荔枝核总黄酮对肝纤维化模型大鼠肝细胞损伤的改善作用

目的:研究荔枝核总黄酮(TFL)对肝纤维化模型大鼠肝细胞损伤的改善作用。方法:大鼠每周前3天ip给予二甲基亚硝胺(DMN),每天1次,连续30 d以复制肝细胞损伤模型。60只大鼠随机均分为正常对照(等容生理盐水)组、模型(等容生理盐水)与TFL高、低剂量(200、100 mg/kg)组,除正常对照组外其他各组复制模型的同时ig给药,每天1次,连续45 d。采用HE染色,显微镜下观察大鼠肝细胞并分析肝组织病理分期(S1~S4);采用Masson染色与免疫组化染色,显微镜下观察大鼠肝细胞并统计肝组织纤维化程度与Bcl-2、Bax表达水平;检测大鼠血清中天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)活性。结果:模型组大鼠肝细胞病理分期以S3、S4期为主,TFL高、低剂量组以S1、S2期为主。与正常对照组比较,模型组大鼠肝组织纤维化程度升高,Bcl-2、Bax表达增强,血清中AST、ALT活性增强;与模型组比较,TFL高、低剂量组大鼠肝组织纤维化程度降低,Bcl-2表达增强,Bax表达减弱,血清中AST、ALT活性减弱;以上差异均具有统计学意义(P<0.05)。结论:TFL对肝纤维化模型大鼠肝细胞损伤具有一定改善作用,其机制可能与上调Bcl-2、下调Bax表达有关。     

荔枝核总黄酮对肝纤维化大鼠肝脏NLRP3表达的影响

目的 研究荔枝核总黄酮(total flavones from Lychee seed, TFL)对二甲基亚硝胺(dimethylnitrosamine, DMN)所诱发的肝脏纤维化大鼠肝脏中核苷酸结合寡聚化结构域样受体蛋白3(nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)表达的影响。方法 研究将Wistar雄性大鼠随机分为正常组、模型组、扶正化瘀组和荔枝核总黄酮高、中、低剂量组。ELISA法测定血清中羟脯氨酸、透明质酸、Ⅳ型胶原、Ⅲ型前胶原和层粘连蛋白的水平;HE染色和Masson染色观察大鼠肝脏组织的病理变化;RT-qPCR测定肝脏组织中硫氧还蛋白结合蛋白(thioredoxin-interacting protein, TXNIP)、核苷酸结合寡聚化结构域样受体蛋白3、半胱氨酸天冬氨酸蛋白酶-1(cysteine-aspartic proteases-1,caspase-1)、白细胞介素-1β(interleulin-1β,IL-1β)的mRNA表达水平。结果 荔枝核总黄酮减轻大鼠肝...     

荔枝核总黄酮对大鼠肝纤维化TGF-β1及 CTGF表达的影响

目的 研究荔枝核总黄酮（TFL）在胆总管结扎诱导的大鼠肝纤维化模型中的治疗效果及其作用机制.方法 雄性SD大鼠100只,随机分为假手术组、模型组、水飞蓟素（SIL）组、TFL大剂量组（200 mg·kg^-1·d^-1）和TFL小剂量组（100 mg·kg^-1·d^-1）.采用胆总管结扎制备肝纤维化大鼠模型.4周后处死大鼠,放射免疫法（RIA）检测大鼠血清中透明质酸（HA）、层粘蛋白（LN）及Ⅲ型前胶原（PCⅢ）的水平;HE染色和Masson染色观察大鼠肝组织病理改变;采用酶联免疫吸附法（ELISA）检测大鼠血清中结缔组织生长因子（CTGF）的水平;免疫组化检测CTGF 及转化生长因子β1（TGF-β1）在肝组织内的表达.结果 与模型组大鼠比较, SIL和TFL大剂量治疗后大鼠血清中HA、LN和PCⅢ的水平显著降低（P〈 0.01）;肝组织内纤维化程度降低,肝小叶结构趋于正常;TFL大剂量给药组和SIL组血清CTGF水平均显著低于模型组（P〈0.05）;2组大鼠肝组织CTGF及TGF-β1表达明显降低（P 〈 0.05）.结论 TFL能有效地减轻胆总管结扎诱导的肝纤维化大鼠的肝损伤及纤维化程度,其机制可能与抑制肝内CTGF 和TGF-β1 表达有关.     

