Qi And Chinese Herbal Medicine Combined With Bmscs Transplantation Intervention In The Experimental Study Of Lumbar Intervertebral Disc Degeneration

ObjectivesTo observation the therapeutic effect of Yiqi Huayu Recipe combining BMSCs on the lumbar intervertebral disc degeneration model of rats, which can provides new ideas and methods for the prevention and control of degenerative lumbar intervertebral disc degeneration; To observation the influence of Yiqi Huayu Recipe medicated serum on proliferation of BMSCs, and analyze the mechanism of Yiqi Huayu Recipe combining BMSCs on the lumbar intervertebral disc degeneration.Methods1.16 Sprague-Dawley male rats were carried out transperitoneal ventral surgery and anular stab was performed using 23-gauge needle to make a full-thickness stab incision in the L4-5 and L6-S1 anterior-lateral disc. The L3-4 and L5-6 level was used as a control. Execute a certain number of rats to collect specimens after 2 weeks,4 weeks,6 weeks, and 8 weeks separately(n=8). Finally, we analyze disc degeneration states by hematoxylin-eosin staining, Alcian blue staining and immunohisto-chemical studies.2. whole bone marrow culture method was used to isolate and purify Sprague-Dawley rats BMSCs that were incubated in L-DMEM containing 10% medicated serum,10% rat serum,5% FBS+5% medicated serum,5% FBS+5% rat serum, and 10% FBS and serum-free media. Morphology and proliferation of BMSCs were observed by MTT method at 24h,48h and 72h in vitro.3.128 Sprague-Dawley male rats, just at the time of establishing disc degeneration models, L4-5 level was injected with BMSCs-PBS complex, L5-6 level was injected with only PBS, L6-S1 level is a control, which just be treated with needling. The animals are randomly divided into two groups, group A ones were fed with Yiqi Huayu Recipe decoction by gastric perfusion 0.85 g/kg daily, and group B ones were fed with distilled water 1ml/100g. execute a certain number of rats to collect specimens after 2 weeks,4 weeks,6 weeks, and 8 weeks separately (n=8). Finally, we analyze disc degeneration states by hematoxylin-eosin staining, immunohisto-chemical studies, Alcian blue staining and DMB spectrophotometer method.Results1. Full-thickness anulus stab incision with 23-gauge needles defined gauges is effective to establish a rat model of intervertebral disc degeneration. After 8 weeks, histologic scores, collagen type II and proteoglycans analysis are significant difference.2. The OD values of the Rats serum and medicated serum training groups are higher than FBS training group at different time points, and OD values are increasing with rats serum and medicated serum concentrations increased; The OD values in different concentrations medicated serum training group higher than ratsserum training group at different time points, they have the remarkable significance difference at 72h point.3. Histological examination and biochemical tests showed that compare with the control group, pure BMSCs transplantation group and Yiqi Huayu Recipe decoction combined with BMSCs transplantation group have less histological score and more collagen typeⅡand proteoglycan content. Yiqi Huayu Recipe decoction with BMSCs transplantation group is better than the other. compared with the control group, pure Yiqi Huayu Recipe decoction group's collagen typeⅡand proteoglycan content increased, but the difference was not statistically significant.Conclusions●Full-thickness anulus stab incision with 23-gauge needles approach results in a slowly progressive intervertebral disc degeneration in rat model. This model is available for studying the mechanism of intervertebral disc degeneration and treatment.●Rat serum group and medicated serum group are both suitable for BMSCs microenvironment to live and proliferation, medicated serum group is better than the other..●Both Yiqi Huayu Recipe decoction combined with BMSCs transplantation and pure BMSCs transplantation supporting its potential use as a treatment of intervertevral disc degeneration, Yiqi Huayu Recipe decoction with BMSCs transplantation group is better than the other.