1.Mechanism of Shenfu Xiongze Prescription in Regulating Autophagy Level to Intervene in Myocardial Remodeling in Rats via AMPK/mTOR Signaling Pathway
Xueqing WANG ; Wei ZHONG ; Liangliang PAN ; Caihong LI ; Man HAN ; Xiaowei YANG ; Yuanwang YU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):136-144
ObjectiveTo explore the mechanism by which the Shenfu Xiongze prescription regulates autophagy in rats with myocardial remodeling through the adenosine monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signaling pathway. MethodsA rat model of myocardial remodeling induced by isoprenaline (ISO) was established. Rats were divided into the blank group,the model group,the low-,medium-, and high-dose groups of Shenfu Xiongze prescription,and the captopril group, 6 rats in each group. Except for the blank group,the rat model of myocardial remodeling was established in the other groups by intraperitoneal injection of 2.5 mg·kg-1 ISO for 3 consecutive weeks. At the same time of modeling, the low-,medium-, and high-dose groups of Shenfu Xiongze prescription were administered the corresponding doses of Shenfu Xiongze prescription solution (8.4,16.8,and 33.6 g·kg-1),and the captopril group was administered captopril solution (25 mg·kg-1). As for the blank group and the model group, the same volume of normal saline was given. The treatment was continued for 3 weeks. Echocardiography was used to observe the cardiac structure and function,and the heart weight index was detected. Masson staining and hematoxylin-eosin (HE) staining were used to observe the pathological morphology changes of myocardial tissue. The levels of interleukin-6 (IL-6) and B-type natriuretic peptide (BNP) in serum were detected by enzyme-linked immunosorbent assay (ELISA). The expression of type Ⅰ collagen (Collagen Ⅰ),type Ⅲ collagen (Collagen Ⅲ),and microtubule-associated protein 1 light chain 3 (LC3) proteins in myocardial tissue was determined by immunohistochemistry. Autophagy was observed by transmission electron microscopy. The mRNA expression of Collagen Ⅰ,Collagen Ⅲ,α-smooth muscle actin (α-SMA),LC3,yeast Atg6 homolog protein (Beclin-1),AMPK,and mTOR in myocardial tissue was detected by quantitative real-time polymerase chain reaction (real-time PCR). The protein expression of Collagen Ⅰ,α-SMA,transforming growth factor-β1 (TGF-β1),LC3,Beclin-1,p62, phosphorylation(p)-AMPK,p-mTOR,AMPK,and mTOR was detected by Western blot. ResultsCompared with the normal group,rats in the model group exhibited significantly decreased values of ejection fraction (EF) and left ventricular fractional shortening (FS) (P<0.01), significantly increased values of left ventricular end-diastolic diameter (LVIDd) and left ventricular end-systolic diameter (LVIDs) (P<0.01). Additionally, the model group also showed increased degrees of inflammatory infiltration and fibrosis of myocardial tissue, significantly elevated levels of serum IL-6 and BNP (P<0.01), significantly increased mRNA and protein levels of Collagen Ⅰ,Collagen Ⅲ,α-SMA,and mTOR (P<0.01),and markedly decreased mRNA and protein levels of LC3,Beclin-1,and AMPK (P<0.05,P<0.01). Compared with the model group, the low-,medium-, and high-dose groups of Shenfu Xiongze prescription presented significantly elevated EF and FS values (P<0.01) and lowered LVIDd and LVIDs (P<0.05). In these groups, the inflammation and fibrosis were alleviated significantly. They also exhibited decreased serum levels of IL-6 and BNP (P<0.01), significantly reduced protein expression of Collagen Ⅰ, α-SMA, TGF-β1, p62, and p-mTOR (P<0.01), significantly decreased mRNA expression of Collagen Ⅰ, Collagen Ⅲ, α-SMA, and mTOR (P<0.01), and significantly increased mRNA and protein levels of LC3, Beclin-1, and AMPK (P<0.05,P<0.01). ConclusionThe Shenfu Xiongze prescription can improve the myocardial remodeling induced by ISO in rats by regulating the autophagy level,enhance cardiac function,and reduce inflammatory and fibrotic levels. This effect may be achieved through the AMPK/mTOR signaling pathway.
