Study On The Proliferation And Collagenic Synthesis Of NIH- 3T3 Cell With Three Chinese Herbal Ingredients

For studying the effects of dracohodin perchlorate (DP), masticinic acid (MA) and myrrh extracts (ME), the indications including the safe medication range, the cell growth curve, the phase change of cell cycle, the collagen in the cell suspension, and the MMPs/TIMPs (MMP-1, MMP-2, MMP-9, TIMP-1, TIMP-2), were assayed through the drug incubation experiments. It was better to research the mechanism of improving the wound healing with Fangfu Shengjisan.NIH-3T3 cell was chosed as the experimental object. The cell toxicity and cell growth effects of DP, MA and ME was detected by using MTT method. The cell cycle of NIH-3T3 cell was detected by using FCM. The Hyp kit was used to detect the collagenic content of NIH-3T3 cell. The secretions of MMP-1, MMP-2, MMP-9, TIMP-1 and TIMP-2 of NIH-3T3 cell were detected by using ELISA kits. The results showed that: (1) The safe medication range of DP, MA and ME effecting on NIH-3T3 cell were respectively 0.3125μg/L~40μg/L, 125mg/L~8g/L, 1.95mg/L~2g/L. During the Chinese herbal ingredients intervened NIH-3T3 cell within 48 hours, DP could inhibit the proliferation of NIH-3T3 cell, wich was significantly different to the control group (P<0.01), however MA and ME could significantly improve the proliferation of NIH-3T3 cell, there were significant differences between them and the control group (P<0.01). In 48 hours after the Chinse herbal ingredients intervention, the Chinse herbal ingredients could not affect the proliferation of NIH-3T3 cell. (2) With prolonging the intervention time of the traditional Chinese herbal compoments, there was a statistical significance that DP could singnificantly increase the apoptosis rate and decrease the survival rate. Comparing with the control group respectively, MA and ME could significantly decrease the apoptosis rate and increase the survival rate (P<0.01). (3) The collagenic synthesis could be significantly improved by DP after intervened for 24 hours and decreased after 48 hours, what were significantly different with the control group (P<0.01). Within 24 hours after MA intervention, the collagenic protein was significantly synthetized more than the control group, but after 48 hours the collagenic protein was lower synthetized, so there was no difference to the control group. ME notably inhibited the collagenic synthesis after 24 hours and was notably different to the control group (P<0.01), but intervening the NIH-3T3 cell for 48 hours, ME started to notably improve the collagnic synthesis, so there was no difference with the control group (P>0.01). (4) Except for no significant difference appeared between the secretory volume of MMP-2 and the intervention time, there were significant differences between the the secretory volume of MMP-1 and the intervention time comparing all the drug groups with the control group (P<0.01), the same as MMP-9, TIMP-1 and TIMP-2.Conclusion: 48 hours after the three Chinese herbal ingredients at EC50 concentration intervening NIH-3T3 cell, DP could inhibit the proliferation and collagenic synthesis of NIH-3T3 cell, improve the apoptosis of NIH-3T3 cell, furthermore inhibit the expressions of MMP-1, TIMP-1 and TIMP-2, and improve the expressions of MMP-2, MMP-9. MA could improve the continuouse collagenic synthesis and the proliferation of NIH-3T3 cell, inhibit the apoptosis of NIH-3T3 cell, furthermore improve the expressions of MMP-2, MMP-9, and inhibit the expression of MMP-1, TIMP-1 and TIMP-2. ME could improve the proliferation and the collagenic synthesis of NIH-3T3 cell, inhibit the apoptosis of NIH-3T3 cell, furthermore improve the expression of MMP-2 and MMP-9, and inhibit the expressions of MMP-1, TIMP-1 and TIMP-2.