荔枝核总黄酮对肝纤维化大鼠肝组织TLR4/NF-κB信号通路的影响

目的探讨荔枝核总黄酮（TFL）对胆管阻塞型肝纤维化大鼠肝组织Toll样受体4（TLR4）、核因子κB（NF κB）表达的影响。方法雄性SPF级SD大鼠80只,随机分为正常对照组、模型组、TFL小剂量组、TFL大剂量组。除正常对照组外,其余各组均采用胆总管结扎法（CBDL）制备肝纤维化大鼠模型。造模后第2天开始,正常对照组、模型组给予0.9%氯化钠溶液灌胃（5 mL&#8226;kg－1）,TFL小剂量组给予TFL 100 mg&#8226;kg－1、TFL大剂量组给予TFL 200 mg&#8226;kg－1灌胃,均每天1次,共4周,第4周末处死大鼠,留取肝脏组织。苏木精 伊红（HE）染色、Masson染色观察大鼠肝组织病理学改变,免疫组化法（IHC）检测TLR4、NF κB在4组大鼠肝组织内的表达。结果与模型组比较,TFL大剂量组肝组织纤维化程度明显减轻,胶原表达明显减少（P＜0.01）;正常对照组、模型组、TFL小剂量组、TFL大剂量组的大鼠肝组织TLR4免疫组化评分分别为（0.4±0.5）,（3.3±0.6）,（3.1±0.5）,（1.4±0.5）分;NF κB免疫组化评分分别为（0.80±0.77）,（5.12±0.89）,（4.68±0.70）,（3.47±0.61）分,模型组TLR4、NF κB表达明显高于正常对照组和TFL大剂量组（P＜0.01）,与TFL小剂量组比较,差异无统计学意义。 结论TLR4和NF κB高表达可能与胆管阻塞型大鼠肝纤维化的发生发展有关,大剂量荔枝核总黄酮可能通过抑制大鼠肝组织TLR4、NK κB的表达来改善胆管阻塞型大鼠肝纤维化程度。     

荔枝核总黄酮对两种肝纤维化大鼠模型的作用比较

目的探讨荔枝核总黄酮(TFL)对四氯化碳(CCl₄)诱导肝纤维化大鼠和胆管结扎(BDL)肝纤维化大鼠的作用及其机制。方法大鼠40只,随机分正常对照组、CCl₄模型对照组、CCl₄+TFL组、BDL模型对照组、BDL+TFL组,每组8只。采用40%CCl₄诱导建立肝纤维化大鼠模型,双重胆管结扎法复制BDL模型。CCl₄+TFL组和BDL+TFL组灌胃给予TFL(300 mg·kg^-1·d^-1),正常对照组、CCl₄组和BDL组灌胃0.9%氯化钠溶液(300 mg·kg^-1·d^-1)。检测血清中碱性磷酸酶(ALP)、丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、总胆红素(T-BiL)。Sirius red染色测算胶原面积;免疫组织化学法检测抗α-平滑肌肌动蛋白(α-SMA)、CollagenⅠ在肝脏组织中的表达;Western blotting法检测α-SMA、...     

鬼针草总黄酮抗大鼠肝纤维化的实验研究

目的:观察鬼针草总黄酮(totalflavonesofBi-denspilosaL,TFB)的抗肝纤维化作用。方法:将大鼠随机分为六组:正常对照组,模型组,TFB160、80、40mg.kg-1组和秋水仙碱0.1mg.kg-1阳性药对照组。除正常对照组外,其余各组采用皮下注射四氯化碳(CCl4)诱导肝纤维化模型。于造模第9周起,给药组分别灌胃相应的受试药物,正常对照组和模型组灌胃等容量的生理盐水,疗程10周。实验结束后,取大鼠血清测ALT、AST、透明质酸(HA)、Ⅲ型前胶原肽(PCⅢ)、Ⅳ型前胶原酶(CⅣ);取大鼠肝、脾称重,计算肝、脾指数;同时取固定部位肝组织,测定组织中MDA、Hyp含量和GSH-Px活性;另取部分肝组织做病理组织学和电镜学检查。结果:TFB160、80mg.kg-1能显著降低肝纤维化大鼠肝、脾指数,并降低血清中ALT、AST活性及HA、PCⅢ、CⅣ和肝组织中MDA、Hyp含量,且升高肝组织中GSH-Px活性(P<0.05)。病理组织学和电镜检查结果显示TFB160、80mg.kg-1组肝脏组织结构明显改善,肝纤维化增生程度减轻。结论:TFB对CCl4所致大鼠肝纤维化有明显治疗作用,其机制可能与抑制氧自由基的生成有关。     

