| ObjectiveInvestigate the relationship between osteoporosis and skeletal muscle from the perspective of mitochondrion,which to provide the objective reasonable quantification basis for the skeletal muscle as a therapeutic target,and provide a scientific basis for chinese medicine theory of "kidney controlling bones,spleen controlling muscle and spleen-renal interrelationship" in the treatment of osteoporosis.MethodThe clinical experiment26 female patients were selected from the first hospital affiliated to Guangzhou university of CM between May 2008 and December 2008,who were diagnosed for lumbar intervertebral disc herniation or lumbar spinal stenosis to need surgery.They were divided into Osteoporosis group(n=13) and non-osteoporosis group(n=13).Mitochondrion were extracted from the muscle tissue that was collecte during the operation,and cytochrome coxidase(COX) and the mitochondrial permeability transition pore(MPPT) were detected. The animal experiment52 female non-pregnant SD rats were randomly divided into normal control and operation group,normal control group were 16,and operation group were 36.The operation group were ovariectomized,while sham operation was performed in normal control group.After 3 monthes,the two groups were randomly selected 10 rats to measured the BMD.After establishing modle,the operation group of 36 were divided into 3 groups:ovariectomized,estrogen supplemented and traditional chinese herbs,the rats of traditional chinese herbs were given the intragastric administration with herbs of reinforcing the kidney and invigorating the spleen according to 4.8g/kg(using the quantity is equal to clinical dosage 10 times),one day at a time.The rats of estrogen supplemented were given nilestriol according to 1mg/kg,once a week.the others were given distilled water,one day at a time.After treating 3 months, mitochondrion were extracted from the muscle tissue that was collecte during the operation,and cytochrome coxidase(COX),the mitochondrial permeability transition pore(MPPT) and reactive oxygen species(ROS) were detected.ResultThe clinical experiment①In the optical delnsity(OD) value of MPPT,OD value of non-osteoporosis group were 0.182±0.047 at 0 min and 0.181±0.046 at 30min.OD value of Osteoporosis group were 0.143±0.071 at 0 min and 0.141±0.071 at 30min. Over time,OD value of the two groups showed a downward trend,there was a statistically significant difference between OD value of 30 min and of 0 min in two group(p＜0.05),that showed MPPT activity,but there was no a statistically significant difference between OD value of two group at the same time(p＞0.05).①In the permeability of MPPT,the ratio of OD value of non-osteoporosis group were 0.994±0.007,osteoporosis group were 0.985±0.006,there was a statistically significant difference between two group(p＜0.05).In the COX, non-osteoporosis group were 0.338±0.142,osteoporosis group were 0.124±0.093,there was a statistically significant difference between two group(p＜0.05).③In the correlation analysis of MPPT and COX,there was positive correlation between the ratio of OD value of MPPT and COX(r=0.405,P=0.04＜0.05),the regression equation was Y(COX)=388.8-647.402X(the ratio of OD value of MPPT).The animal experiment①35 rats were all involved into the result analysis.②In the optical delnsity(OD) value of MPPT,the change of MPPT of each group showed a downward trend.There was a statistically significant difference between OD value of 30 min and of 60 min in normal control group(p＜0.05). There was a statistically significant difference between OD value of 60 min and of other time in ovariectomized group(p＜0.05).There was also a statistically significant difference between OD value of 30 min and of 0 min in ovariectomized group(p＜0.05).There was no a statistically significant difference in all time in estrogen supplemented group(p＞0.05). There was a statistically significant difference between OD value of 60 min and of other time in traditional chinese herbs group(p＜0.05),but there was no a statistically significant difference between OD value of 30 min and of 0 min(p＞0.05).