Mechanism Research Of Xinfeng Capsule On Cardiac And Pulmonary Function Based On KEAP1-NRF2/Are Signaling Pathways In Sjogren’s Syndrome Patients

1ObjectiveIn the traditional Chinese medicine "glove, lung and spleen correlation" under the guidance of the theory, in the heart, lung certification certification as the breakthrough point, analysis, summary of sjogren’s syndrome (SS) in TCM pathogenesis of heart and lung function damage, from the perspective of clinical observation of heart and lung function in patients with SS changes, peripheral blood ROS and RNS, SOD, T-AOC, MDA, GSH, TRX and B, T lymphocyte attenuation factor (B and T lymphocyte attenuator, BTLA), CD19+, the expression of CD24+regulatory B cells (Treg) frequency, peripheral blood IL-18, IL-35, TNF alpha, TNF-gamma, IL-4, the change of IL-10and the patient’s quality of life and fresh air capsule on the impact;System from the point of view of animal experiment to observe the water quantity changes of rats, changes in body mass, spleen index, the index of submandibular gland, heart, lung and histological changes, then observe the changes of SS cardiopulmonary function of rats, peripheral blood IL-18, IL-35, TNF alpha, TNF-gamma, ROS, MDA, SOD, T-AOC, change detection of SS in the rat heart and lung tissue Keapl Nrf2-ARE signaling pathways and gamma GCS, the change of the channel proteins, HO-1evaluation of fresh air capsule (XFC) on the impact of Chinese medicine, discusses XFC to improve the mechanism of action of SS in the rat heart and lung function.2Methods2.1Theoretical studySystem for collecting and organizing SS heart and lung lesions related literature, screening and summarizes the SS to the pathogenesis of heart and lung lesions caused by domestic and foreign research present situation, the evaluation and analysis Keapl Nrf2-ARE pathway activation pathways, SS heart, reduced lung function from the perspective of theory research and Keap1Nrf2-ARE signaling pathways.Compliance with the basic theory of TCM in this paper, the traditional Chinese medicine "heart" and "spleen" and "lung" and "spleen" relationship, from the perspective of physiological pathology and etiology of "mist","lung and spleen", explore the spleen wet only strengthen the law mechanism for the treatment of SS cardiopulmonary function decrease.2.2clinical research2.2.1observation of64cases of heart and lung function in patients with sjogren’s syndrome, oxidative stress index, cell factor and quality of life, anxiety and depression.2.1.2cytokines in patients with sjogren’s syndrome, BTLA, CD19+, the expression of CD24+regulatory B cells (Treg) the change of frequency, and the correlation study: select20healthy control group, all from the first affiliated hospital of anhui institute of traditional Chinese medical examination center.Group match, age, gender and SS are for good health, no obvious organic disease.By leaf source of Shanghai biological technology co., LTD. Experiment enzyme-linked immunosorbent (ELISA) kits healthy controls and patients with sjogren’s syndrome serum IL-18, IL-35, TNF alpha, TNF-gamma, IL-4, IL-10, ROS and RNS, SOD, T-AOC, the change of MDA, GSH, TRX, the Beckman coulter XL--ESPIC MCL type flow cytometry instrument testing healthy controls and patients with serum BTLA, CD19+T cells and the CD24+cells in B cell expression of frequency, and at the same time using automatic biochemistry analyzer determine the general laboratory indicators:SS patients blood sedimentation (ESR), hypersensitive c-reactive protein (CRP), immunoglobulin G (IGg).Using SPSS17.0software for statistical analysis of oxidative stress, quality of life, heart and lung function change, cytokines and BTLA in regulatory B cells expressing the relationship between frequency and other indicators.2.2.3clinical efficacy research:64patients according to random number table can be divided into air capsule group (32cases) and hydroxychloroquine control group (32cases), the quality of life brief scale SF-36(Short form-36), depression self rating scale (SDS), self-evaluation of anxiety scale (SAS) and grade of TCM symptom score, echocardiography and pulmonary function test were used respectively to patients’ cardiopulmonary function change, observe two groups before and after treatment the quality of life, anxiety, depression and the TCM symptom integral value, cardiopulmonary function change, BTLA and oxidative stress, the improvement of related indicators2.3Experimental StudyRats were randomly divided into normal control (NC) group, model