Experimentlal Study Of Caffeic Acid Combined Hydrogen-rich Normal Saline To Treat Acute Radiation Disease Of Bone Marrow

ObjectiveAccording to Applying the treatment of the Caffeic acid(CFA) combined withhydrogen rich normal saline(HNS) to mice which have acute radiation disease(ARD) ofbone marrow, we observed their effect on marrow hematopoietic recovery of radiationmice, in order to investigate the mechanism of Caffeic acid combined with HNS treatingARD of bone marrow and explore a new way for the clinical treatment ARD of bonemarrow.MethodsWe construct the ARD of bone marrow mice model by simulating the ionizingradiation environment in this study. The research was divided into3parts. The part I:Study on the role of CFA treatment ARD of bone marrow.180SPF inbred line male andfemale BALB/c mice were randomly divided into9groups,20rats in each group: theSham group (no radiation, no filling medicine), the CFA negative control group (afterirradiation carboxymethyl cellulose gavage), the CFA positive control group(intraperitoneal injection of G-CSF+IL-11after irradiation), the CFA prevention group(irradiation and as7days before the start of CFA in low, middle, high dose administered,nameing the CFA low dose prevention group, the CFA middle dose prevention group andthe CFA high dose prevention group), The CFA treatment group (according to begin theday after irradiation and CFA in low, middle, high dose administered). The CFA treatmentgroup (according to CFA in low, middle, high dose administered, naming the CFA low dosetreatment group, the CFA middle dose treatment group and the CFA high dose treatmentgroup). The part II: Study on the role of HNS treatmenting ARD of bone marrow.80SPFinbred BALB/c mice were randomly divided into4groups according to the above method,10male and10female of each group: the HNS Sham group (without irradiation, onlyintraperitoneal injection of saline), the HNS negative control group (irradiation+ intraperitoneal injection of saline), The HNS prevention group (irradiation+7days beforeexposure intraperitoneally injected HNS), the HNS treatment group (irradiation+the dayafter intraperitoneal injection of hydrogen-rich normal saline). The part III: Study on therole of CFA combined with HNS treatment ARD of bone marrow. Taking merit-based theCFA high doses prevention group from the first part, and taking merit-based the the HNSprevention group constructed the combination group,20in each group. Body weight ofmice was measured every week, each taking12respectively after irradiation d4, d14fromeyeball blood, hematopoietic positive regulatory factors as G-CSF, TPO, IL-1β andhematopoietic negative regulator factor as TNF-α levels and oxidative damage markers:MDA, SOD content and activity were measured, and the content of DNA damagemarker:8-OHdG; in d14,From the left femur, BMNC parallel counting was measured,CFU-GM and CFU-MK in vitro and were counted; In d14, we removed spleen, weightedit and measured SI and CFU-S counts. The remaining mice in each group were irradiatedday d0, after irradiation d4, d7, d14, d21, blood samples were collected after venous plexusd28eye peripheral WBC, HGB, PLT count.Results1. Three experimental parts all show that the mice WBC, HGB, PLT significantlydecreased after4.5Gy X-ray irradiation. There was a significant difference (P <0.01)compared with the Sham group, and previous irradiation decreased by more than1/3,indicating a successful bone marrow type ARD model was made.2. After irradiation d4, d7, d14, d21, d28, WBC, HGB, PLT counting of the partⅠofexperiment: The CFA negative control group <The CFA prevention and treatment groups<The CFA positive control group (P <0.01). Between The CFA prevention group andtreatment group, the CFA high dose prevention group had the strongest effect(P<0.01); ThePart II: The HNS negative control group <The HNS group <The HNS prevention group(P<0.05); The part III: The HNS prevention group <The CFA high dose prevention group <the combination group (P <0.01).3. After irradiation d14, BMNC, CFU-GM, CFU-MK, CFU-S and SI in mice of thepart I of experiment: the CFA negative control group <the CFA prevention and treatmentgroup (P <0.01). Between the CFA prevention group and treatment group, the CFA highdose prevention group had the strongest effect(P <0.01). The part II: the HNS negativecontrol group <the HNS treatment group <the HNS prevention group (P<0.05); The partIII: The HNS prevention group <the CFA high dose prevention group <The combinationgroup (P <0.01). 