1.Effects of acupuncture and moxibustion at Feishu(BL13)on inflammatory responses and intestinal short-chain fatty acids in asthma model rats
Ling REN ; Jingying ZHOU ; Yitian LAI ; Yizhuo QU ; Guoshan ZHANG ; Yanping YANG
Journal of Acupuncture and Tuina Science 2024;22(2):111-120
Objective:To observe the effects of acupuncture and moxibustion at Feishu(BL13)on inflammatory responses and intestinal short-chain fatty acids(SCFAs)in rats with asthma. Methods:Fifty-six Sprague-Dawley rats were randomly divided into a normal group(16 rats)and a modeling group(40 rats).Rats in the modeling group were subjected to establishing asthma models using ovalbumin(OVA).Model evaluation was conducted using 4 rats from each group.The remaining rats that successfully developed asthma were then randomly divided into a model group,an acupuncture group,and a moxibustion group,with 12 rats in each group.Rats in the acupuncture group received acupuncture treatments,and those in the moxibustion group received moxibustion treatments,both at Feishu(BL13)for 30 min.Following the treatments,the rats were exposed to atomization excitation with a 1%OVA solution for 20 min daily for 14 consecutive days.At the end of the experiment,inflammatory markers in the rats'peripheral blood were analyzed using a biochemical method.In addition,inflammatory cells in the bronchoalveolar lavage fluid(BALF)were counted using Wright-Giemsa staining.The lung tissue of rats was examined under a light microscope after staining with hematoxylin-eosin to observe morphological or pathological changes.Furthermore,real-time fluorescence quantitative polymerase chain reaction was utilized to measure the mRNA expression of inflammatory factors in the lung tissue.Lastly,the concentration of SCFAs in the rat's feces was determined using gas chromatography-hydrogen flame ionization. Results:The levels of eosinophils(Eos),neutrophils(Neu),and lymphocytes(Lym)in the peripheral blood,as well as Eos and Neu in the BALF,and the expression of interleukin(IL)-4,IL-5,IL-13,IL-33,and thymic stromal lymphopoietin(TSLP)mRNAs in the lung tissue were all found to be significantly increased(P<0.05 or P<0.01);the lung tissue structure displayed severe injuries;the levels of acetic acid,propionic acid,isobutyric acid,butyric acid,and valeric acid in the feces decreased significantly in the model group(P<0.05 or P<0.01).Compared with the model group,the peripheral blood levels of Eos,Neu,and Lym,as well as Eos in the BALF,and the mRNA expression levels of IL-4 and IL-5 in the lung tissue decreased significantly in both the acupuncture group and the moxibustion group(P<0.05 or P<0.01).This reduction was accompanied by alleviated pathological damage in the lung tissue.Additionally,there were significant increases in the levels of acetic acid,propionic acid,isobutyric acid,and butyric acid in the feces in both the acupuncture group and the moxibustion group(P<0.05 or P<0.01).In the acupuncture group,the expression levels of Lym in the BALF and IL-13 mRNA in the lung tissue decreased significantly(P<0.05 or P<0.01).In the moxibustion group,the mRNA expression levels of IL-33 and TSLP in the lung tissue also reduced significantly(P<0.05 or P<0.01).Furthermore,the level of valeric acid in the feces increased notably in the moxibustion group(P<0.01).Compared with the acupuncture group,it was found that the mRNA levels of IL-5 and IL-13 in the lung tissue,as well as the acetic acid level in the feces,were significantly higher in the moxibustion group(P<0.05 or P<0.01). Conclusion:Both acupuncture and moxibustion were effective in reducing abnormal inflammation and regulating intestinal SCFAs in asthma model rats.Acupuncture demonstrated superiority in inhibiting pro-inflammatory factors,particularly IL-5 and IL-13,while moxibustion exhibited better regulation on intestinal metabolites SCFAs,especially acetic acid.
2.Research progress of thioredoxin system
Zihua LIU ; Peng ZHOU ; Xiaojuan LI ; Dongrong TANG ; Wen WANG ; Yitian REN ; Jianlin WANG
International Journal of Biomedical Engineering 2021;44(3):256-261
The thioredoxin system is composed of thioredoxin (Trx), thioredoxin reductase (TR) and reduced nicotinamide adenine dinucleotide phosphate. Trx is an important antioxidant molecule that can resist cell death caused by various stresses and plays a prominent role in redox reactions. TR is a protein containing selenium (selenocysteine), mainly in three forms, i.e. TR1, TR2 and TR3. TR1 mainly distributed in the cytoplasm, TR2 mainly distributed in the mitochondria, and TR3 mainly distributed in the testes. TR can regulate cell growth and apoptosis. After the cell becomes cancerous, the expression of TR increases to promote cell growth and metastasis. Trx system is closely related to neurodegenerative diseases, parasitic infections, acquired immunodeficiency syndrome, rheumatoid arthritis, hypertension, myocarditis and so on. The Trx system can remove the reactive oxygen species (ROS) in the body, keep the inside and outside of the cell in a balanced state, and it interacts with the thioredoxin interacting protein (TXNIP), which plays an important role in the regulation of glucose metabolism and tumor treatment. The Trx system is an important target for drug treatment of many diseases. In this paper, the research progress of the thioredoxin system was reviewed.
