1.Protective Effect and Molecular Mechanism of Hirudin on Kidney of Diabetic Rats
Yiqian HE ; Jun XIE ; Bingjia ZHAO ; Xiaochun LIANG ; Ling QU
Medical Journal of Peking Union Medical College Hospital 2024;15(6):1372-1381
To investigate the protective effect of hirudin on kidneys of diabetic rats and its molecular mechanism. Eighteen Sprague-Dawley rats were randomly divided into control group ( Compared with the control group, the diabetic model group showed significantly elevated random blood glucose and renal function parameters during the 6-week treatment (both Hirudin improved renal pathological changes in diabetic rats, and its mechanism may be related to the decrease of fibrosis-related factors and inhibition of the activation of the JAK2/STAT3 pathway.
2.Serum-free culture of the MKN-45 gastric cancer cell line and preparation of stem cells
Yiqian JIANG ; Xiaoxiang GUAN ; Qingmin GUO ; Xiaoping XU ; Suhong AN ; Yiyang HE ; Juncai LIANG ; Lijuan HAN
Journal of Medical Postgraduates 2016;29(4):380-383
[Abstract ] Objective Difficulties with the preparation of gastric cancer stem cells (CSCs) have been a main obstacle to the studies of gastric cancer .This article addresses the technology of the serum-free medium suspension cultivation of the MKN-45 gastric cancer cell line and screening of stem cells from the cell line based on the biomarkers of gastric CSCs . Methods MKN-45 cells were cultured in serum-free medium for 8 weeks and those in the logarithmic phase cultivated with hoechst 33342 followed by detection of the side cells by flow cytometry .When the side population cells reached 25%, all the cell microspheres were collected , hatched with CD133 and CD44, and subjected to fluorescence-activated cell sorting.The CDl33 +and CD44 +cells were selected as gastric CSCs . Results About 40%of the MKN-45 gastric CSCs were alive , prolif-erated, and formed floating cell balls .Side population cells constitu-ted 3.4% of the MKN-45 cells and 26.9% of the cell balls.The CDl33 +and CD44 +cells made up 11.2% of the MKN-45 cells and 90.3%of the cell balls. Conclusion Cell balls rich in CSCs can be successfully obtained by serum-free medium suspension culti-vation and CSCs can be screened out with hoechst 33342 and surface markers , which may serve as an experimental ground for the stud-ies of gastric CSCs .
3.Targeted metabolomics reveals the aberrant energy status in diabetic peripheral neuropathy and the neuroprotective mechanism of traditional Chinese medicine JinMaiTong
Zhao BINGJIA ; Zhang QIAN ; He YIQIAN ; Cao WEIFANG ; Song WEI ; Liang XIAOCHUN
Journal of Pharmaceutical Analysis 2024;14(2):225-243
Diabetic peripheral neuropathy(DPN)is a common and devastating complication of diabetes,for which effective therapies are currently lacking.Disturbed energy status plays a crucial role in DPN pathogenesis.However,the integrated profile of energy metabolism,especially the central carbohy-drate metabolism,remains unclear in DPN.Here,we developed a metabolomics approach by targeting 56 metabolites using high-performance ion chromatography-tandem mass spectrometry(HPIC-MS/MS)to illustrate the integrative characteristics of central carbohydrate metabolism in patients with DPN and streptozotocin-induced DPN rats.Furthermore,JinMaiTong(JMT),a traditional Chinese medicine(TCM)formula,was found to be effective for DPN,improving the peripheral neurological function and alleviating the neuropathology of DPN rats even after demyelination and axonal degeneration.JMT ameliorated DPN by regulating the aberrant energy balance and mitochondrial functions,including excessive glycolysis restoration,tricarboxylic acid cycle improvement,and increased adenosine triphosphate(ATP)generation.Bioenergetic profile was aberrant in cultured rat Schwann cells under high-glucose conditions,which was remarkably corrected by JMT treatment.In-vivo and in-vitro studies revealed that these effects of JMT were mainly attributed to the activation of adenosine monophosphate(AMP)-activated protein kinase(AMPK)and downstream peroxisome proliferator-activated receptor gamma coactivator 1-alpha(PGC-1α).Our results expand the thera-peutic framework for DPN and suggest the integrative modulation of energy metabolism using TCMs,such as JMT,as an effective strategy for its treatment.
