The Mechanism Research Of Xinfeng Capsule On The Cardiac Function Of Adjuvant Arthritis Rats Based On The Neuro-endocrine-immune Network

1 ObjectiveUnder the guidance of CCInese medicine theory, summarize and analyze cardiac function changes in rheumatoid arthritis and the CCInese medicine impact on it from the perspective of neuro-endocrine-immune network; study cardiac function and NEI network indicators changes of rats with adjuvant arthritis, as well as the impact of Strengthening the spleen-Resolving dampness-Promoting collaterals method-xinfeng capsule (XFC) on it by animal experiments, reveal the regulation & control as well as the possible mechanisms of XFC on cardiac function of AA rats.2. Methods2.1 TheoryThrough literature study, summarize and analyze the relationsCIp between NEI network and cardiac function, spleen, as well as RA; explain the main pathogenesis of RA cardiac function changes; explain changes of RA cardiac function and meridians of CCInese medicine XFC on them from the perspective of NEI network. 2.2 Experimental StudyA total of 60 sprague dawley(SD)male rats were randomly divided into 5 groups:normal control (NC) group, model control (MC) group, tripterygium wilfordii polyglycoside tablet (TPT)group, metho-trexate (MTX) group and XFC group, and there are 12 rats in each group; excluding NC group, the other groups are prepared by AA rats. The first 19 days after inflammation began to give the appropriate drugs treatment for 30 days. During the experiment observated the rats body mass, paw swelling and arthritis index (AI); and the end day of the experiment detected cardiac function of AA rats:after the end day of the experiment detected NEI network indexes:using enzyme-linked immunosorbent assay (ELISA)to detect the level of 5-hydroxytryptamine,5-HT), dopamine(DA), brain natriuretic peptide (BNP), endothelin-1(ET-1), calcitonin gene related peptide(CGRP), interleukin-6 (IL-6), interleukin-10 (IL-10), interleukin-17 (IL-17) and tumor necrosis factor-α(TNF-α); using flow cytometry detected the level of CD4+CD25+reguLatory T cells (Treg) in blood; using immunoCIstochemistry detected the expression of spleen Foxp3 protein, reverse transcription-polymerase chain reaction (RT-PCR) detected the expression level of heart tissue BNPmRNA and spleen Foxp3mRNA; using HE staining and transmission electron microscopy detected the heart morphology and using western blot detected the expression levels of matrix metalloproteinase-9 (MMP9) and tissue inCIbitor of matrix metalloproteinase-1 (TIMP1) protein, using RT-PCR detected the expression levels of MMP9mRNA, TIMP1mRNA.3. Results3.1 Theoretical results3.1 Theoretical results 3.1.1 Close RelationsCIp between NEI network and changes of RA cardiac functionThe three systems support and control each other to keep the dynamic balance of body. So, any pathology is closely related to NEI network. RA cardiac function changes means the development of RA disease, during the process of wCIch, NEI network has some effect.3.1.2 According to CCInese medicine, pleen is an important backbone to regulate NEI network, heart and spleen as mother-cCIld relationsCIp, impact each other on physiological functions and pathological changesSpleen is the central part to study Visceral Phase theory. Spleen can be seen as nervous systems,endocrine systems,immune systems ect these systems are closely related functional groups. Spleen is pertaining to earth, wCIle heart is pertaining to fire. They are like mother and baby. If the baby is ill, it will endanger the mother, or it will weak the strength of mother. Both can endanger the heart because of the disorder of spleen.3.2 Experimental results3.2.1 Cardiac function in AA ratsCompared with NC group rats, MC group rats'heart rate (heart rate,HR), cardiac index (heart indexe, CI), left ventricular end systolic pressure(left ventricle end-systolic pressure, LVESP), left ventricular end diastolic pressure (left ventricular end diastolic pressure, LVEDP) significantly become CIgher (P<0.05), the maximum change rate of left ventricular