3.Clinical characteristics and prognosis of male dermatomyositis patients with positive anti-melanoma differentiation associated gene 5 antibody
Yitian SHI ; Fenghong YUAN ; Ting LIU ; Wenfeng TAN ; Ju LI ; Min WU ; Zhanyun DA ; Hua WEI ; Lei ZHOU ; Songlou YIN ; Jian WU ; Yan LU ; Dinglei SU ; Zhichun LIU ; Lin LIU ; Longxin MA ; Xiaoyan XU ; Yinshan ZANG ; Huijie LIU ; Tianli REN
Chinese Journal of Rheumatology 2024;28(1):44-49
Objective:To investigate the clinical features and prognosis of male with anti-melanoma differentiation-associated gene 5 (MDA5) autoantibody.Methods:The clinical data of 246 patients with DM and anti-MDA5 autoantibodies hospitalized by Jiangsu Myositis Cooperation Group from 2017 to 2020 were collected and retrospectively analyzed. Chi-square test was performed to compared between counting data groups; Quantitative data were expressed by M ( Q1, Q3), and rank sum test was used for comparison between groups; Single factor survival analysis was performed by Kaplan-Meier method and Log rank test; Cox regression analysis were used for multivariate survival analysis. Results:①The male group had a higher proportion of rash at the sun exposure area [67.1%(47/70) vs 52.8%(93/176), χ2=4.18, P=0.041] and V-sign [50.0%(35/70) vs 30.7%(54/176), χ2=8.09, P=0.004] than the female group. The male group had higher levels of creatine kinase [112(18, 981)U/L vs 57 (13.6, 1 433)U/L, Z=-3.50, P<0.001] and ferritin [1 500 (166, 32 716)ng/ml vs 569 (18, 14 839)ng/ml, Z=-5.85, P<0.001] than the female group. The proportion of ILD [40.0%(28/70) vs 59.7%(105/176), χ2=7.82, P=0.020] patients and the red blood cell sedimentation rate[31.0(4.0, 101.5)mm/1 h vs 43.4(5.0, 126.5)mm/1 h, Z=-2.22, P=0.026] in the male group was lower than that of the female group, but the proportion of rapidly progressive interstitial lung disease (PR-ILD) [47.1%(33/70) vs 31.3%(55/176), χ2=5.51, P=0.019] was higher than that of the female group. ②In male patients with positive anti-MDA5 antibodies,the death group had a shorter course of disease[1.0(1.0, 3.0) month vs 2.5(0.5,84) month, Z=-3.07, P=0.002], the incidence of arthritis [16.7%(4/24) vs 42.2%(19/45), χ2=4.60, P=0.032] were low than those in survival group,while aspartate aminotransferase (AST)[64(22.1, 565)U/L vs 51(14,601)U/L, Z=-2.42, P=0.016], lactate dehydrogenase (LDH) [485(24,1 464)U/L vs 352(170, 1 213)U/L, Z=-3.38, P=0.001], C-reactive protein (CRP) [11.6(2.9, 61.7) mg/L vs 4.95(0.6, 86.4) mg/L, Z=-1.96, P=0.050], and ferritin levels [2 000(681, 7 676) vs 1 125 (166, 32 716)ng/ml, Z=-3.18, P=0.001] were higher than those in the survival group, and RP-ILD [95.8%(23/24) vs 22.2%(10/45), χ2=33.99, P<0.001] occurred at a significantly higher rate. ③Cox regression analysis indicated that the course of disease LDH level, and RP-ILD were related factors for the prognosis of male anti-MDA5 antibodies [ HR (95% CI)=0.203(0.077, 0.534), P=0.001; HR (95% CI)=1.002(1.001, 1.004), P=0.003; HR (95% CI)=95.674 (10.872, 841.904), P<0.001]. Conclusion:The clinical manifestations of male anti-MDA5 antibody-positive patients are different from those of female. The incidence of ILD is low, but the proportion of PR-ILD is high. The course of disease, serum LDH level, and RP-ILD are prognostic factors of male anti-MDA5 antibody-positive patients.
