1.Ameliorative Effect of Wendantang Combined with Danshenyin and Dushentang on Ischemic Heart Disease with Phlegm-stasis Syndrome in Mice Based on Circulating Monocytes
Fenghe YANG ; Ziqi TIAN ; Zhiqian SONG ; Shitao PENG ; Wenjie LU ; Tao LIN ; Chun WANG ; Zhangchi NING
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):22-32
ObjectiveTo investigate the ameliorative effect of Wendantang combined with Danshenyin and Dushentang (WDD) on mice with ischemic heart disease (IHD) presenting phlegm-stasis syndrome based on the inflammatory phenotype and differentiation of circulating monocytes. MethodsA model of IHD with phlegm-stasis syndrome was established using left anterior descending coronary artery ligation supplemented with a high-fat diet. Eighty model mice were randomly assigned to the model group, WDD low-dose group (WDD-L), WDD medium-dose group (WDD-M), WDD high-dose group (WDD-H), and atorvastatin calcium tablet group, with 16 mice in each group. An additional 16 C57BL/6J mice were designated as the sham-operation group. The WDD groups received intragastric administration at doses of 8.91, 17.81, 35.62 g·kg-1, and the atorvastatin calcium tablet group received the corresponding drug at 1.3 mg·kg-1, twice daily. The sham-operation and model groups were given the same volume of pure water by gavage each day. After 5 consecutive weeks of administration, the cardiac index was calculated. Cardiac function was assessed by echocardiography. Myocardial histopathology was examined by hematoxylin-eosin (HE) staining. Serum N-terminal pro-B-type natriuretic peptide (pro-BNP) content was measured by enzyme-linked immunosorbent assay (ELISA). Hemorheological parameters were analyzed using an automated hemorheology analyzer. Serum levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) were determined using an automated biochemical analyzer. Changes in circulating monocytes were detected by flow cytometry. Mouse bone marrow mononuclear cells were isolated in vitro and divided into blank group, model serum group, WDD-L drug-containing serum group, WDD-M drug-containing serum group, and WDD-H drug-containing serum group. CD36 expression and macrophage differentiation in each group were assessed by flow cytometry. The mechanism by which WDD mediates circulating monocyte differentiation was further explored using CD36 knockdown/overexpression RAW264.7 cell lines. ResultsCompared with the sham-operation group, the model group showed a significantly increased cardiac index (P0.01), significantly decreased fractional shortening (FS) (P0.01), and significantly increased left ventricular end-diastolic internal diameter (LVDD) and left ventricular end-systolic internal diameter (LVDS) (P0.01). Cardiomyocytes exhibited marked deformation and necrosis with inflammatory cell infiltration. Serum pro-BNP levels were significantly elevated (P0.01), and whole-blood viscosity (BV) at high, medium, and low shear rates was significantly increased (P0.01). Compared with the model group, the WDD groups showed significantly reduced cardiac index (P0.05, P0.01), significantly increased FS (P0.05, P0.01), significantly decreased LVDD and LVDS (P0.01), markedly improved cardiomyocyte morphology, significantly reduced inflammatory infiltration, significantly decreased serum pro-BNP levels (P0.01), and significantly decreased BV at high, medium, and low shear rates (P0.01), with the most pronounced improvement observed in the WDD-M group. Compared with the sham-operation group, TC, TG, and LDL levels were significantly increased in the model group (P0.05, P0.01), while HDL levels were significantly decreased (P0.05). After WDD-H treatment, TC, TG, and LDL levels were significantly reduced and HDL levels were significantly increased in mice (P0.05, P0.01). Compared with the sham-operation group, classical monocytes in blood and bone marrow and intermediate monocytes in blood were significantly increased in the model group (P0.01), whereas intermediate monocytes in bone marrow and non-classical monocytes in blood were significantly decreased (P0.01). After WDD administration, all circulating monocyte subsets in blood and bone marrow were significantly alleviated (P0.05, P0.01), with the WDD-M group showing the optimal effect. In vitro, compared with the blank group, CD36 expression on bone marrow monocytes and the proportion of differentiated macrophages were significantly increased in the model serum group (P0.01), and CD36 expression was significantly upregulated on RAW264.7 cells (P0.01). Compared with the model serum group, all drug-containing serum groups exhibited significantly reduced CD36 expression on bone marrow monocytes and significantly reduced macrophage differentiation (P0.01). WDD downregulated CD36 expression in both CD36 knockdown and overexpression RAW264.7 cell lines (P0.05, P0.01), with the strongest regulatory effect observed in the WDD-M drug-containing serum group. ConclusionWDD can significantly improve the manifestations of phlegm-stasis syndrome in IHD mice and reduce the proportion of classical circulating monocytes. Its mechanism may be related to the inhibition of CD36 expression on classical circulating monocytes.
