| Background and ObjectiveCholesterol gallstone are a common and prevalent clinical condition in surgery.Studies have demonstrated that human intestinal flora is a major factor in the formation of cholesterol gallstone,and many applied investigations have been conducted in recent years both domestically and internationally.These stones can be broadly divided into pigment cholesterol gallstone,cholesterol stones,and mixed stones,with cholesterol stones being the most prevalent.The purpose of this study is to apply Qingrelidan granules and ursodeoxycholic acid to explore the efficacy of high cholesterol fat diet-induced cholesterol gallstone mouse model and the regulatory role of intestinal flora,which will provide support for cholesterol gallstone prevention and treatment.In this experiment,ursodeoxycholic acid was used for western medicine and Qingrelidan granules for Chinese medicine to intervene in the formation of cholesterol gallstone in mice model induced by high cholesterol lithogenic diet.To provide experimental basis and theoretical foundation for further research on the mechanism of cholesterol gallstone and the therapeutic mechanism of Qingrelidan granules.MethodsRandomly dividing forty SPF-grade male C57BL/6 mice at 6 weeks of age into five groups,eight were placed in the NC group,eight in the GS group,and eight in the QRLD prevention group,eight in the ursodeoxycholic acid prevention group.eight in the QRLD treatment group,eight in the negative control(NC group),eight in the negative control(GS group),eight in the QRLD prevention group(QRLD.P group),eight in the QRLD treatment group(QRLD.T group)and eight in the ursodeoxycholic acid Prevention group(UDCA.P group).The negative control was fed with normal feed,and the negative control,QRLD.T,QRLD.T group and UDCA.P were fed with high cholesterol feed to prepare the mouse models.In the QELD.P and UDCA.P,equal doses of gavage were given for 10 weeks(Qingrelidan granules for the group of Qingrelidan granules and ursodeoxycholic acid tablets for the group of ursodeoxycholic acid);in the QRLD.T,equal doses of 0.9% saline gavage were given for 1-4 weeks and continuous gavage with Qingrelidan granules for 5-10 weeks;in the negative control and the negative control of mice,equal doses of 0.9% saline gavage were given for 1-10 weeks.The normal and negative controls were gavaged with equal amounts of 0.9% saline for 1-10 weeks.The mice were monitored weekly for changes in general status and body weight.After 10 weeks of modeling,mice were executed and serum,gallbladder tissue,liver tissue,distal ileum tissue and feces were collected.Serum biochemical parameters and pathological damage of gallbladder,liver and distal ileum tissues,as well as analysis of fecal flora,were examined.HE staining method was employed to observe morphological alterations in the gallbladder,liver,and distal ileum of mice from each group,with the negative control as a control.The m RNA expression levels of CYP7a1 and CYP8b1 in mouse liver were measured by q-PCR method.The mouse fecal samples were sequenced with 16 SRNA for colony sequencing.Results3.1.Body weight changes: The negative control group body weight was significantly higher than the negative control at the conclusion of the modeling(P<0.05).Additionally,the Qingrelidan granules prevention group,Qingrelidan granules treatment group,and ursodeoxycholic acid prevention group body weight had a decreasing trend,yet no statistical difference was seen(P>0.05).3.2 Comparison of serum TC,TBA and TG contents.Comparison of serum contents of total cholesterol(TC),total bile acid(TBA)and total triglyceride(TG): Compared with the negative control,the m RNA expression of CYP7a1 and CYP8b1 in the liver of mice in the negative control was significantly lower(P<0.05);compared with the negative control,the levels of TC in the serum of mice in the QRLD Prevention group,the QRLD treatment group and the ursodeoxycholic acid prevention group decreased significantly(P<0.05);compared with the negative control,the levels of TBA in the serum of mice in the QRLD prevention group,the QRLD treatment group and the ursodeoxycholic acid prevention group decreased significantly(P<0.05).Compared with the negative control,the serum TBA content of mice in the treatment group decreased(P<0.05);compared with the QRLD treatment group,the serum TBA content of mice in the QRLD prevention group decreased significantly(P<0.05),compared with the negative control,the serum TBA content of mice in the QRLD prevention group increased(P<0.05);compared with the negative control the serum TG content of mice in the