Lingguizhugan Decoction (LGZG) demonstrates significant efficacy in treating various cardiovascular diseases clinically, yet its precise mechanism of action remains elusive. This study aimed to elucidate the potential mechanisms and effects of LGZG on isoproterenol (ISO) continuous stimulation-induced chronic heart failure (CHF) in mice, providing direct experimental evidence for further clinical applications. In vivo, continuous ISO infusion was administered to mice, and ventricular myocytes were utilized to explore LGZG?s potential mechanism of action on the ?1-adrenergic receptor (?1-AR)/Gs/G protein-coupled receptor kinases (GRKs)/?-arrestin signaling deflection system in the heart. The findings reveal that LGZG significantly reduced the messenger ribonucleic acid (mRNA) expression of hypertrophy-related biomarkers [atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP)] and improved cardiac remodeling and left ventricular diastolic function in mice with ISO-induced CHF. Furthermore, LGZG inhibited the overactivation of Gs/cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signaling and downregulated the downstream transcriptional activity of cAMP-response element binding protein (CREB) and the expression of the coactivator CBP/P300. Notably, LGZG downregulated the expression of ?-arrestin1 and GRK 2/3/5 while upregulating the expression of ?1-AR and ?-arrestin2. These results suggest that LGZG inhibits Gs/cAMP/PKA signaling and ?-arrestin/GRK-mediated desensitization and internalization of ?1-AR, potentially exerting cardioprotective effects through the synergistic regulation of the ?1-AR/Gs/GRKs/?-arrestin signaling deflection system via multiple pathways.
Animals ; Heart Failure/genetics* ; Signal Transduction/drug effects* ; Drugs, Chinese Herbal/pharmacology* ; Mice ; Male ; G-Protein-Coupled Receptor Kinases/genetics* ; Mice, Inbred C57BL ; Humans ; Isoproterenol ; Arrestins/genetics* ; Chronic Disease

AIM:To establish methods for the isolation,identification,primary culture and evaluation of gas-tric smooth muscle cells(GSMC)from rat and mouse.METHODS:Thirty SD rats and thirty C57BL/6 mice were random-ly assigned into three groups,with 10 rats or mice in each group.Three parallel experiments were conducted to isolate pri-mary GSMC under aseptic conditions.The isolated cells were identified through morphological observation,immunofluo-rescence and Western blot.Flow cytometry was employed to analyze the purity of the cells.Additionally,trypan blue ex-clusion test was utilized to evaluate the viability of the cells after resuscitation.RESULTS:Isolated rat and mouse GSMC exhibited a fusiform or polygonal morphology.Immunofluorescence and Western blot results demonstrated the expression of α-smooth muscle actin(α-SMA).Flow cytometry showed that the positive expression rate of α-SMA was 99.6%in rat GSMC and 97.4%in mouse GSMC.Moreover,the viability rates of rat and mouse GSMC after cryopreservation were found to be greater than 97%.CONCLUSION:A stable and reliable method for the isolation,culture and evaluation of primary GSMC from rat and mouse has been established.