2.Ameliorative Effect of Wendantang Combined with Danshenyin and Dushentang on Ischemic Heart Disease with Phlegm-stasis Syndrome in Mice Based on Circulating Monocytes
Fenghe YANG ; Ziqi TIAN ; Zhiqian SONG ; Shitao PENG ; Wenjie LU ; Tao LIN ; Chun WANG ; Zhangchi NING
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(3):22-32
ObjectiveTo investigate the ameliorative effect of Wendantang combined with Danshenyin and Dushentang (WDD) on mice with ischemic heart disease (IHD) presenting phlegm-stasis syndrome based on the inflammatory phenotype and differentiation of circulating monocytes. MethodsA model of IHD with phlegm-stasis syndrome was established using left anterior descending coronary artery ligation supplemented with a high-fat diet. Eighty model mice were randomly assigned to the model group, WDD low-dose group (WDD-L), WDD medium-dose group (WDD-M), WDD high-dose group (WDD-H), and atorvastatin calcium tablet group, with 16 mice in each group. An additional 16 C57BL/6J mice were designated as the sham-operation group. The WDD groups received intragastric administration at doses of 8.91, 17.81, 35.62 g·kg-1, and the atorvastatin calcium tablet group received the corresponding drug at 1.3 mg·kg-1, twice daily. The sham-operation and model groups were given the same volume of pure water by gavage each day. After 5 consecutive weeks of administration, the cardiac index was calculated. Cardiac function was assessed by echocardiography. Myocardial histopathology was examined by hematoxylin-eosin (HE) staining. Serum N-terminal pro-B-type natriuretic peptide (pro-BNP) content was measured by enzyme-linked immunosorbent assay (ELISA). Hemorheological parameters were analyzed using an automated hemorheology analyzer. Serum levels of total cholesterol (TC), triglycerides (TG), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) were determined using an automated biochemical analyzer. Changes in circulating monocytes were detected by flow cytometry. Mouse bone marrow mononuclear cells were isolated in vitro and divided into blank group, model serum group, WDD-L drug-containing serum group, WDD-M drug-containing serum group, and WDD-H drug-containing serum group. CD36 expression and macrophage differentiation in each group were assessed by flow cytometry. The mechanism by which WDD mediates circulating monocyte differentiation was further explored using CD36 knockdown/overexpression RAW264.7 cell lines. ResultsCompared with the sham-operation group, the model group showed a significantly increased cardiac index (P<0.01), significantly decreased fractional shortening (FS) (P<0.01), and significantly increased left ventricular end-diastolic internal diameter (LVDD) and left ventricular end-systolic internal diameter (LVDS) (P<0.01). Cardiomyocytes exhibited marked deformation and necrosis with inflammatory cell infiltration. Serum pro-BNP levels were significantly elevated (P<0.01), and whole-blood viscosity (BV) at high, medium, and low shear rates was significantly increased (P<0.01). Compared with the model group, the WDD groups showed significantly reduced cardiac index (P<0.05, P<0.01), significantly increased FS (P<0.05, P<0.01), significantly decreased LVDD and LVDS (P<0.01), markedly improved cardiomyocyte morphology, significantly reduced inflammatory infiltration, significantly decreased serum pro-BNP levels (P<0.01), and significantly decreased BV at high, medium, and low shear rates (P<0.01), with the most pronounced improvement observed in the WDD-M group. Compared with the sham-operation group, TC, TG, and LDL levels were significantly increased in the model group (P<0.05, P<0.01), while HDL levels were significantly decreased (P<0.05). After WDD-H treatment, TC, TG, and LDL levels were significantly reduced and HDL levels were significantly increased in mice (P<0.05, P<0.01). Compared with the sham-operation group, classical monocytes in blood and bone marrow and intermediate monocytes in blood were significantly increased in the model group (P<0.01), whereas intermediate monocytes in bone marrow and non-classical monocytes in blood were significantly decreased (P<0.01). After WDD administration, all circulating monocyte subsets in blood and bone marrow were significantly alleviated (P<0.05, P<0.01), with the WDD-M group showing the optimal effect. In vitro, compared with the blank