| Acute spinal cord injury(ASCI) manifests injury on white and graymatter in pathology, which results in primary injury and secondary lesion.The secondary lesion is a complicate cascade reaction several minutes todays after primary injury, and the immune inflammation reaction afterASCI plays a important role in secondary lesion. The immuneinflammation reaction after ASCI can result in the reside and adjacentnerve cells death and apoptosis, at the same time the micro environment ininjury spinal cord exits negative factors which can inhabit neural stem cellproliferation, and it is the main reason that recover bad after ASCI. Withstudy further on the mechanism of ASCI, the secondary lesion is realizedcould be intervened. It can protect residue spinal cord and promote thefunction of spinal cord recovery. The tetrandrine(TET) is the effectiveingredients of Radix Stephaniae Tetrandrae. A lot of research prove it canagainst inflammation and lipid peroxidation, prevent cell injury, improvemicrocirculation, and antagonize the Ca2+.Our study discuss if the TET could promote the function of spinal cord recovery through adjust theinflammation factors, inhabit the cell apoptosis and infect on neural stemcell proliferation, and provide theory proof for TET treat ASCI.PART ONE: EFFECT OF TETRANDRINE ON CELL APOPTOSISFOLLOWING ACUTE SPINAL CORD INJURY IN RATSObjective:To observe the influence of tetrandrine on cell apoptosisfollowing spinal cord injury in rats.Methods:114SD rats were randomly divided into three groups:sham group(group A),control group(group B),methylprednisolone(MP)group(group C) and tetrandrine(Tet) group(group D).The modified Allenmethod was adopted to make the models of T9spinal cord injury in B andC and D group. BBB score was recorded at8h,1d,3d,,7d,14d,28drespectively. The samples were taken from the spinal cord injury sites at8h,1d,3d,7d,14d,28d tested by HE staining for morphology and detectedthe expression levels of Bcl-2、Bax protein by immunohistochemistryassay. The semi-quantitative analysis for apoptosis nerve cells can begained by the flow cytometry (FCM).Results:The BBB score of group C and D were significantly higherthan that of group B at7,14and28day time point. HE stain showed thespinal cord of group C and D has fewer necrosis than that of group B. Thenumber of Bcl-2postive cells in group C and D were significantly higherthan those of group B at each time points. However the number of Baxpostive cells in group C and D were significantly lower than those of groupB at each time points.Conclusions: Tetrandrine can prevent neurons from apoptosis andpromote the nerve function recover by inhibiting the expression of Bax and promoting the expression of Bcl-2.PART TWO: EFFECT OF TETRANDRINE ON RELEASE OFINFLAMMATORY FACTOR FOLLOWING ACUTE SPINALCORD INJURY IN RATSObjective:To observe the influence of tetrandrine on release ofinflammatory factor following spinal cord injury in rats.Methods:78SD rats were randomly divided into:shamgroup,control group, Tet group.The modified Allen method was adopted tomake the models of T9spinal cord injury in control group and Tet group.The samples were taken from the spinal cord injury sites at6h,12h,1d,3d,5d,7ddetected the contents of TNF-α,IL-1β,IL-6,IL-10,NF-kBby using enzyme-linked immunosorbent assay.Results: The expression of TNF-α, IL-1β, IL-6, IL-10, NF-kB ininjury control group and Tet group at6h,12h,1d,3d,5d,7d After ASCI washigher than that in sham operation group, has obviously increased at6hafter ASCI, peaked at24h-72h, began to decrease at5day after ASCI,.Theexpression of IL-10in Tet group was higher than the control group,but theexpression of TNF-α, IL-1β, IL-6, NF-kB is lower than the control group.Conclusion:The content of TNF-α,IL-1β,IL-6,IL-10,NF-kB wasincrease in the spinal cord. Tetrandrine is able to improve local microenvironment and promote the nerve function recover by increasing theexpression of IL-10and decreasing the expression of TNF-α,IL-1 β,IL-6,NF-kB in the spinal cord injury.PART THREE: EXPERIMENTAL STUDY ON EFFECT OFTETRANDRINE IN THE ENDOGENOUS NEURAL STEM CELLPROLIFERATION AND DIFFERENTIATION AFTER SPINALCORD INJURY IN RATSObjective: Discuss the effects of tetrandrine on the endogenousneural stem cell proliferation and differentiation after spinal cord injury inrats.Methods:78rats were randomly divided into3groups: injuriedgroup (n=36), Tet-treated group (n=36),sham-operated group (n=6).Injuried group and Tet-treated group damage spinal cord with Allen ’scombat modeling method.30minutes after damage, Tet was injected with adosage22.5mg/kg in Tet group at30minutes,24h and48h after ASCI, andthe same dosement of saline was injected in injuried group as control. Thesamples were dissected from the spinal cord injury sites at1d,3d,1w,2w,3w,4w and detected the expression of BrdU and nestin byimmunofluorescence staining.Results: A little nestin positive cells and brdU positive cells werefound in injuried group and Tet-treated group at1day after injury. A largenumber of positive cells were found in both groups at one week after injuryand reached the peak which lasted for2weeks and then decreasedgradually. The expression of Nestin positive cells and brdU positive cells decreased significantly at4weeks after injury. Compared with the shamoperation group, the number of Nestin positive cells and brdU positive cellswas more in injuried group and Tet-treated group at1d,3d,1W,2W,3W afterinjury. The expression was higher in Tet-treated group than injuried groupat1d,3d,1w,2w,3w after injury and had no difference at4weeks afterinjury.Conclusion: It was concluded that tetrandrine could increase thenumber of Nestin positive cells, brdU positive cells and endogenous neuralstem cells.It is beneficial to the recovery of spinal cord injury in rats. |
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