control group (MC), hydroxychloroquine (HCQ) sulfate group, astragalus polysaccharide (APS) group, fresh air capsule (XFC) group, each group of12.In addition to the normal control group, to each animal right rear foot plantar intradermal Freund’s complete adjuvant and rat submandibular gland homogenate mixture of0.2ml/inflammatory, only made dry syndrome (Sjogren syndrome, SS) model.19day,30days in a row.Rats to observe changes of water quantity, quality, then we observed the changes of spleen index, the index of submandibular gland, lung histomorphology changes, then observe the changes of SS cardiopulmonary function of rats, enzyme-linked immunosorbent method to detect cytokine interleukin (IL)-18, interleukin (IL)-35, tumor necrosis factor (TNF)-alpha, tumor necrosis factor (TNF)-gamma, active oxygen free radicals (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), the change of total antioxidant capacity (TAOC), immunohistochemical staining to observe the active oxygen free radicals (ROS) and reactive nitrogen free radicals (RNS), glutathione (GSH) and thioredoxin (TRX) protein expression, real-time fluorescent quantitative PCR (realtime fluores cence-policy PCR, RTFQ PCR) to detect Keapl, Nrf2, motorcycle mRNA expression of the protein, Western Blot method for determining rat myocardial tissue gamma glutamine cysteine synthetase (GCS) gamma, heme oxygenase1(HO1) the change of protein, fresh capsule (XFC) on the impact evaluation of traditional Chinese medicine, discusses XFC to improve the mechanism of action of SS in the rat heart and lung function.3Results3.1The theoretical research results3.1.1"pixu (spleen deficient) is to reduce SS heart function as the basis of the":the spleen as the source of qi and blood biochemical, pixu (spleen deficient), deficiency of qi and blood, spleen and the main operation, and the transport disorder, pixu (spleen deficient) phlegmy wet condensation, qi deficiency is promoting powerless, spirit not cloth, aggravate phlegm wet of drinking water, drinking water condensation heart, heart palpitations, chest tightness, not mei, heartache.3.1.2"spleen wet t2dm method" can improves cardiac function in SS levels: according to the SS heart function to reduce the pathogenesis of TCM of the spleen and stomach weakness, lack of qi and blood, the endogenous dampness ", the characteristics of clinical use "spleen wet t2dm" treatment, use of XFC for SS and heart function damage, has obtained the good effect, XFC by improving the sjogren’s syndrome systemic symptoms and signs, symptoms and signs of further improve the core.3.1.3"pixu (spleen deficient)" is the SS as the basis of the reduction in lung function: cause pixu (spleen deficient) and qi and blood deficiency, spleen earth cannot bear the lung, alongwith time cause lung gas, pulmonary dysfunction XuanSu, cough, spontaneous sweating, shortness of breath, etc;Pixu (spleen deficient) transport function disorder, stagnant water, wet coke, phlegmy wet endogenous, gather and ShengTan into drink, dry in the lungs, lung failure section, visible phlegm, chest full sultry, throat phlegm in the audio, chest pain, deferment not more, even palpitations;Phlegm and blood stasis caused by pixu (spleen deficient) alternating knot, lung collaterals block, see more texture disorder, nodules, pulmonary interstitial hyperplasia of fibre, etc.3.1.4"spleen wet t2dm method" lung function can improve SS level:according to the SS reduced lung function and pathogenesis of TCM with the spleen and stomach weakness, lack of qi and blood, endogenous dampness and phlegm and blood stasis and knot "and" factors ", the characteristics of clinical employs "supporting" therapeutic principle, adopt "spleen wet t2dm" treatment, use of XFC for SS and pulmonary function injury, has obtained the good effect, XFC by improving the sjogren’s syndrome systemic symptoms and signs, so as to further improve the pulmonary symptoms and signs, improve the dispersion function and ventilation function.3.2clinical research results3.2.1cardiopulmonary function in patients with SS, oxidative stress index, cytokines and laboratory index and correlation analysis64SS patients and20cases of NC group, doppler ultrasound abnormality rate SS group of doppler ultrasound heart function change detection results heart function parameter E peak, FS%, E/A ratio were significantly lower (P<0.01);A peak, LADd significantly increased (P<0.01).Cardiac function to reduce the42cases, the abnormal rate was60.93%, the cardiac function in