| Abelmoschus manihot (L.) Medic is a well-known and widely distributed traditional herbal medicine in China, which was first recorded in Jiayou Ben Cao. In the past, Abelmosehus Manihot was served as "an important drug in treatment of various malignant sore, cell ?Lites and burns". Modern researches have indicated that total flavone of A. manihot (L.) Medic (TFA) has protective effects on heart and brain injury caused by ischemia. This present paper is to investigate the possible effects and mechanisms of TFA on ischemic injury in vivo and in vitro.Part One:TheoryAfter ischemia occurs, anaerobic glycolysis takes the place of aerobic metabolism, thus large amounts of glycolysis products generate, they further lead to tissue injury which is charactered by tissue edema, inflammatory infiltration and muscle fiber necrosis. Angiogenesis is a critical biological process, which play a key role in repairing the ischemia tissue and improving the function. Under the pathological conditions like inflammation and trauma, the genes of angiogenesis-related growth factors express and induce endothelial cells of capillaries and ven ? Les rapidly into the division cycle, and proliferated vase ? Lar endothelial cells which participate in angiogenesis play a important role in treating ischemia cardio-cerebrovasc ? Lar disease, limb ischemia, wound repair, as well as tissue engineering. Recently, increasing studies have been focused on the effects of inducing agents and factors associated with angiogenesis on ischemia. And mountains of animal experiments and clinical trials proved that therapeutic angiogenesis may be a promising therapeutic strategy. Chinese traditional medicine treatment of ischemia disease has a long history, and it has lots of good quality, including rich clinical experience and favorable price. A certain dosage of TFA has a significant effect on increasing the proliferation, migration, and differentiation, so it has the capacity to promote angiogenesis. Moreover, endothelial cells are derived from endothelial progenitor cells (EPCs) which are one of three types of stem cells found in bone marrow. To improve the antioxidant ability of EPCs, inhibit apoptosis and promote EPCs to transform into mature endothelial cells are important mechanisms for the treatment of ischemic injury.Part Two:Experimental StudyTo study anti-inflammation effect of TFA on limb ischemia model of rats. After 4 weeks of treatment with TFA, the limb ischemia rats were sacrificed. Using flow cytometry to assess the alteration of reg ? Latory T cells (Tregs) in model group and TFA group. In addition, expression level of tumor necrosis factor (TNF) was measured by enzyme linked immunosorbent assay (ELISA) and RT-qPCR. These res ? Lts shown that TFA decreased the expression of TNF to inhibit cascade reactions induced by inflammation. The res ? Lts of this section provide evidences of the protective effect of TFA on ischemia issue through reg ? Lation of Tregs.After isolation, c?Lture and characterization, treat EPCs with TFA, study the protective effect of TFA on oxidative damage induced by reactive oxygen species (ROS). The levels of apoptosis-related proteins were increased in the EPCs following hydrogen peroxide (H2O2) treatment. However, western blotting res ? Lts revealed that TFA down-reg ? Lated phosphorylated caspase-3 and up-reg ? Lated BAK. These res ? Lts suggested that TFA possessed the ability of anti-apoptosis. Therefore, we hypothesized that TFA co ? Ld protect issue from ischemia in overall level though its anti-apoptosis effect on EPCs.To study the effects of TFA on limb ischemia model of rats. Pathological slides revealed that TFA ameliorated limb injury level caused by ischemia. Biochemical assays res?Lts which were in accordance with that in vitro shown that the level of ROS in serum decreased, and the level of phosphorylated caspase-3, while the level of Bcl-2 significantly down-reg?Lated. These res ? Lts indicated that TFA played a positive role in suppressing limb aschemia.To investigate the protective effects of TFA on myocardial ischemia/reperfusion injury model of rats. This study found that TFA ameliorated myocardial ischemia/reperfusion injury by inhibiting inflammatory reactions induced by oxidative stress and suppressing activation of inflammasome NLRP3.This present study suggests:1. In limb ischemia model of rats, the treatment with TFA significantly depresses the inflammatory responses of injury sites which indicated by down-reg ? Lation of inflammatory factor TNF and improving the ratio of Tregs.2. TFA protect EPCs though its anti-apoptosis effect.3. In vivo, TFA effectively improve the situation of limb ischemia injury by inhibiting apoptosis of VECs and promoting angiogenesis.4. TFA inhibits the activation of NLRP3 in myocardial ischemia/reperfusion injury model of rats. |
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