ObjectiveTaking the cuproptosis/oxidative stress pathway as the entry point， this study investigated the effect and mechanism of Liangyi Paste on hepatic lipid deposition in naturally aged mice fed with a high-fat diet. MethodsAfter adaptive feeding， 80 ten-week-old male C57BL/6 mice were used. Thirty of them were randomly divided into three groups （10 mice per group）： The 12-month-old control group （12MCON）， the 15-month-old control group （15MCON）， and the 15-month-old group with a high-fat diet （15MHFD）. The 12MCON and 15MCON groups were continuously fed a standard diet， while the 15MHFD group started receiving a high-fat diet at 12 months of age. Tissue samples were collected at the corresponding time points for each group. The remaining 50 mice were randomly divided into five groups （10 mice per group）： the 20-month-old control group （20MCON）， the model group， and the low-， medium-， and high-dose Liangyi Paste groups （2.91 ， 5.82 ， 11.64 g·kg^-1·d^-1， respectively）. The 20MCON group was continuously fed a standard diet， while the other groups started receiving a high-fat diet at 15 months of age. At 18 months of age， the Liangyi Paste groups were administered the corresponding doses of Liangyi Paste by gavage， while the 20MCON and model groups were given an equal volume of saline by gavage. After 8 weeks of continuous gavage （when the mice reached 20 months of age）， tissue samples were collected. Hepatic TG levels were measured using assay kits； liver histology and lipid deposition were observed via hematoxylin-eosin （HE） and oil red O staining； reactive oxygen species （ROS） were detected by enzyme-linked immunosorbent assay （ELISA）； Cu²⁺， superoxide dismutase （SOD）， and malondialdehyde （MDA） levels were measured by colorimetry； mRNA and protein expression of genes related to cuproptosis and oxidative stress pathways were analyzed by Real-time polymerase chain reaction（Real-time PCR） and Wes automated protein expression system. ResultsCompared with 12MCON， the 15MCON group showed significantly increased hepatic TG， Cu²⁺， ROS， and MDA levels （P<0.01）， decreased SOD （P<0.01）， hepatocyte swelling， and disordered arrangement. The mRNA and protein levels of ferredoxin 1 （FDX1）， dihydrolipoamide S-acetyltransferase （DLAT）， heat shock protein 70 （HSP70）， dihydrolipoamide dehydrogenase （DLD）， pyruvate dehydrogenase E1 subunit-β （PDHB）， nuclear factor erythroid 2-related factor 2 （Nrf2）， and peroxisome proliferator-activated receptor γ （PPARγ） were significantly elevated （P<0.05， P<0.01）. Compared with 15MCON group， the 15MHFD and 20MCON groups exhibited further increases in TG， Cu²⁺， ROS， and MDA （P<0.01）， reduced SOD （P<0.01）， and aggravated hepatocyte swelling and disorder. There were increased lipid droplets with mild vacuolization in the 15MHFD group， and no significant lipid deposition was observed in the 20MCON group. FDX1， DLAT， HSP70， DLD， PDHB， Nrf2， and PPARγ mRNA and protein levels were significantly increased （P<0.05， P<0.01）. Compared with 20MCON group， the model group demonstrated markedly elevated TG， Cu²⁺， ROS， and MDA （P<0.01）， reduced SOD （P<0.01）， severe hepatic steatosis， and upregulated expression of FDX1， DLAT， HSP70， DLD， PDHB， Nrf2， and PPARγ mRNA and proteins （P<0.05， P<0.01）. All abnormalities were significantly reversed after Liangyi Paste treatment. ConclusionLiangyi paste can ameliorate hepatic lipid deposition in naturally aged mice with a high-fat diet by modulating the cuproptosis/oxidative stress pathway.

