A pte MV, et al. Activation of pancreatic stellate cells in human and experimental pancreatic fibrosis. Am J Pathol,1999,155(4):1087-1095
6.孙蕴伟,顾柏炜,朱颖,袁耀宗.促纤维化细胞因子在纤维化胰腺组织中的表达[J].上海医学,2004,27(5)：340-343.
7. Read LC,Lemmy AB,Hoearth GS,et al.The gastrointestinaltract is one of the most responsive target tissues for IGF-1 and its potent analogs. In:Spencer EM,eds. Modernconcepts of Insulin-like growth factors. New York:Elsevier,1998:225-234.
8.Gomez G,Lee H,He Q, et al. Acute pancreatitis signals activation of apoptosis-associated and survivalgenes in mice. Exp Biol Med,2001,226(7):692-700.
9. Warzecha Z, Dembinski A, Ceranowicz P, et al. IGF-lstimulates production of interleukin-10 and inhibi tsdevelopment of caerulein-inducedpancreatitis. J PhysiolPharmacol,2003, 54 (4):575-590.
10. Apte MV, Phillips PA, Fahmy RG, et al.Does alcohole directly stimulate pancreatic fibrogenesis studies with pancreatic stellate cells[J]. Gastroenterology,2000,118(4):780-794.
11.Casini A, Galli A, Pignalosa P, et al. Collagen type Ⅰ synthesized by pancreatic periacinar stellate cell(PSC) co-localizes with lipid peroxidation-derived aldehydes in chronic alcoholic pancreatitis. J Pathol,2000,192 (1):81-89.
12. Manning AM,et al. J Inflammation,1995,4:5283-296.
13.王彩花,高志强,叶斌,谢传,等.大黄素干预大鼠慢性胰腺炎胰腺纤维化进展的研究[J].中华医学杂志,2006,86(36)：2552-2555.
14.吴恺,张汝玲,王兴鹏.甘草酸对大鼠胰腺纤维化的干预作用及其机制[J].中国药理学通报,2003,19(12)：1424-7.
15.McCloy R. Chronic pancreatitis at Manchester, UK. Focus on antioxidant therapy. Digestion1998,59:36-38.
16.李铁强.酒客乐对慢性胰腺炎大鼠胰腺纤维化和酒精性胰腺损害的防治[D],湖北中医学院,2006.
17.王艳明.健脾疏肝活血法抗慢性胰腺炎纤维化的机理研究[D]黑龙江中医药大学,2005.
18.刘杰波.黄芩苷对细菌脂多糖致星形胶质细胞损伤保护机制的研究[J].湖南中医药导报.2004：10(3)：79.
19.陈忻,董晓丹,张楠,等.黄芩苷对局灶性脑缺血大鼠的保护作用[J].广州中医药大学学报,2006,23(6)：506-508.
20.杜肇宗等.兔血小板脂加氧同工酶的分析及黄芩苷的抑制作用[J].南京铁道医学院学报,1987,6(2)：9.
21.苏宁,周乐全,等.黄芩苷下调TGF-β1、PKC表达减少肾小管Ⅳ-C沉积的作用[J].中药新药与临床药理,2008,19(4)：272-274.
22.邝枣园,符林春,罗海燕,等.从受体角度探讨黄芩苷对肺炎衣原体感染细胞的干预作用[J].广州中医药大学学报,2005,22(3)：210-213.
23.刘煜德,邝枣园,吴伟,等.黄芩苷抗肺炎衣原体致动脉内皮细胞损伤研究[J].辽宁中医杂志,2006,33(12)：1645-1648.
24.杨婧,傅颖媛.黄芩苷、黄连素体外杀灭阴道毛滴虫的实验研究[J].中国中医药信息杂志,2006,13(4)：37-38.
25.姜英子,金将杰,金成赞,等.黄芩苷对小鼠急性肝损伤的保护作用[J].延边大学医学学报,2003,26(4)：255-257.
2.Kloppel G,Maurer R,HofmannE,et al. Solid-cystic (papillary-cystic) tumours within and outside the pancreas in men:report of two patients [J].Virchows Arch A Pathol Anat Histopathol.1991,418 (2):179-83.
3. Sarles H, de Caro A,Multinger L, et al. Giant pancreatic stones in teetotal women due to absence of the "stone protein"? [J]. Lancet.1982,2 (8300):714-5.
4. Bachem MG, Scheider E, Gross H, et al. Identification culture and characterization of pancreatic stellate cell in rats and humans [J]. Gastroenterology,1998,115 (2):421-432.
5.Shek FW,Williams EJ,Zhou X,et al.Autocrine up-regulation of CTGF by TGF-β1 in pancreatic stellate cell cultures:Implications for progressive fibrosis in chronic pancreatitis [J]. pancreatology,2001,1 (2):136
6.孙蕴伟,顾柏炜,朱颖,等.促纤维化细胞因子在纤维化胰腺组织中的表达[J].上海医学,2004,27(5)：340-343.
