1.Mechanism of Shenqi Dihuangtang in Blocking Renal Fibrosis Injury in Diabetic Kidney Disease Mediated by Epithelial-mesenchymal Transition Through Inhibiting TGF-β1/Smad Signaling Axis
Liangjing LIU ; Haolan LIU ; Xiaoling MAO ; Min YU ; Weitong YAN ; Chao LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):32-45
ObjectiveThis paper aims to study the potential active compound components and action mechanism of Shenqi Dihuangtang in the treatment of diabetic kidney disease (DKD) through network pharmacology and in vivo experimental verification. MethodsUltra-high-performance liquid chromatography-Q-exactive orbitrap mass spectrometry (UPLC-Q-Exactive Orbitrap MS) technology was used to clarify the main active chemical components of Shenqi Dihuangtang, and it was combined with network pharmacology methods such as gene ontology (GO) functional annotations and Kyoto encyclopedia of genes and genome (KEGG) to predict the potential action mechanism of Shenqi Dihuangtang in treating DKD. Subsequently, the DKD model of db/db male mice was established, and the mice were randomly divided into model group, low-dose Shenqi Dihuangtang group (6.10 g·kg-1), medium-dose Shenqi Dihuangtang group (12.19 g·kg-1), high-dose Shenqi Dihuangtang group (24.38 g·kg-1), and daplizin group (1.25 mg·kg-1). During the same period, C57BL/6J male mice were selected into normal group and received drug intervention for 8 weeks, respectively. During this period, the body weight and fasting blood glucose (FBG) of the mice were dynamically monitored, and oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) were performed at the end of dosing. The levels of serum creatinine (SCr), blood urea nitrogen (BUN), uric acid (UA), albumin (ALB), and total protein (TP) were measured by an automatic biochemical analyzer, and 24-hour urine protein was measured by a urine protein quantitative kit. Hematoxylin-eosin (HE), periodic-acid Schiff (PAS), and Masson staining were employed to observe the renal histopathology. The expression of nephrotic protein Nephrin was observed by immunohistochemistry. Western blot was used to detect the expression of epithelial-mesenchymal transition (EMT)-related proteins such as TGF-β1, Smad2/3, α-smooth muscle actin (α-SMA), neural-cadherin (N-Cadherin), and snail protein. ResultsUPLC-Q-Exactive Orbitrap MS identified 384 active compounds in the aqueous extract of Shenqi Dihuangtang. According to oral bioavailability≥30% and the five drug-like principles, 44 key active ingredients were screened out, and 169 intersection targets highly correlated with DKD were matched. Among them, there was a significant interaction relationship between tumor necrosis factor(TNF), interleukin(IL)-6, protein kinase B(Akt)1, Caspase-3, Jun proto-oncogene (JUN), hypoxia inducible factor-1α(HIF-1α), B cell lymphoma-2(Bcl-2), matrix metallopeptidase-9(MMP-9), IL-1β, and TGF-β1. GO functional annotations were significantly enriched in cellular components such as membrane rafts, membrane microdomains, and collagen-containing extracellular matrix, molecular functions such as DNA-binding transcription factor binding, R-Smad binding, and Smad protein binding, as well as biological processes such as reactions to lipopolysaccharides(LPS), reactions to bacteria-derived molecules, and wound healing. The KEGG pathway was significantly enriched in lipids and atherosclerosis, TGF-β signaling pathway, phosphatidylinositol 3 kinase (PI3K)/Akt signaling pathway, etc. In vivo experimental results showed that the high-dose Shenqi Dihuangtang group could significantly reduce FBG levels in db/db mice (P<0.01), improve OGTT (P<0.01) and ITT (P<0.01) levels, reduce SCr (P<0.01), BUN (P<0.01), UA (P<0.01) and 24-hour BUN (P<0.01), and increase ALB (P<0.01) and TP (P<0.01) levels. Pathological staining confirmed that the high-dose Shenqi Dihuangtang group could significantly reduce the glomerular mesangial matrix area and collagen deposition (P<0.01) and upregulate the positive expression rate of Nephrin (P<0.01). Western blot results showed that the high-dose Shenqi Dihuangtang group significantly downregulated the expression of TGF-β1 (P<0.01) and Smad2/3 (P<0.01) signal molecules and inhibited the protein levels of α-SMA (P<0.01), N-Cadherin (P<0.01), and Snail (P<0.01). ConclusionShenqi Dihuangtang can inhibit the TGF-β1/Smad signaling axis and block the renal EMT process, thereby improving DKD renal fibrosis damage. Further analysis of its key active components and clinical transformation pathways is needed in the future.
2.Study on the General Pharmacology of Potentill Discolor Bung
Jiaju YAN ; Haifeng SUN ; Ping YAN ; Weitong YAN
China Pharmacist 2017;20(8):1466-1468
Objective: To study the effects of potentill on all systems of experimental animals to provide experimental evidence for the comprehensive safety evaluation of potentill.Methods: Mice and rats were given potentill orally to determine the influence on all systems, and the general pharmacological activities of potentill were recorded.Results: The general behavior of mice was normal after the adminisration of potentill , the test samples showed no synergetic hypnotic effect with nembutal at subthreshold dose, and had no influence on the spontaneous activity and body coordination ability of mice (P>0.05).The test samples showed no obvious influence on the cardiovascular system and respiratory system of rats (P>0.05).Conclusion: Potentill meets the general requirements of pharmacology at the prescribed dose.
3.Determination of ginsenoside Rg_1 and Re in Shenqi Granula by HPLC
Yan SUN ; Weitong ZHANG ; Haibin CHEN ; Yuning WANG ; Xiaobing ZHOU
Chinese Traditional and Herbal Drugs 1994;0(07):-
Objective To establish an HPLC method for the determination of ginsenosides Rg1 and Re in Shenqi Granula.Methods Chromasil C18 column(250 mm?4.6 mm)was used with acetonitrile-0.05% phosphoric acid solution(21∶79)as mobile phase.The flow rate was 1 mL/min and the detected wavelength was 203 nm.Results Ginsenosides Rg1 and Re could be baseline separated with in 30 min.The average recovery rates were 99.60% and 98.5%,corresponding RSD were 1.93% and 2.31% for ginsenoside Rg1 and Re,respectively(n=5).Conclusion This method is fast and accurate and can be used for quality control of Shenqi Granula.

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