4. After irradiation d4and d14, G-CSF, TPO, IL-1β of the partⅠof experiment: TheCFA negative control group <The CFA prevention and treatment groups <The CFApositive control group (P <0.01). Between The CFA prevention group and treatment group,the CFA high dose prevention group had the strongest effect (P <0.05). In d4andd14,TNF-α of the CFA negative control> the CFA positive control group> the CFAhigh-dose prevention group (P <0.01); The partⅡ: In d4and d14, G-CSF, TPO and IL-1βof the HNS negative control group <the HNS treatment group <the HNS preventiongroup (P <0.05) and TNF-α of the HNS negative control group> the HNS treatment group> the HNS prevention group (P <0.05). The part III: In d4and d14,G-CSF, TPO, IL-1β ofthe HNS prevention group <the CFA high dose prevention group <the combination group(P <0.05) and TNF-α of the HNS prevention group> the CFA high doses prevention group> the combination group(P <0.05). In the Part I and II, In d4, IL-1β of each irradiatedgroup was significantly higher than the Sham group(P <0.01). However, no statisticallysignificant difference among the irradiated groups(P>0.05). In d14, IL-1β of the CFAnegative control group <the CFA prevention and treatment groups <the CFA positivecontrol group (P <0.05). Between CFA prevention group and treatment group, the CFAhigh dose prevention group had the strongest effect(P <0.05).5. In d4and d14, MDA and8-OHdG of the partⅠof experiment: the CFA negativecontrol> the CFA positive control group> the CFA prevention and treatment group (P<0.01) and SOD of the negative control group CFA <the CFA positive control group> theCFA prevention and treatment group (P <0.01). There was a significant difference in theCFA positive control group, the CFA prevention and treatment groups, and the high doseof CFA prevention group had the strongest effect (P <0.01). the partⅡ: In d4and d14,MDA and8-OhdG of the HNS negative control group> the HNS treatment group> theHNS prevention group (P <0.01) and SOD of the HNS negative control group <the HNStreatment group <the HNS prevention group (P <0.01). the part III: In d4and d14, MDAand8-OHdG of the combination group <the HNS prevention group <the CFA high doseprevention group (P <0.01) and SOD of the combined group>the HNS prevention group>the CFA high dose prevention group (P <0.05).6. In the CFA prevention and treatment groups, HNS prevention and treatment groupand combined group，the G-CSF、TPO and IL-1β on fourteenth day increased comparedto the fourth day, while the TNF-α、MDA、SOD and8-OHdG on fourteenth day werelower than the fourth day, the difference was sta tistically significant(P <0.01). 7. The CFA prevention and treatment groups for blood cell count, BMNC, CFU-GM,CFU-MK, CFU-S, SI, hematopoietic regulatory factors, MDA, SOD and8-OHdG showedsignificant dose-dependent and the effect will become stronger with the dose increasing,the difference was statistically significant (P <0.05).ConclusionsCaffeic acid and hydrogen-rich normal saline can prevent and treat the ARD of bonemarrow, and have effect on hematopoietic recovery. The above two kinds of effects have asynergistic effect. The mechanism of action may be:1. CFA and hydrogen-rich normal saline can promot bone marrow by increasing thehematopoiet ic positive regulatory factors as G-CSF, TPO, IL-1β, and suppressing thenegative regulat ory of hematopoietic factors as TNF-α of the radiation mouse;2. CFA and hydrogen-rich normal saline can promote bone marrow recovery byprotectivingand stimulating the bone marrow stem/progenitor cell proliferation;3. CFA and hydrogen-rich normal saline can protect spleen and promot the spleenextramedullary hematopoiesis;4. CFA and hydrogen-rich normal saline can protect bone marrow cells by reducingoxidative damage and DNA damage, enhance the activity of SOD.