鬼针草总黄酮抗大鼠免疫性肝纤维化作用及部分机制研究

目的探讨鬼针草总黄酮(TFB)抗大鼠免疫性肝纤维化的作用及机制。方法SD大鼠随机分为正常组、模型组、TFB大、中、小剂量组和秋水仙碱阳性药对照组,采用腹腔注射猪血清诱导肝纤维化模型。造模后,给药组分别灌服相应的受试药物,正常组和模型组灌服等量的生理盐水。ELISA法测定大鼠血清透明质酸(HA)、层粘蛋白(LN)、Ⅲ型前胶原酶(PCⅢ)、Ⅳ型胶原酶(CⅣ)含量;取固定部位肝组织做病理组织学检查;免疫组织化学技术测定肝组织中α-SMA蛋白的表达;RT-PCR技术测定肝组织中TGF-β1mRNA的表达情况。结果TFB能明显降低肝纤维化大鼠血清中HA、LN、PCⅢ、CⅣ含量;病理组织学检查结果显示TFB组肝脏组织结构明显改善,肝纤维化程度减轻;TFB大、中剂量组明显抑制肝组织中α-SMA蛋白和TGF-β1mRNA的表达。结论TFB对猪血清所致大鼠免疫性肝纤维化有明显治疗作用,其机制可能与降低TGF-β1mRNA表达进而抑制肝星状细胞活化有关。     

鬼针草总黄酮对肝纤维化大鼠细胞因子的影响

目的观察鬼针草总黄酮(totalflavones of Bidenspilo-saL,TFB)对肝纤维化大鼠细胞因子的影响。方法将大鼠随机分成正常组、对照组、TFB160、80、40mg·kg-1组和秋水仙碱(Col)0.1mg·kg-1阳性药对照组。除正常组外,其余各组采用四氯化碳(CCl4)诱导肝纤维化模型。于造模wk9起,给药组分别灌胃相应的受试药物,正常组和对照组灌胃等容量的生理盐水,疗程10wk。实验结束后,股动脉采血,放免法测定血清中肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)的含量,同时取固定部位肝脏组织,免疫组化技术测定TFB对肝组织中TGF-β1和NF-κB蛋白表达的影响,RT-PCR技术测定TFB对肝组织中TGF-β1 mRNA表达的影响。结果TFB160、80mg·kg-1能明显降低肝纤维化大鼠血清TNF-α、IL-1β含量,抑制肝纤维化大鼠肝组织中NF-κB、TGF-β1蛋白和TGF-β1 mRNA表达。结论TFB通过降低肝纤维化大鼠炎症细胞因子而抗肝纤维化。     

赖氨大黄酸与大黄酸对大鼠胆汁淤积性肝纤维化的疗效比较

目的探讨赖氨大黄酸(rhein lysinat,RHL)和大黄酸对胆汁淤积性肝纤维化模型大鼠肝脏的保护作用,比较RHL与大黄酸对大鼠胆汁淤积性肝纤维化的疗效。方法采用胆总管结扎(bile duct ligation,BDL)的方法制造了大鼠胆汁淤积性肝纤维化模型,将大鼠随机分为5组:赖氨酸组、RHL组、大黄酸组、模型组、对照组。检测大鼠血清中天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)、总胆汁酸(TBA)和总胆红素(TBIL)的含量,HE染色观察肝脏组织病理学改变,Masson三色染色观察肝脏纤维结缔组织改变,使用酶标仪通过试剂盒检测肝脏羟脯氨酸(hydroxyproline,Hyp)含量,免疫组化染色观察α平滑肌肌动蛋白(α-smooth muscleactin,α-SMA)的分布与表达,Western blotting检测α-SMA相对表达量的变化。结果与对照组相比较,模型组大鼠生长速率、肝脏与体质量比、血清中AST、ALT、TBA和TBIL的含量升高(P<0.05),肝小叶的结构被破坏,纤维异常增生,Hyp的含量升高,免疫组化染色显示α-SMA分布于胞膜和胞质且表达增多,Western blotting结果显示模型组大鼠α-SMA相对表达量升高,提示肝星形细胞活化增多;与模型组相比,用药组大鼠生长率、肝脏与体质量比虽无显著变化,但是血清中AST、ALT、TBA、TBIL水平明显降低;用药组大鼠肝脏组织中纤维成分经治疗后减少,Hyp含量降低,用药后大鼠肝脏组织中α-SMA表达含量亦相应减少(P<0.05),提示肝星形细胞活化减少,且RHL组较大黄酸组降低显著。结论 RHL和大黄酸通过抑制肝星形细胞激活、肝脏纤维变性、蓄积来发挥对胆汁淤积性肝纤维化大鼠肝脏的保护作用,且RHL的作用强于大黄酸,提示RHL有望成为防治胆汁淤积性肝纤维化的药物。     

