柚皮素对慢性胰腺炎小鼠胰腺纤维化及胰腺星状细胞活化、增殖和凋亡的影响  

The effects of naringenin on pancreatic fibrosis in chronic pancreatitis mouse model and activation, proliferation and apoptosis of pancreatic stellate cells

作　　者：吕彦玮 王丽娟 黄仁前 林曦 韩超 胡良皞 李兆申 Lyu Yanwei;Wang Lijuan;Huang Renqian;Lin Xi;Han Chao;Hu Lianghao;Li Zhaoshen(Department of Gastroenterology,First Affiliated Hospital of Naval Medical University,Shanghai 200433,China)

机构地区：[1]海军军医大学第一附属医院消化内科,上海200433

出　　处：《中华胰腺病杂志》2022年第3期185-190,共6页Chinese Journal of Pancreatology

基　　金：国家自然科学基金(81770635)。

摘　　要：目的研究柚皮素对CP小鼠胰腺纤维化及胰腺星状细胞(PSCs)的活化、增殖与凋亡的影响。方法18只C57BL/6小鼠随机分为对照组、CP组及柚皮素组,每组6只。采用腹腔注射雨蛙素法构建CP模型,柚皮素组于造模后第4周予柚皮素200 mg/kg灌胃,每天1次,直至处死小鼠的前一天;对照组和CP组予等容积0.5%羧甲基纤维素钠溶剂灌胃。造模期间行最后一次雨蛙素注射5 d后处死小鼠,取胰腺组织行苏木精-伊红染色及天狼星红染色,并进行病理学评分及胶原纤维沉积程度检测。用不同浓度(0、5、10、20、50、100、150、200μmol/L)柚皮素干预人PSCs株24 h,使用CCK-8法检测细胞存活率。用外源转化生长因子-β1(TGF-β1)重组蛋白(2 ng/ml)单纯刺激PSCs(TGF-β1刺激组),以及TGF-β1(2 ng/ml)刺激后1 h加低(50μmol/L)、中(100μmol/L)、高(150μmol/L)浓度柚皮素干预PSCs 36 h,采用蛋白质免疫印迹法检测PSCs活化相关蛋白FN和COL1A1、细胞增殖标志物p21以及抗凋亡蛋白Bcl-xL和促凋亡蛋白Bax、Bid的表达水平。结果CP组和柚皮素组小鼠胰腺组织病理评分[(7.33±1.15)、(4.67±1.15)分]和胶原纤维阳性区域所占百分比[(46±4)%、(28±2)%]均显著高于对照组[0分、(4±2)%],柚皮素组又显著低于CP组,差异均有统计学意义(P值均<0.05)。对照组,TGF-β1刺激组,TGF-β1刺激后低、中、高浓度柚皮素干预组FN蛋白表达量分别为0.02、0.76、0.67、0.34、0.07,COL1A1蛋白表达量分别为0.51、1.71、1.34、0.84、0.11,TGF-β1刺激组显著高于对照组,TGF-β1刺激后低、中、高浓度柚皮素干预组则显著低于TGF-β1刺激组,差异均有统计学意义(P值均<0.05)。上述5组p21蛋白表达量分别为0.87、1.18、1.27、1.22、1.00,TGF-β1刺激组显著高于对照组,仅TGF-β1刺激后高浓度柚皮素干预组显著低于TGF-β1刺激组,差异均有统计学意义(P值均<0.05);上述5组Bcl-xL蛋白表达量分别为2.09、2.21、2.38、2.50、2.12,BaxObjective To study the effects of naringenin on pancreatic fibrosis in the mouse model of chronic pancreatitis(CP)and its effects on the activation,proliferation and apoptosis of pancreatic stellate cells(PSCs).Methods Eighteen C57BL/6 mice were randomly divided into control group,CP group and naringenin group,with 6 mice in each group.The CP mouse model was established by intraperitoneal injections of caerulein.Naringenin group was given naringenin(200 mg/kg/day)by gavage once a day from the first day of the fourth week of modeling process to the day before the killing;the control group and CP group were treated by gavage with an equivalent amount of drug solvent containing 0.5%sodium carboxymethyl cellulose(CMC-Na).Mice were killed 5 days after the last caerulein injection,and their pancreatic tissues were collected for hematoxylin-eosin staining and Sirius Red staining,pathological scoring and collagen sedimentation detection.Naringenin with different concentrations(0,5,10,20,50,100,150,200μmol/L)were used to intervene HPSC for 24 hours,and CCK-8 method was used to detect the cell activity.TGF-β1 recombinant protein(2 ng/ml)was used to induce PSCs for 1 hour(TGF-β1 stimulation group),and naringenin with low(50μmol/L),middle(100μmol/L)and high(150μmol/L)concentration was used to intervene for 36 hours after TGF-β1 stimulation,respectively.Western Blotting was used to detect the expression of PSC activation related proteins FN and COL1A1,cell proliferation marker p21,anti-apoptotic protein Bcl-xL,pro-apoptotic protein Bax and Bid.Results The pathological scores of pancreatic tissue[(7.33±1.15),(4.67±1.15)]and the percentage of collagen positive areas[(46±4),(28±2)%]in CP group and naringenin group were higher than those in the control group[0,(4±2)%].However,these indexes in the naringenin group were lower than those in CP group,and the differences were all statistically significant(all P value<0.05).The relative expression of FN in control group,TGF-β1 stimulation group and low,medium and high nari

关 键 词：柚皮素 胰腺炎 慢性 纤维化 胰腺星状细胞 细胞增殖 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]