abnormal rate is highest for EF, followed by E, A.Compared with NC group, FVC, FEV1, FEV1/FVC, PEF, MEF25, MEF50, MEF75significantly decreased (P<0.05or P<0.01).Reduced lung function of the45cases, accounted for70.31%of all cases, the abnormal rate of lung function FEF75is the highest, followed by FEF50, FEF25, etc.Lung function parameters of FVC, FEV1, FEV1/FVC, PEF, MEF25, MEF50, significantly lower MEF75, FEF25in lung function parameters, FEF50, FEF75reflect the state of small airway, FEV1, MMF, PEF reflects ventilation function, explanation of lung function in patients with SS reduce characteristics on ventilatory function is given priority to, accompanied by a certain degree of small airway disorders.SS patients oxidative stress, cytokines, laboratory results show that the SS patients peripheral blood lymphocyte percentage, BTLA expression frequency, BTLA significantly decreased (P<0.01), CD19+, expression of CD24+frequency was significantly increased (P<0.05);SS patients oxidation index MDA and ROS values significantly higher (P<0.01), anti-oxidation index SOD, TRX values were significantly lower (P<0.01or P<0.05);SS patients serum cytokine IL-1beta, inflammatory index [ESR and CRP] a significant rise in value and IgA (P<0.01), IL4value decreased significantly (P<0.05).Correlation analysis showed that patients with SS heart function parameters of FS, LADd, E peak and the antioxidant index TRX has significant negative correlation (P<0.05);LADd and IL-4was significantly negative correlation with disease activity index (P<0.05), CRP was significantly positive correlation (P<0.01);The FS and ESR were significantly negative correlation (P<0.05).Lung function parameters in FEV1and IL-17were positively correlated, MEF25-75and IL-10was significantly positively related.Show SS patients BTLA expression with the change of heart and lung function decline, the rise of CD19+CD24+Brge and inflammatory factor.The heavier disease, disease activity, the more the more obvious the cardiopulmonary function.3.2.2air capsule to improve SS cardiopulmonary function, oxidative stress index, cytokines and laboratory indexes and the influence of quality of life XFC can obviously improve EF%in patients with SS, E peak;Compared with before treatment, XFC can obviously improve FEV1in patients with SS, MEF50, MEF25and PEF;Frequency of XFC can obviously increase BTLA expression, BTLA lymphocyte percentage, SOD, TRX value, IL-4, reduce CD19+, expression of CD24+frequency, MDA and ROS values, IL-1beta, ESR, CRP, IgA;And on the rise, SOD, TRX BTLA expression frequency, IL-4, reduce CD19+, expression of CD24+frequency, MDA and ROS values, IL-1beta, ESR, CRP, IgA is better than that in control group.XFC can obviously reduce the amount of SS symptoms in patients with grade total score, improve GanYan dry mouth, dry eyes, body tired fatigue, less food, stay, heart palpitations, chest tightness, shortness of breath symptoms, and in reducing symptoms classifying total score, improve GanYan dry mouth, dry eyes, body tired fatigue, less food, stay is better than that in control group;SF-36each dimension score increases, SAS and SDS score.3.3experiment results3.3.1SS rats drinking, spleen index, the index of submandibular gland, body quality and XFC influence on itCompared with NC group, the MC group rats significantly increased water quantity, submandibular gland index, spleen index (P<0.01or P<0.05), body mass was significantly decreased (P<0.01).Compared with MC group, XFC group after the treatment, the HCQ group rats drinking, submandibular gland index, spleen index decreased significantly (P<0.05), and significantly increased body mass in rats (P<0.05).Compared with HCQ group, group XFC water decreased obviously (P<0.05or P<0.01).3.3.2rainfall distribution on10-12SS rats serum cytokine changes and XFC to its effectsCompared with NC group, the MC group of TNF alpha, IFN-gamma, IL-18levels are higher, and cut down the content of IL-35(P<0.01,P<0.05). Compared with MC group, after dosing XFC group, HCQ IL-35, and TNF alpha, IFN-gamma, IL-18decreased (P<0.05, P<0.01).3.3.3SS rats oxidative stress index change and XFC influence on itCompared with NC group, content of MDA and ROS MC group were increased, whereas T AOC, SOD, GSH, TRX protein expression were decreased (P<0.01, P<0.05).Compared with MC group, XFC group after the treatment, the contents of SOD, T AOC HCQ, GSH, TRX protein expression increased, and MDA and ROS decreased (P<0.05, P<0.01).3.3.4SS in the rat heart tissue morphological