1.Enhancement of distribution of dermal multipotent stem cells to bone marrow in rats of total body irradiation by platelet-derived growth factor-AA treatment
Zhaowen ZONG ; Yongchuan REN ; Yue SHEN ; Yonghua CHEN ; Xinze RAN ; Chunmeng SHI ; Tianmin CHENG
Chinese Journal of Radiological Medicine and Protection 2011;31(4):433-436
Objective To observe whether dermal multipotent stem cells (dMSCs) treated with platelet-derived growth factor-AA ( PDGF-AA )could distribute more frequently to the bone marrow in rats of total body irradiation (TBI).Methods Male dMSCs were isolated and 10 μg/L PDGF-AA was added to the culture medium and further cultured for 2 h.Then the expression of tenascin-C were examined by Western blot, and the migration ability of dMSCs was assessed in transwell chamber.The pre-treated dMSCs were transplanted by tail vein injection into female rats administered with total body irradiation, and 2 weeks after transplantation, real-time PCR was employed to measure the amount of dMSCs in bone marrow.Non-treated dMSCs served as control.Results PDGF-AA treatment increased the expression of tenascin-C in dMSCs, made (1.79 ± 0.13) × 105 cells migrate to the lower chamber under the effect of bone marrow extract, and distributed to bone marrow in TBI rats, significantly more than ( 1.24 ± 0.09) ×105 in non-treated dMSCs (t = 8.833, P < 0.0l ).Conclusions PDGF-AA treatment could enhance the migration ability of dMSCs and increase the amount of dMSCs in bone marrow of TBI rats after transplantation.
2.Re-endothelialization after placement of drug-eluting stents in patients with CHD
Minghua LUO ; Huaimin GUAN ; Jinhong XIE ; Yushan CHEN ; He WANG ; Chengjie QIU ; Wenjie DONG ; Yonghua ZONG
The Journal of Practical Medicine 2016;32(5):724-727
Objective To investigate the characteristics of coronary vessel re-endothelialization after placement of drug-eluting stents (DES), and to provide clinical evidence for the double anti-platelet treatment. Methods Optical coherence tomography (OCT) was performed in 43 patients in 1 year after DES implantation. Characteristics of re-endothelialization and percentage of neointimal coverage of stent struts were evaluated by OCT. Results The rate of stent struts intimal coverage was 90.70%, and the remain was lack of endothelial coverage; The ratio of neointimal thickness (NIT) between 0-99, 100-199 and above 200 microns was 19.92%, 37.55% and 42.53%, respectively. The rate of neointimal coverage was higher and the degree of neointimal hy-perplasia was more extensive in patients with DM and in patients with ACS than those of patients without DM and of patients with stable angina pectoris. Conclusion One year after stent placement, most of the stent struts were covered with neointima and few struts obtained poor coverage of endothelial. DM and ACS may be impact factors for the progress of re-endothelialization after DES placement.
3.Impact of Alcohol Septal Ablation for Different Coronary Septal Branches on Cardiac Function in Experimental Canines
He WANG ; Junmeng WANG ; Dan SUN ; Yonghua ZONG ; Wenjie DONG ; Jinhong XIE ; Yushan CHEN ; Minghua LUO ; Huaimin GUAN
Chinese Circulation Journal 2016;31(2):170-174
Objective: To compare the percutaneous transluminal septal myocardial ablation (PTSMA) and percutaneous transluminal septal tunnel myocardial ablation (PTSTMA) on cardiac function in experimental canines.
Methods: According to CAG determined coronary septal branches, a total of 25 hybrized canines were divided into 2 groups:PTSMA group, n=13 canines with the bigger septal branches and PTSTMA group, n=12 canines with the smaller or uneven septal branches. Alcohol ablation model was established. Electrocardiograph (ECG) at before and after the operation, biomarkers for myocardial injury, echocardiography and hemodynamic changes were recorded. The animals were scariifes at 1 week after operation, the pathological changes in ventricular septal were observed by HE and Masson staining.
Results: Myocardial infarction (MI) could be induced by either PTSMA or PTSTMA and the thickness of septal was decreased. LVEDd, LVEF and hemodynamic indexes were similar between 2 groups. The alcohol volume used in operation, EKG and echocardiography ifndings were similar between 2 groups, P>0.05. Pathological staining indicated that there was a well-demarcate between the ablation focal and normal myocardium, merging area had neutrophiles invasion, infarcted cells were partially having the ghost cell sample and they were gradually replaced by ifbrous tissue. There was nest-like necrosis in ablated lumen and the normal vessel wall disappeared. PTSMA group had vessel lumen conifguration in septal branch and the necrosis limited inside the lumen;while in PTSTMA group, the vessel wall of was discontinued and some necrosis materials move out to from lumen.
