1.Mechanism of Jianfu mixture in the treatment of erectile dysfunction based on network pharmacology analysis, molecular docking and in vitro experimental validation
Yantao YANG ; Chao YU ; Zhihang ZHANG ; Yujiong PAN ; Xiaofeng HE ; Min XU
Journal of Pharmaceutical Practice and Service 2026;44(6):296-305
Objective To explore the molecular mechanism of Jianfu mixture in the treatment of erectile dysfunction (ED) by network pharmacology and molecular docking techniques, and validate its core targets and mechanisms through in vitro experiments. Methods The active components and corresponding molecular targets of Jianfu mixture were searched by searching TCMSP and Batman-TCM databases, and the disease targets of ED were searched by using GeneCards database. Find the intersection of drug ingredient target and disease target. The interaction between intersected targets was described and analyzed by String database, and the analysis results were visualized by Cytoscape software to determine the core target and the corresponding active components. GO functional enrichment analysis and KEGG pathway enrichment analysis were performed for intersection targets; the core target within the intersection were found through MCODE plug-in on Cytoscape software and molecular docking was performed with the corresponding active ingredients. An endothelial dysfunction model was established by transfecting HUVECs with si-eNOS. Intervene with different concentrations of the Jianfu mixture for the model cells for 24 h. QPCR was used to detect mRNA expression of core targets (MAPK1, MAPK3, JUN, ESR1, MAPK8); Western blot was used to analyze protein expression (eNOS, JUN, p-JUN, MAPK, p-MAPK) and phosphorylation levels. Results 144 effective active components and 168 active components target-disease targe intersection of Jianfu mixture were obtained. GO analysis revealed 200 5 biological processes, 151 molecular functions, and 63 cellular components. KEGG analysis yielded 181 pathways. 5 core targets including MAPK1, MAPK3, JUN, ESR1 and MAPK8 were screened out. The active components such as β-sitosterol, kaempferol, astapterocarpan had good binding affinity with the core target. In vitro experiments confirmed successful construction of the endothelial dysfunction model (eNOS expression significantly decreased after si-eNOS transfection). Jianfu mixture dose-dependently inhibited mRNA expression of MAPK1, MAPK3, JUN, ESR1, and MAPK8. Additionally, it reduced phosphorylation levels of JUN and MAPK, indicating inhibition of the JNK/c-Jun and ERK/MAPK signaling pathways to improve endothelial function. Conclusion Jianfu mixture treats ED by suppressing abnormal activation of multi-target signaling pathways (MAPK/JUN/ESR1), reducing endothelial apoptosis, and promoting NO synthesis. This mechanism aligns with the traditional Chinese medicine principle of “activating blood circulation, resolving stasis, tonifying Qi, and strengthening cardiovascular function.” The study provided molecular-level evidence for the therapeutic efficacy of Jianfu mixture in ED management.
2.Mechanism of Jianfu mixture in the treatment of erectile dysfunction based on network pharmacology analysis, molecular docking and in vitro experimental validation
Yantao YANG ; Chao YU ; Zhihang ZHANG ; Yujiong PAN ; Xiaofeng HE ; Min XU
Journal of Pharmaceutical Practice and Service 2026;44(6):296-305
Objective To explore the molecular mechanism of Jianfu mixture in the treatment of erectile dysfunction (ED) by network pharmacology and molecular docking techniques, and validate its core targets and mechanisms through in vitro experiments. Methods The active components and corresponding molecular targets of Jianfu mixture were searched by searching TCMSP and Batman-TCM databases, and the disease targets of ED were searched by using GeneCards database. Find the intersection of drug ingredient target and disease target. The interaction between intersected targets was described and analyzed by String database, and the analysis results were visualized by Cytoscape software to determine the core target and the corresponding active components. GO functional enrichment analysis and KEGG pathway enrichment analysis were performed for intersection targets; the core target within the intersection were found through MCODE plug-in on Cytoscape software and molecular docking was performed with the corresponding active ingredients. An endothelial dysfunction model was established by transfecting HUVECs with si-eNOS. Intervene with different concentrations of the Jianfu mixture for the model cells for 24 h. QPCR was used to detect mRNA expression of core targets (MAPK1, MAPK3, JUN, ESR1, MAPK8); Western blot was used to analyze protein expression (eNOS, JUN, p-JUN, MAPK, p-MAPK) and phosphorylation levels. Results 144 effective active components and 168 active components target-disease targe intersection of Jianfu mixture were obtained. GO analysis revealed 200 5 biological processes, 151 molecular functions, and 63 cellular components. KEGG analysis yielded 181 pathways. 5 core targets including MAPK1, MAPK3, JUN, ESR1 and MAPK8 were screened out. The active components such as β-sitosterol, kaempferol, astapterocarpan had good binding affinity with the core target. In vitro experiments confirmed successful construction of the endothelial dysfunction model (eNOS expression significantly decreased after si-eNOS transfection). Jianfu mixture dose-dependently inhibited mRNA expression of MAPK1, MAPK3, JUN, ESR1, and MAPK8. Additionally, it reduced phosphorylation levels of JUN and MAPK, indicating inhibition of the JNK/c-Jun and ERK/MAPK signaling pathways to improve endothelial function. Conclusion Jianfu mixture treats ED by suppressing abnormal activation of multi-target signaling pathways (MAPK/JUN/ESR1), reducing endothelial apoptosis, and promoting NO synthesis. This mechanism aligns with the traditional Chinese medicine principle of “activating blood circulation, resolving stasis, tonifying Qi, and strengthening cardiovascular function.” The study provided molecular-level evidence for the therapeutic efficacy of Jianfu mixture in ED management.
