【摘要】：目的：觀察膽石六號湯劑對C57小鼠膽固醇結(jié)石和豚鼠膽色素結(jié)石形成的防治作用，并初探其作用機(jī)制。 方法： （1）對C57小鼠膽固醇結(jié)石的影響將50只雌性小鼠隨機(jī)分為空白對照組（n=10）、模型組（n=15）、膽石六號湯劑組（n=15）和�；切苋パ跄懰彡栃越M（n=10）。除空白對照組外，其余各組采用高脂飲食誘發(fā)法建立膽固醇結(jié)石模型。造模60天，第61天開始膽石六號湯劑組給予膽石六號湯劑21g·kg-1·d-1，陽性組給予�；切苋パ跄懰�12mg·kg-1·d-1灌胃（ig）治療。ig給藥30天后觀察各組小鼠的成石率、血脂含量、肝臟超氧化物歧化酶（SOD）活性及丙二醛（MDA）含量。 （2）對豚鼠膽色素結(jié)石的作用及機(jī)制研究40只雌性豚鼠隨機(jī)分為空白對照組（n=10）、模型組（n=15）和膽石六號湯劑組（n=15）。除空白對照組外，其余2組采用飲食誘發(fā)法建立膽色素結(jié)石模型。同時(shí)膽石六號湯劑組給予膽石六號湯劑7.7g·kg-1·d-1ig30天。之后觀察各組的成石率、血清膽色素含量、肝臟SOD活性及MDA含量、膽囊收縮素a受體（CCKaR）、5-羥色胺3a受體（5-HT3aR）基因和蛋白的表達(dá)情況。 結(jié)果： （1）對C57小鼠膽固醇結(jié)石模型的影響膽石六號湯劑給藥劑量為21g·kg-1·d-1，模型組成石率為85.71%，顯著高于空白對照組、膽石六號湯劑組和陽性組（P0.05）。膽石六號湯劑組血清低密度脂蛋白、谷草轉(zhuǎn)氨酶、谷丙轉(zhuǎn)氨酶、間接膽紅素、總膽汁酸含量低于模型組（P0.05），小鼠肝臟SOD活性升高和MDA含量下降，與模型組比較（P0.05）。 （2）對豚鼠膽色素結(jié)石模型的作用及機(jī)制研究空白對照組僅有1只有結(jié)石發(fā)生，成石率為10.00%；模型組成石率為91.67%。膽石六號湯劑組成石率為26.67%，較模型組明顯降低。膽石六號湯劑組血清總膽紅素、直接膽紅素和間接膽紅素含量低于模型組（P0.05），同時(shí)肝臟SOD活性明顯高于模型組，而MDA含量明顯低于模型組。RT-PCR檢測模型組中CCKaR、5-HT3aR基因的表達(dá)明顯低于空白組和膽石六號湯劑組（P0.05）。免疫組化結(jié)果顯示模型組5-HT3aR蛋白相對于空白組顯著下降，而膽石六號湯劑組陽性表達(dá)相對于模型組顯著上升（P0.05）。模型組CCKaR蛋白的表達(dá)相對于空白組有所下降，膽石六號湯劑組陽性表達(dá)相對于模型組有上升趨勢，但是差異沒有統(tǒng)計(jì)學(xué)意義（P0.05）。 結(jié)論： （1）膽石六號湯劑可降低小鼠膽固醇結(jié)石模型成石率，，改善血清和肝臟的生化相關(guān)指標(biāo)，從而發(fā)揮防治膽結(jié)石形成的作用。 （2）膽石六號湯劑可降低豚鼠膽色素結(jié)石模型成石率，其機(jī)制與其抗氧化應(yīng)激有關(guān)。 （3）膽石六號湯劑顯著影響了膽色素結(jié)石豚鼠膽組織CCKaR、5-HT3aR基因及蛋白的表達(dá)，其防治膽色素結(jié)石的作用一定程度上是通過增加它們的表達(dá)實(shí)現(xiàn)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.
【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R575.62

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