7.朱颖,孙蕴伟,乔敏敏,等.小柴胡汤对大鼠胰腺纤维化的治疗作用[J].胃肠病学,2006,11(1)：25-29.
8.王彩花,高志强,叶斌,等.大黄素干预大鼠慢性胰腺炎胰腺纤维化进展的研究[J].中华医学杂志,2006,86(36)：2552-2555.
9.吴恺,张汝玲,王兴鹏.甘草酸对大鼠胰腺纤维化的干预作用及其机制[J].中国药理学通报,2003,19(12)：1424-1427.
10.李铁强.酒客乐对慢性胰腺炎大鼠胰腺纤维化和酒精性胰腺损害的防治[D].湖北中医学院,2006.
11.王艳明.健脾疏肝活血法抗慢性胰腺炎纤维化的机理研究[D].黑龙江中医药大学,2005.
12.刘萍,李倩,刘兆平,等.黄芩苷对脑缺血再灌注损伤大鼠神经细胞凋亡及相关基因表达的影响[J].中国新药与临床杂志,2007,26(2)：109-112.
13.刘平,隗义双,周乾坤,等.黄芩苷对实验性脑出血大鼠脑损伤的保护作用[J].中国中医药信息杂志,2008,15(4)：34-35.
14.张喜平,程琪辉,陆贝,等.黄芩甙和奥曲肽对重症急性胰腺炎大鼠心脏损伤保护作用的比较[J].中国中西医结合急救杂志,2007,14(2)：114-118.
15.苏宁,周乐全,匡忠生,等.黄芩苷下调TGF-β1、PKC表达减少肾小管Ⅳ-C沉积的作用[J].中药新药与临床药理,2008,19(4)：272-274.
16.杨鹤梅,李素婷,梅立新,等.黄芩茎叶总黄酮对纤维化大鼠肝脏星形细胞活化的影响[J].中国中医基础医学杂志,2006,12(1)：42-44.
17.孙海燕,黄民,关溯.黄芪总苷对实验性急性胰腺炎的保护作用[J].时珍国医国药,2006,17(10)：1980-1981.
18. Zhang XP, Zhang L, He JX, et al. Experimental study of therapeutic efficacy of Baicalin in rats with severe acute pancreatitis [J]. Gastroenterology,2007,13 (5):717-724.
19.Inoue T,Jackson EK.Strong antiprolifentive effects of baicalein in cultured rat hepatic stellate cells [J].Eur J Pharmcol,1999,378(1):129-35.
20.李海燕,李夏,金向群,等.黄芩苷对大鼠慢性酒精性肝损伤的影响[J].中国实验方剂学杂志,2008,14(6)：95-96.
21.冯友根,郭丽敏.黄芩苷防治糖尿病并发症的作用与临床应用[J].医药导报,2004,23(8)：572-573.
22.matsumura N, Ochi K, Ichimura M, et al. Study on free radicals and pancreatic fibrosis-pancreatic fibrosis induced by repeated injections of superoxide dismutase inhibitor [J]. Pancreas,2001,22:53-57.
23.Meng M, Li YQ, Yan MX, et al. Effects of epigallocatechin gallate on diethyldithiocarbamate-induced pancreatic fibrosis in rats[J].Biol Pharm Bull,2007,30 (6):1091-6.
24.王兴鹏,龚自华,吴凯.三硝基苯磺酸诱导大鼠慢性胰腺炎模型的建立[J].中华病理学杂志,2003,32(3)：267-269.
25.吴其夏,余应年,卢建,等.新编病理生理学.中国协和医科大学出版社,1999,174-190.
26.Heikkila RE,Cabbat FS,Cohen G. In vivo inhibition of superoxode dismutase in mice by diethyldithiocarbamate. J Biol Chem,1976,251:2182-2185.
27. Sasaki T. Studies on oxygen-derived free radicals in acute pancreatic damage in rats-pancreatic damage experimentally induced by diethyldithiocarbamate. Nippon Shokakibyo Gakkai Zasshi,1990,87:1436-1443.
28.张汝玲,吴恺,龚自华,等.三硝基苯磺酸诱导大鼠胰腺纤维化的病理学演变.胰腺病学,2002,2(3)：137.
29.张宇,李进京,孙红岩,等.黄芩叶对损伤胰腺的保护作用[J].中药材,2007,30(11)：1433-1434.
30. Masamune A,Shimosegawa T. Signal transduction in pancreatic stellate cells. J gastroenterol.2009,44(4):249-60.
31.杨鹤梅,李素婷,梅立新,等.黄芩茎叶总黄酮对纤维化大鼠肝脏星形细胞活化的影响[J].中国中医基础医学杂志,2006,12(1)：42-44.
32. Masamune A, Kikuta K, Watanabe T, et al. Fibrinogen induces cytokine and collagen production in pancreatic stellate cells.Gut,2009,58(4):550-9.
33. Aoki H, Ohnishi H, Hama K, et al Existence of autocrine loop between interleukin-6 and transforming growth factor-betal in activated rat pancreatic stellate cells [J].J cell Biochem,2006,99(1):221-228.
