Roles Of Six Constituents From Atractylodes Lancea Rhizome Water Extract In Reducing The Triptolide-induced Toxicities

Triptolide(TP), is one of the major active component in Tripterygium wilfordii Hook F(TWHF). It has several pharmacological activities such as anti-cancer, anti-senil dementia, anti-rheumatoid and anti-inflammatory etc. However, some research showed that, the effective dose of TP was similar with toxical dose. This fact greatly limited the application of TP. How to reduce the triptolide-induced toxicity and keep its pharmacological activities has become a hot topic in research.Atractylodes lancea rhizome is the dried roots of compositae plants Atractylodes lancea(Thunb.)DC. It has pharmacological activities such as anti-inflammatory, anti-tumor and protecting liver etc. Our previous study found that its water extract could reduce TP,s toxicity by inducing CYP3 A protein levels, and it also enhanced the anti-inflammatory effects of TP. To find the active components in the water extract, six commercially available constituents(atractyloside A, vanillic acid, syringic acid, protocatechuic acid, protocatechuic aldehyde, and limonoids) were further tested. The major results were summarized as follows:1. Six constituents from A. lancea water extract and their toxicities in C57BL/6 mice. Privous studies reported that those six constituents could be found in the water extract. Then the toxical effects of six constituents were determined by blood chemistry, viscera indexes, and histopathology. In comparison with control, no significant changes were observed in C57BL/6 mice treated with those six constituents.2. Regulation of the six constituents on mouse hepatic P450 s. Western blotting results showed that atractyloside A could induce CYP3 A, CYP1A2, CYP2 C and CYP2E1 protein expression; vanillic acid could induce CYP3 A, CYP1A2 and CYP2 D protein expression; syringic acid could induce CYP3 A, CYP1A1, CYP1A2, CYP2 D and CYP2E1 protein expression; protocatechuic acid could induce CYP1A1, CYP1A2, CYP2 B and CYP2E1 protein expression; protocatechuic aldehyde could induce CYP3 A, CYP1A2, CYP2 B and CYP2 C protein expression; and limonoids could induce CYP3 A, CYP1A1, CYP1A2, CYP2 A, CYP2 D and CYP2E1 protein expression. Real-time PCR results showed that liver Cyp3a11、Cyp1a2 m RNA levels significantly increased in the mice treated with atractyloside A and limonoids; liver Cyp3a11 m RNA levels significantly increased in the mice treated with vanillic acid、syringic acid and protocatechuic aldehyde; protocatechuic acid could increase the liver Cyp1a2 m RNA levels.3. Toxicity-reducing effects of the six constituents when co-treated with TP in C57BL/6 mice. In TP-treated mice, significant toxicity on liver, kidneys, spleen, and testis were observed. But the toxicity was no observed in co-treated groups. This result hinted that six constituents could protect TP-induced toxicity. But, toxicity-reducing effect of A. lancea water extract co-treated with TP was better than six constituents.4. Anti-inflammation effects of six constituents when co-treated with TP in C57BL/6 mice. The results showed that, in the six constituents treated mice, lower swelling degree and higher inhibition rate of ear edema were recorded compared with control mice. The anti-inflammatory effects in co-treated mice appeared to be better in comparison with those in TP-treated mice.5. Anti-inflammation mechanism of six constituents in C57BL/6 mice. MTT results showed that six constituents had little effect on macrophage proliferation. Meanwhile, the content of NO, and the gene transcription of IL-1? and IL-6 was remarkably inhibited by the six constituents.