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Sinomenine increases osteogenesis in mice with ovariectomyinduced bone loss by modulating autophagy

World Journal of Stem Cells 2024年第5期16卷 486-498页

作者： Hai-Xiang Xiao Lei Yu Yu Xia Kai Chen Wen-Ming Li Gao-Ran Ge Wei Zhang Qing Zhang Hong-Tao Zhang De-Chun Geng Department of Orthopedics The Fourth Affiliated Hospital of Soochow UniversitySuzhou Dushu Lake HospitalMedical Centre of Soochow UniversitySuzhou 215006Jiangsu ProvinceChina Department of Orthopedics Jingjiang People’s Hospital Affiliated to Yangzhou UniversityJingjiang 214500Jiangsu ProvinceChina Department of Orthopedics The First Affiliated Hospital of Soochow UniversitySuzhou 215006Jiangsu ProvinceChina Department of Orthopedics Hai’an People’s HospitalHai’an 226600Jiangsu ProvinceChina Department of Orthopedics The Affiliated Huai’an Hospital of Xuzhou Medical UniversityHuai’an Second People’s HospitalXuzhou 223002Jiangsu ProvinceChina Department of Orthopaedics The First Affiliated Hospital of Soochow UniversitySuzhou 215006Jiangsu ProvinceChina

BACKGROUND A decreased autophagic capacity of bone marrow mesenchymal stromal cells(BMSCs)has been suggested to be an important cause of decreased osteogenic differentiation.A pharmacological increase in autophagy of BMSCs is a potential therapeutic option to increase osteoblast viability and ameliorate osteoporosis.AIM To explore the effects of sinomenine(SIN)on the osteogenic differentiation of BMSCs and the underlying mechanisms.METHODS For in vitro experiments,BMSCs were extracted from sham-treated mice and ovariectomized mice,and the levels of autophagy markers and osteogenic differentiation were examined after treatment with the appropriate concen-trations of SIN and the autophagy inhibitor 3-methyladenine.In vivo,the therapeutic effect of SIN was verified by establishing an ovariectomy-induced mouse model and by morphological and histological assays of the mouse femur.RESULTS SIN reduced the levels of AKT and mammalian target of the rapamycin(mTOR)phosphorylation in the phosphatidylinositol 3-kinase(PI3K)/AKT/mTOR signaling pathway,inhibited mTOR activity,and increased autophagy ability of BMSCs,thereby promoting the osteogenic differentiation of BMSCs and effectively alleviating bone loss in ovariectomized mice in vivo.CONCLUSION The Chinese medicine SIN has potential for the treatment of various types of osteoporosis,bone homeostasis disorders,and autophagy-related diseases.

关键词： Sinomenine Osteogenesis Autophagy ovariectomy Osteoporosis

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The sphingosine-1-phosphate/RhoA/Rho associated kinases/myosin light chain pathway in detrusor of female rats is down-regulated in response to ovariectomy

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Chinese Medical Journal 2020年第10期133卷 1203-1210页

作者： Wei Zhang Xiao-Dong Liu Jia-Wen Wang Ling-Feng Meng Yao-Guang Zhang Jian-Ye Wang Peking University Fifth School of Clinical Medicine Beijing 100730China Department of Urology Beijing HospitalNational Center of GerontologyBeijing 100730China.

Background:Dysuria is one of the main symptoms of genitourinary syndrome of menopause,which causes serious disruption to the normal life of peri-menopausal women.Studies have shown that it is related to decrease of detrusor contractile function,but the exact mechanism is still poorly understood.Previous results have suggested that the sphingosine-1-phosphate(S1P)pathway can regulate detrusor contraction,and this pathway is affected by estrogen in various tissues.However,how estrogen affects this pathway in the detrusor has not been investigated.In this study,we detected changes of the S1P/RhoA/Rho associated kinases(ROCK)/myosin light chain(MLC)pathway in the detrusor of ovariectomized rats in order to explore the underlying mechanism of dysuria during peri-menopause.Methods::Thirty-six female Sprague-Dawley rats were randomly divided into SHAM(sham operation),OVX(ovariectomy),and E groups(ovariectomy+estrogen),with 12 rats in each group.We obtained bladder detrusor tissues from each group and examined the mRNA and protein levels of the major components of the S1P/RhoA/ROCK/MLC pathway using quantitative real-time polymerase chain reaction and Western blotting,respectively.We also quantified the content of S1P in the detrusor using an enzyme linked immunosorbent assay.Finally,we compared results between the groups with one-way analysis of variance.Results::The components of the S1P pathway and the RhoA/ROCK/MLC pathway of the OVX group were significantly decreased,as compared with SHAM group.The percent decreases of the components in the S1P pathway were as follows:sphingosine kinase 1(mRNA:39%,protein:45%)(both P0.05).The percent decreases of the components in the RhoA/ROCK/MLC pathway were as follows:ROCK2(protein:41%,mRNA:36%)(both P<0.0

