Studies Of Apoptosis And Expression Of α-Mannosidase In Rat Brain By Contacting Oxytropis Kansuensis

Swainsonine contained in Oxytropis kansuensis can be used as anticancer drugs in recentresearch. It can restrain the tumour metastasis and promote the apoptosis of cancer cell.Moreover, it can also strengthen immune system. But whether it can induce apoptosis of braintissue was not mentioned. Swainsonine inhibited the activity of-mannosidase by combiningthe-mannosidase, however the sbustrate released from the enzyme and accumulated in thecells, then the synthetization and metabolization of glycoprotein were affected. Other than, theeffection of-mannosidase by swainsonine in the genes and protein level were not reported.In this chronic poisoning experiment, SD rats were used as model animal, O. kansuensiswas used as the source of swainsonine. The apoptosis of brain tissue was detected by thetransmission electron microscope and TUNEL kit. Expression of apoptosis related proteinsand some types of-mannosidase were detected by Western blotting and Real-timequantitative PCR. The results were as follows:1. Reconstruction of chronic O. kansuensis poisoning model in SD rats64female SD rats were divided into control group and experimental group Ⅰ, Ⅱ and Ⅲ.Different amounts of grass powder (15%,30%,45%respectively, SW content were0.003%,0.006%,0.009%) were mixed with feeds for three experimental groups. SD rats were feededuntil119d. Hematology index was tested. The results showed the increase of WBC, MO, NE,RDW and MPV, while the decrease of LYM, RBC, HGB and HCT. Interestingly, MCV rosefirst then declined, but PLT is opposite during poisoning period, some significant differencesbetween the experimental groups (P＜0.05or P＜0.01). This study suggests swainsonine oflow-dose can cause anemia of SD rats in long-term, but it can enhance immunity and promotebone marrow function.2. Apoptosis detection in brain of the chronic poisoning SD ratsSD rats were feeded with O. kansuensis45%contained in diet for119d, materials wererespectively used for the detection of electron microscope and test in TUNEL kit. Theapoptotic body was not found, but some inflammatory reaction and edema were detected,vascular endothelial cells were severely damaged in the result of electron microscope. Strongpositive result was not found, weak positive result appeared in lesions of the purkinje cells,but it is not obvious. Toxicity test in vivo showed that swainsonine had no obvious influence on the apoptosis of neurons in brain of SD rats.3. Expression of apoptosis related protein in brain of the chronic poisoning SD ratsBrain tissues were adopted from control group and experimental group Ⅰ, Ⅱa ndⅢ inthe35,63,91and119d. Expression of Bax, Bcl-2, Fas, FasL, Caspase-3, Caspase-8,Caspase-9and Caspase-12were detected by Western Blotting and Real-time quantitativeRCR. The results showed that swainsonine had effect on the expression of apoptosis relatedprotein in SD rats brain in the genes and protein level. The performance of swainsonine canactivate some key elements in the mitochondria apoptosis way and endoplasmic reticulumapoptosis, but the influence on external cell apoptosis way needs further research.4. Expression of-mannosidase in brain of the chronic poisoning SD ratsExpression of protein about MAN2A1and MAN2C1were tested, and expression ofmRNA about MAN2A1, MAN2B1and MAN2C1were detected. The results show thatswainsonine had influence on the expression of-mannosidase in the genes and protein levels.The expression of MAN2A1and MAN2B1were inhibited during the earlier stage, theexpression of MAN2C1were inhibited in the earlier stage but increased during the laterperiod. Swainsonine had the influence of glycoprotein synthesis and metabolism, affecting thebody’s immune system.