ObjectiveThe study aims to investigate the intervention effect of modified Xiangsha Liujunzitang （M-XSLJZ） on intestinal-derived lipopolysaccharide （LPS）-activated Kupffer cell inflammation in rats with hyperlipidemia spleen deficiency syndrome. MethodsSeventy male SD rats were randomly divided into seven groups （n=10）： blank control （CON）， high-fat diet without spleen deficiency （HFD）， high-fat diet with spleen deficiency （SD-HFD）， M-XSLJZ low-， medium-， and high-dose groups （XS-L， XS-M， XS-H）， and western medicine control （R）. Spleen deficiency was induced in SD-HFD， XS-L， XS-M， XS-H， and R groups via irregular diet combined with exhaustive swimming for 15 days. The CON group received a standard diet， while other groups were fed a high-fat diet for 10 weeks to establish the hyperlipidemia model. After successful modeling， rats were treated for 8 weeks： M-XSLJZ was administered at 3.51， 7.02， 14.04 g·kg^-1 in XS-L， XS-M， and XS-H groups， respectively. The R group received 9×10^-4 g·kg^-1 of a reference drug. D-xylose excretion rate was measured by the phloroglucinol method. Blood lipids were assessed using an automated biochemical analyzer. Hematoxylin-eosin （HE） staining was used to evaluate the pathological conditions of the liver， and oil red O staining was used to observe the lipid deposition in the liver. The levels of LPS， portal vein serum LPS， LPS-binding protein （LBP）， serum interleukin-6 （IL-6）， IL-1β， and tumor necrosis factor-α （TNF-α） were detected by enzyme-linked immunosorbent assay （ELISA）. Immunofluorescence was used to evaluate CD86 expression and CD68/TLR4 co-localization in the liver. Protein levels of TLR4， MyD88， NF-κB p65， and p-NF-κB p65 in Kupffer cells were analyzed via Western blot automated protein analysis. Hepatic IL-6， TNF-α， and IL-1β mRNA and protein levels were measured using Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. ResultsCompared with the CON group， the SD-HFD group showed a decrease in D-xylose excretion （P<0.01）. TC， TG， HDL-C， and LDL-C increased （P<0.05， P<0.01）. A large number of hepatic lipid vacuoles and orange-red lipid droplet deposition appeared in the liver. Ileal LPS， portal LPS， and LBP increased （P<0.05， P<0.01）. The levels of serum IL-6， TNF-α， and IL-1β increased （P<0.01）. The expression of CD86 was upregulated （P<0.01）， and the co-expression of CD68 and TLR4 was enhanced. The protein levels of TLR4， MyD88， and p-p65 in Kupffer cells increased （P<0.01）. The mRNA and protein levels of IL-6， TNF-α， and IL-1β increased （P<0.05， P<0.01）. Compared with the HFD group， the SD-HFD group exhibited decreased D-xylose excretion （P<0.01）， higher HDL-C， LDL-C （P<0.05）， increased portal LBP and LPS （P<0.05）， increased serum IL-6 and TNF-α （P<0.01）， upregulated CD86 （P<0.01）， enhanced CD68/TLR4 co-expression， and higher TNF-α mRNA/protein （P<0.05）. Compared with the SD-HFD group， all M-XSLJZ treatment groups showed reduced TC， TG， and LDL-C （P<0.05， P<0.01）. XS-H and R groups displayed improved hepatic lipid deposition. XS-H and R groups had lower ileal LPS， portal LPS， and LBP levels （P<0.05， P<0.01）. All M-XSLJZ treatment groups exhibited reduced serum IL-6， IL-1β， and TNF-α （P<0.01）. The XS-H group showed downregulated CD86 （P<0.01） and weakened CD68/TLR4 co-expression. The XS-H group had reduced TLR4， MyD88， and p-NF-κB p65 in Kupffer cells （P<0.01）. XS-H and R groups showed lower IL-6， TNF-α， and IL-1β mRNA/protein （P<0.05， P<0.01）. ConclusionM-XSLJZ may exert its lipid-lowering effects by inhibiting intestinal-derived LPS and alleviating Kupffer cell inflammation in the liver.

