1.Modified Taohe Chengqitang Regulates Notch/eNOS Signaling Pathway to Inhibit Hepatic Sinusoidal Capillarization
Chang SHAO ; Xiguang SUN ; Jiaxin HUANG ; Linmao YE ; Xiaofan LIANG ; Yi HE ; Junjie ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(19):42-54
ObjectiveTo investigate whether modified Taohe Chengqitang (JTCT) treats liver fibrosis by inhibiting hepatic sinusoidal capillarization, and to verify whether its specific mechanism is related to the Homologous genes responsible for the notches along the edges of Drosophila wings(Notch) signaling pathway and endothelial nitric oxide synthase (eNOS) signaling pathway. MethodsFifty C57BL/6 mice were randomized into 5 groups: Control, model (5 mL·kg-1·3 d-1), low-dose JTCT, medium-dose JTCT, and high-dose JTCT (JTCT-L,JTCT-M,JTCT-H). Except the control group, the other groups were intraperitoneally injected with 20% carbon tetrachloride (CCl4) olive oil solution every 3 days for 4 weeks to induce liver fibrosis. Meanwhile, JTCT-L, JTCT-M, and JTCT-H groups were administrated with JTCT by gavage at doses of 16.6, 33.3, and 66.6 g·kg-1·d-1, respectively, for 4 consecutive weeks (the control and model groups received equal volumes of normal saline by gavage). Primary liver sinusoidal endothelial cells (LSECs) were isolated from rat livers through in situ perfusion of collagenase, Percoll density gradient centrifugation, and selective removal of Kupffer cells. After verification of cell phenotype, LSECs were used for experiments. For cells at the logarithmic growth phase, the small interfering RNA(siRNA) kit was used to knock down the expression of soluble guanylate cyclase (sGC) through transient transfection. Subsequently, the Notch agonist Jagged-1 was applied. Cell counting kit-8 (CCK-8) was used to examine the cytotoxicity of JTCT on LSECs. Real-time PCR and Western blot were employed to measure the expression of molecules in the Notch and eNOS signaling pathways at mRNA and protein levels, respectively. Intracellular Ca2+ was labeled with the fluorescent probe Fluo-4 acetoxymethyl ester(Fluo-4/AM). ResultsThe animal experiment showed that compared with the control group, the model group exhibited hepatic steatosis, inflammatory cell infiltration, hepatocyte degeneration and necrosis, lobular structural disorder, and fibrous tissue hyperplasia in the mouse liver tissue, significantly elevated serum levels of hydroxyproline (Hyp), alanine transaminase (ALT), and aspartate transaminase (AST) (P<0.01), significantly upregulated expression levels of Notch1, Hes family bHLH transcription factor 1(Hes1), Cluster of Differentiation 31(CD31), and von Willebrand factor(vWF) (P<0.01), and significantly downregulated phosphorylate(p)-eNOS/eNOS ratio and sGC expression (P<0.01). Compared with the model group, all the JTCT groups showed significantly reduced serum Hyp, ALT, and AST levels (P<0.01), downregulated expression of Notch1, Hes1, CD31, and vWF (P<0.05, P<0.01), and upregulated p-eNOS/eNOS ratio and sGC expression (P<0.05, P<0.01) in the liver tissue, with the JTCT-H group showing the most significant changes (P<0.01). The cell experiment showed that compared with the LSEC-1day group, the LSEC-5day group exhibited significantly increased expression of Notch1, Hes1, CD31, and vWF (P<0.01), significantly decreased p-eNOS/eNOS ratio and sGC expression (P<0.01), and elevated intracellular Ca2+ concentration (P<0.01). Compared with the LSEC-5day group, the JTCT group showed decreased expression of Notch1, Hes1, CD31, and vWF (P<0.01), significantly increased p-eNOS/eNOS ratio (P<0.01), and significantly reduced intracellular Ca2+ concentration (P<0.01). No significant differences were observed in the expression of Notch1, Hes1, eNOS, p-eNOS, CD31, vWF, sGC, or intracellular Ca2+ concentration between the Jagged1 group and the Jagged1+JTCT group. Compared with the LSEC-5day group, the sGC knockout group (sGC-KO) showed significantly increased expression of CD31 and vWF (P<0.01). There was no significant difference in CD31 or vWF expression between the sGC-KO group and the sGC-KO+JTCT group. ConclusionJTCT inhibits the activation of the Notch signaling pathway and restores the activity of the eNOS signaling pathway to suppress sinusoidal capillarization of LSECs, thereby treating liver fibrosis.
2.Exploring the Pathogenesis of Lung Cancer from Xuanfu and Its Corresponding Syndrome Differentiation and Treatment Based on YE Tianshi's"Acrid"Theory
Weiling WANG ; Yongjun ZHANG ; Linmao YE
Journal of Zhejiang Chinese Medical University 2023;47(12):1406-1412
[Objective]To explore the pathogenesis of lung cancer from the perspective of Xuanfu,and to analyze the theoretical basis and clinical significance of YE Tianshi's acrid theory in the treatment of lung cancer.[Methods]According to the theory of Xuanfu,this paper discussed its influence on the occurrence,development and metastasis of lung cancer from three aspects:the loss of opening and closing of Xuanfu,the loss of"Xuanfu-collaterals"and the loss of"Xuanfu Qi-liquid".By reading YE Tianshi's A Guide to Clinical Practice with Medical Records,YE Tianshi's"acrid"theory and its differentiation and treatment experiences for cancer were summarized,and demonstrated it with a medical record.[Results]Xuanfu opening and closing loss,collaterals loss,Qi and liquid loss,the body's healthy Qi deficiency,Qi stagnation,blood stasis,phlegm and toxin interaction,led to the occurrence and development of lung cancer;YE Tianshi was good at using acrid drugs in the treatment of cancer,and advocated that acrid drugs could dispel wind,move Qi,dissolve phlegm,break concretions and tonify deficiency.In the elderly patients mentioned in the medical records,lung cancer recurred due to deficiency of Qi and stagnation of Xuanfu,phlegm and blood stasis.Therefore,the treatment of acridity was given,and lung cancer was alleviated.[Conclusion]YE Tianshi's"acrid"theory is suitable for different stages of lung cancer treatment by regulating the opening and closing of Xuanfu,which is worthy of clinical promotion.
3.Study on Preparation and Pharmaceutical Character of Capreomycin Sulfate Liposomes by Three Kinds of Methods
Chunhui ZHAO ; Dongkai WANG ; Xiang LI ; Linmao YE ; Xiaoling HE
China Pharmacy 2007;0(31):-
OBJECTIVE:To prepare and characterize capreomycin sulfate liposomes(CSL).METHODS:Capreomycin sulfate was entrapped into liposomes using active loading that pH gradient methods,ammonium sulfate gradient methods and sodium acetate gradient methods respectively followed by lyophilization technique.The liposome was characterized by entrapment efficiency,particle size,? potential and the stability.RESULTS:The entrapment efficiency of CSL pre-and post-lyophilization prepared by three methods were 65.7% and 65.2 %,20.1% and 18.6%,34.6% and 32.4%,with particle size of 136 and 145 nm,144 and 153 nm,142 and 159 nm,? potential of —20.2 and —19.5 mV,—24.4 and —22.9 mV,—18.7 and —17.8 mV respectively.No obvious changes were found in all the indexes in the stability test.CONCLUSIONS:The pH gradient technique is suitable for preparing CSL in 3 kinds of methods.

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