| BackgroundDiminished Ovarian Reserve(DOR)refers to a disease in which the number and/or quality of available oocyte in the ovaries decreases.Among them,the DOR that occurs with age is a physiological change,and when the ovarian function decreases too early and too quickly compared to healthy women of the same age,it is a pathological DOR.In recent years,the incidence and detection rate of DOR have increased year by year,and there is a trend of younger age,if not intervented,DOR will seriously affects women’s physical and mental health and fertility,but also leads to cardiovascular and cerebrovascular diseases.The risk of osteoporosis is significantly increased.Although bone loss is affected by genetics,environment and other factors,for women,the weakening of ovarian function is the most important factor.The incidence of Osteoporosis in women over 50 years old in China is 32.1%,and the fracture risk of menopausal women before the age of 45 is 1.5-3 times that of normal age menopauses,and the risk of fracture in POI patients is higher.DOR women are younger,often without obvious chest and back pain,spinal deformation,short height and other typical symptoms of bone loss,which is also the reason why they do not pay attention to their bone mass problems,resulting in many patients in the initial examination found that bone mass has different degrees of reduction,and even reached the diagnostic criteria of osteoporosis.As a high-risk group for Osteoporosis,women with DOR need our attention and actively seek effective measures to prevent and treat this type of disease,so as to delay the progression of such diseases and improve the quality of life for this group of women.On the basis of inheriting Professor Cai Lianxiang’s theory of "essence and blood",the mentor,Director Huang Yuxiao has long been committed to the research of "kidney store essence and main reproduction" theory and the prevention,treatment,and the management of DOR.She has been using Kidney Tonifying and Essence Filling Decoction to treat DOR year-round and the clinical effect is quite obvious and stable.This study intends to explore the effect of Kidney Tonifying and Essence Filling Decoction on the improvement of bone metabolism related DOR,so as to expand the clinical application of this method and provide theoretical basis and evidence support for clinical research on the management of chronic diseases with ovarian dysfunction.Objective1.To observe and compare the changes in bone density,serum bone metabolism indicators,serum endocrine hormones,symptom scales,and traditional Chinese medicine syndrome scores of patients with DOR before and after intervention with Kidney Tonifying and Essence Filling Decoction.2.To observe the effects of Kidney Tonifying and Essence Filling Decoction on bone density and femur microstructure in mice with cisplatin-induced Diminished Ovarian Reserve.Methods1.Clinical study on the effect of Kidney Tonifying and Essence Filling Decoction on bone metabolism in patients with Diminished Ovarian ReserveCollect 30 patients who visited the clinic from December 2021 to December 2022,met the diagnosis of DOR in Western medicine,and were diagnosed with liver kidney yin deficiency according to traditional Chinese medicine,with a bone mineral density T values between-2.50 to-1.0 in the lumbar spine or left hip joint as the research subjects.Starting from the 5th day of menstruation,the Kidney Tonifying and Essence Filling Decoction was orally given for a total of 3 months as a course of treatment.Observe and compare the changes in bone density,serum bone metabolism indicators,serum endocrine hormones,symptom scales,and traditional Chinese medicine syndrome scores of patients with DOR before and after treatment,and evaluate the safety of treatment.2.Experimental study on the effect of Kidney Tonifying and Essence Filling Decoction on bone quality in mice with Diminished Ovarian Reserve model52 mice with regular estrous cycles were selected from 60 aged 6-8 weeks SPF grade female C57BL/6J mice fed adaptively and observed for 12 consecutive days for inclusion in the experiment,and according to the random number table method,16 mice were divided into blank group and 36 mice in chemotherapy group.Each mouse in the chemotherapy group was injected intraperitoneally with cisplatin solution(concentration 2mg/mL)0.2mL/d for 7 consecutive days to replicate the DOR model,and 0.2mL of 0.9%normal saline was injected intraperitoneally in the blank group for 7 consecutive days.After the modeling,6 mice in each group were randomly selected to measure serum FSH,E2 and AMH levels to verify the modeling.After successful modeling,the remaining 30 mice in the chemotherapy group were randomly divided into three groups:model group,estradiol valerate group,and Kidney Tonifying and Essence Filling Decoction group,with 10 mice in each group.From the 8th day,Each mouse in the Kidney Tonifying and Essence Filling Decoction group and estradiol valerate group was gavaged with corresponding drug solution of 0.2mL/d,and each mouse in the model group and blank group were gavaged with 0.2mL of 0.9%normal saline.After 21 days of continuous gavage,mouse femur samples