Objective Based on apolipoprotein a-Ⅰ(Apoa-Ⅰ)gene knockout mice,the role and mechanism of phosphotidylcholine(PC)in improving cholesterol reverse transport were explored.Methods Thirty Apoa-Ⅰ-/-mice were randomly divided into an Apoa-Ⅰ-/-group,Apoa-Ⅰ-/-+HFD group,and Apoa-Ⅰ-/-+HFD+PC group using the random number table method;30 C57BL/6J mice were randomly divided into a WT group,WT+HFD group,and WT+HFD+PC control groups,with 10 mice in each group.The Apoa-Ⅰ-/-group and WT groups were fed basic feed,while the other groups were fed high-fat feed for 8 weeks to establish a hyperlipidemia model.From the 9th week,the WT+HFD+PC group and Apoa-Ⅰ-/-+HFD+PC group were given PC 2.5 g/(kg·d),while the remaining mice were given physiological saline by gavage for a total of 4 weeks of intervention.The serum lipid levels of the mice were detected using a fully automated analyzer.Hematoxylin and eosin and Oil red O staining were used to observe pathological and morphological changes,and the COD-PAP method was used to detect cholesterol levels in mouse liver tissue.The ELISA method was used to detect LCTA levels in mouse serum,and RT-qPCR and Western Blot method were used to detect the mRNA and protein expression of cholesterol ATP binding cassette transporter A1(ABCA1),ATP binding cassette transporter G1(ABC A1),lecithin cholesterol acyltransferase(LCAT),hepatic lipase(HL),scavenger receptor class B type Ⅰ(SR-B1),and low-density lipoprotein receptor(LDL-R)in liver tissue.Results Compared with the WT group,the serum lipid levelsof WT+HFD group mice were significantly increased(P＜0.01),LCAT levels were significantly reduced(P＜0.05),hepatic fat vacuoles were obvious,hepatic lipid deposition was significant,and liver tissue TC levels were significantly increased(P＜0.01).The mRNA and protein expression of ABCA1,ABCG1,LCAT,SR-B1,HL,and LDL-R were significantly reduced(P＜0.05,P＜0.01).Compared with the WT+HFD group,serum lipid levels in the WT+HFD+PC group were significantly reduced(P＜0.05,P＜0.01),LCAT levels were significantly increased(P＜0.05),hepatic fat vacuoles were significantly reduced,hepatic lipid deposition was alleviated,and liver tissue TC levels were significantly reduced(P＜0.05);mRNA and protein expression of ABCA1,LCAT,SR-B1,HL and LDL-R were significantly increased(P＜0.05,P＜0.01).The serum levels of TC,TG,and LDL-C were significantly increased,while the levels of LCAT、HDL-C were significantly reduced(P＜0.05,P＜0.01)in the Apoa-Ⅰ-/-+HFD group mice.Hepatocytes underwent balloon-like transformation,liver lipid deposition was significantly aggravated,and liver tissue TC levels were significantly increased(P＜0.05).The mRNA and protein expression of ABCA1,LCAT and HL were significantly reduced(P＜0.05,P＜0.01).Compared with the WT+HFD+PC group mice,the Apoa-Ⅰ-/-+HFD+PC group mice showed a significant increase in serum lipid levels(P＜0.05,P＜0.01),LCAT levels were significantly reduced(P＜0.05),significant hepatic lipid vacuoles,significant hepatic lipid deposition,and a significant increase in TC levels in liver tissue(P＜0.05).Their mRNA and protein expression of ABCA1,ABCG1,LCAT,SR-B1,and HL were also significantly reduced(P＜0.05,P＜0.01).Conclusions Phosphatidylcholine can improve dyslipidemia by interfering with Apoa-Ⅰ and thus regulating cholesterol reverse transport.

