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Study on the treatment of rheumatoid arthritis with hydrogen water

Source : QingsiyuEditor : QingmaiView : 50Time : 2020-07-27 14:12
This is the abstract of a postgraduate thesis from Nanjing General Hospital. It is helpful for us to understand hydrogen treatment of rheumatoid arthritis. If you are interested, you can check the full paper.
As an effective antioxidant and anti-inflammatory substance, hydrogen may have therapeutic effect on diseases with inflammatory reaction as pathological basis. Rheumatoid arthritis is an autoimmune disease. Although the molecular mechanism of rheumatoid arthritis is not fully understood, the pathological inflammatory reaction caused by the dysfunction of autoimmune system is its core process. Clinical use of a variety of means to block the inflammatory response can indeed alleviate the disease, is the basic strategy for the treatment of the disease. However, the anti-inflammatory tools used in the past often had high price or side effects. Hydrogen, as an extremely safe substance with almost no toxic and side effects, has inherent advantages in the treatment of inflammatory diseases. Japanese scholars have also carried out preliminary clinical trials, and preliminary results show that hydrogen is effective for rheumatoid arthritis. However, the basic theoretical basis and preclinical evidence of hydrogen therapy for rheumatoid arthritis are still lacking, which is not conducive to the understanding and further study of the disease. This thesis will enrich the preclinical research and lay an important foundation for the clinical research of hydrogen treatment of rheumatoid arthritis.
reference
Meng Jia,. Molecular hydrogen decelerates rheumatoid arthritis progression through inhibition of oxidative stress.[J]. American journal of translational research,2016,8(10).
Study on the role and mechanism of hydrogen in the treatment of rheumatoid arthritis [D]. Meng Jia. Second Military Medical University. 2017
Key research results: the pictures are from the papers published abroad.
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A. Normal animals, B, arthritis animals, C, arthritis after hydrogen water treatment.
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Pathological results: A, normal, B, arthritis, C, hydrogen water treatment.
Here is a summary and comments from qingsiyu.
1、 Research background and purpose
Rheumatoid arthritis (RA) is a systemic autoimmune disease characterized by chronic synovial inflammation. The incidence rate of rheumatoid arthritis in Europe and America is as high as 0.5-2.0%, and the incidence rate in China is 0.32-0.38%. The pathogenesis of rheumatoid arthritis is complex and affected by many factors, which lead to immune imbalance and excessive inflammatory reaction, which lead to the destruction of joint structure.
Oxidative stress is involved in the pathogenesis of rheumatoid arthritis and has great influence on the pathological mechanism of rheumatoid arthritis. The oxidative damage of rheumatoid arthritis is mainly caused by reactive oxygen species and reactive nitrogen, which are harmful free radicals produced in the process of metabolism. Oxidative stress can increase the levels of oxidative markers such as malondialdehyde (MDA), 8-hydroxy-2 '- deoxyguanosine (8-OHdG) and protein carbonyl in patients with rheumatoid arthritis. Oxidative stress can also activate the classic nuclear factor - κ B (NF - κ b) signaling pathway. Oxidative stress can also change the redox state of glutathione (GSH), and induce the inactivation of NF - κ B inhibitor (I κ b), and then ectopic into the nucleus.
Hydrogen is a colorless and odorless gas widely existing in nature. It has selective antioxidant activity and can selectively neutralize reactive oxygen species. However, the specific mechanism of hydrogen selective antioxidation is still unclear, and the upstream molecules that play a pivotal role in the regulation of hydrogen are not understood. Nuclear factor kappa B (NF - κ b), mitogen activated protein kinases (MAPKs), transforming growth factor β 1 (TGF - β 1), which are closely related to inflammation, mitogen activated protein kinases (MAPKs), and transforming growth factor β 1 (TGF - β 1), a potent chemokine of fibroblasts, These factors are known to play an important role in hydrogen antioxidant stress pathway. Keap1 / Nrf2 / are signaling pathway is an important pathway in the body's antioxidant stress response, and is a hot channel in the field of antioxidant research in recent years. Hydrogen can play its selective antioxidant effect through Nrf2, but which way does hydrogen induce Nrf2 to produce effect? This specific mechanism is still a hot and difficult point in hydrogen medical research. Hydrogen is also used in the treatment of rheumatoid arthritis because of its selective antioxidant effect. Foreign scholars reported that in patients with rheumatoid arthritis, the disease activity Score-28 (DAS28) and inflammatory indexes in blood were significantly improved after hydrogen treatment. However, the antioxidant effect of hydrogen on fibroblast like synoviocytes (FLS) and its effects on FLS proliferation, apoptosis and related signal pathways have not been reported. In addition, the effect of hydrogen on FLS has not been reported, and whether it is related to MAPK, NF - κ B and other signal factors is not clear. Whether Nrf2, an important antioxidant signal channel in oxidative stress disease, is also involved in the inhibition process of rheumatoid arthritis disease by hydrogen should be further studied.