Conclusion: Both PTSMA and PTSTMA were effective for alcohol septal ablation in different coronary septal branches, the impacts on cardiac function and hemodynamic changes were similar in experimental canines.
4.Effects of Zhenfang Baiwanzi Decoction combined with alteplase injection on serum inflammatory factors and vascular endothelial active substances in the patients with acute ischemic stroke and syndrome of wind-phlegm invading collaterals
Fang ZONG ; Yonghua CHEN ; Yuqiang DAI
International Journal of Traditional Chinese Medicine 2023;45(9):1075-1080
Objective:To explore the effects of Zhenfang Baiwanzi Decoction combined with alteplase injection on serum inflammatory factors and vascular endothelial active substances in patients with acute ischemic stroke (AIS).Methods:Randomized controlled trial. A total of 87 patients with AIS admitted to Mengcheng County Hospital of Traditional Chinese Medicine between June 2019 and July 2022 were selected as the observation subjects by prospective cohort study, and they were divided into treatment group (44 cases) and control group (43 cases) according to the last number of medical records. The control group was given thrombolytic therapy with alteplase injection on the basis of routine therapy, and the treatment group was given Zhenfang Baiwanzi Decoction on the basis of the control group. Both groups were treated for 2 weeks. TCM symptoms were scored before and after treatment, and National Institutes of Health Stroke Scale (NIHSS) was used to evaluate the degree of neurological deficit, and Activity of Daily Living Scale (ADL) was applied to evaluate the quality of life. The levels of serum IL-6, CRP, TNF-α, monocyte chemoattractant protein-1 (MCP-1), VEGF, NOS and endothelin-1 (ET-1) were detected by ELISA. The adverse reactions were recorded and the clinical efficacy was evaluated.Results:The total effective rate was 93.18% (41/44) in treatment group and 76.74% (33/43) in control group ( χ2=4.62, P=0.032). After treatment, the scores of hemiparalysis, skin insensitivity, deviated mouth and eye, inhibited speech and headache and dizziness in treatment group were significantly lower than those in the control group ( t=3.38, 3.77, 2.69, 2.60, 2.36, P<0.01 or P<0.05). The NIHSS score was significantly lower than that of the control group ( t=7.53, P<0.01) while the ADL score was significantly higher than that of the control group ( t=2.99, P<0.01). After treatment, the levels of serum IL-6, CRP, TNF-α and MCP-1 in treatment group were significantly lower than those in the control group ( t=6.07, 5.70, 5.30, 3.36, P<0.01), and the levels of VEGF [(364.54±33.04)ng/L vs. (346.86±29.63)ng/L, t=2.63] was significantly lower than that of the control group ( P<0.05), NOS [(20.77±3.12) μmol/L vs. (29.46±5.36) μmol/L, t=9.27] and ET-1 [(85.41±7.09) ng/L vs. (94.11±9.38) ng/L, t=4.89] were significantly lower than those in the control group ( P<0.01). Conclusion:Zhenfang Baiwanzi Decoction combined with alteplase injection can improve the clinical symptoms, reduce levels of inflammatory factors, protect the vascular endothelial function, and enhance the efficacy of patients with AIS.
