课题背景：广西自然科学基金项目（0640153）。目前课题研究成果已能够成功而熟练地分离、提取骨髓基质干细胞，并且成功地进行原代细胞与传代细胞培养，初步证实三七总皂苷能抑制酒精诱导兔骨髓基质干细胞的成脂分化,并可以促进其成骨分化。

应用要点：通过检测含酒精培养液培养的骨髓基质干细胞内三酰甘油含量与碱性磷酸酶活性的变化，观察三七总皂苷对骨髓基质干细胞向脂肪细胞分化的抑制作用及向成骨细胞分化的诱导作用。结果表明，在酒精作用后的骨髓基质干细胞继续培养过程中,加入50，100 mg/L两种浓度三七总皂苷可促进骨髓基质干细胞的成骨表达,而抑制成脂表达。

相关链接：研究证明,酒精能够促使骨内脂肪堆积, 抑制成骨分化,导致成骨不足, 骨小梁稀疏、变细, 骨密度降低, 最终可发生骨质疏松或骨坏死。三七总皂苷是三七的主要活性成分。已有研究证实，三七总皂苷能阻断受体操纵性钙通道、增强一氧化氮的扩张血管作用、改善微循环、具有抗缺血再灌注损伤的作用。而三七总皂苷对酒精性骨坏死的防治作用研究尚少。

摘要
目的：三七总皂苷能阻断受体操纵性钙通道，增强一氧化氮的扩张血管作用，改善微循环，具有抗缺血再灌注损伤的作用，关于三七总皂苷对酒精性骨坏死防治作用的研究尚少。实验拟观察三七总皂苷对酒精诱导兔骨髓基质干细胞成脂分化的抑制作用。
方法：实验于2006-11/2007-08在广西医科大学完成。①实验材料：三七总皂苷由江苏康宝制药有限公司提供（国药准字Z32020670）。4周龄大白兔由广西医科大学实验动物中心提供，清洁级，实验过程中对动物处置符合动物伦理学标准。②实验方法：采用密度梯度离心法与贴壁法相结合体外分离、培养兔骨髓基质干细胞。取第3代细胞,并随机分为3组：空白组：细胞在含10%胎牛血清DMEM培养基中培养,不加酒精和三七总皂苷；模型组:每次更换培养液时加酒精0.09 mol/L；50 mg/L三七总皂苷组:每次更换培养液时, 加0.09 mol/L酒精与三七总皂苷50 mg/L；100 mg/L三七总皂苷组:每次更换培养液时, 加酒精0.09 mol/L与三七总皂苷100 mg/L。③实验评估：检测细胞内三酰甘油含量与碱性磷酸酶活性。
结果：①干预14 d时,模型组细胞内碱性磷酸酶活性降低,明显低于三七总皂苷干预组和空白组（P < 0.05）。三七总皂苷干预两组细胞内碱性磷酸酶活性,与空白组比较差异无统计学意义（P > 0.05）。②干预21 d时,模型组脂滴最多,三七总皂苷干预两组较模型组明显减少(P < 0.05),空白组未见明显脂滴出现。③三七总皂苷干预组和空白组三酰甘油含量差异无统计学意义 (P > 0.05) ,三七总皂苷干预组和空白组细胞内三酰甘油含量低于模型组细胞内三酰甘油含量(P < 0.05)。
结论：三七总皂苷能抑制酒精诱导的兔骨髓基质干细胞成脂分化,而促进成骨分化,可能有治疗酒精性骨股头缺血性坏死的作用。
关键词：三七总皂苷;抑制;酒精;骨股头缺血性坏死;成脂分化;成骨分化

王大伟，潘华，李红波，王换新，陈跃平，顾国龙.三七总皂苷对酒精诱导骨髓基质干细胞分化影响的实验研究[J].中国组织工程研究与临床康复，2008，12(8):1410-1413 [www.zglckf.com/zglckf/ejournal/upfiles/08-8/8k-1410(ps).pdf]

Effect of Tongxinluo on neurocyte lineage derived from rat embryonic neural stem cells

Abstract

AIM：The total saponins of panax notoginseng can blockade the nature channel of calcospherite, increase the function about nitrogen monoxide of eurysma the vessel, and improve microcirculation and ischemia/reperfusion injury. The researches on the total saponins of panax notoginseng function on the prevention and treatment of alcohol-induced osteonecrosis are few. This study investigated the inhibitory effect of total saponins of panax notoginseng on differentiation of rabbit bone marrow stromal stem cells into adipocytes induced by alcohol.

METHODS: Experiments were conducted at Guangxi Medical University from November 2006 to August 2007. ①The total saponins of panax notoginseng was provided by Jiangsu Kangbao Pharmaceutic CO., LTD. (batch number: Z32020670). Rabbits aged 4 weeks were offered by Experimental Animal Center at Guangxi Medical University（cleanness grade）. The method we disposed in the experimental animal is consonant with animal ethical standard. ②The primary rabbit bone marrow stromal stem cells were isolated and cultured in vitro by density gradient centrifugation and adherence screening method. The 3rd passage cells were divided into 3 groups. Cells were cultured in DMEM medium containing 10% fetal bovine serum without alcohol or total saponins of panax notoginseng in a blank group. 0.09 mol/L alcohol was treated when culture medium was changed in a model group. 0.09 mol/L alcohol and 50 mg/L total saponins of panax notoginseng were treated when the culture medium was changed in a 50 mg/L total saponins of panax notoginseng group. 0.09 mol/L alcohol and 100 mg/L total saponins of panax notoginseng were treated when the culture medium was changed in a 100 mg/L total saponins of panax notoginseng group. ③The alkaline phosphatase activity and triglyceride level in the cells was measured.

RESULTS: ①At day 14, the alkaline phosphatase activity in cells in the model group was reduced, which was significantly lower than that in the other groups (P < 0.05). There was no significant difference in alkaline phosphatase activity in each group (P > 0.05). ②At day 21, the number of adipocytes in the model group was the most. The number of adipocytes was decreased in the total saponins of panax notoginseng groups (P < 0.05), and no adipocytes were found in the blank group. ③There was no significant difference between the two total saponins of panax notoginseng groups and blank group (P > 0.05). The triglyceride levels in the cells in the model group were the highest (P < 0.05).

CONCLUSION: Total saponins of panax notoginseng inhibit differentiation of bone marrow stromal cells into adipocytes induced by alcohol and enhance differentiation of bone marrow stromal stem cells into osteoblasts, which may prevent the development of alcohol-induced osteonecrosis of the femoral head.

Wang DW, Pan H, Li HB, Wang HX, Chen YP, Gu GL.Influence of total saponins of panax notoginseng on the alcohol-induced differentiation of bone marrow stromal stem cells.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 2008;12(8):1410-1413(China) [www.zglckf.com/zglckf/ejournal/upfiles/08-8/8k-1410(ps).pdf]