1.Construction and evaluation of spleen-deficiency hyperlipidemia mouse models
Lijuan CHEN ; Xinxue GAO ; Jin WU ; Ying DU ; Meijun LYU ; Guoyuan SUI ; Lianqun JIA ; Guowei PAN
Chinese Journal of Tissue Engineering Research 2025;29(29):6237-6242
BACKGROUND:Traditional Chinese medicine has unique advantages in preventing and treating spleen-deficiency and hyperlipidemia.In basic studies,models of spleen-deficiency and hyperlipidemia are commonly found in rats,pigs,and other animals.This has limitations for medical research that can only use mouse models.It is urgent to establish and evaluate mouse models of spleen-deficiency and hyperlipidemia to support basic research on traditional Chinese medicine in preventing and treating spleen-deficiency and hyperlipidemia.OBJECTIVE:To establish a mouse model of spleen-deficiency hyperlipidemia.METHODS:Totally 24 C57BL/6J mice were randomly divided into normal group(n=12)and spleen-deficiency hyperlipemia group(n=12).Mice in normal group were fed basic diet.Mice in the spleen-deficiency hyperlipemia group were prepared with a diet disorder+fatigue internal injury+high-fat feeding method to establish a spleen-deficiency high-fat model.In the first 2 weeks,the mice were forced to swim to their endurance limit on a single day and were only fed cabbage,with free access to water.They were also gavaged with refined lard+high-fat feed on two-day intervals.After 2 weeks,the mice were fed a high-fat diet every day and the diet continued until 12 weeks.The mice were fed with a high-fat diet for 4 and 12 weeks,and their body weight,food intake,gripping strength,fecal water content,small intestinal charcoal propulsion rate,serum D-xylose and gastrin levels,spleen index and thymus index,blood lipid level,total body fat mass,body fat percentage,and liver lipid deposition were tested.RESULTS AND CONCLUSION:(1)Compared with the normal group,the body weight,fecal water content,total body fat mass,body fat percentage,triglyceride and total cholesterol levels of the mice in the spleen-deficiency hyperlipemia group fed with high-fat diet for 4 and 12 weeks were increased(P<0.05);the daily food intake,gripping force,and D-xylose level of the mice fed with high-fat diet for 4 and 12 weeks were decreased(P<0.05);the spleen index of the mice fed with high-fat diet for 4 weeks was increased(P<0.05);the small intestinal carbon propulsion rate,gastrin level,spleen index,and thymus index of the mice fed with high-fat diet for 12 weeks were decreased(P<0.05);the low-density lipoprotein cholesterol level of the mice fed with high-fat diet for 12 weeks was increased(P<0.05).(2)The results of liver oil red O staining showed that the lipid deposition in the spleen-deficiency hyperlipemia group after 4 weeks of high-fat diet feeding was slightly more than that in the normal group,and the lipid deposition in the high-fat diet feeding for 12 weeks was significantly more than that in the normal group.(3)The results show that a stable spleen deficiency and hyperlipidemia mouse model can be prepared by the compound method of eating disorders,exhaustion,and high-fat feeding.