AIM:This study aims to investigate the effects of baicalin on the hypoxia-inducible factor-1α(HIF-1α)/solute carrier family 7 member 11(SLC7A11)/glutathione peroxidase 4(GPX4)axis and the ferroptosis induced by oxidized low-density lipoprotein(ox-LDL)in RAW264.7 macrophage-derived foam cells.METHODS:RAW264.7 cells were categorized into five groups:control,ox-LDL,baicalin+ox-LDL,ferrostatin-1(Fer-1;ferroptosis inhibitor)+ox-LDL,and baicalin+Fer-1+ox-LDL.To induce foam cell formation,RAW264.7 macrophages were exposed to 100 μg/mL ox-LDL for 24 h.Oil red O staining was employed to visualize lipid droplet formation in each group.The ultrastructure of the mitochondria was examined using transmission electron microscopy.Fluorescence microscopy was utilized to assess the fluorescence intensity of intracellular reactive oxygen species(ROS),lipid peroxides,and Fe2+.A colorimetric assay facilitated the measurement of malondialdehyde(MDA)and glutathione(GSH)levels.Additionally,Western blot analy-sis was conducted to quantify protein levels of HIF-1α,SLC7A11,and GPX4.RESULTS:The model group exhibited foam cell formation,abundant lipid droplets,significant swelling of mitochondrial structures,and observable shortening or disappearance of cristae.There was a marked increase in intracellular fluorescence intensity of ROS,lipid peroxides,and Fe2+,alongside elevated MDA levels and decreased GSH levels.HIF-1α protein expression was significantly increased,while SLC7A11 and GPX4 protein expressions were notably decreased(P<0.05).In comparison to the model group,both the baicalin+ox-LDL and Fer-1+ox-LDL groups demonstrated a significant reduction in lipid droplets,improved mitochon-drial structures,decreased fluorescence intensity of ROS,lipid peroxides,and Fe2+,as well as lower MDA levels and higher GSH levels.Additionally,HIF-1α expression significantly decreased,while SLC7A11 and GPX4 expressions sig-nificantly increased(P<0.05).Furthermore,the baicalin+Fer-1+ox-LDL group showed a more pronounced reduction in lipid droplets,near-normal mitochondrial structures,lower fluorescence intensity of ROS,lipid peroxides,and Fe2+,de-creased MDA levels,and increased GSH levels compared to the baicalin+ox-LDL group;HIF-1α,SLC7A11,and GPX4 protein expressions were also significantly reduced(P<0.05).CONCLUSION:Baicalin modulates the HIF-1α/SLC7A11/GPX4 axis,thereby inhibiting ox-LDL-induced ferroptosis in macrophage-derived foam cells.

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.

Objective To explore the effect of Yinchenhao Decoction plus Zexie Decoction on ferroptosis mediated by IRE1 signaling pathway of endoplasmic reticulum stress in NASH mice.Methods Fifty C57BL/6J mice were randomly divided into blank control group,model group,Yinchenhao decoction group,Zexia decoction group and Hefang group,with 10 mice in each group.Except the blank control group,the other groups were fed with high-fat diet.After 20 weeks,Yinchenhao Tang group,Zexia Tang group and Hefang group were respectively given the corresponding drug by intragastric administration,once a day for 8 weeks.HE staining,oil red O staining and Masson staining were used to observe the pathological changes of liver tissue,automatic biochemical analyzer was used to detect blood lipid and liver function indexes,and RT-qPCR was used to detect the mRNA levels of TFR1,FPN,XBP1,Xbp1s and XBP1-dependent UPR target genes.The expressions of GRP78,P-IRE1α,GPX4,TFR1 and FPN proteins in liver were detected by Western Blot,the contents of Fe2+and MDA were detected by colorimetry,SOD activity was detected by WST-8,ROS content was detected by ELISA,and TG content in mouse liver tissue was detected by GPO-PAP.Results Compared with the model group,the NAS scores of Yinchenhao Tang group,Zexia Tang group and Hefang group were significantly reduced,and the effect of Hefang group was more significant.Compared with the blank control group,the liver lipid deposition of model mice was obvious,and the liver lipid deposition of Yinchenhao Tang group,Zexia group and Hefang group were improved to varying degrees,and the effect of Hefang group was more