TCM Combined With Bone Marrow Origin Stem Cell Transplantation For Cerebral Ischemia-reperfusion Injury In Rat Experimental Study

Purpose:Observe the treatment efficiency of transplantation allogeneic Bone Mesenchymal StemCell(BMSC) or Bone Marrow Mononuclear Cell(BMNC) through subarachnoid combinedwith traditional Chinese medicine(TCM) for the ischemic cerebrovascular disease rats.Explore the mechanisms of bone marrow origin stem cell transplatation combined with TCMin the treatment of ischemic stroke. Analyse the significance of the nerve growth factor(NGF)in cerebrospinal fluid (CSF). Therefore that provides a reliable and valuabletheoretical basis and scientific evidence for the comprehensive treatment plan in clinicallyapplicating TCM with BMSC and BMNC transplantation in the treatment of ischemiccerebrovascular disease.Materials and Method：BMSCs and BMNCs were separate and cultivated.140adult Wistar rats were dividedrandomly into7groups(n=20):group1:normal group, group2:pretent surgery group,group3:model group, group4:BMNC group, group5: BMNC&TCM group, group6:BMSCgroup, group7: BMSC&TCM group. Group3,4,5,6,7were performed (2hours) middlecerebral arterial occlusion (MCAO).100μl0.01M PBS(group3),BMNCS (group4、5，2×107)or BMSCS(group6、7，2×107) were injected into rats’ subarachnoid after24h ofMCAO corresponded.Group5and7were treated united with TCM on the transplantantionday(po.Qd.). Recovery standards of MCAO rat neurological function were analyzed withmNSS at the4th day and the28th day after transplantation. GDNF level in MCAO rat brainand CSF were analyzed with Elisa at the4th day and the28th day after transplantation. Results：1.The mNSS score of model group is decreased at the28thday4th day’s(P<0.05). Theresult suggest that cerebral ischemia-reperfusion can injure neurological function in rat.TheMCAO rat have the self-healing capability.Compared with model group,The mNSS scorechange of group4isn’t statistical significance at the4thand28thday(P>0.05). BMNCtransplantation doesn’t express the neurological repair capability. The mNSS score of group5decreased at the28thday(P<0.05). The mNSS score of group6and group7decreased atthe4thand28thday(P<0.05). The result suggest that group5,6,7express the neurologicalrepair capability. Compared with BMSC group, the mNSS score of group7decreased at the4thday(P<0.05). Compared with BMNC&TCM group, the mNSS score of group7decreasedat the4thday(P<0.05). The result suggest that BMSC transplantation combined with TCMexpress the advanced neurological repair capability compared with BMSC transplantation orBMNC transplantation combined with TCM.2.The GDNF level in brain of model group is increased at the28th day (P<0.05).Thelevel change of group3isn’t statistical significance at the4th day(P>0.05).The MCAO rat’scapability of adjusting environment within the brain is limited.The GDNF levels of group4,6are higher than the model group’s (P<0.05) at the4th and28th day.That express the raisedcapacity. The raised capacity of group BMSC are better than group BMNC at the28th day.The GDNF levels of group5,7are higher than the group4,6’s (P<0.05) at the4th and28th day.Compared with group7, The GDNF levels of group5is higher at the4th day(P<0.05).Itisn’t statistical significance at the28th day.3. The GDNF level in Cerebrospinal fluid(CSF) is lower than that in brain,but theStatistical results between groups are Convergence.Conclusion:1.Transplantantion of BMNCs express the neurological repair capability. Thetransplantantion united with TCM inprove the capability of neurological repair after cerebralischemia-reperfusion.The MCAO rats don’t appear adverse reactions after transplantantthrough subarachnoid.The Transplanting Methods through subarachnoid is a safe andeffective way to transplant stem cell. 2.Transplanting bone marrow origin stem cell through subarachnoid will increase theGDNF level in brain of MCAO rats. The transplantantion united with TCM inprove thecapability of the enhance.The enhance capability appear earlier,more powerful andprotracted.That reflect the advantage of transplantantion SC united with TCM.3.The test results in CSF could reflect the the metabolism of brain after SCtransplanting.That is helpful to realize the biological functions of SC and brainmicroenvironment after SC transplanting.