Study On The Chemical Composition And Biological Activity Of Steamed Codonopsis Lanceolata

Codonopsis lanceolata(family Campanulaceae), a vines herbaceous perennial plants, is also called the mountains carrots, sweet potato, goat milk. It has been used in Asian countries for several effects including tonify deficiency, promoting lactation and detoxification, as precious medicinal herbs. C. lanceolata is also as materials of practical in folk, with "ShanCai King" reputation. It has been reported that C. lanceolata had a protective effect against alcoholic fatty liver. Also, C. lanceolata exerted better anti-tumor, anti-obesity, anti-inflammatory and antidiabetics effects. Chemical analyses of this herb have revealed that the bioactive constituents are saponins.The fresh roots of C. lanceolata were steamed in an autoclave at 105°C for 1 h and then dried at room temperature(25°C) to obtain steamed C. lanceolata(SCL). The SCL powders were ultrasound-assisted extraction with 10 volumes of methanol at the temperature of 40°C for 90 min. After extracting for 3 times, the obtained filtrate was concentrated under reduced pressure in a rotary evaporator to give a crude extract and make up solution to the animal experiment and detecte. In the present study, under the optimal chromatographic and MS conditions, one polyacetylene(lobetyolin) and six saponins were identified by comparing the retention time and by matching the empirical molecular formula with that of the reported known saponins,for its future development lay the foundation.The present study was designed to evaluate the anti-tumor effect in vivo of SCL in H22tumor-bearing mice. The results showed that SCL could not only inhibit the tumor growth, but also prolonged the survival time of H22 tumor-bearing mice. Besides, the levels of serum cytokines including interferon gamma(IFN-γ), tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), and interleukin-2(IL-2) were enhanced by SCL administration. The observations of Hoechst 33258 staining demonstrated that SCL was able to induce tumor cell apoptosis. Finally,immunohistochemical analysis showed that SCL treatment increased Bax expression and decreased Bcl-2 and VEGF expression of H22 tumor tissues in a dose-dependent manner, and400mg/Kg dosage group showed a significant effect. Taken together, the findings in the present investigation indicated that SCL inhibited the tumor growth in vivo at least partly via improving the immune functions, inducing apoptosis, and inhibiting angiogenesis.In addition, we establish cisplatin-induced kidney injury model to evaluate the protective effect of SCL against cisplatin-induced kidney injury in mice. The results showed that theactivities of BUN, CRE, TNF-α and IL-1β in serum, MDA level in kidney tissue, decreased significantly in the SCL-treated groups compared with the model group. H&E and Hoechst33258 examination revealed that SCL can inhibit cell apoptosis. Finally, immunohistochemical analysis showed that SCL treatment increased Bax expression and decreased Bcl-2 expression in a dose-dependent manner. On the whole, our date suggest that SCL can protect cisplatin-induced kidney injury in mice.The purpose of this study is to investigate the hepatoprotective effect of SCL on ethanol induced liver injury in mice. Biochemical markers and enzymatic antioxidants from serum, liver tissue were determined. The results showed that the activities of ALT, AST, and TG in serum,MDA level in liver tissue, decreased significantly in the SCL-treated groups compared with the alcohol group. On the contrary, the GSH level was increased markedly. Histopathological examination revealed that SCL(400mg/Kg) pretreatment noticeably prevented alcohol-induced hepatocyte apoptosis and fatty degeneration.Finally, the theses investigated whether syringin, the main active material, could protect against cisplatin-induced acute renal injury by using a preclinical mice model. The author demonstrated that syringin pretreatment noticeably decrease the activities of CRE, BUN, TNF-αand IL-1β in serum, SOD, GSH and CAT level in renal tissue. Furthermore, H&E, Hoechst33258 and AB-PAS examination showed that syringin improve the cisplatin-induced renal damage. Syringin treatment increased Bax, caspase-3 expression and decreased Bcl-2expression by immunohistochemical analysis. The results showed that syringin has a protective effect on cisplatin caused acute renal injury in mice.To sum up, in this thesis, our findings had shown that SCL exerted the anti-tumor effect in H22 bearing tumor mice and had protective effects on acute alcoholic liver damage. Moreover,SCL and syringin can protect cisplatin-induced acute kidney injury in mice.