5.Effect of M1 macrophage polarization regulated by berberine combined with curcumin on atherosclerosis
Yushan CHEN ; Tingting WANG ; Xinyi HAN ; Chengjun HUA ; Boyuan JIN ; Shasha SHANG ; Yonghua ZONG ; Yazhou LIANG
The Journal of Practical Medicine 2024;40(14):1915-1921
Objective To explore the effect and mechanism of berberine combined with curcumin on ath-erosclerosis(AS)by mediating M1 macrophages polarization.Methods M1-type macrophages were obtained from mouse mononuclear macrophages(RAW264.7)induced by lipopolysaccharide(LPS,100 ng/mL)and interferon(IFN)-γ(20 ng/mL).A cell model was established.The cells were divided into a control group,model group,berberine group,curcumin group and berberine plus curcumin group.Concentrations of berberine and curcumin were detected by CCK-8 assay.The expression levels of M1-type macrophage markers iNOS,TNF-α,CXCL9 and p-STAT6/STAT6 in macrophage supernatant were detected by ELISA.Levels of iNOS,TNF-α and CXCL9 mRNA were detected by RT-PCR.Levels of iNOS,STAT6 and p-STAT6 proteins in each group were detected by Western blot.After down-regulation of STAT6 level by siRNA technology,expression of p-STAT6 protein was detected by Western blot.Expression levels of iNOS,TNF-α,CXCL9 and p-STAT6 were detected by ELISA.Results In the polarization of M1 macrophages induced by LPS and IFN-γ,berberine(25 μmol/L)and curcumin(20 μmol/L)were the best concentrations as compared with other drug concentration groups,and neither alone nor combined use could significantly inhibit the viability of RAW264.7 cells(P<0.05).As compared with the normal group,iNOS,TNF-α and CXCL9 mRNA and protein levels were increased in the model group,while P-STAT6/STAT6 levels were decreased,with statistical differences(P<0.05).As compared with the model group,iNOS,TNF-α and CXCL9 mRNA and protein levels in the berberine group,curcumin group,and berberine plus curcumin group were decreased,while P-STAT6/STAT6 levels were increased,and the changes were more obvious in berberine plus curcumin group,with statistical difference(P<0.05).After transfection of STAT6 siRNA in M1 macrophages in the berberine plus curcumin group,P-STAT6 levels were down-regulated,while expressions of iNOS,TNF-α and CXCL9 were up-regulated,with statistical differences(P<0.05).Conclusions Both berberine and curcumin can inhibit the activity of M1-type macrophages and reduce inflammatory response.The action of berberine combined with curcumin is more advantageous than that of either drug alone,which may be the main mechanism of action through activation of STAT6.
6.Berberine regulates polarization of macrophages:a mechanistic study based on PI3K/Akt1 signaling pathway
Yushan CHEN ; Tingting WANG ; Zhonghua MENG ; Shasha SHANG ; Junmeng ZHENG ; Yonghua ZONG ; Chunying SI ; Yazhou LIANG ; Huaimin GUAN
Chinese Journal of Geriatric Heart Brain and Vessel Diseases 2024;26(6):694-698
Objective To explore the effect and possible mechanism of berberine on the macro-phage polarization of human myeloid leukemia monocytic cell line THP-1 induced by oxidized low-density lipoprotein(ox-LDL).Methods THP-1 cells were induced into macrophages by PMA,and then according to different concentrations of berberine,the cells were divided into con-trol group,and 5,10,20,40 and 50 μmol/L berberine groups.After intervention for 24 or 48 h,CCK8 assay was used to detect cell viability for optimal concentration and time of berberine treat-ment.PMA-induced THP-1 macrophages were assigned into blank group,model group(ox-LDL),berberine group,inhibitor group(phosphatidyl inositol 3-kinase(PI3K)inhibitor LY294002)and berberine+inhibitor group(berberine+LY294002).The contents of inducible nitric oxide syn-thase(iNOS)and TGF-β1 were detected by ELISA.qPCR was employed to measure the mRNA expression of TNF-α,arginase 1(Arg1),PI3K and protein kinase B Akt1,and Western blotting was applied to detect the protein levels of Akt1 and phosphorylated protein kinase B antibody(p-Akt1).Results In 24 h after intervention,the macrophage activity was significantly lower in the 40 and 50 μmol/L berberine groups than the control group(P<0.05),and after 48 h,the ac-tivity in all the 5 doses of berberine groups was obviously lower than that in the control group[(0.89±0.02)%,(0.82±0.03)%,(0.71±0.02)%,(0.62±0.03)%and(0.53±0.02)%vs(1.01±0.01)%,P<0.05].Berberine treatment of 20 μmol/L for 24 h had little effect on cell viability,and the dose and the time were regarded as the best concentration and time.Compared with the blank group,iNOS content and TNF-α mRNA level were increased in the model group,while TGF-β1 content,mRNA levels of Arg1,PI3K and Akt1,and p-Akt1/Akt1 protein levels were de-creased(P<0.05).iNOS content and TNF-α mRNA level were decreased,while TGF-β1 content,mRNA levels of Arg1,PI3K and Akt1 and protein levels of p-Akt1/Akt1s were increased in the berberine group than the model group(P<0.05).Compared with the berberine group,iNOS con-tent and TNF-α mRNA level were increased,while mRNA levels of Arg1,PI3K and Akt1 and protein levels of p-Akt1/Akt1 were decreased in the berberine+inhibitor group(P<0.05).Con-clusion Berberine can inhibit the inflammatory response of THP-1 macrophages induced by ox-LDL by activating PI3K/Akt1 pathway,and inhibit the M1 polarization and promote the M2 polarization of macrophages.