关键词： Myosin light chains ovariectomy Rats Rho-associated kinases Sphingosine-1-phosphate Urinary bladder

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Impact of ovariectomy and estradiol-17β(E_(2))replacement on the brain steroid levels of the Indian stinging catfish Heteropneustes fossilis

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Aquaculture and Fisheries 2022年第5期7卷 474-483页

作者： Surabhi Mishra Radha Chaube Zoology and Fisheries Department Dolphin(PG)College of Science and AgricultureChunni KalanFatehgarh SahibPunjab140406India Department of Zoology Institute of ScienceBanaras Hindu UniversityVaranasi221005India

In the present study,effects of ovariectomy(Ovx)and estradiol-17β(E2)replacement on the brain steroid levels were investigated in the resting and prespawning phases of the female catfish Heteropneustes fossilis.The study showed that Ovx resulted in significant decreases in plasma levels of estradiol-17β(E2)and its precursors,and the effect varied with the reproductive stage and duration.Our study showed that E2 supplementation reversed the effect of Ovx in 3-week Ovx fish group,either restored or increased according to the dose.In contrast,E2 levels increased in the brain after Ovx in the resting stage and produced a biphasic effect of significant decreases on week 1 and 2,and significant increases on week 3,4 and 5 in the pre-spawning stage.The brain E2 levels significantly inhibited in E2 supplementation groups.The testosterone(T)levels showed biphasic effects,an initial decrease(up to 3 weeks Ovx)and an increase later(week 4,5)in the resting stage.In the pre-spawning stage,the T levels increased significantly after Ovx.The E2 supplementation significantly inhibited the T levels,more severely in the low dose group in the resting and pre-spawning stages.Brain 17-hydroxyprogesterone(17-OHP4)levels increased(week 1,2,3)but decreased on week 4 and 5 of Ovx in the resting stage and increased throughout Ovx in the pre-spawning stage except in the 5th week(though compared to other steroids,level was very low;almost negligible).The E2 replacement significantly decreased the steroid levels further down compared to the Ovx control group.Brain progesterone(P4)and pregnenolone(P5)levels decreased initially(week 1 and 2 Ovx)and recovered later.The E2 replacement significantly decreased P4 levels in the Ovx fish in the pre-spawning stage and in the lower dose group in the resting stage.The P5 levels remained inhibited after the E2 supplementation in the resting stage but unchanged or increased it in the pre-spawning stage.Brain cortisol(F)levels were inhibited initially(1 and 2 weeks and later increased after OVX in the rest

关键词： Catfish Neurosteroids ovariectomy Estrogen replacement Seasonal variations

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艾灸“肾俞”早期干预去卵巢合D-半乳糖注射AD样大鼠效应中的特异性

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World Journal of Acupuncture-Moxibustion 2020年第4期30卷 277-284页

作者：杜艳军陶一鸣田青曾鹏吴文辉肖佳欢湖北中医药大学针灸骨伤学院武汉430061 华中科技大学同济医学院武汉430000 武汉大学中南医院武汉430000

Objective:To observe the effect of the early intervention with moxibustion at "Shènshū(肾俞BL23)" on learning and memory of Alzheimer’s disease(AD) rats modeled by ovariectomy and D-Galactose(D-Gal)injection and the effect on Ca2+/CaMKII/CREB signaling pathway so as to provide the evidence for the acupoint selection in the early intervention with acupuncture and moxibustion for AD.Methods:Sixty female SD rats at the age of 3 months were selected and randomly divided into a normal group,a sham-operation group,a model group,a "BL23" group,a "Wèishū"(胃俞BL21) group,and a non-acupoint group,10 rats in each group.Except in the normal group and the sham-operation group,bilateral ovaries were removed in the other 4 groups.3 days after the operation,D-Gal was injected intraperitoneally at a dose of 150 mg/kg/d for 90 days to establish AD rat model.In the sham-operation group,only the peripheral fat of ovarian was removed and 0.9% normal saline was injected intraperitoneally of the same dose for 90 days.In the "BL23" group,the "BL21" group and the non-acupoints group,since the 2 nd day after ovariectomy,moxibustion was applied at 9:00 a.***.every day in the rats.The rats were fixed on a fixed table,the mild moxibustion with self-prepared moxa stick was applied to bilateral "BL23""BL21" and the non-acupoint of each rat,the ignited end of moxa stick was 2 cm to 3 cm far from skin surface.The infrared thermometer was adopted to measure the temperature of the epidermis of each acupoint from time to time so as to maintain the local temperature to be(41 ± 0.5)℃.Mild moxibustion lasted for 10 min each time,consecutively for 5 days a week and at an interval of 2 days.The total consecutive intervention duration was 12 weeks.In the normal group and the shamoperation group,no moxibustion intervention was performed,but the rats were restricted in a same way during the experiment.At the end of modeling and intervention,water maze test was adopted to detect the learning and memory abilities of rats in each group.The western blotting was use

