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| Bcl2促进UMR-106细胞BMP-2、OPG表达及补肾健脾活血方对其影响 |
| Bcl2 promotes expression of BMP-2 and OPG and the effect of nourishing kidney, invigorating spleen, and promoting blood flow recipe on the protein expression in UMR-106 cells |
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| DOI：10.3969/j.issn.1006.7108.2018.07.001 |
| 中文关键词: 补肾健脾活血方 UMR-106细胞 B淋巴细胞瘤-2 过表达/沉默 骨形态发生蛋白2 骨保护素 |
| 英文关键词:Nourishing kidney, invigorating spleen, and promoting blood flow recipe UMR-106 Bcl2 overexpress/silencing BMP-2 OPG |
| 基金项目:国家自然科学基金项目（81373653，81674004 ，81673786）；广东省科技计划项目（2016ZC0096，2016A020216024）；广东省自然科学基金项目（2015A030313367）；广州市荔湾区科技计划项目（201704041） |
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| 中文摘要: |
| 目的 基于构建Bcl2沉默和过表达腺病毒观察Bcl2对成骨细胞骨形态发生蛋白2（bone morphogenetic protein-2，BMP-2）、骨保护素（osteoprotegerin，OPG）蛋白的调节作用及补肾健脾活血方对其影响。方法 将培养的 UMR-106细胞分为空载腺病毒组（沉默Bcl2空载腺病毒 NC-bcl2，过表达Bcl2空载腺病毒 WT-bcl2）、 Bcl2腺病毒组（沉默Bcl2腺病毒 sh-bcl2，过表达Bcl2腺病毒 Ad-bcl2）,空载腺病毒+空白血清组（NC-bcl2，WT-bcl2）、空载腺病毒+补肾健脾活血方含药血清组（NC-bcl2+ME，WT-bcl2+ME）、腺病毒+空白血清组（sh-bcl2，Ad-bcl2）、腺病毒+补肾健脾活血方血清组（sh-bcl2+ME，Ad-bcl2+ME），用RT-qPCR检测Bcl2、OPG mRNA的变化、应用免疫印迹( Western Blot)检测Bcl2、OPG、BMP-2的变化。结果 ①荧光倒置显微镜观察包装腺病毒转染UMR-106细胞；②在正常培养基中，与NC-bcl2比较，sh-bcl2 可以降低Bcl2、OPG、BMP-2蛋白的表达（P均<0.05），与WT-bcl2比较，Ad-bcl2可以提高Bcl2、OPG、BMP-2蛋白的表达（P均<0.05）；③与NC-bcl2比较，RT-qPCR显示sh-bcl2可以降低Bcl2 mRNA的表达，但OPG mRNA无统计学意义；④在大鼠血清干预中，与空载腺病毒比较，补肾健脾活血方含药血清均可以增加BMP-2、OPG蛋白表达（P均<0.05）；在sh-bcl2干预的细胞中，补肾健脾活血方含药血清提高BMP-2、OPG蛋白的表达（P均<0.05）；在Ad-bcl2干预的细胞中，中药干预后，BMP-2、OPG未见明显变化。结论 Bcl2可以影响BMP-2、OPG蛋白的表达，补肾健脾活血方可能通过作用于Bcl2影响成骨细胞成骨相关蛋白的表达。 |
| 英文摘要: |
| Objective By the construction of the silencing and overexpression adenovirus of Bcl2, to obverse the effect Bcl2 on bone morphogenetic protein-2 (BMP-2) and osteoprotegerin (OPG), and the influence of nourishing kidney, invigorating spleen, and promoting blood flow recipe (Recipe). Method UMR-106 cells were divided into vector group (silencing vector Bcl2, NC-bcl2, and overexpression vector Bcl2, WT-bcl2), Bcl2 adenovirus group (silencing Bcl2, sh-bcl2, and overexpression Bcl2, Ad-bcl2), vector adenovirus+blank serum group (NC-bcl2 and WT-bcl2), vector adenovirus+Recipe serum group (NC-bcl2+ME and WT-bcl2+ME), adenovirus+blank serum group (sh-bcl2 and Ad-bcl2), adenovirus+Recipe serum group (sh-bcl2+ME and Ad-bcl2+ME). Bcl2 and OPG mRNA were detected with RT-qPCR. The portion expressions of Bcl2, OPG, and BMP-2 were detected with Western blotting. Results 1) Adenovirus fluorescent in UMR-106 cells was observed with fluorescent inverted microscope. 2) In normal medium, compared with NC-bcl2, sh-bcl2 reduced the protein expressions of Bcl2, OPG, and BMP-2 (P<0.05). Compared with WT-bcl2, the protein expressions of Bcl2, OPG, and BMP-2 were significantly improved in Ad-bcl2 (P <0.05). 3) Compared with NC-bcl2, the mRNA expression of bcl2 decreased in sh-bcl2. However, mRNA expression of OPG had no significance. 4) Compared with vector adenovirus in rat serum intervention, the protein expressions of BMP-2 and OPG increased in Recipe serum (P <0.05). The protein expressions of BMP-2 and OPG increased in the Recipe serum and sh-bcl2 intervention (P<0.05). The protein expressions of BMP-2 and OPG had no significance in Ad-bcl2 intervention. Conclusion Bcl2 could affect the protein expressions of BMP-2 and OPG, and Recipe may affect the osteoclast-related protein expressions through Bcl2. |
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