由Hedgehog信号通路探讨左归丸对卵巢切除大鼠骨质疏松症的影响  

To explore the mechanism of Zuogui Pill on osteoporosis induced by ovariectomy in rats based on Hedgehog signaling pathway

作　　者：王雨荷 刘红[1] 史婧儒 赵宏艳[1] 李艳[1] 王少君[1] 潘静华[1] 汪文来[2] 张立石[2] 刘梅洁[1] WANG Yuhe;LIU Hong;SHI Jingru;ZHAO Hongyan;LI Yan;WANG Shaojun;PAN Jinghua;WANG Wenlai;ZHANG Lishi;LIU Meijie(Beijing Key Laboratory of Traditional Chinese Medicine Basic Research on Prevention and Treatment for Major Diseases,Experimental Research Center,China Academy of Chinese Medical Sciences,Beijing 100700;Institute of Basic Theory for Chinese Medicine,China Academy of Chinese Medical Sciences,Beijing 100700)

机构地区：[1]中国中医科学院医学实验中心中医药防治重大疾病基础研究北京重点实验室,北京100700 [2]中国中医科学院中医基础理论研究所,北京100700

出　　处：《中国骨质疏松杂志》2023年第6期781-785,791,共6页Chinese Journal of Osteoporosis

基　　金：国家自然科学基金面上项目(82074290);全国中医药创新骨干人才培训项目(ZX2020002)。

摘　　要：目的由Hedgehog信号通路探讨补肾方药左归丸对卵巢切除所致骨质疏松症(osteoporosis,OP)大鼠的作用机理。方法选用雌性SD大鼠,随机分为空白对照组、假手术组、造模组。造模组切除双侧卵巢,假手术组不切除卵巢,仅切除卵巢周围少量脂肪组织。12周后,将造模组随机分为卵巢切除组(OVX组)、阳性对照组和左归丸组。阳性对照组灌服戊酸雌二醇混悬液,左归丸组灌服左归丸水煎液,OVX组灌服等体积纯水,给药13周后取材。通过Micro-CT检测各组骨密度(bone mineral density,BMD),免疫组化法(immunohistochemistry,IHC)检测各组胫骨骨髓基质细胞中Ihh、Ptch、Smo、Gli1、OPG、RANKL的蛋白表达。结果假手术组骨密度(bone mineral density,BMD)高于OVX组、阳性对照组、左归丸组,且OVX组BMD较阳性对照组、左归丸组更低;OVX组Hedgehog信号通路关键因子Ihh、Ptch、Smo、GLi1蛋白表达水平以及RANKL/OPG比值较假手术组明显增高;阳性对照组和左归丸组Ihh、Ptch、Smo、GLi1蛋白表达水平及RANKL/OPG比值均低于OVX组,但仍高于假手术组。结论左归丸可抑制卵巢切除所致的Hedgehog信号通路异常活跃,使其关键因子Ihh、Ptch、Smo、GLi1蛋白表达水平明显降低;进而降低RANKL/OPG比值,抑制破骨细胞(osteoclast,OC)活性,减少骨量流失。Objective To explore the mechanism of Zuogui Pill,a traditional Chinese medicine for tonifying the kidney,in treating ovariectomized osteoporosis in rats based on the Hedgehog signaling pathway.Methods Female SD rats were randomized into 3 groups:the blank control group,the sham operation group,and the model group.The ovaries of the model group rats were removed.In the sham operation group,only a small amount of fat tissue around the ovary was removed.After 12 weeks,the model group was randomly divided into 3 groups:the OVX group,the positive control group,the Zuogui Pill group.The positive control group was perfused with estradiol valerate suspension,the Zuogui Pill group was perfused with Zuogui Pill decoction,and the other groups were perfused with equal volume pure water.After 13 weeks,the rats were anesthetized and killed,their tibia and femur were taken.The Bone Mineral Density(BMD)of each group was detected by micro CT;The expression of the key factors of the Hedgehog signaling pathway,OPG and RANKL in bone marrow stromal cells of tibia was detected by immunohistochemistry(IHC).Results BMD in the sham operation group was higher than that in OVX group,the positive control group and the Zuogui pill group,and BMD in OVX group was lower than that in the positive control group and the Zuogui pill group.The expression levels of Ihh,Ptch,Smo,GLi1 and the ratio of RANKL/OPG in the OVX group were higher than that in the sham operation group;The protein expression levels of Ihh,Ptch,Smo,GLi1 and the ratio of RANKL/OPG in the positive control group and the Zuogui pill group were lower than those in the OVX group,but still higher than those in the sham operation group.Conclusion Zuogui Pill can inhibit the abnormal activity of Hedgehog signaling pathway caused by ovariectomy,and significantly reduce the expression level of key factors Ihh,Ptch,Smo and GLi1;Further,it can reduce the ratio of RANKL/OPG,inhibit the activity of osteoclasts(OC)and prevent bone loss.

关 键 词：中医中药 骨质疏松症 左归丸 HEDGEHOG信号通路 骨密度 

分 类 号：R228[医药卫生—中医基础理论] R285[医药卫生—中医学]