4.Exploration of the Mechanism of Yangxue Qingnao Granules in the Treatment of Hypertension Based on Network Pharmacology and Molecular Docking
Jiawen SHI ; Lei HAO ; Yu WANG ; Zhipeng HUO ; Yiqian ZHANG ; Zhaohui SONG ; Yi HE
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(8):1206-1214
Objective To explore the mechanism of Yangxue Qingnao Granules(Siwu Decoction modified)in the treatment of hypertension based on network pharmacology and molecular docking.Methods The chemical composition analysis results of Yangxue Qingnao Granules in the early stage of the research group were used as the basis for the screening of active compounds.The oral bioavailability≥30%and drug-likeness≥0.18 were used as the screening conditions,and the blood components were supplemented in combination with the literature.TCMSP,chemical professional database and SWISS database were used to predict the targets of potential active compounds of Yangxue Qingnao Granules.Hypertension-related targets were obtained through GeneCards and DiGSeE databases.The intersection of the targets related to hypertension disease and the targets of the potential active compounds of Yangxue Qingnao Granules(common targets)is the potential target of Yangxue Qingnao Granules for the treatment of hypertension.The potential targets were matched with the potential active compounds of Yangxue Qingnao Granules to obtain the antihypertensive active compounds of Yangxue Qingnao Granules.PPI analysis was performed on the potential targets of serum brain granules in the treatment of hypertension through the STRING database,and the core targets were screened according to the degree value.The David database was used to analyze the GO function and KEGG pathway enrichment of the core targets.The core targets with the top six degrees were selected as the docking target proteins,and molecular docking verification was performed with the antihypertensive active compounds.Results A total of 32 potential active compounds,161 active compound targets and 1 539 hypertension-related targets were obtained.After intersection,88 potential targets(common targets)of Yangxue Qingnao Granules in the treatment of hypertension were obtained,involving 29 antihypertensive active compounds.PPI analysis screened 14 core targets:PPARG,ACHE,IL4,CCL2,JUN,NOS3,APP,IL1B,CAT,PTGS2,CASP3,TP53,TNF,IL6,involving 158 GO entries and 13 signaling pathways.Five key active ingredients,chlorogenic acid,rosmarinic acid,paeoniflorin catechinic acid and aloe emodin,were obtained by molecular docking,which were combined with PTGS2,CASP3,TNF,CAT,TP53 and IL6,respectively.Conclusion Yangxue Qingnao Granules may act on core targets such as PTGS2 and CASP3 through key active components such as chlorogenic acid and rosmarinic acid,regulate key pathways such as TNF signaling pathway,MAPK signaling pathway and Toll-like receptor signaling pathway,and play a role in the treatment of hypertension through anti-inflammatory effects.
5. Preparation of chaperone-antigen peptide vaccine derived from human gastric cancer stem cells and its immune function
Yiqian JIANG ; Qingmin GUO ; Xiaoping XU ; Juncai LIANG ; Yiyang HE ; Suhong AN ; Fang SU ; Chaoyang LI ; Changxin HUANG
Chinese Journal of Oncology 2017;39(2):109-114
Objective:
To explore the method of extracting chaperone antigen peptide complexes from gastric cancer stem cells and its immune function.
Methods:
Gastric cancer stem cells and gastric cancer cells were screened by low temperature ultrasonic lysis. After salting out and dialysis, the lysate supernatant was processed with SDS-PAGE to analyze the expression of chaperone antigen peptide complexes, and then was separated and purified with CNBr-activated SepharoseTM 4B. Reverse high pressure liquid chromatography (HPLC), SDS-PAGE and Western blotting were used to analyze the purity and nature of the acquired albumen. Lymphocyte proliferation assay and lymphocytotoxicity assay were used to ditermine the immunological activity of the chaperone-antigen peptide complexes.
Results:
The chaperone antigen peptide complexes of gastric cancer stem cells were prepared and identified successfully, of which the main components were the antigen peptides of HSP60, HSP70, HSP90 and HSP110. 0.75 μg and 1.00 μg HSP70-antigen peptide and 1.00 μg HSP90-antigen peptide activated lymphocytes significantly. Their
6.Protective Effect and Molecular Mechanism of Hirudin on Kidney of Diabetic Rats
Yiqian HE ; Jun XIE ; Bingjia ZHAO ; Xiaochun LIANG ; Ling QU
Medical Journal of Peking Union Medical College Hospital 2024;15(6):1372-1381
To investigate the protective effect of hirudin on kidneys of diabetic rats and its molecular mechanism. Eighteen Sprague-Dawley rats were randomly divided into control group ( Compared with the control group, the diabetic model group showed significantly elevated random blood glucose and renal function parameters during the 6-week treatment (both Hirudin improved renal pathological changes in diabetic rats, and its mechanism may be related to the decrease of fibrosis-related factors and inhibition of the activation of the JAK2/STAT3 pathway.