胆石六号汤剂抗胆结石形成作用及机制研究

发布时间：2018-11-26 10:13

【摘要】：目的：观察胆石六号汤剂对C57小鼠胆固醇结石和豚鼠胆色素结石形成的防治作用，并初探其作用机制。方法：（1）对C57小鼠胆固醇结石的影响将50只雌性小鼠随机分为空白对照组（n=10）、模型组（n=15）、胆石六号汤剂组（n=15）和牛磺熊去氧胆酸阳性组（n=10）。除空白对照组外，其余各组采用高脂饮食诱发法建立胆固醇结石模型。造模60天，第61天开始胆石六号汤剂组给予胆石六号汤剂21g·kg-1·d-1，阳性组给予牛磺熊去氧胆酸12mg·kg-1·d-1灌胃（ig）治疗。ig给药30天后观察各组小鼠的成石率、血脂含量、肝脏超氧化物歧化酶（SOD）活性及丙二醛（MDA）含量。（2）对豚鼠胆色素结石的作用及机制研究40只雌性豚鼠随机分为空白对照组（n=10）、模型组（n=15）和胆石六号汤剂组（n=15）。除空白对照组外，其余2组采用饮食诱发法建立胆色素结石模型。同时胆石六号汤剂组给予胆石六号汤剂7.7g·kg-1·d-1ig30天。之后观察各组的成石率、血清胆色素含量、肝脏SOD活性及MDA含量、胆囊收缩素a受体（CCKaR）、5-羟色胺3a受体（5-HT3aR）基因和蛋白的表达情况。结果：（1）对C57小鼠胆固醇结石模型的影响胆石六号汤剂给药剂量为21g·kg-1·d-1，模型组成石率为85.71%，显著高于空白对照组、胆石六号汤剂组和阳性组（P0.05）。胆石六号汤剂组血清低密度脂蛋白、谷草转氨酶、谷丙转氨酶、间接胆红素、总胆汁酸含量低于模型组（P0.05），小鼠肝脏SOD活性升高和MDA含量下降，与模型组比较（P0.05）。（2）对豚鼠胆色素结石模型的作用及机制研究空白对照组仅有1只有结石发生，成石率为10.00%；模型组成石率为91.67%。胆石六号汤剂组成石率为26.67%，较模型组明显降低。胆石六号汤剂组血清总胆红素、直接胆红素和间接胆红素含量低于模型组（P0.05），同时肝脏SOD活性明显高于模型组，而MDA含量明显低于模型组。RT-PCR检测模型组中CCKaR、5-HT3aR基因的表达明显低于空白组和胆石六号汤剂组（P0.05）。免疫组化结果显示模型组5-HT3aR蛋白相对于空白组显著下降，而胆石六号汤剂组阳性表达相对于模型组显著上升（P0.05）。模型组CCKaR蛋白的表达相对于空白组有所下降，胆石六号汤剂组阳性表达相对于模型组有上升趋势，但是差异没有统计学意义（P0.05）。结论：（1）胆石六号汤剂可降低小鼠胆固醇结石模型成石率，，改善血清和肝脏的生化相关指标，从而发挥防治胆结石形成的作用。（2）胆石六号汤剂可降低豚鼠胆色素结石模型成石率，其机制与其抗氧化应激有关。（3）胆石六号汤剂显著影响了胆色素结石豚鼠胆组织CCKaR、5-HT3aR基因及蛋白的表达，其防治胆色素结石的作用一定程度上是通过增加它们的表达实现的。
[Abstract]:Objective: to observe the preventive and therapeutic effects of cholelithiasis No. 6 decoction on cholesterol gallstones in C 57 mice and the formation of pigment gallstones in guinea pigs, and to explore its mechanism. Methods: (1) 50 female mice were randomly divided into control group (n = 10), model group (n = 15), gallstone No. 6 decoction group (n = 15) and taurine ursodeoxycholic acid positive group (n = 10). Cholesterol calculi were induced by high fat diet in all groups except blank control group. The model was made for 60 days, and the gall stone No. 6 decoction group was given 21 g kg-1 d-1 on the 61st day. The positive group was treated with (ig) by intragastric administration of 12mg kg-1 d-1. The rate of stone formation, the content of blood lipid, the activity of liver superoxide dismutase (SOD) and the content of malondialdehyde (MDA) were observed 30 days after the administration of ig. (2) study on the effect and mechanism of pigment gallstone in guinea pigs; 40 female guinea pigs were randomly divided into three groups: blank control group (n = 10), model group (n = 15) and cholelithiasis No.6 decoction group (n = 15). In addition to the blank control group, the other two groups were induced by diet to establish the model of pigment gallstone. At the same time, the group of Ganshi No.6 decoction was given 7. 7 g kg-1 d-1ig30. Then the lithogenesis rate, serum bile pigment content, liver SOD activity and MDA content, and the expression of (CCKaR), 5-hydroxytryptamine 3a receptor (5-HT3aR) gene and protein of cholecystokinin a receptor were observed. Results: (1) the effect on cholesterol gallstone model in C57 mice was significantly higher than that in the blank control group. The dosage of Ganshi No. 6 decoction was 21g kg-1 d-1, and the model composition stone rate was 85.71g, which was significantly higher than that of the control group. Ganshi No.6 decoction group and positive group (P0.05). The contents of serum low density lipoprotein, glutamic oxaloacetic transaminase, alanine aminotransferase, indirect bilirubin and total bile acid in the Ganshi No.6 decoction group were lower than those in the model group (P0.05). The activity of SOD and the content of MDA decreased in the liver of mice. Compared with the model group (P0.05). (2) study on the effect and mechanism of pigment gallstone model in guinea pigs; in the blank control group, there was only one stone, the stone formation rate was 10.00, and the model composition stone rate was 91.67. The percentage of stone composition of the decoction was 26.67, which was significantly lower than that of the model group. The content of serum total bilirubin, direct bilirubin and indirect bilirubin were lower in the Ganshi No.6 decoction group than in the model group (P0.05). Meanwhile, the SOD activity of liver was significantly higher than that of the model group, while the content of MDA was lower than that of the model group. RT-PCR was used to detect CCKaR, in the model group. The expression of 5-HT3aR gene was significantly lower than that of blank group and cholelithiasis 6 decoction group (P 0.05). Immunohistochemical results showed that the 5-HT3aR protein in the model group was significantly lower than that in the blank group, while the positive expression in the Ganshi No. 6 decoction group was significantly higher than that in the model group (P0.05). The expression of CCKaR protein in the model group was lower than that in the blank group, and the positive expression in the Ganshi No. 6 decoction group had an upward trend compared with the model group, but the difference was not statistically significant (P0.05). Conclusion: (1) Ganshi No.6 decoction can reduce the rate of cholesterol stone formation in mice and improve the biochemical indexes of serum and liver so as to play a role in preventing and treating gallstone formation. (2) Ganshi No. 6 decoction can reduce the rate of stone formation in guinea-pig model of pigment gallstone, and its mechanism is related to its antioxidant stress. (3) Ganshi No. 6 decoction significantly affected the expression of CCKaR,5-HT3aR gene and protein in the gallbladder tissues of guinea pigs with gallbladder pigment stone, and its effect on the prevention and treatment of gallstone was achieved by increasing their expression to some extent.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R575.62

【参考文献】