4.Effects of Moxibustion at "Feishu (BL13)" and "Tianshu (ST25)" on Airway Remodeling,Immune Cell Differentiation and Intestinal Short-chain Fatty Acids in Bronchial Asthma Model Rats
Ling REN ; Jingying ZHOU ; Yitian LAI ; Yizhuo QU ; Guoshan ZHANG
Journal of Traditional Chinese Medicine 2024;65(21):2240-2249
ObjectiveTo explore the possible mechanism of moxibustion at "Feishu (BL13)" and "Tianshu (ST25)" in treatment of bronchial asthma. MethodsA total of 48 SD rats were randomly divided into normal group (n=12) and modeling group (n=36). The bronchial asthma rat model was established by sensitization with ovalbumin (OVA) injection and aerosol provocation. Thirty-two successfully modeled rats were further randomly divided into four groups including model group, Feishu group, Tianshu group, and Feishu-Tianshu group, with 8 rats in each group. Rats in the normal group and the model group were tied and fixed without intervention, while those in the Feishu group received moxibustion at bilateral of "Feishu (BL13)" for 30 minutes; rats in the Tianshu group received moxibustion at bilateral "Tianshu (ST25)" for 30 minutes, and those in the Feishu-Tianshu group received moxibustion at both "Feishu (BL13)" and "Tianshu (ST25)" bilaterally for 15 minutes each. One hour after the intervention, 1% OVA solution was aerosolized for 20 minutes in all groups except the normal group, which was given the same volume of 0.9% sodium chloride solution instead of OVA solution for aerosol stimulation. The above interventions were performed once daily for 14 days. Behavioral observations were performed after modeling and during the interventions. The samples were collected 24h after the last intervention. HE and Masson staining were used to observe pathological morphological changes of lung tissues, and the percentage of collagen fiber deposition area was counted. The levels of leukocyte differentiation antigen11b (CD11b), leukocyte differentiation antigen 40 (CD40), leukocyte differentiation antigen 86 (CD86), and programmed death ligand 2 (PD-L2) in serum, as well as the expression of interleukin-8 (IL-8), interleukin-11 (IL-11), interleukin-27 (IL-27) in lung tissue, were detected by enzyme-linked immunosorbent assay (ELISA). Western blot was used to detect the expression of matrix metalloproteinase-9 (MMP-9), matrix metalloproteinase inhibitor 1 (TIMP-1) and transforming growth factor β (TGF-β) proteins in lung tissue. The content of six short-chain fatty acids (SCFAs) including acetic acid, propionic acid, isobutyric acid, butyric acid, valeric acid, and hexanoic acid in feces was detected by gas chromatography-mass spectrometry. ResultsCompared to the normal group, the rats in the model group gradually showed mental depression or restlessness, dull hair, slow activity, reduced food intake, unformed stool, accompanied by symptoms of shortness of breath and wheezing. The pathological results showed severe abnormalities in lung tissue structure in the model group, including extensive infiltration of inflammatory cells around the bronchi, thickening of the airway smooth muscle layer, and substantial deposition of collagen fibers. Significant increases were observed in the levels of serum CD11b, CD40, CD86, and PD-L2, levels of IL-8, IL-11, and IL-27 in the lung tissue, as well as protein expression levels of MMP-9, TGF-β, and TIMP-1 in lung tissue, while the fecal levels of acetic acid, propionic acid, isobutyric acid, butyric acid, valeric acid, and n-caproic acid significantly decreased (P<0.05 or P<0.01). Compared to those in the model group, the spirit, hair, activity, drinking and eating condition, shortness of breath, and wheezing symptoms of rats in the Feishu group, Tianshu group, and Feishu-Tianshu group were improved; the stool was basically formed, and the pathological morphology of lung tissue were improved; the levels of serum CD11b, CD40, CD86 and PD-L2, the levels of IL-8 and IL-27 in the lung tissue, the percentage of collagen fiber deposition area, and the TGF-β protein expression notably decreased; content of IL-11 and MMP-9 in the lung tissue and protein expression of T1MP-1 in Feishu group and Feishu-tianshu group significantly decreased; content of six SCFAs in the Feishu-Tianshu group increased (P<0.05 or P<0.01). Compared to those in the Feishu group, the percentage of collagen fiber deposition area and TIMP-1 protein expression in lung tissue in the Feishu-Tianshu group significantly decreased, while the fecal levels of acetic acid and butyric acid notably increased (P<0.05 or P<0.01). Compared to those in the Tianshu group, the serum level of CD40 in the Feishu-Tianshu group was significantly reduced, and the percentage of collagen fiber deposition area, the content of IL-11, and the protein expressions of MMP-9, TGF-β and TIMP-1 in the lung tissue notably decreased, while the fecal levels of acetic acid, propionic acid, and butyric acid significantly increased (P<0.05 or P<0.01). ConclusionMoxibustion at "Feishu (BL13)" and "Tianshu (ST25)" exhibits a favorable therapeutic effect on airway remodeling in bronchial asthma rats, and the combined application of "Feishu (BL13)" and "Tianshu (ST25)" acupoints demonstrates a synergistic effect. The mechanism may be related to the regulation of intestinal SCFAs content, influencing the differentiation of immune cells, and reducing airway inflammation.