1.Immunomodulatory effects of seaweed polysaccharide in aged mice
Zhen HUANG ; Xiuwen CHI ; Zhenhua SHU ; Junsheng SUN
International Journal of Laboratory Medicine 2015;(13):1801-1803,1806
Objective To explore the immunomodulatory effects of seaweed polysaccharide(PSS)in aged mice induced by D-ga-lactose (D-gal).Methods D-gal was injected intraperitoneally to establish the aged mice model,meanwhile the aged mice was intra-gastricly administrated by PSS.Peritoneal macrophages were collected,and macrophage secretion of nitric oxide (NO),nitric oxide synthase (NOS)levels and expression levels of inducible nitric oxide synthase (iNOS)mRNA were detected.The spleen indexs of the agd mice were calculated,and effects of PSS on spleen microscopic structure of mouse were observed.The changes of spleen cell cycle in aged model mice were detected by flow cytometry assay.Results PSS could enhance macrophage synthesis of NO and NOS of the aged mice and up-regulate the expression of iNOS mRNA levels.And the spleen index of the aged mice increased obviously, the hyperplasia of spleen capsule was obvious.Moreover,PSS could increase the percentage of S phase and G2/M phase cells of the aged mice spleen.Conclusion PSS could enhance the immune function of aged mice induced by D-gal,which is worthy of further study,which development.
2.Preparation and Characterization of Polycolnal Antibody of Galectin-7
Zhen HUANG ; Xiuwen CHI ; Qingwen WEI ; Zhenhua SHU ; Junsheng SUN ; Qian WANG
Journal of Modern Laboratory Medicine 2015;(4):60-62,66
Objective To prepare the mouse anti recombinant human Galectin-7 antibody and the antibody was characterized in bladder cancer.Methods The gene coding for Galectin-7 was amplified by PCR from the cDNA of human foreskin cells and cloned into prokaryotic expression vector pET28a.Then the recombinant plasmid pET28a/Galectin-7 was transformed into E.coli BL21 (DE3)and expressed under IPTG induction.The recombinant Galectin-7 was purified through Ni2+-NT agarosegel column and the purified Galectin-7 used as imunogen to imunize the mouse.The titer and specificity of the anti-Galectin-7antibody from the mouse were analyzed by ELISA,Western blot and immunohistochemistry,respectively.Results The recombinant Galectin-7 was successfully expressed and purified,and the polyclonal ani-Galectin-7 antibody was suc-cessfully prepared.The titer of the antiserum was 1∶32 000 by ELISA.Western blot analysis showed this antiboday reacted specifically with Galectin-7.Immunohistochemistry analysis showed the antibody could recognize the native Galectin-7 in the human bladder cancer tissue.Conclusion The preparation recombinant Galectin-7 protein was as immunogen in rabbits.It was successful to produce high titer and high specificity of anti Galectin-7 polycolonal antibody.
3.Dendrobium Polysaccharides Mitigate CSE-induced Inflammatory Damage of Human Bronchial Epithelial Cells via TLR4/NF-κB Pathway
Lei WANG ; Yajuan WANG ; Zengyang HE ; Fengqing XU ; Wenbin WANG ; Tongsheng WANG ; Junsheng SHU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(23):64-71
ObjectiveTo observe the effects of Dendrobium polysaccharides on the secretion of inflammatory cytokines and Toll-like receptor 4 (TLR4)/nuclear factor (NF)-κB pathway in 16HBE cells exposed to cigarette smoke extract (CSE). MethodThe 16HBE cells were classified into the control, CSE, and CSE+ Dendrobium polysaccharides (100, 200, 400 mg·L-1) groups. The cell-counting kit-8 (CCK-8) assay was employed to measure the cell viability, and a microscope was used to observe the cell morphology. The enzyme-linked immunosorbent assay was employed to measure the levels of interleukin (IL)-8, IL-1β, IL-4, IL-13, and transforming growth factor (TGF)-β in cell culture supernatants. Real-time PCR was carried out to determine the mRNA levels of Toll-like receptor 4 (TLR4), nuclear factor-κB (NF-κB), and IL-4. Western blot was employed to determine the protein levels of interleukin-4 receptor (IL-4R), TLR4, myeloid differentiation primary response protein 88 (MyD88), NF-κB, phosphorylated nuclear factor-κB (p-NF-κB), and nucleoproteins nuclear factor-κB (NEs-NF-κB). The immunofluorescence assay was employed to measure the nuclear translocation of NF-κB. ResultCompared with the control group, the CSE group showed elevated levels of IL-8, IL-1β, IL-4, IL-13, and TGF-β in the cell culture supernatants (P<0.05, P<0.01), up-regulated expression levels of TLR4, MyD88, NF-κB, p-NF-κB, NEs-NF-κB, and IL-4 (P<0.01), and significant nuclear translocation of NF-κB. Compared with the CSE group, Dendrobium polysaccharides increased the cell survival rate, recovered the cell activity, lowered the levels of IL-8, IL-1β, IL-4, IL-13, and TGF-β, down-regulated the expression of TLR4, MyD88, NF-κB, p-NF-κB, NEs-NF-κB, and IL-4 (P<0.05, P<0.01), and reduced the nuclear translocation of NF-κB. ConclusionDendrobium polysaccharides showed significant protective effects on the 16HBE cells exposed to CSE by inhibiting the TLR4/NF-κB signaling pathway.