荔枝核中鞣质类型分析及其提取方法研究

目的研究荔枝核鞣质提取工艺,分析提取物中鞣质的类型。方法采用颜色-沉淀反应对荔枝核鞣质进行鉴别;以提取物中鞣质含量为指标,采用水浸泡,70%甲醇、乙醇、丙酮浸泡,70%丙酮加热回流及超声等提取方法提取得浸膏,用干酪素法测鞣质含量。结果不同溶剂提取工艺中,鞣质提取率高低顺序为丙酮加热回流>丙酮超声>丙酮浸泡>甲醇浸泡>乙醇浸泡>水浸泡。结论荔枝核鞣质为缩合鞣质,70%丙酮加热回流提取物中鞣质含量最高,该方法可为提取荔枝核中鞣质的提取方法。     

Delayed progression of diabetic cataractogenesis and retinopathy by Litchi chinensis in STZ-induced diabetic rats

Context: The study was carried out to evaluate the effect of the aqueous fruit pericarp extract of Litchi chinensis (APLC) on parameters which leads to diabetic cataractogenesis and retinopathy in the streptozotocin-induced diabetic rats.

Objective: The objective of the study is to evaluate the APLC for in vivo antioxidant activity and its role in inhibiting the polyol pathway and formation of advanced glycation end products (AGEs).

Materials and methods: The diabetic animals were treated with L. chinensis for a period of 12 weeks. At the end of 12 weeks, the animals were killed and the biochemical pathways involved in the pathogenesis of cataract such as oxidative stress by protein content, superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), and polyolpathway by aldose reductase (AR) in lens homogenates, alterations in protein carbonyl content (PCO) and AGEs in both serum and lens the APLC-treated diabetic rats were compared against diabetic control rats. Cataract progression due to hyperglycemia was monitored by slit lamp bio microscope and classified into four stages. Fundoscope test and retinal histopathology were done for assessing retinopathy.

Results: Statistically significant reduction in glucose, and elevation of protein content, SOD, CAT, and GSH levels and decreased levels of AR and PCO in lens homogenate and significant reduction in AGEs serum and lens homogenate were observed. Slit lamp examination, fundoscope, and histopathology showed improvement in retinal changes in APLC-treated rats compared to diabetic control animals.

Conclusion: The treatment with APLC found to delay the progression of diabetic cataractogenesis and retinopathy, which might be due to its antioxidant activity, because of the presence of active phytochemicals in APLC.     

五味子粗多糖提取物对大鼠肝纤维化的防治作用研究

目的:研究五味子粗多糖提取物对复合病因诱导的大鼠肝纤维化的防治作用。方法:实验分为6组,即正常对照(生理盐水)、模型(生理盐水)、秋水仙碱(0.1mg·kg-1)和五味子粗多糖提取物高、中、低剂量(400、200、100mg·kg-1)组。腹腔注射CCl4复制大鼠肝纤维化模型;灌胃给药。检测大鼠血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)的活性,肝匀浆丙二醛(MDA)和羟脯氨酸(HOP)的含量以及超氧化物岐化酶(SOD)的活性;形态学上观察肝组织病理学变化,用细胞图像分析系统(CIAS)对肝纤维化进行半定量。结果:与正常对照组比较,Massion染色显示模型组肝纤维化明显,有大量胶原纤维存在,肝小叶结构紊乱,血清ALT和AST显著升高,肝中MDA和HOP含量显著升高,SOD活性显著下降(P<0.01)。与模型组比较,各用药组大鼠血清ALT和AST显著降低;肝中MDA含量下降,SOD活性升高,具有显著性差异(P<0.01),并呈剂量依赖效应;肝HOP含量及胶原面积百分比显著下降(P<0.01)。结论:五味子多糖可减轻肝损害、肝脂质过氧化和肝纤维化的作用。     