group, CD36 expression on bone marrow monocytes and the proportion of differentiated macrophages were significantly increased in the model serum group (P<0.01), and CD36 expression was significantly upregulated on RAW264.7 cells (P<0.01). Compared with the model serum group, all drug-containing serum groups exhibited significantly reduced CD36 expression on bone marrow monocytes and significantly reduced macrophage differentiation (P<0.01). WDD downregulated CD36 expression in both CD36 knockdown and overexpression RAW264.7 cell lines (P<0.05, P<0.01), with the strongest regulatory effect observed in the WDD-M drug-containing serum group. ConclusionWDD can significantly improve the manifestations of phlegm-stasis syndrome in IHD mice and reduce the proportion of classical circulating monocytes. Its mechanism may be related to the inhibition of CD36 expression on classical circulating monocytes.
3.Exploring Synergistic Mechanism of Processing with Vinegar on Olibanum in Treatment of Ulcerative Colitis Based on Intestinal Flora
Wenjie LU ; Shitao PENG ; Zhiqian SONG ; Zhenli LIU ; Chun WANG ; Zhangchi NING
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(15):182-191
ObjectiveTo explore the improving effects and its synergistic mechanism of Olibanum before and after processing with vinegar on glycodesoxycholic acid(GDCA) intervention in mice with ulcerative colitis(UC) based on the perspective of intestinal flora. MethodC57BL/6J male mice were randomly divided into the normal group, model group, GDCA group, Olibanum group(1.5 g·kg-1) and vinegar-processed Olibanum(1.5 g·kg-1) group, with 6 mice in each group. Mice in the normal group drank water freely, and mice in the other groups were given 2% dextran sulfate sodium(DSS) periodically to establish a UC mouse model. During the modeling, GDCA group, Olibanum group and vinegar-processed Olibanum group were intervened by intraperitoneally injection of GDCA(0.05 g·kg-1). From the 13th day after modeling, Olibanum group and vinegar-processed Olibanum group were given the corresponding doses of drugs by gavage, once a day, for 36 d. During this period, the body mass of mice was recorded and the disease activity index(DAI) was assessed. On day 48, faeces were collected for 16S rRNA and metagenomic sequencing to analyse changes in intestinal flora. On the 49th day, hematoxylin-eosion(HE) staining was used to observe the colon histological lesions, enzyme-linked immunosorbent assay(ELISA) was used to determine serum levels of tumour necrosis factor-α(TNF-α), interleukin(IL)-1β and IL-6, and Spearman correlation analysis was used to explore the correlation between differential bacterial species and inflammatory factor levels. ResultCompared with the normal group, the model group showed a significant decrease in body weight(P<0.01), a significant increase in DAI(P<0.05), and a significant increase in TNF-α, IL-1β and IL-6 levels(P<0.01), and there was partial infiltration of inflammatory cells in the colon. Compared with the model group, mice in the GDCA group showed a significant decrease in body weight, a significant increase in DAI and levels of TNF-α, IL-1β and IL-6(P<0.01), and severe disruption of colonic crypt structure, extensive infiltration of inflammatory cells, and a significant decrease in goblet cells. Compared with the GDCA group, both the Olibanum and vinegar-processed Olibanum groups showed a significant recovery in body weight, a significant decrease in DAI and levels of TNF-α, IL-6 and IL-1β(P<0.05, P<0.01), and the modulating effect of vinegar-processed Olibanum was significantly better than that of Olibanum. Alpha diversity showed that Chao1 index of UC mice significantly increased(P<0.01) and Shannon index decreased significantly(P<0.05) in UC mice after GDCA intervention. Beta diversity showed that the microbial community structure of the 5 groups had significant changes, Olibanum and vinegar-processed Olibanum could modulate the changes in the structure of the intestinal flora in UC mice after GDCA intervention. Microbial sequencing results indicated that, compared with the normal group, the Firmicutes/Bacteroidetes ratio in the model group was significantly higher(P<0.05), and the relative abundance of 3 genera and 5 species of flora changed significantly(P<0.05, P<0.01). Compared with the model group, the Firmicutes/Bacteroidetes