2.Construction and evaluation of spleen-deficiency hyperlipidemia mouse models
Lijuan CHEN ; Xinxue GAO ; Jin WU ; Ying DU ; Meijun LYU ; Guoyuan SUI ; Lianqun JIA ; Guowei PAN
Chinese Journal of Tissue Engineering Research 2025;29(29):6237-6242
BACKGROUND:Traditional Chinese medicine has unique advantages in preventing and treating spleen-deficiency and hyperlipidemia.In basic studies,models of spleen-deficiency and hyperlipidemia are commonly found in rats,pigs,and other animals.This has limitations for medical research that can only use mouse models.It is urgent to establish and evaluate mouse models of spleen-deficiency and hyperlipidemia to support basic research on traditional Chinese medicine in preventing and treating spleen-deficiency and hyperlipidemia.OBJECTIVE:To establish a mouse model of spleen-deficiency hyperlipidemia.METHODS:Totally 24 C57BL/6J mice were randomly divided into normal group(n=12)and spleen-deficiency hyperlipemia group(n=12).Mice in normal group were fed basic diet.Mice in the spleen-deficiency hyperlipemia group were prepared with a diet disorder+fatigue internal injury+high-fat feeding method to establish a spleen-deficiency high-fat model.In the first 2 weeks,the mice were forced to swim to their endurance limit on a single day and were only fed cabbage,with free access to water.They were also gavaged with refined lard+high-fat feed on two-day intervals.After 2 weeks,the mice were fed a high-fat diet every day and the diet continued until 12 weeks.The mice were fed with a high-fat diet for 4 and 12 weeks,and their body weight,food intake,gripping strength,fecal water content,small intestinal charcoal propulsion rate,serum D-xylose and gastrin levels,spleen index and thymus index,blood lipid level,total body fat mass,body fat percentage,and liver lipid deposition were tested.RESULTS AND CONCLUSION:(1)Compared with the normal group,the body weight,fecal water content,total body fat mass,body fat percentage,triglyceride and total cholesterol levels of the mice in the spleen-deficiency hyperlipemia group fed with high-fat diet for 4 and 12 weeks were increased(P<0.05);the daily food intake,gripping force,and D-xylose level of the mice fed with high-fat diet for 4 and 12 weeks were decreased(P<0.05);the spleen index of the mice fed with high-fat diet for 4 weeks was increased(P<0.05);the small intestinal carbon propulsion rate,gastrin level,spleen index,and thymus index of the mice fed with high-fat diet for 12 weeks were decreased(P<0.05);the low-density lipoprotein cholesterol level of the mice fed with high-fat diet for 12 weeks was increased(P<0.05).(2)The results of liver oil red O staining showed that the lipid deposition in the spleen-deficiency hyperlipemia group after 4 weeks of high-fat diet feeding was slightly more than that in the normal group,and the lipid deposition in the high-fat diet feeding for 12 weeks was significantly more than that in the normal group.(3)The results show that a stable spleen deficiency and hyperlipidemia mouse model can be prepared by the compound method of eating disorders,exhaustion,and high-fat feeding.
3.Exploration of the effects of Huayu qutan formula on atherosclerosis in ApoE-/- mice based on EETs-mediated pyroptosis
Na ZHAO ; Guoyuan SUI ; Jiawei MENG ; Meijun LYU ; Lianqun JIA
China Pharmacy 2024;35(12):1437-1442
OBJECTIVE To explore the effects and mechanism of Huayu qutan formula on atherosclerosis (AS) in ApoE-/- mice. METHODS Thirty ApoE-/- mice were randomly divided into model group, Huayu qutan formula group [20 g/(kg·d)], rosuvastatin group [1.55 mg/(kg·d)], with 10 mice in each group. Another 10 C57BL/6J mice were selected as normal control group. ApoE-/- mice were given high-lipid diet for 12 weeks to induce AS model. After modeling, each group was given relevant medicine or normal saline intragastrically, once a day, for 4 consecutive weeks. After the last administration, the levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL- C), 5,6-epoxyeicosatrienoic acid (5, 6-EET), 8,9-EET, 11,12-EET, 14,15-EET, interleukin-1β (IL-1β) and IL-18 in serum were detected; mRNA expressions of inhibitor of nuclear factor κB (IκB), nuclear factor κB (NF-κB), NOD-like receptor thermal protein domain associated protein 3 (NLRP3), Caspase-1, IL-1β and IL-18 in the aortic tissue of mice were detected; protein expression levels of IκB, NF- κB, NLRP3, apoptosis-associated speck-like protein containing CARD (ASC), Caspase-1, gasdermin D (GSDMD), IL-1β and IL-18 in the aortic tissue of mice were detected. The morphological changes of the aortic tissue were observed. RESULTS Compared with model group, the serum levels of TC, TG, LDL-C, IL-1β and IL-18, the mRNA expressions of IκB, NF-κB, NLRP3, Caspase-1, IL-1β and IL-18 in aortic tissue, and the protein expressions of IκB, NF-κB, NLRP3, ASC, Caspase-1, GSDMD, IL-1β and IL-18 were all decreased significantly in Huayu qutan formula group and rosuvastatin group (P<0.05), while the serum levels of 5,6-EET, 8,9-EET, 11,12-EET and 14,15-EET were increased significantly (P<0.05). The aortic atherosclerotic plaques were alleviated significantly. CONCLUSIONS Huayu qutan formula can play role of anti-AS through EETs-mediated pyroptosis.

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