2.Xuebijing injection inhibits TLR4 signaling pathway through up-regulating ADAM17 activity in EA.hy926 cells
Yingjie ZHANG ; Jing WANG ; Hongfei QI ; Yalong KANG ; Jing DONG ; Yuanwang YU ; Haifang WANG
Immunological Journal 2024;40(4):359-364
This study was performed to investigate the inhibitory effects of Xuebijing injection(XBJ)on lipopolysaccharide(LPS)-induced inflammatory signals on EA.hy926 vascular endothelial cells and the underlying mechanism,and to provide a theoretical basis for the treatment of sepsis with XBJ.WST-1 assay was used to detect the effects of XBJ on the cell viability;Western blot analysis was performed to detect the protein expression levels of IκBα,p-p65,p-ERK,p-JNK,p-p38,p-AMAD17 in cell lysates and the content of sTLR4 fragment in the concentrated culture supernatants.ADAM17 sheddase activity in cells was detected by using a commercial available kit.Data showed that all of the TLR4-mediated inflammatory signals were significantly inhibited by the treatment of XBJ(P<0.01).ADAM17 phosphorylation and shedding activity were induced by XBJ treatment,simultaneously the sTLR4 contents in the culture media were increased.XBJ-induced shedding of TLR4 was suppressed by the preteatment of 10 μmol/L TAPI-1(an ADAM17 inhibitor).Taken together,XBJ can induce the shedding of TLR4 from cell membrane by up-regulating ADAM17 shedding activity,thereby inhibiting the activation of TLR4-mediated intracellular inflammatory signals in EA.hy926 cells.
3.The protective effect of Danggui Buxue decoction on radiation injury in mice
Chinese Journal of Comparative Medicine 2015;(10):55-58
Objective To observe the protective effect of Danggui Buxue Decoction on 60 Co-γray irradiated mice. Methods The experimental mice were randomly divided into 5 groups ( the normal control group , model group , Danggui Buxue Decoction of low , middle and high dose groups ) .The mouse except the normal control group were irradiated by 60Co-γray and established the radiation-damage model.Peripheral blood neutrophil count (NEUT), the number of bone marrow nucleated cells ( BMC ) , the activity of serum superoxide dismutase ( SOD ) and the serum levels of malondialdehyde ( MDA) were measured .Results Radiation injury in mice of NEUT , BMC and SOD were significantly decreased (P <0.01), MDA were increased (P <0.01).Danggui Buxue Decoction can increase the levels of NEUT , BMC and SOD in mice after radiation injury , decrease the content of MDA .And protective effect of high dose group was better (P <0.01).Conclusions Danggui Buxue Decoction has protective effect on radiation injury in mice .And there is a correlation between the protection effect and dose .
4.Comparison of the experimental mice of different parts of the bone marrow biopsy
Chinese Journal of Comparative Medicine 2015;(7):64-65
Objective To explore the best sampling position of bone marrow biopsy in mice.Methods Mice were sacrificed by cervical dislocation, then take the skull, sternum, tibial, femoral and iliac bone, making pathological section.Observed and photographed under light microscope. Comparison of the distribution of the essence and the hematopoietic microenvironment in different parts.Select site which can best reflect the hematopoietic function of bone marrow.Results A large number of hematopoietic cells in ilium marrow sections visible.The cells are evenly distributed. Blood and megakaryocytes were clearly visible.The arrangement of the structure of the scaffolds for tissue closely.The number of fat cells less.Bone marrow hematopoietic cells in the skull, sternum, tibial, femoral were not clear and not active.And there are more fat cells.Conclusions As the best sampling position of bone marrow biopsy, is ilium.
5.Effect of Xianhua Huogudan Granules on hemorrheology and blood lipid of steroid-induced avascular necrosis of femoral head
Chuandao SHI ; Yuanwang YU ; Yongjun WEN ; Wenpen JA ; Xiauhang YANG ; Jianjun DANG
Chinese Traditional Patent Medicine 1992;0(09):-
AIM: Study on the effects of Xianhuahuogudan Granule (XHHGDG) (Radix et Rhizowa Salviae uiltiorrhizae, Rhizoma Drynariae,etc) in treatment of steroid-induced avascular necrosis of femoral head in order to provide basis for clinical use. METHODS: 40 full-grown well New Zealand rabbits were divided randomly into four groups:normal group,model group,MaShGuPian group and XHHGDG group.Except the normal group, The other three groups were administered intramuscularly with steroid.From the 9 th week after production of models,the normal group and the model group were treated with normal saline,XHHGDG group with XHHGDG aqueous solution,and MaShGuPian group with MaShGuPian aqueous solution.All of the rabbits were killed 14 weeks after baseline for the research material. After making pathomorphology of bone,blood viscosity and content of blood lipid will be acquired. RESULTS: The blood viscosity and content of blood lipid were lower significantly in the XHHGDG group than those in the model group.Compared with model group,XHHGDG group significantly showed followings:the small bone trabeculas were fuller;most of bone cells were normal.Although of adipose cell existed in the margin of marrow,the cells for producing blood were enriched;numbers of empty bone lacuna were 16.4%(P

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