ratio in the GDCA group was significantly higher(P<0.05), the relative abundance of 7 pathogenic bacterial genera and four species was significantly increased(P<0.05, P<0.01), and the relative abundance of three beneficial bacterial genera and Bacteroides_intestinalis was significantly decreased(P<0.05, P<0.01). Olibanum group and vinegar-processed Olibanum group could modulate the Firmicutes/Bacteroidetes ratio, the relative abundance of pathogenic bacteria and beneficial bacteria, and the vinegar-processed Olibanum group was significantly superior to Olibanum group in terms of modulating the Firmicutes/Bacteroidetes ratio, the relative abundance of the three genera and five species of bacteria(P<0.01, P<0.05). Correlation analysis showed that the relative abundance of Bacteroides_intestinalis was negatively correlated with the levels of TNF-α, IL-6 and IL-1β, the relative abundance of Prevotella_sp_CAG873, Bacteroides_sp_CAG927, Bacteroidales_bacterium_52_46 and Bacteroidales_bacterium was positively correlated with TNF-α, IL-6 and IL-1β levels. ConclusionGDCA can exacerbate UC colonic inflammation, and Olibanum and vinegar-processed Olibanum have an ameliorative effect on GDCA-mediated UC, with the vinegar-processed Olibanum showing a stronger ameliorative effect, the mechanism may be related to the regulation the abundance and structure of intestinal beneficial and pathogenic bacteria, and the reduction of inflammatory factor levels.
4.Exploring Synergistic Mechanism of Vinegar-processed Olibanum in Treatment of Ulcerative Colitis Based on Bile Acid-regulated "Gut-liver" Crosstalk
Shitao PENG ; Zhenli LIU ; Zhiqian SONG ; Chun WANG ; Zhangchi NING
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(21):59-68
ObjectiveTo investigate the synergistic mechanism of vinegar-processed Olibanum on ulcerative colitis(UC) via the bile acids regulating "gut-liver" crosstalk. MethodRats were randomly divided into normal group, model group, Olibanum group and vinegar-processed Olibanum group. UC model of rats was induced by intracolonic instillation of 2,4,6-trinitrobenzenesulfonic acid(TNBS). Ultra high performance liquid chromatography-triple quadrupole-mass spectrometry(UPLC-QQQ-MS) was used to perform the qualitative analysis of 30 bile acids in the colon of rats. Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) and Western blot were used to detect changes in the expression of farnesoid X receptor(FXR), fibroblast growth factor 15(FGF15) and FGF receptor 4(FGFR4) in "gut-liver" crosstalk at mRNA and protein levels. And with the help of HcoEpiC cell model intervened by conjugated bile acids, simulating the UC state, and according to the different modes of intervention, they were divided into the blank group, conjugated bile acid group, Olibanum group, vinegar-processed Olibanum group and 3-O-acetyl-9,11-dehydro-β-boswellic acid(ADHBA) group. The effect of Olibanum before and after processing with vinegar and the main differential component ADHBA on the mRNA expression of FXR and FGF19 were explored by Real-time PCR. ResultCompared with the normal group, the levels of conjugated bile acids in the model group increased significantly(P<0.01), and the mRNA and protein expressions of "gut-liver" crosstalk factors FXR, FGF15 and FGFR4 decreased significantly(P<0.05, P<0.01). Compared with the model group, the content of conjugated bile acids in the Olibanum group and vinegar-processed Olibanum group was significantly decreased(P<0.01), the mRNA and protein expressions of FXR, FGF15 and FGFR4 were significantly elevated(P<0.05, P<0.01), and vinegar-processed Olibanum exhibited superior effects than Olibanum. In cellular experiments, a significant decrease in mRNA expression of FXR and FGF19 was observed in the conjugated bile acid group when compared with the blank group(P<0.01). Compared with the conjugated bile acid group, the mRNA expressions of FXR and FGF19 were significantly higher in the Olibanum, vinegar-processed Olibanum and ADHBA groups(P<0.05, P<0.01), and the effect of vinegar-processed Olibanum was more favorable. ConclusionVinegar-processed Olibanum may enhance the ameliorating effect on UC by enhancing the down-regulation of conjugated bile acids in the colon and the up-regulation of FXR-FGF15/19-FGFR4 "gut-liver" crosstalk pathway, and ADHBA may be the main material basis for the synergy.