QRLD prevention group increased(P<0.05);compared with the negative control,the serum TBA content of mice in the QRLD prevention group increased(P<0.05).3.3 Serumcontents of high density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)compared with the negative control,HDL-C in the serum of mice in the prevention group of clearing heat and bile granules,the prevention group of ursodeoxycholic acid and the treatment group of clearing heat and bile granules were reduced(P<0.05).3.4 Serum AST and ALT content: The serum levels of TC,TBA and TG in the mice in the negative control significantly increased(P<0.05)when compared to the negative control;conversely,the serum levels of TC in the serum of mice in the QRLD prevention group,the QRLD treatment group and were all significantly higher than those of the negative control.The levels of TBA in the serum of mice in the QRLD prevention group,the QRLD treatment group,and the ursodeoxycholic acid prevention group all significantly decreased(P < 0.05)when compared to the negative control,with the latter group exhibiting a significant decrease(P<0.05).The serum TBA content of mice in the treatment group was significantly lower than that of the negative control(P < 0.05);similarly,the serum TBA content of mice in the QRLD prevention group was significantly lower than that of the QRLD treatment group(P < 0.05).In contrast,the serum TBA content of the negative control was not significantly different.The QRLD prevention group saw a marked rise in the number of mice(P < 0.05);additionally,the serum TG content of the QRLD prevention group was found to be higher than that of the negative control(P < 0.05),and the serum TBA content of the QRLD prevention group was also found to be higher than that of the negative control(P < 0.05).Significance was found in the serum of mice in the prevention group of clearing heat and bile granules,the prevention group of ursodeoxycholic acid,and the treatment group of clearing heat and bile granules when compared to the negative control.The serum of mice in the prevention group of clearing heat and bile granules,ursodeoxycholic acid,and the treatment group of clearing heat and bile granules all showed a significantly reduced HDL-C(P<0.05)when compared to the negative control(P<0.05).The serum AST content of mice in the negative control significantly increased(P < 0.05)when compared to the negative control,while the serum AST content of mice in the prevention group of fever and bile granules,ursodeoxycholic acid,and the treatment group of fever and bile granules decreased significantly(P < 0.05).Compared with the negative control,the serum ALT of mice in the negative control,the prevention group of fever and bile granules,the treatment group of fever and bile granules,and the prevention group of ursodeoxycholic acid had a tendency to increase,among which the increase was most obvious in the negative control,but there was no statistical difference(P>0.05).3.5 Serum TNF-α content: compared with the negative control,the serum TNF-α content of tumor necrosis factor in mice in the negative control,the prevention group of clearing heat and bile granules and the treatment group of clearing heat and bile granules all decreased significantly(P<0.05).3.6 Comparison of m RNA expression of CYP7a1 and CYP8b1 in mice liver: compared with the negative control,the m RNA expression of CYP7a1 and CYP8b1 in the liver of mice in the negative control was significantly lower P<0.05,compared with the negative control,the m RNA expression of CYP7a1 and CYP8b1 in the liver of mice in the prevention group of clearing fever and bile granules,the prevention group of ursodeoxycholic acid and the treatment group of clearing fever and bile granules m RNA expressions in the liver of mice in the treatment groups of Qingrelidan granules,ursodeoxycholic acid prevention group and Qingrelidan granules had a tendency to increase,but there was no statistical difference(P>0.05).Conclusions1.Granules of Qingrelidan can reduce the difference between the intestinal flora of mice in the negative control and the negative control,reduce the proportion of harmful bacteria,increase the proportion of lactobacillus with cholesterol-lowering effects,and normalize the structure of the intestinal flora of mice induced by high-cholesterol diet2.Qingrelidan granules can improve the liver function of gallstone mice,reduce the serum levels of TC,TG,TBA,LDL and HDL in gallstone mice,and up-regulate the gene expression of CYP7a1 and CYP8b1 m RNA,the rate-limiting enzymes that convert cholesterol into bile acids. |
PDF Full Text Request