左归丸联合温和灸调节骨质疏松症模型大鼠OPG/RANKL轴的作用研究

Effects of Zuogui Pills combining mild-warm moxibustion on OPG/RANKL of osteoporosis model rats

目的:观察左归丸联合温和灸对骨质疏松症模型大鼠OPG/RANKL轴的影响。方法:雌性SD大鼠48只,随机分为对照组、模型组、左归丸组、左归丸+温和灸组,每组12只。除对照组外,其余大鼠采用手术完整摘除双侧卵巢法进行造模,对照组仅开腹后缝合,饲养3个月后每组大鼠随机取1只作病理组织学观察判断造模成功。左归丸组予左归丸灌胃,左归丸+温和灸组予左归丸灌胃+"长强穴"艾灸治疗,对照组和模型组予0.9%氯化钠溶液灌胃。治疗90d后,骨密度测定观察大鼠股骨近端及第2腰椎骨密度变化,RT-PCR检测大鼠骨组织VPAC2、OPG和RANKL mRNA的表达水平。结果:(1)与对照组比较,各组大鼠平均股骨近端密度、第2腰椎骨密度均显著减少(P<0.05,P<0.01)。与模型组比较,左归丸组、左归丸+温和灸组大鼠平均股骨近端密度、第2腰椎骨密度均显著增加(P<0.05,P<0.01);与左归丸组比较,左归丸+温和灸组大鼠平均股骨近端密度、第2腰椎骨密度均增加(P<0.05)。(2)与对照组比较,模型组大鼠骨组织VPAC2 mRNA、RANKL mRNA表达显著上调,OPG mRNA表达显著下调,OPG/RANKL比值显著降低(P<0.05,P<0.01);与模型组比较,左归丸组、左归丸+温和灸组VPAC2 mRNA、RANKL mRNA显著下调,OPG mRNA表达显著上调,OPG/RANKL比值显著升高(P<0.05,P<0.01);与左归丸组比较,左归丸+温和灸组大鼠骨组织OPG mRNA表达显著上调,RANKL mRNA表达显著下调,OPG/RANKL比值显著升高(P<0.05)。结论:左归丸联合温和灸对骨质疏松症模型大鼠的骨密度下降具有一定的改善作用,能通过上调OPG mRNA,下调大鼠骨组织VPAC2、RANKL mRNA的表达,达到治疗骨质疏松症的目标。

Objective： To investigate the effect of Zuogui Pills combined with mild-warm moxibustion on osteoporosis model rats. Methods： Forty-eight female SD rats were randomly divided into four groups： control group（Group C）, model group（Group M）, Zuogui Pills group（Group Z）, and Zuogui Pills combining mild-warm moxibustion group（Group Z＋W）. Expect for the control group, rats from Group M, Group Z and Group Z＋W were completely excised bilateral ovaries to establish osteoporosis model, while Group C only had laparotomy and suture. One rat from each group was randomly picked for histopathology observation to decide whether the model was successfully established 3 months after the surgery. Group Z was administered with Zuogui Pills orally, Group Z＋W was administered with Zuogui Pills orally and mild-warm moxibustion on changqiang（GV1）, while group C and group M were given the same amount of saline. After 3 months＇ treatment, all rats were taken for bone densitometry of proximal femur and the second lumbar vertebra. RT-PCR was used to observe the mRNA expression of vasoactive intestinal polypeptide receptor 2（VPAC2）, osteoprotegerin（OPG） and receptor activator for nuclear factor-κB ligand（RANKL） of rat bone tissues. Results：（1）Compared with Group C, bone density reduced in every other groups（P〈0.05, P〈0.01）. After treatment, compared with group M, bone density in both Group Z and Group Z＋W decreased（P〈0.05, P〈0.01）. Compared with Group Z, bone density in Group Z＋W increased（P〈0.05）.（2）Compared with Group C, VPAC2 mRNA expression in group M increased（P〈0.05, P〈0.01）, OPG mRNA expression reduced, RANKL mRNA expression increased and OPG/RANKL reduced（P〈0.01）. Compared with Group M, expression of VPAC2 mRNA reduced in both Group Z and Group Z＋W（P〈0.05, P〈0.01）, OPG mRNA expression increased, RANKL mRNA expression reduced while OPG/RANKL increased（P〈0.05, P〈0.01）. Compared with Group Z, OPG mRNA expression in Group Z＋W increased, RANKL mRNA expression reduced and OPG/RANKL increased（P〈0.05）. Conclusion： Zuogui Pills combined with mild-warm moxibustion has a certain effect on decreased bone density of osteoporosis model rats, could increase expression of OPG mRNA, and reduce expression of VPAC2, RANKL mRNA in rat bone tissues to treat osteoporosis.