5.Comparison of Alkaloids and Polysaccharides in Different Parts of Wild Aconitum in Nanyang
Meiling SUN ; Dou ZHANG ; Shuo YANG ; Shitao PENG ; Xiaohua SONG ; Peng LIU ; Hong DU ; Fei LI ; Huihua HU
World Science and Technology-Modernization of Traditional Chinese Medicine 2018;20(12):2231-2236
Objective: Comparison of alkaloid and polysaccharides content in different parts of whole plant of wild aconitum in Nanyang, and to explore the medicinal value of aboveground part and fibrous root of wild aconitum in Nanyang. Methods: The contents of three monoester alkaloids, three diester alkaloids in wild aconitum, radix aconiti carmichaeli, fibrous root and aboveground parts in Nanyang were determined by HPLC method; The content of crude polysaccharides was determined by sulfuric-phenol method, crude polysaccharides content from different tissues of wild aconitum in Nanyang in different harvest time was compared, including wild aconitum in Nanyang, wild radix aconiti carmichaeli in Nanyang, fibrous root, aboveground part. Results: Among the 4 parts of the wild Aconitum in Nanyang, the contents of 3 mono ester alkaloids rang from high to low: wild aconitum in Nanyang> wild radix aconiti carmichaeli in Nanyang> fibrous root> aboveground part (P < 0.05), the contents of three diester alkaloids rang from high to low: wild radix aconiti carmichaeli in Nanyang> fibrous root> wild aconitum in Nanyang> aboveground part (P < 0.05), the order of total alkaloid content from high to low is: wild radix aconiti carmichaeli in Nanyang> fibrous root> wild aconitum in Nanyang> aboveground part (P < 0.05), The content of crudepolysaccharides in different tissues of wild aconitum in Nanyang were in the sequence of wild radix aconiti carmichaeli in Nanyang> wild aconitum in Nanyang> aboveground part> fibrous root (P < 0.05) . Conclusion: In this study, an attempt was made to explore the differences of 6 kind salkaloid content in different parts of whole plant of wild aconitum in Nanyang, which provided scientific basis for the medicinal value of the fibrous roots and the aboveground parts.
6.Development of Portable Battlefield Multifunction Diagnosis and Treatment Bed
Liming TANG ; Shitao QI ; Tiebing LIU ; Min WU ; Peng SUN
Chinese Medical Equipment Journal 1989;0(02):-
Objective To develop an improved product of the existing battlefield diagnosis and treatment bed by making the product smaller,the structure firmer,the function more perfect and the use more convenient,thus to benefit the use under the battlefield situations,and to improve the logistics support ability.Methods '08F,304'steel materials were adopted as prop-up parts of the bed.The fold type structure design,infusion shelf and mosquito net pole functions could be exchanged with each other.The special oxygen bottle clip was equipped.Results It had such advantages as lighter weight(half of the former),enlarged strength,diverse functions,simple operation and easy taking.As far as safety and reliability and operation flexibility concerned,it was far improved compared with the former one.The treatment bed met the requirement of diagnosis,treatment and nursing for patients under battlefield conditions.National patent of the product was declared.Conclusion The portable battlefield diagnosis and treatment bed is used under the field situation,so some more complicated structures and functions of the normal beds are omitted.Meanwhile,reasonable function combination and material and structure design are key factors in development.[Chinese Medical Equipment Journal,2008,29(2):10-11,14]

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