changes and XFC to its effects Were showed:compared with NC group, the MC group rat myocardial cells arranged disorder, has obvious myocardial cell edema.XFC arrange fair treatment group myocardial cells, myocardial cell edema is not obvious, the lesion degree is lighter than other treatment group;HCQ group of myocardial cell arranged a little disorder, mild edema myocardial cells.3.3.5SS rats lung tissue morphological changes and XFC to its effectsWere showed:compared with NC group, the MC group of rat lung tissue in bronchioles lungs partial necrosis and shedding, a large number of lymphocyte infiltration, part of the alveolar structure, wide alveolar septa, see a large number of lymphocytes, within a small amount of neutrophil infiltration, interstitial vascular engorgement.XFC group in bronchioles lungs most intact columnar epithelium mucosa cilia, mucous membrane and submucous see inside a few lymphocytes and foam cells, most of the alveolar structure is complete, no damage, part of the alveolar interval see small focal foam cells and lymphocytes;HCQ group in bronchioles lungs most of diamond film ciliated columnar epithelium in good condition, a few, a few thin bronchial submucosal focal lymphocytic infiltration, alveolar structure most in good condition, a small number of alveolar interval see focal lymphocytic, foam cells infiltration and interstitial vascular engorgement.3.3.6heart tissue Keapl Nrf2/ARE the influence of the change of the signal path and XFCCorrelation analysis showed that quality of heart index (HI) and ROS were positively correlated (P<0.05);Left ventricular systolic pressure (LVSP), left ventricular diastolic pressure (LVEDP) and the Keapl Nrf2/ARE signaling pathways was significantly positive correlation (P<0.01), and significantly negative correlated to the TAC (P<0.05, P<0.01);Left ventricular pressure up and down the maximum rate (+dp/dt Max) with Keap1Nrf2/ARE signaling pathways was significantly negative correlation (P<0.05), and TNF-gamma show significant positive correlation (P<0.05), the difference was statistically significant.Compared with NC group, the MC model group Keap1, Maf, Nfr2mRNA, gamma, GCS, HO-1were higher (P<0.01).Compared with MC group, XFC group, HCQ Keapl, Maf, Nfr2mRNA, gamma, GCS, HO-1were lower (P<0.01);Compared with HCQ group, fresh air capsule group Keap1, Maf reduced significantly (P<0.01).3.3.7Keap1Nrf2/lung tissue ARE the influence of the change of the signal path and XFCCorrelation analysis showed that lung function parameters FEF25and negatively correlated with T-AOC (P<0.05);FEF50and Maf, Nfr2mRNA, TNF alpha show obvious negative correlation (P<0.05);FEF75, PEF and Keapl, Maf, Nfr2mRNA, T AOC, TNF alpha was significantly negative correlation (P<0.01, P<0.05), the difference was statistically significant.Compared with NC group, the MC model group Keapl, Maf, Nfr2mRNA, gamma, GCS, HO-1were higher (P<0.01).Compared with MC group, XFC group, HCQ Keapl, Maf, Nfr2mRNA, gamma, GCS, HO-1were lower (P<0.01);Compared with HCQ group, fresh air capsule group Keap1, Maf, gamma GCS reduced significantly (P<0.01).4Conclusions4.1sjogren’s syndrome in patients with heart and lung function decline and integral decline and the quality of life all dimensions with anxiety depression, sjogren’s syndrome in patients with cardiopulmonary function and had significant effect on the quality of life;Heart and lung function in patients with sjogren’s syndrome and CD19+4.2, the expression of CD24+and BTLA level, oxidative stress, such as closely related, prompt B cells involved in sjogren’s syndrome in the pathogenic process, increase the patient’s symptoms and signs, make SS cardiopulmonary function and quality of life in patients with each dimension integral to drop, the anxiety depression score;4.3air capsule can effectively improve heart and lung function in patients with SS, improve patients’ quality of life, improve clinical symptoms, reduce CRP, ESR and other inflammatory indexes, the improvement of anxiety depression;4.4air capsules improve cardiopulmonary function in patients with SS, the effective mechanism is increase peripheral blood BTLA expression frequency, lower CD19+, expression of CD24+frequency, increase SOD, TRX, IL-10expression, inhibition of IL-17, MDA and ROS expression, reduce the number of immune complex deposition, reduce SS mouth in patients with dry eye symptoms, reduce the lung tissue inflammation reaction, improve the patients’ cardiopulmonary function, improve the patients quality of life.