[Objective]To clarify the therapeutic effect of five groups of ascending and descending drugs on ulcerative colitis(UC),and to explore its different characteristics and potential regulatory pathways.[Methods]Sixty-four C57BL/6J male mice were randomly divided into normal control group,model control group,positive control group and five groups of ascending and descending drug groups,with 8 mice in each group.The UC model of mice was induced by dextran sulfate sodium(DSS).Except for the normal control group,the other groups of mice freely drank 2.5%DSS solution every day,and the mice in each drug group were given corresponding doses of drugs by gavage.During the modeling period,the state of the mice was observed every day,and the body weight,food intake,water intake and fecal morphology of the mice were recorded.At the end of the experiment,the disease activity index(DAI)score and colonic histopathological changes of mice in each group were compared.Transcriptome sequencing was used to analyze the differentially expressed genes in the colon of mice in each intervention group.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression level of node genes in key signaling pathway.[Results]Compared with normal control group,the DAI score of the mice in model control group was significantly increased(P<0.0001),the body weight was significantly decreased(P<0.05),the colon was significantly shortened(P<0.05),and a large number of inflammatory cell infiltration was observed.Compared with model control group,the diet of UC mice in Citri Reticulatae Pericarpium-Aurantii Fructus Immaturus group increased gradually;Platycodonis Radix-Armeniacae Semen Amarum group still had formed feces after modeling;the expression of estrogen signaling pathway-related genes closely related to pro-inflammatory response was down-regulated in Scutellariae Radix-Pinelliae Rhizoma Praeparatum Cum Zingibere Et Alumine group;the DAI index of UC mice in Atractylodis macrocephalae Rhizoma-Poria cocos group was significantly lower than that in other groups,and the liver index of UC mice in Atractylodis macrocephalae Rhizoma-Paeoniae Radix Alba group was significantly lower than that in model control group(P<0.01).The G protein-coupled receptors(GPCRs)signaling pathway was up-regulated in the Atractylodis macrocephalae Rhizoma-Poria cocos group and Atractylodis macrocephalae Rhizoma-Paeoniae Radix Alba group.[Conclusion]The five groups of ascending and descending drug pairs had different therapeutic characteristics in the treatment of UC induced by DSS.Among them,Citri Reticulatae Pericarpium-Aurantii Fructus Immaturus group,Atractylodis macrocephalae Rhizoma-Poria cocos group and Atractylodis macrocephalae Rhizoma-Paeoniae Radix Alba group had obvious improvement effects on colon tissue pathological damage and submucosal collagen deposition.The protective effect on colonic goblet cells was relatively obvious in Atractylodis macrocephalae Rhizoma-Poria cocos group and Atractylodis macrocephalae Rhizoma-Paeoniae Radix Alba group.The reduction of apolipoprotein A1(ApoA1)was most significant in Atractylodis macrocephalae Rhizoma-Poria cocos group,and the down-regulation of CXC chemokine ligand 10(CXCL10)gene expression was most obvious in Platycodonis Radix-Armeniacae Semen Amarum group.In general,the protective effect of Atractylodis macrocephalae Rhizoma-Poria cocos drug pair on UC was more comprehensive and worthy of further study.

AIM:To establish methods for the isolation,identification,primary culture and evaluation of gas-tric smooth muscle cells(GSMC)from rat and mouse.METHODS:Thirty SD rats and thirty C57BL/6 mice were random-ly assigned into three groups,with 10 rats or mice in each group.Three parallel experiments were conducted to isolate pri-mary GSMC under aseptic conditions.The isolated cells were identified through morphological observation,immunofluo-rescence and Western blot.Flow cytometry was employed to analyze the purity of the cells.Additionally,trypan blue ex-clusion test was utilized to evaluate the viability of the cells after resuscitation.RESULTS:Isolated rat and mouse GSMC exhibited a fusiform or polygonal morphology.Immunofluorescence and Western blot results demonstrated the expression of α-smooth muscle actin(α-SMA).Flow cytometry showed that the positive expression rate of α-SMA was 99.6%in rat GSMC and 97.4%in mouse GSMC.Moreover,the viability rates of rat and mouse GSMC after cryopreservation were found to be greater than 97%.CONCLUSION:A stable and reliable method for the isolation,culture and evaluation of primary GSMC from rat and mouse has been established.