正交试验优选荔枝核总皂苷提取工艺

目的:优选荔枝核总皂苷的提取工艺。方法:以荔枝核总皂苷的含量和提取物收率为指标,以提取溶剂、溶剂用量、提取次数和提取时间为考察因素,采用正交试验法优选最佳提取工艺。结果:最佳提取工艺是以70%乙醇为提取溶剂,溶剂用量为1g药材粉末加入8mL溶剂,提取1次,每次1h。结论:此工艺简便、易行,稳定性好,可为进一步利用荔枝核提供理论参考。     

糖皮质激素对淤胆型药物性肝病的疗效

目的比较激素及非激素在治疗严重的淤胆型药物性肝病(DILD)中的作用。方法淤胆型DILD诊断依据为Za-kim分类标准,疗效标准为治疗后血清胆红素降至正常,疗效的显著性比较用t检验进行分析。结果1996～2006年10年间收治的严重淤胆型DILD78例,依据是否应用激素治疗将其分为激素治疗组及非激素治疗组。激素治疗组以激素使用前的总胆红素为基数,非激素治疗组以入院时的总胆红素为基数;以住院期间平均每天总胆红素升高或下降占基数的百分比表示治疗反应。激素治疗组为52例,治愈率为92.31%。非激素治疗组为26例,治愈率为69.23%,在治疗有效的病例中,激素治疗组开始治疗时总胆红素为(354.36±137.68)μmol/L,比非激素治疗组的(254.77±74.51)μmol/L明显为高(P<0.05)。激素治疗组总胆红素每天平均下降速率为3.56%±2.11%,明显快于非激素治疗组的2.12%±1.02%(P<0.05)。结论对于胆汁淤积型DILD,糖皮质激素可以有效减轻淤胆症状,使黄疸消退。     

Aqueous extract of Artemisia iwayomogi Kitamura attenuates cholestatic liver fibrosis in a rat model of bile duct ligation

Cholestatic liver fibrosis, characterized by excessive accumulation of extracellular matrix (ECM) proteins, is associated with bile acid-induced oxidative stress and lipid peroxidation. We evaluated the therapeutic or protective effect of an aqueous extract of Artemisia iwayomogi Kitamura (WAI) in a rat bile duct ligation (BDL)-induced hepatic fibrogenesis model. After BDL, rats were treated once daily with 25 or 50 mg/kg of WAI for 2 weeks. The serum bilirubin, aspartate transaminase, alanine transaminase, malondialdehyde, and liver hydroxyproline levels were drastically increased in the BDL group. WAI administration significantly reduced these markers and restored BDL-induced depletion of glutathione content and glutathione peroxidase activity. Cholestatic liver injury and collagen deposition were markedly attenuated by WAI treatment, and these changes were paralleled by significantly suppressed gene and protein expression of fibrogenic factors, including hepatic alphasmooth muscle actin, platelet-derived growth factor, and transforming growth factor β. Our data suggest that WAI may have antifibrotic properties via both improvement of antioxidant activities and inhibition of ECM protein production in the rat model of BDL.     

Antifibrotic effect ofStephania tetrandra on experimental liver fibrosis induced by bile duct ligation and scission in rats

We examined the antifibrotic effect of a methanol extract from Stephania tetrandra (ST) on experimental liver fibrosis. Liver fibrosis was induced by bile duct ligation and scission (BDL/S) in rats. In BDL/S rats, activity levels of aspartate transaminase (AST), alanine transaminse (ALT), alkaline phosphatase (ALP), concentration of total bilirubin in serum, and hydroxyproline content of the liver were significantly increased. The ST treatment (either 100 mg/kg/day or 200 mg/kg/day, p.o. for 4 weeks) in BDL/S rats reduced the serum AST, ALT and ALP activity levels significantly (p< 0.01). Similarly, when compared to the control group, the concentration of hydroxyproline in the livers of the BDL/S rats treated with 100mg or 200mg ST treated rats decreased by 40% and 33% respectively, when compared to the BDL/S control group (p<0.01). The morphological characteristics of fibrotic liver that were observed in the BDL/S control group, improved in the ST treated BDL/S group. In the fibrotic liver of BDL/S rats treated with ST, a marked reduction in the numbers of alpha smooth muscle cell actin positive stellate cells was observed. These results indicate that doses of either 100 or 200 mg/kg/day of methanol extract from S. tetrandra, had an antifibrotic effect in rats with liver fibrosis induced by bile duct ligation and scission.