34.孙蕴伟,章永平,乔敏敏,等.转化生长因子β1对胰腺星状细胞中结缔组织生长因子基因的调控作用.中华医学杂志,2004,85(15)：1240-1242.
35. Vogelmann R,Rut D, Wagner M, et al.Effects of fibrogenic mediators on the development of pancreatic fibrosis in a TGF-betal transgenic mouse model. Am J Physiol Gastrointest Liver Physiol,2001,280:G164-172.
1.钱家鸣.慢性胰腺炎[M].内科学第六版,2004,6(44)：472-473.
2.巫协宁.慢性胰腺炎胰纤维化形成机制新解[M].胃肠病学,1999,4(3)：166-167.
3. Bachem MG, Scheider E, Gross H, et al. Identification, culture, and characterization of pancreatic stellate cells in rats and humans Gastroenterology 1998,115 (2):421-432
4.Apte MV, Haber PS, Darby SJ,et al. Pancreatic stellate cells are activated by proinflammatory cytokines:Implications for Pancreatic fibrogenesis. Gut,1999,44(4):534-541.
5. Haber PS, Keogh GW, A pte MV, et al. Activation of pancreatic stellate cells in human and experimental pancreatic fibrosis. Am J Pathol,1999,155(4):1087-1095
6.孙蕴伟,顾柏炜,朱颖,袁耀宗.促纤维化细胞因子在纤维化胰腺组织中的表达[J].上海医学,2004,27(5)：340-343.
7. Read LC,Lemmy AB,Hoearth GS,et al.The gastrointestinaltract is one of the most responsive target tissues for IGF-1 and its potent analogs. In:Spencer EM,eds. Modernconcepts of Insulin-like growth factors. New York:Elsevier,1998:225-234.
8.Gomez G,Lee H,He Q, et al. Acute pancreatitis signals activation of apoptosis-associated and survivalgenes in mice. Exp Biol Med,2001,226(7):692-700.
9. Warzecha Z, Dembinski A, Ceranowicz P, et al. IGF-lstimulates production of interleukin-10 and inhibi tsdevelopment of caerulein-inducedpancreatitis. J PhysiolPharmacol,2003, 54 (4):575-590.
10. Apte MV, Phillips PA, Fahmy RG, et al.Does alcohole directly stimulate pancreatic fibrogenesis studies with pancreatic stellate cells[J]. Gastroenterology,2000,118(4):780-794.
11.Casini A, Galli A, Pignalosa P, et al. Collagen type Ⅰ synthesized by pancreatic periacinar stellate cell(PSC) co-localizes with lipid peroxidation-derived aldehydes in chronic alcoholic pancreatitis. J Pathol,2000,192 (1):81-89.
12. Manning AM,et al. J Inflammation,1995,4:5283-296.
13.王彩花,高志强,叶斌,谢传,等.大黄素干预大鼠慢性胰腺炎胰腺纤维化进展的研究[J].中华医学杂志,2006,86(36)：2552-2555.
14.吴恺,张汝玲,王兴鹏.甘草酸对大鼠胰腺纤维化的干预作用及其机制[J].中国药理学通报,2003,19(12)：1424-7.
15.McCloy R. Chronic pancreatitis at Manchester, UK. Focus on antioxidant therapy. Digestion1998,59:36-38.
16.李铁强.酒客乐对慢性胰腺炎大鼠胰腺纤维化和酒精性胰腺损害的防治[D],湖北中医学院,2006.
17.王艳明.健脾疏肝活血法抗慢性胰腺炎纤维化的机理研究[D]黑龙江中医药大学,2005.
18.刘杰波.黄芩苷对细菌脂多糖致星形胶质细胞损伤保护机制的研究[J].湖南中医药导报.2004：10(3)：79.
19.陈忻,董晓丹,张楠,等.黄芩苷对局灶性脑缺血大鼠的保护作用[J].广州中医药大学学报,2006,23(6)：506-508.
20.杜肇宗等.兔血小板脂加氧同工酶的分析及黄芩苷的抑制作用[J].南京铁道医学院学报,1987,6(2)：9.
21.苏宁,周乐全,等.黄芩苷下调TGF-β1、PKC表达减少肾小管Ⅳ-C沉积的作用[J].中药新药与临床药理,2008,19(4)：272-274.
22.邝枣园,符林春,罗海燕,等.从受体角度探讨黄芩苷对肺炎衣原体感染细胞的干预作用[J].广州中医药大学学报,2005,22(3)：210-213.
23.刘煜德,邝枣园,吴伟,等.黄芩苷抗肺炎衣原体致动脉内皮细胞损伤研究[J].辽宁中医杂志,2006,33(12)：1645-1648.
24.杨婧,傅颖媛.黄芩苷、黄连素体外杀灭阴道毛滴虫的实验研究[J].中国中医药信息杂志,2006,13(4)：37-38.
25.姜英子,金将杰,金成赞,等.黄芩苷对小鼠急性肝损伤的保护作用[J].延边大学医学学报,2003,26(4)：255-257.