[Objective]To clarify the therapeutic effect of five groups of ascending and descending drugs on ulcerative colitis(UC),and to explore its different characteristics and potential regulatory pathways.[Methods]Sixty-four C57BL/6J male mice were randomly divided into normal control group,model control group,positive control group and five groups of ascending and descending drug groups,with 8 mice in each group.The UC model of mice was induced by dextran sulfate sodium(DSS).Except for the normal control group,the other groups of mice freely drank 2.5%DSS solution every day,and the mice in each drug group were given corresponding doses of drugs by gavage.During the modeling period,the state of the mice was observed every day,and the body weight,food intake,water intake and fecal morphology of the mice were recorded.At the end of the experiment,the disease activity index(DAI)score and colonic histopathological changes of mice in each group were compared.Transcriptome sequencing was used to analyze the differentially expressed genes in the colon of mice in each intervention group.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression level of node genes in key signaling pathway.[Results]Compared with normal control group,the DAI score of the mice in model control group was significantly increased(P<0.0001),the body weight was significantly decreased(P<0.05),the colon was significantly shortened(P<0.05),and a large number of inflammatory cell infiltration was observed.Compared with model control group,the diet of UC mice in Citri Reticulatae Pericarpium-Aurantii Fructus Immaturus group increased gradually;Platycodonis Radix-Armeniacae Semen Amarum group still had formed feces after modeling;the expression of estrogen signaling pathway-related genes closely related to pro-inflammatory response was down-regulated in Scutellariae Radix-Pinelliae Rhizoma Praeparatum Cum Zingibere Et Alumine group;the DAI index of UC mice in Atractylodis macrocephalae Rhizoma-Poria cocos group was significantly lower than that in other groups,and the liver index of UC mice in Atractylodis macrocephalae Rhizoma-Paeoniae Radix Alba group was significantly lower than that in model control group(P<0.01).The G protein-coupled receptors(GPCRs)signaling pathway was up-regulated in the Atractylodis macrocephalae Rhizoma-Poria cocos group and Atractylodis macrocephalae Rhizoma-Paeoniae Radix Alba group.[Conclusion]The five groups of ascending and descending drug pairs had different therapeutic characteristics in the treatment of UC induced by DSS.Among them,Citri Reticulatae Pericarpium-Aurantii Fructus Immaturus group,Atractylodis macrocephalae Rhizoma-Poria cocos group and Atractylodis macrocephalae Rhizoma-Paeoniae Radix Alba group had obvious improvement effects on colon tissue pathological damage and submucosal collagen deposition.The protective effect on colonic goblet cells was relatively obvious in Atractylodis macrocephalae Rhizoma-Poria cocos group and Atractylodis macrocephalae Rhizoma-Paeoniae Radix Alba group.The reduction of apolipoprotein A1(ApoA1)was most significant in Atractylodis macrocephalae Rhizoma-Poria cocos group,and the down-regulation of CXC chemokine ligand 10(CXCL10)gene expression was most obvious in Platycodonis Radix-Armeniacae Semen Amarum group.In general,the protective effect of Atractylodis macrocephalae Rhizoma-Poria cocos drug pair on UC was more comprehensive and worthy of further study.

OBJECTIVE:To investigate the effects of levocarnitine on peripheral blood T cell subsets and inflammatory factors in diabetic patients underwent peritoneal dialysis. METHODS:A total of 100 diabetic patients underwent peritoneal dialysis in se-lected as research objects were divided into conventional therapy group and levocarnitine group according to random number table, with 50 cases in each group. Conventional therapy group received peritoneal dialysis and conventional therapy. Levocarnitine group was additionally given levocarnitine 1.0 g added into 0.9% Sodium chloride injection 20 mL intravenously,3 times a week,on the basis of conventional therapy group for 12 weeks. CD3+,CD4+,the proportion of Th1 and Th2,the contents of TNF-α,INF-γ, IL-10 and IL-4 in supernatant were all detected before and after treatment. The expression of T-bet and GATA-3 were compared be-tween 2 groups. RESULTS:After treatment,CD3+,CD4+,Th2 cells,GATA-3 expression and IL-10,IL-4 levels were significantly increased,the expression of Th1 cells,T-bet content and TNF-αand INF-γwere significantly reduced,and levcarnitine group was sig-nificantly better than conrentional therapy group,with statistically significant (P<0.05). CONCLUSIONS:Levocarnitine can im-prove inflammatory reaction and the expression of related transcription factors by promoting T cell subsets and Th1/Th2 cell subsets.

Objective To explore the relationship of peritoneal transport patterns and the nutritional status in continuous ambu-latory peritoneal dialysis(CAPD) patients .Methods 89 patients who took CAPD in 2010 January to December 2012 in the perito-neal dialysis center of our hospital were selected in this study .According to the ratio of creatinine in peritoneal dialysis solution and in blood(D/Pcr)of the peritoneal equilibrium test(PET) ,all CAPD patients were divided into HA group(high average transport) and LA group(low average transport) .Nutritional status including serum albumin(ALB) ,prealbumin(PA) ,transferrin(TF) ,hemo-globin(Hb) ,Lean body mass(LBM) ,subjective global assessment(SGA)and normalized Protein Equivalent of Nitrogen Appearance (nPNA) were compared in both groups .The relationship of D/Pcr and the nutritional status were analyzed .Results D/Pcr in HA group were significantly higher than in LA group(P<0 .01) .ALB ,PA ,TF ,Hb and SGA in HA group were significantly higher than in LA group(P<0 .01) ,but nPNA in two groups has no statistical differences (P>0 .05) .D/Pcr was negative related to ALB , TF ,Hb and SGA(P<0 .01) ,but not related to nPNA ,PA and LBM (P>0 .05) .Conclusion The nutritional status is related to peritoneal transport patterns in peritoneal HA patients ,it is necessary to strengthen nutritional guidance ,as the nutritional status is worse compared to LA ones .