3.Analysis of the particle size of traditional Chinese medicine ointment in the hospital
Yujiong PAN ; Zhigao HE ; Xin ZHOU ; Qianyuan HUANG
Journal of Pharmaceutical Practice and Service 2025;43(10):519-524
Objective To investigate the particle size of traditional Chinese medicine (TCM) ointments in various hospitals. Methods The powders of 7 varieties of externally-applied TCM ointments from Longhua Hospital, Shanghai University of Traditional Chinese Medicine (our hospital) were subjected to comminution processes. Each variety was separately processed by a universal grinder, a hammer mill, and a jet mill, yielding a total of 21 samples. The particle sizes of these 21 samples from our hospital, along with 6 samples obtained from 6 other hospitals, were measured by employing a Bettersize2 laser particle size analyzer and microscopic examination. Results The volume-based average particle size of 21 samples from our hospital ranged from 3.34 to 52.53 μm, while that of 6 samples from other hospitals ranged from 38.59 to 118.50 μm. Notably, the particle size of samples processed by jet milling could be reduced by 12% to 86% compared with those processed by universal mechanical crushing. Microscopic observation revealed that particles larger than 180 μm were detected in 17 samples from our hospital and all 6 samples from other hospitals. Conclusion Compared with universal grinder sample and hammer grinder, jet mill could reduce the average particle size of powder. However, the increasement in pulverization time and loss of pulverization were substantial. Furthermore, in accordance with the particle size requirements for ointments specified in the current Pharmacopoeia of the People's Republic of China (2020 Edition), particle size testing of TCM ointments remains challenging. Therefore, further data accumulation should be imperative to establish particle size standards for TCM ointments.
4.Mechanism of Qizhenziyin mixture in the treatment of hypogonadism based on network pharmacology analysis and molecular docking
Yujiong PAN ; Zhigao HE ; Shixiu CHEN ; Gui ZHOU ; Xin ZHOU ; Chao YU
Journal of Pharmaceutical Practice and Service 2024;42(1):24-31
Objective To investigate the mechanism of Qizhenziyin mixture in the treatment of hypogonadism by using the network pharmacology approach. Methods The active components of Qizhenziyin mixture were obtained by searching TCMSP ,TCMID and HIT databases.The related targets of candidate compounds were obtained by searching STITCH databases. The potential targets of Qizhenziyin mixture in the treatment of hypogonadism were obtained by mapping the disease genes of hypogonadism with Genecards and DisGeNet databases. The protein interaction platform database (STRING) was used to construct the interaction relationship between action targets. The target protein interaction (PPI) network was constructed by introducing Cytoscape software. The mechanism of Qizhenziyin mixture in the treatment of hypogonadism was explained through the enrichment analysis of GO, KEGG and molecular docking technology. Results A total of 148 drug-disease chemical compounds, 96 drug-disease intersection targets, 1085 disease targets were obtained;the components for treating diseases are: quercetin,kaempferol, luteolin, etc; enrichment analysis of GO revealed 1792 biological processes (BP), 31 cellular components (CC) and 79 molecular functions (MF);the results of KEGG pathway enrichment analysis indicated such as FOXO signaling pathway, prostate cancer, AGE-RAGE signaling pathway in diabetic complications, HIF-1 signaling pathway, etc.The results of molecular docking showed that kaempferol and LEP had the best and stable binding energy. Conclusion The active components of Qizhenziyin mixture may play a role of the treatment of hypogonadism by improving insulin resistance and the expression of testosterone synthetase of Leydig cells.
5.Determination of camphor residue and borneol content in Qingchang suppository by GC
Yujiong PAN ; Zhigao HE ; Xin ZHOU ; Hengzhou ZHANG ; Yuehong YANG ; Jingshan HUANG
Journal of Pharmaceutical Practice 2023;41(9):552-556
Objective To establish a gas chromatography for simultaneous determination of camphor residue and borneolum content in Qingchang Suppository. Methods Gas chromatograph method was used. The chromatographic column was Agilent capillary column(30 m×0.25 mm×0.25 µm). The column temperature was 140 ℃. The sample injection temperature was 250 ℃. The FID detector temperature was 250 ℃. Results Camphor,borneol and isoborneol content showed good linear in the extent of 0.0299~1.497(r=1.000), 0.0205~1.025(r=1.000), 0.0097~0.4830 µg (r=1.000). RSDs of precision,stability and repeatability test results were less than 2%. The recovery was 99.7%, 101.0%, 102.5%. Conclusion This method is simple and quick with accurate result, which could be used for the content determination of Borneol in Qingchang Suppository.

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