Ginsenoside Regulates Treg/Th17 Cell Ratio and Inhibits Inflammation to Treat COPD
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ORIGINAL ARTICLES The First School of Clinical Medicine1, Nanjing University of Chinese Medicine; Department of Pneumology2, Jiangning District Hospital of Chinese Medicine; Department of Pneumology3, Jiangsu Province Hospital of Chinese Medicine, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, China Ginsenoside regulates Treg/Th17 cell ratio and inhibits inflammation to treat COPD YAN-QIU XU1,2, WEN LV2, HONG-JUAN WU2, SUO-FANG SHI3,* Received August 7, 2020, accepted September 18, 2020 *Corresponding author: Suofang Shi, Department of Pneumology, Jiangsu Province Hospital of Chinese Medicine, Affiliated Hospital of Nanjing University of Chinese Medicine, 155 Hanzhong Road, Qinhuai District, Nanjing 210029, China [email protected] Pharmazie 75: 590-594 (2020) doi: 10.1691/ph.2020.0696 Objective: Several studies have suggested an involvement of the immune system in the occurrence and devel- opment of chronic obstructive pulmonary disease (COPD), but the mechanism is still unclear. The aim of this study was to explore the mechanism of ginsenoside in inhibiting inflammation by regulating FOXP3 in COPD. Methods: Eighty COPD patients were selected and 35 healthy people were enrolled in the study to determine clinical efficacy, observation index, and SGRQ scores. Percentage of Treg and Th17 cells were detected by flow cytometry; HE staining was used to detect the effect of ginsenoside therapy on pathological changes of COPD in mice. Additionally, we transfected FOXP3 inhibitor; RT-PCR and western blot were used to detect the inflam- mation related genes and proteins. Results: The basic information of the patients were comparable. The clinical outcome in the treatment group was better than that in the control group, which indicated that ginsenoside has a certain therapeutic effect on COPD patients. The lung function and 6MWT distance results indicated that ginse- noside could stabilize the clinical symptoms of COPD patients and improve their quality of life. Flow cytometry results showed that ginsenoside can increase Treg expression while reducing Th17 cell expression. RT-PCR and western blot results showed that the expression of TNF-α and IL-17 in the model group was significantly increased after treatment, obviously caused by an increased expression of FOXP3. Conclusion: Ginsenoside can inhibit inflammation in COPD by up-regulating FOXP3. 1. Introduction control local inflammation. Studies have shown that the number of Chronic obstructive pulmonary disease (COPD) is a group of lung Treg cells and FOXP3 mRNA are reduced in patients with COPD diseases characterized by airflow limitation. Airflow limitation is (Ito et al. 2019), and FOXP3 expression is increased in the airway not completely reversible and shows a progressive development of smokers with normal lung function and COPD patients. (Brandsma et al., 2017; Hillas et al. 2016). Chronic inflammation In our study, we planned to use patients with stable chronic obstruc- of the airways, lung parenchyma, and pulmonary blood vessels tive pulmonary disease, and randomly assign COPD patients with are typically involved in COPD (Barnes 2017). Inflammatory signed informed consent to a treatment group and a control group, cells such as neutrophils, macrophages, and T lymphocytes all with 40 cases in each group. The control group was recommended contribute to the pathogenesis of COPD. The activation and aggre- to use combined inhalation of tiotropium bromide, salmeterol tica- gation of neutrophils are an important step in the inflammation sone powder inhalation and salmeterol inhalation. The treatment process of COPD (Scoditti et al. 2019; Szucs et al. 2019). The group was given ginsenoside treatment and given the same treat- release of neutrophil elastase, neutrophil cathepsin G, neutrophil ment after 3 mouths. We observed patients’ FOXP3 mRNA, IL-17, protease 3 and matrix metalloproteinase causes chronic mucus lung function, quality of life (CAT), 6-minute walking distance and hypersecretion and destroys the lung parenchyma. Therefore, in other indicators. The purpose of this study was to observe the clin- COPD treatment, it is an effective way to suppress the occurrence ical effect of ginsenosides on COPD patients in the stable phase, and development of inflammation. and to explore the role of ginsenosides in regulating Treg cells Both Treg and Th17 are derived from the common initial CD4+T and Th17 cells by inhibiting inflammation to improve the immune cells. Th17 cells have the effect of promoting inflammation, function of COPD patients and reduce the acute onset of disease. while Treg cells are antagonizing the inflammatory response (Mathur et al. 2019; Tahmasebinia and Pourgholaminejad 2017). 