significant.Masson staining results showed that compared with blank control group,liver fibrosis was obvious in model group,Yinchenhao Decoction group,Zexia group and Hefang group,the degree of liver fibrosis was improved,and the effect of Hefang group was more significant.Compared with the blank control group,the levels of serum TC,HDL-C,LDL-C,AST and ALT in the model group were significantly increased,the contents of TG,Fe2+,MDA and ROS in the liver were significantly increased,and the activity of SOD was significantly decreased.The mRNA expression levels of Xbp1,Xbp1s,Dnajb9,Edem1,Sec61a1,Bip,Chop and TFR1were significantly up-regulated,the mrna expression levels of FPN were significantly down-regulated,and the protein expression levels of GRP78,P-IREα,GPX4 and TFR1 in liver were significantly increased.The expression level of FPN protein decreased significantly.Compared with model group,the serum levels of TG,TC,LDL-C,AST and ALT in the combined formula group were significantly decreased,the contents of TG,Fe2+,MDA and ROS in the liver of the combined formula group were significantly decreased,and the activity of SOD was significantly increased.The mRNA expression levels of Xbp1,Xbp1s,Dnajb9,Edem1,Sec61a1,Bip,Cho and TFR1 in liver were significantly decreased,the mrna expression level of FPN was significantly increased,and the protein expression levels of GRP78,P-IRE1α,GPX4 and TFR1 in liver were significantly down-regulated.The expression level of FPN protein was significantly up-regulated.Conclusion Yinchenhao Decoction plus Zexie Decoction may ameliorate NASH by inhibiting ferroptosis of hepatocytes through IRE1α pathway of endoplasmic reticulum stress.

AIM:This study aims to investigate the effects of baicalin on the hypoxia-inducible factor-1α(HIF-1α)/solute carrier family 7 member 11(SLC7A11)/glutathione peroxidase 4(GPX4)axis and the ferroptosis induced by oxidized low-density lipoprotein(ox-LDL)in RAW264.7 macrophage-derived foam cells.METHODS:RAW264.7 cells were categorized into five groups:control,ox-LDL,baicalin+ox-LDL,ferrostatin-1(Fer-1;ferroptosis inhibitor)+ox-LDL,and baicalin+Fer-1+ox-LDL.To induce foam cell formation,RAW264.7 macrophages were exposed to 100 μg/mL ox-LDL for 24 h.Oil red O staining was employed to visualize lipid droplet formation in each group.The ultrastructure of the mitochondria was examined using transmission electron microscopy.Fluorescence microscopy was utilized to assess the fluorescence intensity of intracellular reactive oxygen species(ROS),lipid peroxides,and Fe2+.A colorimetric assay facilitated the measurement of malondialdehyde(MDA)and glutathione(GSH)levels.Additionally,Western blot analy-sis was conducted to quantify protein levels of HIF-1α,SLC7A11,and GPX4.RESULTS:The model group exhibited foam cell formation,abundant lipid droplets,significant swelling of mitochondrial structures,and observable shortening or disappearance of cristae.There was a marked increase in intracellular fluorescence intensity of ROS,lipid peroxides,and Fe2+,alongside elevated MDA levels and decreased GSH levels.HIF-1α protein expression was significantly increased,while SLC7A11 and GPX4 protein expressions were notably decreased(P<0.05).In comparison to the model group,both the baicalin+ox-LDL and Fer-1+ox-LDL groups demonstrated a significant reduction in lipid droplets,improved mitochon-drial structures,decreased fluorescence intensity of ROS,lipid peroxides,and Fe2+,as well as lower MDA levels and higher GSH levels.Additionally,HIF-1α expression significantly decreased,while SLC7A11 and GPX4 expressions sig-nificantly increased(P<0.05).Furthermore,the baicalin+Fer-1+ox-LDL group showed a more pronounced reduction in lipid droplets,near-normal mitochondrial structures,lower fluorescence intensity of ROS,lipid peroxides,and Fe2+,de-creased MDA levels,and increased GSH levels compared to the baicalin+ox-LDL group;HIF-1α,SLC7A11,and GPX4 protein expressions were also significantly reduced(P<0.05).CONCLUSION:Baicalin modulates the HIF-1α/SLC7A11/GPX4 axis,thereby inhibiting ox-LDL-induced ferroptosis in macrophage-derived foam cells.

