ABSTRACT
AIM:To investigate the protective effect of pinacidil postconditioning on rat myocardium suffering ischemia/reperfusion injury by mitochondrial proteomics .METHODS: Langendorff apparatus was used to establish the model of myocardial ischemia/reperfusion injury .Sprague-Dawley rats were randomly divided into 2 groups:pinacidil post-conditioning group (Pina group) and ischemia/reperfusion injury group (I/R group).After 20 min of perfusion with K-H solution, the perfusion was suspended for 40-min (global ischemia) follow by 60 min of reperfusion in I/R group.In Pina group at the end of 40 min global ischemia , the isolated hearts were perfused with K-H solution containing pinacidil ( 50μmol/L) for 2 min followed 58-min perfusion with regular K-H solution.Total proteins extracted from the mitochondria were applied to the two-dimensional gel electrophoresis (2-DE).The differentially expressed protein spots over 2 times were evaluated by a software .Then they were subjected to in-gel digestion , and analyzed by spectrometry .RESULTS:The expression levels of NDUFA10, NDUFS2 and NDUFV2 were elevated but those of IDHA and ECH 1 were decreased in Pina group compared with I/R group.Interestingly, 2 spots in the 2-DE map were identified as ATPase subunit δ.The ex-pression levels of one spot was elevated , while the other was decreased .CONCLUSION:Pinacidil postconditioning may decrease the degree of increased expression levels of NDUFA 10, NDUFS2 and NDUFV2, promote the expression of IDHA and ECH1, and induce the phosphorylation of ATPase subunit δ, which may be related to the protective mechanism of pinacidil postconditioning .
ABSTRACT
[Objective]To investigate the effect of reconstituted bone xenografts(RBX) on tendon-to-bone healing by means of imaging,histological and biomechanical studies.[Method]Anterior cruciate ligament(ACL) reconstruction was performed bilaterally in 25 skeletally mature rabbits using long digital extensor tendon grafts.RBX were implanted into the treated knee of each rabbit,with the contralateral knees as controls.Every three rabbits were killed at 2,6 and 12 weeks postoperatively for routine histological studies.The samples were processed through Micro CT and subsequent HE and toludine blue staining.The remaining 16 rabbits were sacrificed at 6 and 12 weeks.Their femur-tendon graft-tibia complexes were harvested for subsequent mechanical testing.[Result]At 6 and 12 weeks after operation,the values of BMD in the RBX group were higher than those in the control group(P
ABSTRACT
[Objective]To study the differentiation of adipose-derived adult stem cells(ADASCs)into chondrocytes induced by TGF-?1 in the presence of different concentrations of serum in vitro.[Method]Adipose tissue-derived adult stem cells were isolated from rabbits subcutaneous adipose tissue using enzymatic digestion.The adipose tissue was minced and digested with collagenase type I,and the released cells were collected by density centrifugation and then placed in culture.After being passaged three times,ADASCs were induced to differentiate into chondrocytes by chondrogenic culture media containing 1% or 10% of FBS for 2 weeks.MTT,alkaline phosphatase toluidine blue staining and type Ⅱ collagen immunohistochemistry staining were then tested in both groups.Gray value was analyzed by Leica patho-image analysis system to observe if there was any difference in differentiative status between the two groups.[Result]MTT showed that the cells of 10% FBS group had higher proliferation activity than those of 1% FBS group(P
ABSTRACT
[Objective]To observe the differentiation of adipose derived stem cells(ADSCs)into osteogenic cells in rabbit in vitro.[Method]ADSCs were prepared by collagenase Ⅰ digestion of subcutaneous fat from the nape of Japanese white rabbit after being excised and finely minced.Cells were identified by stro-1 immunocy to chemistry,and collagen Ⅰ immunocytochemistry,alkaline phosphatase assay and Von Kossa stain after osteogenic differentiation was performed.[Result](1)ADSCs were stro-1 positive.(2)The expression of collagen Ⅰ,alkaline phosphatase and calcium deposit of ADSCs were all positive under the influence of osteogenic differentiation medium.[Conclusion]ADSCs have similar characteristics to bone marrow stromal cells,but are much more easier to have high number upon harvest and bring less pain will occur during taking ADSCs than taking BMDCs.
ABSTRACT
Objective To observe the expression of core-binding factor ?1 (Cbf?1) in aging osteoblast, and try to provide new mechanisms of Chinese medicine on bone formation. Methods Rat osteoblast from different age donors (6 months and 16 months) were isolated and cultured by the tissue-fragment-migrating-growth method. The biological characteristic was observed by phase-contrast microscope and identified with mineralizing nodule staining. The expression of Cbf?1 was examined by RT-PCR and immunohistochemistry. Results RT-PCR and immunohistochemistry showed that the expression of Cbf?1 was upregulated by Gubenzhuanggu and Jinkuishenqi, and downregulated by Zhibaidihuang in aging osteoblast, but the effect was different on the Cbf?1 expression in Bushenyijing. Conclusion Buyang Chinese medicine among Bushen Chinese medicine promoted bone formation.