2. Investigations and results The dynamic balance of the two in terms of number and role is to maintain the body’s immune normal dynamics. Balance is of 2.1. Basic information of patients great significance and is an important prerequisite for the body to The 80 patients with COPD in the study were divided into two maintain immune homeostasis (Faschinget al. 2017). Th17/Treg groups according to random number table method; control group imbalance plays an important role in lung immune dysfunction. (n=40) was composed of 25 males and 15 females (Table 1). The There are enough Tregs in the lungs of smokers with normal lung average age was 40.94±3.13 years; the average course of disease function, which can inhibit the destruction of CD8+T cells. Even was 7.59±2.45 years. In the treatment group, there were 29 males smoking does not cause the disease, indicating that the normality and 11 females; the average age was 41.08±3.3 years; the average of regulatory T cells is one of the keys to COPD (Zhou et al. 2019). course of disease was 7.56±2.78 years. No obvious difference was Treg expression is elevated in COPD tissues, and its role may be to observed between the groups. 590 Pharmazie 75 (2020) ORIGINAL ARTICLES Table 1: Basic information about the patients Table 4: Comparison of SGRQ score between the two groups before and after treatment (x±s) Groups Blank Treatment Control Groups Clinical Activity Psychological Total score Males/females 25/10 29/11 25/15 symptoms ability influence Average age 41.78±3.26 41.08±3.37 40.94±3.13 Treatment (n=40) Average course - 7.56±2.78 7.59±2.45 Before treatment 22.18±1.45 23.74±4.42 9.65±1.17 55.57±10.04 After treatment 11.25±3.57*# 12.65±3.31*# 6.71±1.61*# 30.61±8.49*# 2.2. Clinical effi cacy Control (n=40) In order to verify the therapeutic effect of ginsenosides on patients Before treatment 22.87±4.76 23.98±5.62 9.29±1.62 56.14±12.00 with COPD, we judged the therapeutic effect by the clinical symp- After treatment 13.87±4.86* 16.63±5.90* 7.76±1.80* 38.26±12.56* toms and signs of the patients. The results show that there was a statistically significant difference in the comparison of the total Compared with before treatment, *P<0.05; compared with control group, #P<0.05 effective rate between the two groups before and after treatment (P<0.05). The clinical outcome in the treatment group was better than that in the control group (Table 2). This proves that ginseno- 2.5. Ginsenosides treat COPD by increasing the ratio of side has a certain therapeutic effect on COPD patients. Treg to Th17 Compared with the model group, the proportion of Treg in the Table 2: Comparison of clinical efficacy between the two groups after control group and treatment group after treatment was significantly treatment increased (P<0.01); compared with the model group, the propor- Groups Clinical Marked Effective Invalid Total efficiency tion of Th17 cells in the control group and treatment group after control effect (%) treatment was significantly reduced (P<0.01); and the ratio of Treg to Th17 increased significantly (P<0.05, P<0.01) (Table 5). Ginse- Treatment (n=40) 33 (82.5) 3 (7.5) 3 (7.5) 1 (2.5) 39(97.5) noside can increase Treg while reducing Th17 cell expression, and Control (n=40) 25 (62.5) 4 (0.1) 4 (0.1) 7(17.5) 33(82.5) it can treat COPD by increasing the ratio of Treg to Th17. 2.3. Comparison of lung function and 6MWT distance Table 5: Percentage of Treg and Th17 cells in each group(x±s) before and after treatment in two groups Groups Treg cells Th17 cells Treg/Th17 In order to verify the effect of ginsenosides on lung function in proportion/% proportion/% ratio COPD patients, the MASTERSCREEN spirometer was used to Treatment (n=40) measure the forced expiratory volume (FEV1) and forced vital capacity (FVC) at the first second before and after treatment, and Before treatment 1.881±0.695** 2.375±0.362** 0.803±0.307* the FEV1/FVC value was calculated. It can be seen from Table After treatment 3.413±1.618## 1.075±0.385## 3.536±1.837## 3, that FEV1, FVC, FEV1/FVC and 6MWT were increased after Control (n=40) treatment compared to control, and the differences were statisti- cally significant (P<0.05). The improvement of lung function Before treatment 1.874±0.597** 2.298±0.402** 0.812±0.312* indexes and the distance of 6MWT were significantly better than After treatment 2.963±1.010# 1.213±0.629# 2.945±1.471# those of the control group (P<0.05). The above results indicate that Blank (n=35) 3.488±1.106 1.038±0.478 3.846±1.937 ginsenoside treatment can help relieve the clinical symptoms of COPD patients in the stable phase and improve lung function. Compared with before treatment, *P<0.05, **P<0.01; compared with control group, #P<0.05, ##P<0.01 Table 3: Comparison of observation index between two groups of pa- 2.6.