关键词： Alzheimer's disease(AD) Ca^2+/caMKII/CREB Learning and memory ability Moxibustion ovariectomy “Shènshū(肾俞BL23)”

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Prevention of Osteopenia and Dyslipidemia in Rats after ovariectomy with Combined Aspirin and Low-dose Diethylstilbestrol

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Biomedical and Environmental Sciences 2013年第4期26卷 249-257页

作者： LIN Si En HUANG Jian Ping WU Ling Zhi WU Tie CUI Liao Department of Pharmacology Guangdong Medical College Department of Stomatology Guangdong Medical College Guangdong Key Laboratory for Research and Development of Natural Drugs

Objective To study whether effect of aspirin plus low-dose diethylstilbestrol is more effective and safer than high diethylstilbestrol dose alone on prevention of ovariectomy-induced osteopenia and dyslipidemia. Methods Thirty-eight 4-month-old female SD rats were divided into baseline （BAS） group （n=6）, sham operation group （n=8） and ovariectomy （OVX） group （n=24）. The OVX group was further divided into vehicle treatment group （n=8）, diethylstilbestrol （30 ug/kg.d） treatment group （OVX＋D30 group, n=8）, and aspirin （9 mg/kg.d） plus diethylstilbestrol （10 ug/kg.d） treatment group （OVX＋A-D10 group, n=8）. Their left tibiae were collected for the bone histomorphometric analysis in undecalcified sections. Left femurs were collected for the bone mineral density measurement. Results The body weight and serum cholesterol were increased, while uterine weight and cancellous bone mass were decreased in OVX rats compared with the SHAM group. Cancellous bone mass was significantly increased, while body weight and bone resorption parameters were decreased in both A-D10 and D30 treatment group compared with OVX group. The rats treated with A-D10 showed significantly increased in bone formation parameters and decreased in serum triglyceride compared with the D30-treated rats. Conclusion Aspirin plus low-dose diethylstilbestrol can effectively prevent osteopenia by reducing bone resorption, and is thus a better treatment modality for preventing dyslipidemia than high-dose diethylstilbestrol alone.

关键词： Aspirin Diethylstilbestrol ovariectomy Osteoporosis Dyslipidemia Rat

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Effect of ovariectomy on Serum Adiponectin Levels and Visceral Fat in Rats

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2014年第6期34卷 825-829页

作者： Camara Ce 周林远马炎朱琳于东赵要武杨年红 Department of Nutrition and Food Hygiene Hubei Key Laboratory of Food Nutrition and Safety Huazhong University of Science and Technology Department of Nutrition and Food Hygiene Hubei Key Laboratory of Food Nutrition and SafetyHuazhong University of Science and Technology Key Lab of Environment and Health of the Ministry of Education School of Public Health Tongji Medical College Huazhong University of Science and Technology

This study was aimed to examine the effect of ovariectomy on visceral fat, serum adi- ponectin levels and lipid profile. Forty-five female Sprague DawIey rats were divided into three groups （n=15 each）： ovariectomized group （OVX）, ovariectomized plus estrogen-treated group （OVX＋E2）, and sham-operated group （SHAM）. Body weight, abdominal adipose tissues, serum adiponectin and lipid profile were measured and compared among the groups after three-month feeding post-surgery. Signifi- cant increases in body weight and visceral fat were found in ovariectomized rats when compared with sham-operated ones and significant increases were also observed in serum adiponectin, triglyceride and very low density lipoprotein cholesterol levels in ovariectomized rats. Body weight, visceral fat and se- rum adiponectin levels were profoundly reduced in OVX＋E2 group as compared with OVX group. It was concluded that ovarian hormone deficiency induced by ovariectomy leads to significant increases in body weight and visceral fat, along with increased serum adiponectin, triglyceride and very low density lipoprotein cholesterol levels in rats. Attenuation in these changes can be achieved by estrogen supple- mentation.