4.Da Chengqitang in Treatment of Sepsis: A Review
Junsheng SHA ; Nan ZHANG ; Weiyi SUN ; Xiang LI ; Shifa YANG ; Qiang ZHI ; Shu CHEN ; Nan GAO ; Liyan FAN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(17):274-282
Sepsis is a systemic inflammatory syndrome induced by infection and other factors, with the number of patients worldwide exceeding 10 million each year. The pathophysiological mechanism is of this disease complex. Sepsis is often accompanied by endotoxin translocation, gastrointestinal dysfunction, inflammatory cytokine activation, immune dysregulation, coagulation disorder, multiple organ function impairment and many other body imbalances, as well as systemic inflammation, apoptosis, oxidative stress injury and other cell damage mechanisms. This disease causes a heavy medical burden due to the difficult diagnosis and treatment and the poor prognosis. Great progress has been achieved in the diagnosis and treatment of sepsis with traditional Chinese medicine (TCM) and western medicine. The value of western medicine in the diagnosis and treatment of sepsis is limited due to antibiotic resistance, hormone abuse, and high medical costs. Sepsis is classified as a warm disease or typhoid fever in TCM. Da Chengqitang is a classical formula in the Treatise on Typhoid Fever to deal with the excess syndrome of Yang brightness Fu-organ. Modern medicine has proved that Da Chengqitang has the effect of inhibiting oxidative stress, reducing inflammation, and delaying apoptosis by improving gastrointestinal dynamics and regulating intestinal microecology. On the basis of the previous theoretical basis and the rich experience in the medication, medical practitioners have proposed a new therapeutic concept of using Da Chengqitang in combination with western drugs from a holistic view involving both bacteria and toxicity for treating both the symptoms and the root cause, which has a wide range of application. The article reviews the classical research and latest findings of Da Chengqitang in the treatment of sepsis, with a view to clarifying the mechanism and advantages of this formula in the adjuvant treatment of sepsis, exploring its potential efficacy, and providing timely, adequate, and scientific theoretical support for the promotion of this formula in the clinical practice.
5.Traditional Chinese Medicine Regulates Oxidative Stress-related Signaling Pathways to Prevent and Treat Bronchial Asthma: A Review
Xuyang ZHENG ; Zhenhui SHU ; Yi LI ; Ziyue QU ; Weilin LI ; Kexin MA ; Junsheng SHA ; Limin ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(18):260-269
Bronchial asthma (asthma for short) is a common clinical respiratory disease mainly characterized by chronic airway inflammation, with complicated pathogenesis and a long treatment cycle. It is lingering and difficult to be cured, and lack specific drugs. Oxidative stress is a new focus in the research on the pathogenesis of asthma and a potential key target for the treatment. Under physiological conditions, the oxidative and antioxidative systems in the body are in a dynamic balance, and the two antagonize each other to maintain normal life activities. In the case of asthma attack, oxidation products such as reactive oxygen species (ROS), malondialdehyde (MDA), and nitric oxide (NO) are produced excessively, while the content of antioxidants such as superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) is reduced. As a result, the oxidation exceeds the removal of oxidation products, which aggravates oxidative stress. In addition, the overproduction of ROS activates oxidative stress-related signaling pathways to produce pro-inflammatory factors, exacerbating inflammation, which leads to lung and airway tissue damage. In recent years, traditional Chinese medicine has garnering increasing attention because of the unique advantages in the treatment of asthma, especially in regulating redox balance, alleviating oxidative stress in asthma patients, and reducing inflammation. On the one hand, by inhibiting the mitogen-activated protein kinase (MAPK) and transcription factor (NF)-κB signaling pathways, traditional Chinese medicine can reduce the content of oxidation products and pro-inflammatory factors from the source. On the other hand, by activating the nuclear factor-erythroid 2-related factor 2 (NrF2) signaling pathway, traditional Chinese medicine can elevate the levels of antioxidant enzymes and enhance the antioxidant system to neutralize the excessive accumulation of oxidation products. Therefore, the adjustment of redox balance state by traditional Chinese medicine may be a new means and a new direction for the prevention and treatment of asthma in the future. This paper summarizes the oxidative stress-related pathways in the pathogenesis of asthma and reviews the latest research progress in the regulation of oxidative stress-related pathways by Chinese medicine extracts and prescriptions in the treatment of asthma, with a view to providing a fuller, more solid, and more scientific theoretical basis for the clinical and basic research on the prevention and treatment of asthma by traditional Chinese medicine.