Objective Based on apolipoprotein a-Ⅰ(Apoa-Ⅰ)gene knockout mice,the role and mechanism of phosphotidylcholine(PC)in improving cholesterol reverse transport were explored.Methods Thirty Apoa-Ⅰ-/-mice were randomly divided into an Apoa-Ⅰ-/-group,Apoa-Ⅰ-/-+HFD group,and Apoa-Ⅰ-/-+HFD+PC group using the random number table method;30 C57BL/6J mice were randomly divided into a WT group,WT+HFD group,and WT+HFD+PC control groups,with 10 mice in each group.The Apoa-Ⅰ-/-group and WT groups were fed basic feed,while the other groups were fed high-fat feed for 8 weeks to establish a hyperlipidemia model.From the 9th week,the WT+HFD+PC group and Apoa-Ⅰ-/-+HFD+PC group were given PC 2.5 g/(kg·d),while the remaining mice were given physiological saline by gavage for a total of 4 weeks of intervention.The serum lipid levels of the mice were detected using a fully automated analyzer.Hematoxylin and eosin and Oil red O staining were used to observe pathological and morphological changes,and the COD-PAP method was used to detect cholesterol levels in mouse liver tissue.The ELISA method was used to detect LCTA levels in mouse serum,and RT-qPCR and Western Blot method were used to detect the mRNA and protein expression of cholesterol ATP binding cassette transporter A1(ABCA1),ATP binding cassette transporter G1(ABC A1),lecithin cholesterol acyltransferase(LCAT),hepatic lipase(HL),scavenger receptor class B type Ⅰ(SR-B1),and low-density lipoprotein receptor(LDL-R)in liver tissue.Results Compared with the WT group,the serum lipid levelsof WT+HFD group mice were significantly increased(P＜0.01),LCAT levels were significantly reduced(P＜0.05),hepatic fat vacuoles were obvious,hepatic lipid deposition was significant,and liver tissue TC levels were significantly increased(P＜0.01).The mRNA and protein expression of ABCA1,ABCG1,LCAT,SR-B1,HL,and LDL-R were significantly reduced(P＜0.05,P＜0.01).Compared with the WT+HFD group,serum lipid levels in the WT+HFD+PC group were significantly reduced(P＜0.05,P＜0.01),LCAT levels were significantly increased(P＜0.05),hepatic fat vacuoles were significantly reduced,hepatic lipid deposition was alleviated,and liver tissue TC levels were significantly reduced(P＜0.05);mRNA and protein expression of ABCA1,LCAT,SR-B1,HL and LDL-R were significantly increased(P＜0.05,P＜0.01).The serum levels of TC,TG,and LDL-C were significantly increased,while the levels of LCAT、HDL-C were significantly reduced(P＜0.05,P＜0.01)in the Apoa-Ⅰ-/-+HFD group mice.Hepatocytes underwent balloon-like transformation,liver lipid deposition was significantly aggravated,and liver tissue TC levels were significantly increased(P＜0.05).The mRNA and protein expression of ABCA1,LCAT and HL were significantly reduced(P＜0.05,P＜0.01).Compared with the WT+HFD+PC group mice,the Apoa-Ⅰ-/-+HFD+PC group mice showed a significant increase in serum lipid levels(P＜0.05,P＜0.01),LCAT levels were significantly reduced(P＜0.05),significant hepatic lipid vacuoles,significant hepatic lipid deposition,and a significant increase in TC levels in liver tissue(P＜0.05).Their mRNA and protein expression of ABCA1,ABCG1,LCAT,SR-B1,and HL were also significantly reduced(P＜0.05,P＜0.01).Conclusions Phosphatidylcholine can improve dyslipidemia by interfering with Apoa-Ⅰ and thus regulating cholesterol reverse transport.

