| Background and Purpose:Alcohol-induced osteonecrosis of the femoral head (ONFH) is a common diease in the clinical, it has few symptoms in the early period and easy to be ignored. When a patient goes to see a doctor for a painful and uncomfort hip, the lesions of the femoral head always tends to developping into ARCOⅡperiod or more than three, the best period of treatment is losing. the disease can caus physical disability in high rate and seriously threaten mankind health. No means can prevent onset of the disease to our knowledge at present. The clinical analysis indicanted that almost 30% patients had A bulk and long-term drinking. A cock or a rabbit model of alcohol-induced ONFH could be made by given A bulk and long-term drinking. Alcoholism has been confirmed as a definite cause of ONFH. The novel pathogenesis of the alcohol-induced ONFH have indicated that the peroxisome proliferator-activated receptor-γ(PPARγ) mRNA,an adipogenesis transcription factor, in the marrow stromal cells (MSCs) could directly induce the differentiation of MSCs into a large number and volume of adipocyte with A bulk and long-term drinking, at the same time,their osteogenic differentiation was inhibited. It explained the pathogenesis of alcoholic ONFH from the cellular and molecular biology. In the study, we will inject SiRNA adenovirus which were identified, selected and amplified into the femoral head of rabbits and make PPARγspecific silence by RNAi effective technique, prevent MSCs'adipogenic differentiation and keep them osteogenic differentiation. The preventive effects of siRNA adenovirus on alcohol-induced ONFH will be observed by the methods of molecular biology, biochemistry, cytology and histology and an effectual prevention and treatment way of alcohol-induced ONFH may be found by from the genes. Materials and Methods:96 2～3 months old, (weight 2.7±0.5kg) New Zealand species healthy rabbits (offered by the experimental animal center of Henan province) were randomly divided into 4 groups(24 each group). Group normal(N):The rabbits were giver 10% glucose at dosage of 10 ml/(kg·d) by pouring into stomach; Group alcohol(M):As the model of alcohol-induced ONFH, the rabbits were given strong wine,46% alcohol content, in 10 ml/(kg·d) by pouring into stomach; Group alcohol+ siRNA (S):In anesthesia,25μl adenovirus vector packed with pAdeno-X-siPPAR-1 was injected into one of femoral head of the rabbits on 1st day of 1,3,5 month from the beginning of the experiment, the puncture hole was closed with bone wax before the incision was sutured, then the rabbits were given the same alcohol as Group M; Group alcohol+ puncture (CO):The rabbits were given the same treatment as Group S, but no siRNA adenovirus was injected. All of the rabbits'total serum cholesterol (CHO) and triglyceride (TG) levels were measured on the end of 2,4,6 month from the beginning of experiment. Meanwhile the femoral heads of rabbits were cut in batches for the histological pathological changes such as, the average diameter of the max adipocyte, the trabeculace area fraction and the percentage of empty osteocyte lacunae in the femoral heads. The PPARγmRNA,the Osteocalcin mRNA and the PPARγprotein of the rabbits'femoral heads were also measured by RT-PCR and Western-blot technology.1. General conditions:All of the rabbits were alive in the course of experiment. During the experiment, the rabbits in Group N had good appetite, weight gain and flexible action; the rabbits in Group M and Group CO had poor appetite, slow weight growing or losing, poor hair gloss and different mental; under the same conditions as Group M and Group CO, the rabbits in Group S had just a little different mental.2. The content change of serum triglyceride (TG) and total cholesterol (CHO) of the experimental animals in each group and each period(mmol/L):At the end of the 2nd,4th and 6th month after the beginning of the experiment, the TG and CHO content results of Group M,CO,S is higher than Group N and it is getting higher and higher along with the eexperiment. the difference was significantly(P<0.05); From the femoral head histological pathological changes of the rabbits, the percentage of empty osteocyte lacunae of Group M,CO were much more than Group N,S; the average diameter of the max adipocyte of Group M,CO were bigger and more than Group N,S; the trabeculace bone of Group M,CO were thinner and sparse than, the difference was significantly(P<0.05); But, there were no more difference between N and Group S(P>0.05).3. The gene expression results of adipogenic and osteogenic differentiation in the femoral head MSCs of the rabbits:The expression levels of PPAR Y mRNA mensurated of group M, CO were higher than that of group N, S; There were the same results for PPARγprotein. On the contrary, the expression levels of osteocalcin mRNA mensurated of group M, CO were lower than that of group N, S; The differences had statistically significance (P<0.05). The expression levels of PPAR mRNA,PPARγprotein and osteocalcin mRNA of group S were approximate to the results in group N. The differences had no statistically significance (P>0.05).1. Given a bulk and long-term drinking, the fat cells in rabbit femoral head could be increased and enlarge, the empty lacunae could be increased and the trabecular bone area fraction could be decreased. What happened in the rabbit femoral head could cause osteonecrosis of the femoral head. 2. Through injected siRNA adenovirus into the femoral head of rabbit, we could make PPARγ, the key regulatory genes of adipogenic differentiation silence and keep MSCs osteogenic differentiation from adipogenic differentiation. And so so, the normal trabecular bones in the femoral head of rabbit were kept, the fat cells and the empty lacunae were decreased.3. We could prevent and control the alcohol-induced ONFH from disease mechanism by Gene Technology.4.
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