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探讨骨质疏松发病过程中T淋巴细胞对骨髓间充质干细胞(bone marrow-derived mesenchymal stem cells,BMMSC)增殖分化的影响。选用健康雌性小鼠行双侧卵巢切除术(ovariectomy,OVX),建立绝经后骨质疏松模型。选用同一批次健康小鼠行双侧卵巢脂肪组织部分切除,建立假手术组(sham),Micro-CT确立模型成功建立。将sham组、OVX组、sham+anti-TNFα组、OVX+anti-TNFα组中T淋巴细胞与BMMSC共培养,ELISA检测sham组与OVX组T淋巴细胞上清液中TNF-α表达的差异,MTT法检测四组共培养体系中BMMSC生长曲线;成骨诱导后碱性磷酸酶和钙化结节茜素红染色法检测BMMSC成骨能力差异;RT-PCR检测小鼠BMMSC成骨相关基因Runx2、碱性磷酸酶(alkaline phosphatase,ALP)的表达。结果显示,与sham组相比,OVX组中BMMSC的增殖受到了抑制,成骨分化减弱(P<0.05),OVX anti-TNF-α刺激组较OVX组增殖显著升高(P<0.05),成骨分化能力显著增强(P<0.05)。以上结果证明,在雌激素缺乏下的T淋巴细胞能影响BMMSC增殖及成骨分化能力,这可能与T淋巴细胞表达TNF-α增强相关。
To investigate the effect of T lymphocytes on the proliferation and differentiation of bone marrow-derived mesenchymal stem cells (BMMSCs) during the pathogenesis of osteoporosis. Healthy female mice were selected to undergo ovariectomy (OVX) to establish a model of postmenopausal osteoporosis. Select the same batch of healthy mice bilateral ovarian adipose tissue partial resection, the establishment of sham group (sham), Micro-CT model established successfully established. T lymphocytes from sham group, OVX group, sham + anti-TNFα group and OVX + anti-TNFα group were co-cultured with BMMSC. The difference of TNF-αexpression in T lymphocyte supernatant between sham group and OVX group was detected by ELISA. MTT method was used to detect the growth curve of BMMSC in four co-culture systems; osteogenic differentiation of BMMSCs was detected by alkaline phosphatase and calcified nodules alizarin red staining after osteogenic induction; Runx2, Alkaline phosphatase (ALP) expression. The results showed that compared with the sham group, the proliferation of BMMSC was inhibited and osteogenic differentiation was decreased in OVX group (P <0.05). Compared with OVX group, the proliferation of OVX anti-TNF-α group was significantly increased (P <0.05) Osteogenic differentiation was significantly enhanced (P <0.05). The above results demonstrate that T-lymphocytes under estrogen deficiency can affect the proliferation and osteogenic differentiation of BMMSCs, which may be related to the increased expression of TNF-α by T lymphocytes.