酒精对人脐静脉间充质干细胞治疗肾虚血瘀型骨质疏松症大鼠的影响

Effect of Alcohol on Human Umbilical Vein Mesenchymal Stem Cells in the Treatment of Kidney Deficiency and Blood Stasis Osteoporosis in Rats

目的:探讨酒精对人脐静脉间充质干细胞(hUV-MSCs)治疗肾虚血瘀型骨质疏松症(OP)大鼠的影响。方法:取雄性SD大鼠30只,采用酒精腹腔注射法制备肾虚血瘀型OP模型,造模成功后将SD大鼠随机分为酒精组、治疗组和对照组,每组10只。酒精组给予酒精[浓度20%,10 ml/(kg·次),1次/周]治疗,对照组在酒精组基础上给予hUVMSCs(1次/周,10^(7)个细胞/次)治疗,治疗组停止酒精注射但给予hUV-MSCs(1次/周,10^(7)个细胞/次)治疗,取SD大鼠10只腹腔注射法生理盐水作为假手术组,给予生理盐水[1次/周,10 ml/(kg·次)]治疗,均连续治疗8周。HE染色检测大鼠骨组织病理损伤(骨小梁面积),TUNEL染色检测观察骨组织细胞凋亡(TUNEL阳性细胞数),µCT检测骨体积分数(BVF)、骨小梁厚度(Tb.Th)和骨小梁分离距离(Tb.Sp),ELISA检测血清碱性磷酸酶(ALP)和抗酒石酸酸性磷酸酶(TRAP)水平。结果:酒精组、对照组、治疗组大鼠骨组织骨小梁面积、BVF、Tb.Th均低于假手术组(P<0.05),骨组织TUNEL阳性细胞数、Tb.Sp及血清ALP、TRAP水平均高于假手术组(P<0.05);治疗组与对照组大鼠骨组织骨小梁面积、BVF、Tb.Th均高于酒精组(P<0.05),骨组织TUNEL阳性细胞数、Tb.Sp及血清ALP、TRAP水平均低于酒精组(P<0.05);治疗组大鼠骨组织骨小梁面积、BVF、Tb.Th均高于对照组,骨组织TUNEL阳性细胞数、Tb.Sp及血清ALP、TRAP水平均低于对照组(P<0.05)。结论:酒精能明显降低hUV-MSCs治疗肾虚血瘀型OP大鼠的效果。

Objective:To investigate the effect of alcohol on human umbilical vein mesenchymal stem cells(hUV-MSCs)in the treatment of kidney deficiency and blood stasis osteoporosis(OP)in rats.Method:Thirty male SD rats were injected intraperitoneally with alcohol to establish the OP model of kidney deficiency and blood stasis.After making model,SD rats were randomly divided into alcohol group,treatment group and control group,with 10 rats in each group.The alcohol group was given alcohol[concentration 20%,10 ml/(kg·time),once a week],the control group was given hUV-MSCs(once a week,10^(7) cells/time)on the basis of alcohol group,the treatment group stopped alcohol injection but was given hUV-MSCs(once a week,10^(7) cells/time),10 SD rats were injected intraperitoneally with normal saline as sham-operation group,and were given normal saline[once a week,10 ml/(kg·time)],all were treated for 8 weeks.HE staining was used to detect pathological injury of bone tissue(bone trabecular area)in rats,TUNEL staining was used to detect the apoptosis of bone tissue cells(TUNEL positive cells),bone volume fraction(BVF),trabecular thickness(Tb.Th)and trabecular separation distance(Tb.Sp)were detected byµCT,the serum levels of alkaline phosphatase(ALP)and tartrate-resistant acid phosphatase(TRAP)were determined by ELISA.Result:The bone trabecular area,BVF and Tb.Th in the alcohol group,control group and treatment group were significantly lower than those in the sham-operation group(P<0.05),and the TUNEL positive cells and Tb.Sp of bone tissues,serum levels of ALP and TRAP were significantly higher than those in the sham-operation group(P<0.05).The bone trabecular area,BVF and Tb.Th in the treatment group and the control group were significantly higher than those in the alcohol group(P<0.05),and the TUNEL positive cells and Tb.Sp of bone tissues,serum levels of ALP and TRAP were significantly lower than those in the alcohol group(P<0.05).The bone trabecular area,BVF and Tb.Th in the treatment group were significantly higher than those in the control group,and the TUNEL positive cells and Tb.Sp of bone tissues,serum levels of ALP and TRAP in the treatment group were significantly lower than those in the control group(P<0.05).Conclusion:Alcohol can significantly reduce the effect of hUV-MSCs in the treatment of kidney deficiency and blood stasis OP rats.