pressure rise and fall(±dp/dtmax) significantly (P<0.05);3.2.2 NEI network changes of AA rats Compared with NC group rats, MC group rats' TNF-α, BNP, IL-17, 5-HT, ET-1 and IL-6 were significantly increased, CGRP, IL-10, CD4+CD25+Treg, and the monitoring index of Foxp3 were significantly-reduced (P<0.05 or P<0.01).CIerarcCIcal cluster analysis of NEI network indicators showed that:all detected targets are divided into two categories, the 8 indicators, including TNF-a,5-HT, BNPmRNA, CGRP, ET-1, IL-6, IL-17, DA,polymerized firstly, in these indicators, the relationsCIp of IL-6, DA, ET-1 is most close; then, the four indicators, including Foxp3mRNA, CD4+CD25+Treg Treg, BNP, IL-10 polymerized, in these four indicators, CD4+CD25+Treg Treg, IL-10, BNP expression are most close.3.2.3 The relationsCIp between parameters of cardiac function of AA rats and indexes of NEI networkThe experimental results show:the parameters of cardiac function of AA rats are closely related to the indexes of NEI network.3.2.4 XFC's effect on cardiac function of AA ratsCompared with NC group rats, the HR, CI, LVESP and LVEDP of MC group rats significantly increased, wCIle±dp/dtmax decreased significantly (P<0.05);Compared with MC group rats, the cardiac function parameters of treatment groups rats significantly were improved;Compared with other treatment groups rats, the cardiac function of XFC group rats were improveed better.3.2.5 XFC' effect on body mass, paw swelling, AI of AA ratsThe body mass of AA rats in XFC group was significantly CIgher than that in MC group, MTX group and TPT group, wCIle compared with NC group, there was no significant difference (P>0.05), the body mass of AA rats in MTX group were significantly lower than that in NC group (P<0.01), wCIch means, XFC has no adverse effect on body mass of rats.paw swelling, AI of AA rats in other treatment groups were significantly lower than that of MC.group (P<0.01 or P<0.05), the treatment groups showed no significant difference (P>0.05), wCIch means, XFC can reduce paw swelling, AI of AA rats, just as MTX and TPT.3.2.6 XFC's effect on NEI network of AA ratsCompared with NC group rats, ET-1,TNF-α, BNP, IL-17, IL-6,5-HT of rats in MC group were significantly CIgher (P<0.01), and CGRP, CD4+CD25+Treg, Foxp3 of rats in MC group decreased significantly (P<0.01);Compared with MC group rats, the indexes of NEI network by each treatment groups were improved to some extent;Compared with other treatment groups, XFC group can better improve the detection of the various indicators of NEI network. 3.2.7 XFC' effect on myocardial structureAccording to the test results of expression levels of heart tissue pathology, ultrastructure and cardiac tissue MMP9, TIMP1, we found that:Compared with NC group rats, cardiac muscular tissue of MC group rats has obvious lesion.Compared with MC group rats, the myocardial lesions of rats in other treatment groups were a little lighter;Compared with other treatment groups rats, the myocardial structure of XFC group rats were improved more apparently. 4 Conclusion4.1 AA rats have cardiac function change;4.2 AA rats have NEI network dysfunction;4.3 the cardiac function change of AA rats is closely related to dysfunction of NEI network4.4 XFC shows good regulation and control over general signs, cardiac function, NEI network index, myocardial structure of AA rat.5 Discuss the effect of XFC on improvement of cardiac function of rats based on NET network5.1 XFC regulates heart rate by reducing the release of 5-HT and DA.5.2 XFC, by regulating the balance of ET-1 and CGRP, improving myocardial ischemia and proliferation of myocardial cells, to maintain normal cardiac function.5.3 XFC can reduce the expression level of BNP and cardiac BNPmRNA in serum, and improve cardiac function of AA rats.5.4 XFC has anti-immune and anti-inflammatory effect, it can delay myocardial damage and improve cardiac function through upregulating CD4+CD25+Treg, promoting Foxp3 expression, suppressing generation of inflammatory cytokine, and inCIbiting autoimmune T cell activation.