were collected,the bone density of the femur was measured and the pathological morphological changes of femur histologis were observed between the groups,so as to evaluate the intervention effect of Chinese medicine on bone quality of mice.Results1.Clinical study on the effect of Kidney Tonifying and Essence Filling Decoction on bone metabolism in patients with Diminished Ovarian Reserve1.1 The total effective rate of bone mineral density after treatment was 79.16%.There was an increasing trend in lumbar bone density,but there was no statistically significant difference before and after treatment(P>0.05);After treatment,the bone density of the left hip joint significantly increased,and the difference before and after treatment was statistically significant(t=2.09,P=0.037).1.2 The total effective rate of traditional Chinese medicine syndrome score was 95.45%.After treatment,the MENQOL scale score and traditional Chinese medicine syndrome score were significantly improved compared to before treatment,and the differences were statistically significant(P<0.001).1.3 After treatment,the serum bE2 levels of DOR patients were significantly improved compared to before treatment,and the differences were statistically significant(P=0.046).1.4 After treatment,the serum bFSH,AMH,VITD-T,OC,CT and other indicators of DOR patients improved,but there was no statistically significant difference before and after treatment(P<0.05).2.Experimental study on the effect of Kidney Tonifying and Essence Filling Decoction on bone quality in mice with Diminished Ovarian Reserve model2.1 Changes in the estrus cycleIn the adaptive feeding stage,the estrus cycle of mice in each group was relatively regular,which was 4-5 days.After 7 days of modeling,the mice in the chemotherapy group had estrous cycle disorder compared with the blank group,indicating that the ovarian function was inhibited.During 21 days of gavage,the estrus cycle of mice in the traditional Chinese medicine group and the western medicine group had a stable trend.2.2 Comparison of serum hormones after modelingAfter seven days of modeling,the overall mean serum FSH of mice in the blank group was(2.52±0.14)mIU/mL,and the overall mean serum FSH of mice in the chemotherapy group was(5.73±0.79)mIU/mL,and the difference between the two groups was statistically significant(t=5.32,P<0.001).Seven days after modeling,the overall mean serum E2 of mice in the blank group was(213.57±35.27)pg/mL,and the overall mean serum E2 of mice in the chemotherapy group was(594.08±222.08)pg/mL,and the difference between the two groups was statistically significant(t=4.15,P=0.008).Seven days after modeling,the median serum AMH of mice in the blank group was 161.03(154.46,205.48)pg/mL,and the median serum AMH of mice in the chemotherapy group was 169.77(155.43,231.65)pg/mL,and the difference between the two groups was statistically significant(t=2.32,P=0.046).The above results prove that the mouse DOR model has been successfully constructed.2.3 Comparison of femoral bone density after treatmentThe average bone density of the blank group was(0.064±0.002)g/cm2,the average bone density of the model group was(0.055±0.006)g/cm2,the average bone density of the western medicine group was(0.060±0.005)g/cm2,and the average bone density of the traditional chinese medicine group was(0.061 ±0.003)g/cm2.Compared with the blank group,the bone density of mice in the model group was reduced,and the difference was statistically significant(P<0.001).Compared with the model group,the bone density of mice in the western medicine group was increased and the difference was statistically significant(P=0.043).Compared with the model group,the bone density of mice in the traditional chinese medicine group was increased and the difference was statistically significant(P=0.011).Compared with the western medicine group,there was no significant difference in bone density between the traditional Chinese medicine group and the western medicine group(P>0.05).2.4 Comparison of femoral microstructures after treatmentCompared with the blank group,the separation degree of femoral trabeculae in the model group was significantly increased,and the number and thickness of bone trabeculae were significantly reduced.Compared with the model group,the number and thickness of bone trabeculae in the traditional chinese medicine group and the western medicine group increased significantly,but they were worse than those in the blank group.The number and thickness of bone trabeculae in the chinese medicine group and the western medicine group were similar.Conclusion1.Kidney Tonifying and Essence Filling Decoction can effectively improve the Bone Mineral Density of the patients with Diminished Ovarian Reserve,and has an improvement trend in serum endocrine hormones and bone metabolism levels.It can alleviate clinical symptoms and improve quality of life for patients.2.Kidney Tonifying and Essence Filling Decoction can effectively improve the bone density of cisplatin-induced DOR mice,improve their bone microstructure,increase the thickness and number of bone trabeculae,and reduce the risk of fracture. |
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