2、 Research contents and methods
CIA mice model was established by subcutaneous injection of xenogeneic type II collagen emulsion on the back and tail of mice. The performance of foot and tail of mice was observed by morphology to evaluate the quality of modeling. The AI score, morphological and pathological observation were used to evaluate the remission of rheumatoid disease in CIA mice model by hydrogen. MH7A rheumatoid synovial fibroblasts were treated with hydrogen peroxide to create an environment of oxidative stress in vitro. The effect of hydrogen on oxidative stress of FLS was evaluated by observing cell proliferation, expression of antioxidant stress system (SOD and GSH) and DNA oxidative damage index (8-OHdG). At the same time, we detected MAPK, NF-kB and TGF - β 1 to evaluate the effect of MAPK channel and TGF - β 1 on oxidative stress response of FLS. Fibroblasts isolated from children's skin were treated with hydrogen peroxide to create an environment of oxidative stress in vitro. The results of apoptosis and necrosis were detected by annexin V / PI, and the production of reactive oxygen species was observed by DHE and DCFH-DA; The expression of MDA, SOD and GSH, DNA oxidative damage index (8-OHdG) were analyzed by ELISA. The effect of hydrogen on oxidative stress of fibroblasts was evaluated. At the same time, we used Western blot to analyze the protein expression of Nrf2, cata and γ - GCS to evaluate whether Nrf2 signaling pathway has effect on oxidative stress response of fibroblasts.
3、 Research results and conclusions
In this study, we obtained the following results:
1. The pathological results showed that hydrogen water treatment was effective.
There were no synovial cell proliferation, inflammatory cell infiltration, pannus formation and bone erosion in the normal control group; there was obvious proliferation of synovial tissue, infiltration of inflammatory cells, pannus production and bone erosion in CIA model group; a small amount of proliferative synoviocytes, a small amount of neutrophils infiltration and slight bone erosion were observed in the hydrogen rich water treatment group. Pathological scores showed that the pathological scores of mice in the hydrogen rich water treatment group were significantly lower than those in the CIA model group.
2. Visual observation showed that hydrogen water treatment was effective.
At 30 days (early stage of the disease), some mice in CIA group developed red and swollen forefeet, and then developed to hind feet. Swelling peaked at day 40 (mid disease). The time of limb redness and swelling of mice in hydrogen rich water treatment group was later than that in CIA model group, and the degree of swelling in hydrogen rich water treatment group was significantly less than that in CIA model group. In CIA model group, the arthritis index (AI) began to increase about 14 days (i.e. in the early stage of the disease), and reached the peak at 21 days, and then decreased gradually. The time of AI index increasing in hydrogen rich water treatment group was the same as that in CIA group, but the increase range was significantly less than that in CIA model group.
3. The survival rate of MH7A cells was detected by MTT assay, which showed that hydrogen was helpful to cell survival, which may be the cellular basis for hydrogen to exert the above effects.
The results were as follows: compared with the control group, the cell proliferation was significantly enhanced after adding H2O2, and the proliferation speed was accelerated with the time. The cell proliferation was inhibited by hydrogen.
4. The results of ELISA showed that hydrogen had an effect on oxidative stress and antioxidant system, which may be the molecular basis of hydrogen. Hydrogen activates antioxidation, reduces oxidative stress, controls inflammatory response and plays a role.
It was found that 8-OHdG increased and SOD and GSH in antioxidant system decreased after treatment with hydrogen peroxide. After adding hydrogen rich medium, the content of 8-OHdG was reduced, and the contents of SOD and GSH were increased. At the same time, hydrogen inhibited the expression of MAPK, NF - κ B and TGF - β 1.
5. For fibroblasts, 24 hours after operation, the results showed that hydrogen peroxide (H2O2) treatment induced oxidative stress model group, compared with other groups, had a higher incidence of apoptosis. The spontaneous apoptosis rate of hydrogen group was lower than that of H2O2 group.
6. The reactive oxygen species (ROS) levels in fibroblasts treated with H2O2 were significantly higher than those treated with H2O2, and the levels of ROS were significantly decreased after hydrogen treatment. After treated with H2O2, 8-OHdG and MDA increased, GSH and SOD decreased. When hydrogen rich medium was added, the contents of 8-OHdG and MDA were decreased, and the contents of GSH and SOD were increased.
7. Western blot analysis showed that Nrf2, γ - GCS and cata of fibroblasts decreased after H2O2 was added. After adding hydrogen, Nrf2 and γ - GCS returned to normal, and the level of cata also increased.
Through the above results, we confirmed that hydrogen can effectively inhibit the occurrence of rheumatoid arthritis by inhibiting excessive oxidative stress. Hydrogen can activate the antioxidant system in vivo and inhibit the expression of MAPK, NF - κ B and TGF - β 1. At the same time, Nrf2 signal channel is activated. Therefore, the mechanism of hydrogen regulating oxidative stress in rheumatoid arthritis is also very complex.
Summary Comments:
This study has proved that hydrogen has therapeutic effect on animal model arthritis from different research levels of cell, molecular biology and animal arthritis model, and has carried out comprehensive research and Analysis on cell survival, various oxidative stress and antioxidant systems, related oxidative stress regulatory molecular signaling pathways, especially the core transcription factor Nrf2 signal channel of oxidative stress. This study provides theoretical evidence for understanding the hydrogen treatment of arthritis, and is conducive to understanding the logical basis of hydrogen treatment of arthritis.
 
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