Experimental Study On The Effect Of Weiganli On Hematopoietic Regulation In Myelosuppressed Mice

Objective and Methods Myelosuppression is one of the commonside-effect for cancer patients when they receiving radiotherapy andchemotherapy.The number of leucoeyte,erythrocyte,and platelet decreaseseverely, what's more,excess myelosuppression may also causeinfeetion,imflamation and bleeding,and the course of radio-andchemotherapy may be hampered,which are limiting factors for improvingthe curative effect of radio-and chemotherapy.Therefore,the prevention ofmyelosuppression has been the focus of many experimental studies.FromTCM perspective,the pathogenesisof myelosuppression is due to theasthenia of spleen and kidney, which are the sources of congenital-Qi,blood, and kidney-Yang.WeiGanLi is a Compound Preparation whichcomposed of Antler, Ginseng,Astragalus,Epimedium,Curculiginis,Fructus,Ganoderma, Cortex Moutan,Licorice and other traditional chinesemedicines,those medicine are of great value in invigorating congenital-Qiand reinforcing kidney-yang.Previous study had demonstrated that thiscompound can efficaciously enhance body immunity.To evaluate the effectof WeiGanLi on bone marrow hematopoiesis,in our study, thesurvival,proliferation,differentiation,apoptosis of the hematopoietie cells, the expressions ofhematopoietic growth factors,anti-apoptosis proteins,matrix metalloproteinase,and the effect of WeiGanLi on bone marrowhematopoiesis was investigated,the mechanism and the characteristics ofWeiGanLi on hematopoietic regulation was also explored,so as to providean experimental basis for WeiGanLi's clinic application.Result The results of this research showed that:1.High dose WeiGanLi can promote the recovery of granulocyte,erythrocyte, Hb and BMC;Medium and low dose WeiGanLi can alsoprovided benefit to the amelioration of the hemogram, especially to therecovery of Pit.2.The result of cell culture in vitro showed that the numbers ofCFU-GM,CFU-E,and BFU-E were obviously increased in high andmedium dose group;In low dose group, the number of CFU-GM andCFU-Meg were also significantly increased.3.The result of flowcytometer showed that WeiGanLi may had less effecton the number of peripheral HSC.However, after WeiGanLi administration,an upward tendency in the number of bone marrow HSC could be seen inhigh dose group;Bone marrow HSC was also significantly increased inmedium dose group and peaked on day 7,then decreased on day 10;Compared with high and medium dose WeiGanLi, low dose WeiGanLi mayhave less effect on stimulating the proliferation of bone marrow HSC.4.The eeU ratio of BMC in G₀/G₁ phase was significantly decreased afterWeiGanLi's administration,whereas the ratio of BMC in S phase and G₂/Mphase was significantly increased,implied that WeiGanLi can active BMCfrom stasis into cell cycling,accelerate the synthesie of DNA and themitosis of BMC,promote the proliferation of hematopoietic cells,which will contribute to the recovery of the hematopoietic function.5.The result of histochemistry showed that the expression of Bcl-2 proteinwas significantly up-regulated,and the expression of Bax protein wassignificantly down-regulated,the ratio of Bcl-2/Bax was increased, whichwill be benefit for the prevention of BMC from apoptosis. The result ofin situ hybridization showed that the change of Bcl-2,Bax protein wascoincided with that ofBcl-2 mRNA,Bax mRNA, which indicated that theadjustion of WeiGanLi on Bcl-2,Bax protein may be happened attranscription and translation level.6. The Level of IL-6,FL was significantly adjusted by Weiganli,but the level of GM-CSF, and the expression of TPO mRNA in bonemarrow stromal cells and spleen cells showed not significant change afterWeiganli administration,this result implied that the recovery of bonemarrow hematopoiesis may attribute to the modulation of Weiganli onmulti-cytokines not on uni-cytokine.7.The result of zymogram electrophoresis showed that the activity ofproMMP-2,proMMP-9 was up-regulated,the activity of MMP-2 wasdown-regulated,and the activity of MMP-9 had not changed after Weiganliadministration,this result implied that the activation of proMMP-2,proMMP-9 and inactivation MMP-2,MMP-9 may play an important role inhematopoiesis.Conclusion1.We first studied Weiganli's function on hematopoietic regulation andexplored it's mechanism by evaluated the survival,proliferation,cell cycle,apoptosis related gene(Bcl-2,Bax),hematopoietic growth factor(IL-6,FL, GFM-CSF,TPO),and bone marrow matrix metalloproteinase(MMP-2MMP-9), the result of our study demonstrated that the myelosuppressionwhich induced by irradiation and chemotherapy agent was significantlyrelieved after Weiganli's administration, Weiganli may play an importantrole in the rehabilitation of hematopoiefie function in myelosuppressedmice.2. As can be showed from the above results, Weiganlipromotehematopoiesis in multiple ways and levels. On one hand promote cellsentering proliferitve cell phase, up-regulate expression of Bcl-2 mRNA andBel-2 protein,down-regulate Bax mRNA and Bax protein, inhibitoccurrence of apoptosis to maintain hemeatopoiesis. On the otherhand,promote the recovery of hematopoiesis by synergistic effect of IL-6,FL and other hematopoietic growth factors.In this study, we first found andposed that the activation of proMMP-2,proMMP-9 and inactivationMMP-2,MMP-9 may play an important role in hematopoiesis.Hypothesisof mechanisms on Weiganli's hematopoietic regulation is first submittedaccording to our studies. Our studies provide useful scientific data oflaboratory for clinic application of Weiganli.