桑椹来源的寡核苷酸显著改善小鼠骨质疏松症

Mulberry-Derived Oligonucleotides Significantly ImprovedOsteoporosis in Mice

本试验旨在探究桑椹来源的寡核苷酸ASON-OP对小鼠骨质疏松症的改善作用。将30只雌性小鼠随机分为5组,分别为假手术组、模型对照组、阳性药物组、ASON-OP组和ASON-NC组(ASON-NC为无义序列,作为阴性对照),每组6只。除假手术组小鼠以外,其他4组小鼠均进行双侧卵巢切除(OVX),构建骨质疏松症小鼠模型。建模成功后,继续饲养1周,而后进行给药处理。假手术组和模型对照组每天灌胃300μL生理盐水;阳性对照组皮下注射特立帕肽,剂量为80μg/(kg BW·d),1周给药5次;ASON-NC组每天灌胃ASON-NC溶液,剂量为2 mg/(kg BW·d);ASON-OP组每天灌胃ASON-OP溶液,剂量为2 mg/(kg BW·d)。连续给药4周后,检测小鼠骨组织微结构、骨转化指标等。结果显示:与模型对照组和ASON-NC组相比,连续4周灌胃寡核苷酸ASON-OP显著增加了OVX小鼠的骨密度(P<0.05);显著提高了OVX小鼠血清成骨因子骨钙素(OCN)含量(P<0.05),显著降低了血清破骨因子Ⅰ型胶原C末端肽(CTX-1)含量(P<0.05);显著提高了OVX小鼠股骨中促进成骨相关基因Runt相关转录因子2(Runx2)、肝/骨/肾型碱性磷酸酶(Alpl)、骨钙素(Bglap)的mRNA相对表达量(P<0.05)。综上所述,桑椹来源的寡核苷酸ASON-OP具有显著的抗骨质疏松症作用。

The objective of this experiment was to explore the improvement effects of ASON-OP,an oligonucleotide derived from the mulberry.Thirty female mice were randomly divided into five groups:sham surgery group,model control group,positive control group,ASON-OP group,and ASON-NC group(ASON-NC is a nonsense sequence,as negative control),with six mice in each group.Except for the mice in the sham surgery group,bilateral ovariectomy(OVX)was performed on mice in the other four groups to induce the osteoporosis mouse model.After successful modeling,continued to feed for 1 week,and then administered the drug.Mice in the sham surgery group and model control group were given 300μL normal saline per day by gavage;mice in the positive control group were injected subcutaneously with teriparatide at the dose of 80μg/(kg BW·d),five times a week;mice in the ASON-NC group were given ASON-NC solution by gavage,2 mg/(kg BW·d);mice in the ASON-OP group were given ASON-OP solution by gavage,2 mg/(kg BW·d).The mice were continuously administered for 4 weeks,and their bone tissue microstructure and bone turnover markers were measured.The results showed as follows:compared to the model control group and ASON-NC group,mice gavaged with oligonucleotide ASON-OP for 4 weeks significantly increased the bone mineral density of OVX mice(P<0.05);it significantly increased the serum content of osteocalcin(OCN)as a bone formation marker of OVX mice(P<0.05),and significantly decreased the serum content of typeⅠcollagen C-terminal telopeptide(CTX-1)as a bone resorption marker(P<0.05);it also significantly increased the mRNA relative expression levels of the bone formation-related genes Runt-related transcription factor 2(Runx2),alkaline phosphatase,liver/bone/kidney(Alpl)and bone gamma-carboxyglutamate protein(Bglap)in the femur of OVX mice(P<0.05).In conclusion,ASON-OP,an oligonucleotide derived from mulberry,exhibits significant anti-osteoporotic effect.