The Anti-osteoporosis Activity Of Flavonoids From Branches Of Cunninghamia Lanceolata

Cunninghamia lanceolata (Lamb.) Hook originates from Taxodiaceae, which is an endemic plant to China. At present, most researches about Cunninghamia lanceolata have focused on planting and ecophysiology in agroforestry, but rather few studies on the medicinal aspect. Cunninghamia lanceolata in our country has been primarily used as timber, but its leaves and branches been treated as waste. In the previous study by our group, we found that its leaves and branches contain a large number of biflavonoids which have good estrogen-like effects. Therefore, on the basis of previous work, this article studied the activities of anti-osteoporosis of total flavonoids and bioflavonoids components from leaves and branches of Cunninghamia lanceolata, and laid a good foundation of development of plant preparations for osteoporosis.In present research work, We studied and determined the effect of anti-osteoporosis of total flavonoids by combining the osteoporosis rat model that induced by retinoic acid. To further clarify the material basis of the anti-osteoporosis effect of total flavonoids, three bioflavonoids compounds which had a big share in total flavonoids were isolated by column chromatography with silica gel, Sephadex LH-20, etc. Combining with ESI-MS and other mass spectrometry, the structures were identified by means of 1-NMR,13C-NMR, they were Sciadopitysin, Isocryptomerin and Amentoflavone. Osteosarcoma cells MG-63-PERE were used to screen the estrogenic effect in vitro, and combined with the above animal models for anti-osteoporosis effect screening, we found that Sciadopitysin and Isocryptomerin had good estrogen-like effects in vitro, but Amentoflavone did not have. While the three all had great effects of anti-osteoporosis in vivo.The extracting process of total flavonoids from leaves and branches of Cunninghamia lanceolata was optimized by central composites design and response surface method with extraction rates of total flavonoids, then the content of total flavonoids and transfer rate were used as the index to study the purification process of total flavonoids, we finally established Preparation of total flavonoids. In addition, extraction rates of Amentoflavone and Sciadopitysin being as the indexes, we used the CCD method to study the extracting process of bioflavonoids.HPLC method was used to assay the contents of Amentoflavone and Sciadopitysin in leaves and branches of Cunninghamia lanceolata from the following areas: Zhejiang Quzhou, Zhejiang Longquan, Zhejiang Lin'an, Zhejiang Qing'an, Zhejiang Jinhua(Baicao Garden and Shuanglong Cave), Hunan Zhangjiajie, Hunan Longshan County, Anhui Chuzhou, etc. The contents of Amentoflavone and Sciadopitysin in leaves and branches of Cunninghamia lanceolata from Zhejiang Lin'an were 0.41% and 0.17% respectively, which were higher than those from other areas.