关键词： ovariectomy estrogen post menopause serum adiponectin visceral fat

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Preventive effect of Peperomia pellucida(L.) Kunth herbs on ovariectomyinduced osteoporotic rats

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Journal of Chinese Pharmaceutical Sciences 2016年第7期25卷 546-551页

作者： Cynthia Astiti Putri I Gusti Agung Ayu Kartika I Ketut Adnyana Department of Clinical Pharmacology and Toxicology School of Pharmacy Bandung Institute of Technology

Osteoporosis is a metabolic skeletal disorder leading to bone fracture which adversely impacts the quality of life. In the present, we aimed to investigate the effect of Peperomia pellucida(L.) Kunth herb juice and ethanolic extract on ovariectomy(OVX)-induced osteoporotic rat model. Osteoporosis was induced by OVX with a double dorsolateral approach on female Wistar rats. Female Wistar rats undergoing sham surgeries(sham-control group) served as controls. The ovariectomized rats were divided into six groups based on administered treatments,(i) CMC Na 0.3%(OVX-control group),(ii) standard drug(ethynil estradiol 4.5 μg/kg),(iii) P. pellucida juice at dose of 50 mg/kg, and(iv) 100 mg/kg,(v) P. pellucida ethanolic extract at dose of 50 mg/kg, and(vi) 100 mg/kg. Treatments were started 1 week after the surgeries and lasted for 6 weeks. Rats treated with 100 mg/kg P. pellucida ethanol extract had significantly decreased serum ALP level and reduced excretion of urine calcium compared with the OVX-control group(P<0.05). These levels were not significantly altered when compared with the sham-control group(P<0.05). Furthermore, 100 mg/kg ethanolic extract-treated group showed improvement on three-dimensional image of the trabecular bone compared with the OVX-control group. Trabecular cavity formation in 100 mg/kg ethanolic extract-treated group was minimal. Ethanolic extract of Peperomia pellucida(L.) Kunth herbs at a dose of 100 mg/kg had preventive effect on OVX-induced osteoporotic rats.

关键词： Osteoporosis ovariectomy Peperomia pellucida(L.)Kunth

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Protective effect of steroidal saponins from rhizome of Anemarrhena asphodeloides on ovariectomy-induced bone loss in rats

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Acta Pharmacologica Sinica 2006年第6期27卷 728-734页

作者： Hua NIAN~2 Lu-ping QIN~2 Wan-sheng CHEN~3 Qiao-yan ZHANG~(2,4)Han-chen ZHENG~2 Yin WANG~5 2 Department of Pharmacognosy,School of Pharmacy,Second Military Medical University,Shanghai 200433.China 3 Department of Pharmacy,Changzheng Hospital,Second Military Medical University,Shanghai 200003,China 5 № 455 Hospital of PLA,Shanghai 200052,China DepartmentofPharmacognosy SchoolofPharmacySecondMilitaryMedicalUniversityShanghai200433China DepartmentofPharmacy ChangzhengHospitalSecondMilitaryMedicalUniversityShanghai200003China No455HospitalofPLA Shang

Aim:To investigate the protective effect of steroidal saponins from Anemarrhenaasphodeloides(ATS)on ovariectomy(OVX)-induced bone loss.Methods:Sprague-Dawley rats were divided into sham and OVX groups.The OVX ratswere treated with vehicle,nylestriol or steroidal saponins extract for 12 weeks.Serum calcium,phosphorus,estradiol(E2),osteocalcin concentration and serumalkaline phosphatase activity were measured.Bone density was assayed by dual-energy X-ray absorptiometry.The undecalcified longitudinal proximal tibial meta-physical(PTM)sections were cut and stained for histomorphometric analysis ofthe bone.Results:In OVX rats,alkaline phosphatase activities in serum weremarkedly increased and concentrations of osteocalcin were decreased by ATStreatment,which had no influence on the body weight.Meanwhile,atrophy of theuterus and descent of bone mineral density(BMD)was suppressed by treatmentwith ATS.In addition,ATS completely corrected the decreased the concentrationof calcium and E2in serum observed in OVX rats.Histological results showedATS prevented decreases in trabecular thickness and increases in trabecular sepa-ration of proximal tibia metaphysis(PTM)in OVX rats.However,it did not alterosteoclast number in OVX rats.Moreover,ATS(300 mg/kg)had a remarkableeffect on promoting bone formation action in OVX rats.Nylestriol treatmentdecreased the bone formation rate and mineral apposition rate.Conclusion:Anadequate supply of steroidal saponins of Anemarrhena asphodeloides preventedOVX-induced bone loss in rats through the promotion of bone formation but notthe inhibition of bone resorption.