③In the permeability of MPPT,in the ratio of OD value of 30min/0min,the ratio of OD value of normal control group were 1.000±0.025,the ovariectomized group were 0.975±0.014,the estrogen supplemented group were 0.993±0.022 and the traditional chinese herbs group were 0.990±0.010.in 60min/0min,the ratio of OD value of normal control group were 1.000±0.047,the ovariectomized group were 0.956±0.016,the estrogen supplemented group were 0.976±0.025 and the traditional chinese herbs group were 0.976±0.025.There was a statistically significant difference that the ratio of OD value of normal control group,of the estrogen supplemented group respectively compared with the ovariectomized group in 30min/0min and 60min/0min(p＜0.05),and there was no a statistically significant difference between the traditional chinese herbs group and other groups in 30min/0min and 60min/0min(p＞0.05).Compared with the ratio of OD value of 30min/0min and of 60min/0min,There was a statistically significant difference in the normal control group(p＜0.05),there was no a statistically significant difference in the other groups(p＞0.05).④In the COX,the value of normal control group were 0.0200±0.0141,the ovariectomized group were 0.0070±0.0051,the estrogen supplemented group were 0.0134±0.0064 and the traditional chinese herbs group were 0.0130±0.0092。There was a statistically significant difference between the normal control group and the ovariectomized(p＜0.05).There was no a statistically significant difference that the value of the estrogen supplemented group,of the traditional chinese herbs group respectively compared with of the ovariectomized group(p＞0.05).⑤In the ROS,the value of normal control group were 1898.89±1078.39,1900.00±1078.76,1911.33±1078.14 and 1911.33±1077.37,the ovariectomized group were 2271.22±1349.76,2282.79±1350.7,2297.22±1376.74 and 2297.00±1375.35,the estrogen supplemented group were 2227.67±1365.95, 2239.89±1363.75,2244.11±1363.55 and 2242.00±1357.36,the traditional chinese herbs group were 1536.25±835.51,1576.38±836.50,1550.86±835.27 and 1550.86±835.27.There was no a statistically significant difference in all groups in all time,but in these time,the ovariectomized group's content were highest,next were the estrogen supplemented group and the normal control group,finally were the traditional chinese herbs group.ConclusionThe change of muscle mitochondria and osteoporois have a relationship of mutual influence.Through drug intervention,The change of muscle mitochondria could be effective in controlling,muscle cell apoptosis could be inhibited,and finally the development of osteoporosis could be controlled effectively.The main conclusions are as follows.①In the permeability of MPPT,the permeability of MMPT of osteoporotic muscle is more greater,it means that there is more apoptosis tostimulate or passes MPPT,and induces the cell apoptosis more quickly.It shows that the damage of muscle mitochondria in osteoporosis compares the non-osteoporosis to be serious②In the COX,the enzymatic activity of COX of osteoporotic muscle is worse than non-osteoporosis,that shows mitochondrial outer membrane integrity of osteoporotic muscle is worse than non-osteoporosis,and apoptosis rate of osteoporotic muscle cell is faster than non-osteoporosis.③In the relationship between the ratio of OD value of MPPT and the enzymatic activity of COX,there is positive correlation between the ratio of OD value of MPPT and COX,but the ratio of OD value of MPPT and the permeability are the inverse relationship.It means that the permeability of MPPT is greater,the smaller the enzymatic activity of COX,as long as maintain the enzymatic activity of COX,we could effectively controll cell premature aging.From another side,it provide a new therapeutic target for the treatment of osteoporosis④In the ROS,there is no significant differences in non-osteoporosis and osteoporosis.The possible reason is that ROS is insensitive to rat skeletal muscle tha is lack of estrogen.But over time,the Content of ROS is increasing that means cells continue to accumulate,and start apoptosis when reached a certain amount. ⑤In the MMPT,COX and ROS,herbs of reinforcing the kidney and invigorating fails to achieve significant results,compared with estrogen.But from its value,we believe that herbs of reinforcing the kidney and invigoratin improves the permeability of MPPT and the enzymatic activity of COX,reduces the the content of ROS.So it is one of drug that research the skeletal muscle as a therapeutic target in the future. |
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