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.

Objective To explore the effect of Yinchenhao Decoction plus Zexie Decoction on ferroptosis mediated by IRE1 signaling pathway of endoplasmic reticulum stress in NASH mice.Methods Fifty C57BL/6J mice were randomly divided into blank control group,model group,Yinchenhao decoction group,Zexia decoction group and Hefang group,with 10 mice in each group.Except the blank control group,the other groups were fed with high-fat diet.After 20 weeks,Yinchenhao Tang group,Zexia Tang group and Hefang group were respectively given the corresponding drug by intragastric administration,once a day for 8 weeks.HE staining,oil red O staining and Masson staining were used to observe the pathological changes of liver tissue,automatic biochemical analyzer was used to detect blood lipid and liver function indexes,and RT-qPCR was used to detect the mRNA levels of TFR1,FPN,XBP1,Xbp1s and XBP1-dependent UPR target genes.The expressions of GRP78,P-IRE1α,GPX4,TFR1 and FPN proteins in liver were detected by Western Blot,the contents of Fe2+and MDA were detected by colorimetry,SOD activity was detected by WST-8,ROS content was detected by ELISA,and TG content in mouse liver tissue was detected by GPO-PAP.Results Compared with the model group,the NAS scores of Yinchenhao Tang group,Zexia Tang group and Hefang group were significantly reduced,and the effect of Hefang group was more significant.Compared with the blank control group,the liver lipid deposition of model mice was obvious,and the liver lipid deposition of Yinchenhao Tang group,Zexia group and Hefang group were improved to varying degrees,and the effect of Hefang group was more significant.Masson staining results showed that compared with blank control group,liver fibrosis was obvious in model group,Yinchenhao Decoction group,Zexia group and Hefang group,the degree of liver fibrosis was improved,and the effect of Hefang group was more significant.Compared with the blank control group,the levels of serum TC,HDL-C,LDL-C,AST and ALT in the model group were significantly increased,the contents of TG,Fe2+,MDA and ROS in the liver were significantly increased,and the activity of SOD was significantly decreased.The mRNA expression levels of Xbp1,Xbp1s,Dnajb9,Edem1,Sec61a1,Bip,Chop and TFR1were significantly up-regulated,the mrna expression levels of FPN were significantly down-regulated,and the protein expression levels of GRP78,P-IREα,GPX4 and TFR1 in liver were significantly increased.The expression level of FPN protein decreased significantly.Compared with model group,the serum levels of TG,TC,LDL-C,AST and ALT in the combined formula group were significantly decreased,the contents of TG,Fe2+,MDA and ROS in the liver of the combined formula group were significantly decreased,and the activity of SOD was significantly increased.The mRNA expression levels of Xbp1,Xbp1s,Dnajb9,Edem1,Sec61a1,Bip,Cho and TFR1 in liver were significantly decreased,the mrna expression level of FPN was significantly increased,and the protein expression levels of GRP78,P-IRE1α,GPX4 and TFR1 in liver were significantly down-regulated.The expression level of FPN protein was significantly up-regulated.Conclusion Yinchenhao Decoction plus Zexie Decoction may ameliorate NASH by inhibiting ferroptosis of hepatocytes through IRE1α pathway of endoplasmic reticulum stress.