关键词： Anemarrhena asphodeloides ovariectomy osteoporosis serum bone mineral density histochemistry

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Tanshinone prevents cancellous bone loss induced by ovariectomy in rats

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Acta Pharmacologica Sinica 2004年第5期25卷 136-142页

作者： Liao CUI2,3, Tie WU3, Yu-yu LIU3, Yi-feng DENG4, Chun-mei AI4, Huai-qing CHEN22Institue of Biomedical Engineering, West China Medical Center, Sichuan University, Chengdu 610041, China 3Department of Pharmacology, 4Guangdong Key Laboratory for Research and Development of Natural Drugs,Guangdong Medical College, Zhanjiang 524023 China 四川大学 Chengdu 610041 Department of Pharmacology Guangdong Medical College Zhanjiang 524023 Department of Pharmacology Guangdong Medical College Zhanjing 524023 Guangdong Medical College Guangdong Key Laboratory for Research and Development of Natural Drugs Zhanjiang 524023

AIM: To investigate the skeletal effects of total tanshinone in ovariectomized rats by analyzing cancellous bonehistomorphometry of fourth lumbar vertebrae (LV4) and proximal tibial metaphyses (PTM). METHODS: Four-month-old Sprague-Dawley female rats were sham-operated and treated with vehicle or ovariectomized and treatedwith either vehicle, total tanshinone (200 mgkg-1d-1, equivalent to 35 gkg-1d-1 of tanshinone II A and 16 mgkg -1d-1 ofcryptotanshinone), or 17a-ethynylestradiol (30 gkg-1d-1 as positive treatment group) starting one day post-sur-gery for 10 weeks. Double in vivo fluorochrome labeling was administered to all rats. The undecalcified longitu-dinal LV4 and PTM sections were cut and stained with Goldners Trichrome (4-m thickness) or unstained (8-mthickness) for the bone histomorphometric analysis. RESULTS: A significant decrease in trabecular bone volume(BV/TV) and trabecular number (Tb.N) and a significant increase in osteoclast surface (OCS/BS) and mineralizingsurface (MS/BS) were found in both LV and PTM of vehicle-treated OVX rats compared with sham controls.Tanshinone completely prevented the decreases in BV/TV and Tb.N and the increase in OCS/BS in the LV4, andpartially prevented the decreases in BV/TV and Tb.N in the PTM of OVX rats. In addition, tanshinone increasedtrabecular thickness (Tb.Th) whereas it did not alter MS/BS. Moreover, tanshinone had no effect on uterine weightand body weight of OVX rats. Estrogen treatment increased BV/TV and Tb.N and decreased OCS/BS, but, alsomarkedly decreased MS/BS and increased uterine weight in OVX rats. CONCLUSION: The current study demon-strated that the adequate supply of tanshinone prevented OVX-induced cancellous bone loss in rats through inhibi-tion of elevated bone resorption.

关键词： tanshinone cryptotanshinone 2-fluoro-17alpha-ethynylestradiol ovariectomy bone and bones osteoclasts osteoporosis histocytochemistry morphology

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The Validity of Osteoarthritis Model Induced by Bilateral Ova-riectomy in Guinea Pig

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2006年第6期26卷 716-719页

作者：戴国锋王韶进李建民刘春梅刘巧慧 Department of Orthopedics Qilu Hospital Shandong University

To evaluate the validity of osteoarthritis model induced by bilateral ovariectomy in guinea pig, 32-month-old female guinea pigs were randomly divided into two groups： a sham operation group （control group） and an ovariectomized group （OVX group）. The animals were killed 6 or 12 weeks after the operation and the degeneration of the knees were assessed microscopically and histologically by scanning electron microscope （SEM）, transmission electron microscope （TEM） and light microscope. The serum levels of estrogen and gestone were detected by immune contest assay. The scoring of articular cartilage histopathology of tibial plateau was performed by histopathological examination. The blood serum levels of estrogen and gestone were decreased significantly in the OVX group as compared with the control group 6 or 12 weeks after the operation. Joint cartilage degeneration as detected by SEM and TEM could be found at the 6th week, but severe degenerative lesions were observed at the 12th week in the OVX group as compared with the control group （P〈0.01） The histopathological score of articular cartilage in tibial plateau in OVX group was higher than that of control group, which was coincident with the changes of estrogen and the ultrastructure （P〈0.01）. The findings suggested that bilateral ovariectomy in guinea pig can induce the severe osteoarthritis that is similar to the aging-induced OA in human. Therefore, the model of the osteoarthritis by bilateral ovariectomy in guinea pig in this study is valid.

关键词： animal model osteoarthritis ovariectomy validity

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