| Objective: To investigate the relationship between the treatment of "activating blood circulation and removing blood stasis" and inflammation and immunity by studying the effect of Tongluo Shenggu Capsule on bone histopathology and TLR4 / NF-?B signaling pathway in rats with hormonal femoral head necrosis Possible mechanism in the prevention and treatment of early femoral head necrosis.Method: 60 rats were divided into 2 groups by random number table method,divided into 40 model groups and 20 negative control groups for modeling.The model was injected intraperitoneally with Lipopolysaccharide(LPS)(10 ?g / kg)twice,with an interval of 24 h each time.Methylprednisolone(40 mg / kg)was injected alternately on both sides of the gluteal muscle 24 h after the last injection,with an interval of 24 h each time.After 4 weeks of feeding,10 rats were randomly selected from the model,and 5 rats were randomly selected from the normal control group for anesthesia for MRI test,and femoral head was taken after sacrifice to test whether the model was successful.After successful modeling,the remaining rats were regrouped into 15 normal control groups,and the remaining model groups were divided into 2 groups,15 experimental groups and 15 positive control groups.The experimental group was gavaged with Tongluo Shenggu capsule for 4 weeks,and the negative control group and positive control group were gavaged with equal volume of normal saline for 4 weeks.After 4 weeks,the rats were sacrificed and the femoral heads were removed for gross specimen observation and histomorphological observation.And WB,PCR detection of bone tissue NF-?B,TNF-?,My D88,TLR4 factor expression.Results: 1.SONFH rat model identification results:(1)After 4 weeks of the experiment,the negative control group was compared with the model: MRI examination showed that the model femoral head showed different shapes,with a small amount of hip effusion,a few The rat model showed mixed signals,and some abnormal signal shadows were seen in some femoral heads.(2)Observation of the general shape of the femoral head The femoral head of the rat model showed that the color of the articular cartilage surface was slightly darker than that of the normal control group and was purple.(3)HE staining results: The trabecular bone and bone marrow of the femoral skull tissue were necrotic.The trabecular bone was loose and the structure was disordered.The number of empty bone lacuna was significantly increased compared with grade I.Or cracked.This indicates that the rat model of hormonal femoral head necrosis has been successfully modeled.2.Gavage 4 weeks after successful modeling,the experimental results :(1)Rat bone tissue empty bone pit rate count results: empty bone pit rate count found that the positive control group,experimental group empty bone pit rate was significantly higher than negative The rate of empty bone lacunae in the control group(P <0.01).The rate of empty bone lacuna in the positive control group was significantly higher than that of the experimental group(P <0.01).(2)Results of WB detection of NF-?B,TNF-?,My D88 and TLR4 protein in rat bone tissue: Compared with the positive control group,the expression levels of NF-?B,TNF-?,My D88 and TLR4 protein in the negative control group and the experimental group were all obvious The difference was statistically significant(P <0.01);compared with the experimental group,the expression of TLR4 and My D88 in the negative control group were not statistically different(P> 0.05).(3)RT-PCR detection of rat bone tissue NF-?B,TNF-?,My D88,TLR4 factor results: compared with the positive control group,the negative control group,the experimental group NF-?B,TNF-?,My D88,TLR4 factor expression Both were significantly reduced,the difference was statistically significant(P <0.01);compared with the experimental group,the expression of NF-?B and My D88 in the negative control group were not statistically different(P> 0.05).Conclusion: Tongluo Shenggu Capsule can improve the pathological morphology of femoral head in SONFH rat model,reduce the rate of empty bone lacuna,reduce the number of osteoclasts,and down-regulate the expression of My D88,TNF-?,TLR4 and NF-?B factors.Its mechanism of action may be through regulating TLR4-My D88-NF-?B signaling channel protein signal,inhibiting the expression of TNF-? inflammatory factor,thereby reducing the inflammatory response and reducing the loss of bone mass,so as to play a preventive and therapeutic role.Experimental results combined with theoretical analysis,the intervention of Tongluo Shenggu Capsule has a protective effect on the femoral head of rats with early femoral head necrosis.Feasibility of prevention and treatment of bone necrosis in Chinese medicine. |
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