Zuogui pills for myelinolysis in a rat model of experimental autoimmune encephalomyelitis

Zuogui pills have been shown to attenuate the inflammatory reaction in a rat model of experimental autoimmune encephalomyelitis （EAE）. The present study attempted to investigate the pathology underlying the influence of Zuogui pills on myelinolysis in EAE rats. Hematoxylin-eosin and Luxol fast blue staining showed that the myelinolysis foci in the cerebrum, cerebellum, brain stem, and the spinal cord of EAE rats were significantly decreased, along with serum myelin basic protein content following treatment with Zuogui pills.

Fourier transform infrared spectroscopy analysis of the active components in serum of rats treated with Zuogui Pill

Objective:To investigate the active components Zuogui Pill,a typical recipe for nourishing kidney essence in the traditional Chinese medicine.Methods:Adult male Sprague Dawley rats were treated with the traditional Chinese herbal medicine Zuogui Pill and the active components found in the serum of the animals were analyzed by Fourier transform infrared(FTIR)spectroscopy.FTIR spectra of serum samples of treated and untreated rats were analyzed and the A2960/A2931 and A1540/A1080 ratios were calculated.Results:A2960/A2931 ratios of the serum samples collected following the administration of Zuogui Pill were significantly higher than those of the normal serum samples.FTIR data were then fitted using a Gaussian equation for wave numbers in the range of 1140e1000 cm
1.ARNA/ADNA ratios in the serum of rats treated with Zuogui Pill were higher than those found in normal rat serum.Conclusion:FTIR spectroscopy could be used as an analytical tool to detect the activecomponents in serum of animals treated with Zuogui Pill.

Regulatory Effects of Zuogui Pill on Apoptosis of Follicles in Rats Injured by 60Co-γRays Based on PI3K/Akt/m TOR Signaling Pathway

[Objectives]To explore the protective effects of Zuogui Pill on ^(60)Co-γ-ray-induced premature aging of rats based on phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/Akt/mTOR)signaling pathway.[Methods]Sixty sexually mature female SD rats were irradiated with ^(60)Co-γ-ray(6.0 Gy,LD 40)for 24 h at one time.These rats were randomly divided into model group,Progynova group[0.18(g·kg)/d],Progynova[0.09(g·kg)/d]+Zuogui Pill high dose[23.625(g·kg)/d)]group,Zuogui Pill high dose[23.625(g·kg)/d)]group,Zuogui Pill medium dose[9.45(g·kg)/d)]group and Zuogui Pill low dose[4.725(g·kg)/d]group.The administration(once a day)lasted 21 d.The rat serum[follicle-stimulating hormone(FSH),luteinizing hormone(LH)and estradiol(E_(2))]were detected by Enzyme-linked immunosorbent assay(ELISA).The morphological changes of ovary were observed by hematoxylin-eosin(HE)staining.The apoptosis rate of granulosa cells was detected by terminal deoxynucleotidyl transferase(TdT)-mediated dUTP nick-end labeling(TUNEL).The protein expression of phosphorylated(p)-PI3K,p-Akt,p-mTOR,B-cell lymphoma-2(Bcl-2),and Bcl-2-associated X protein(Bax)in ovarian tissues were detected by Western blot.[Results]Compared with the normal group,the model group showed significant increase in the serum FSH(P<0.01),significant decrease in serum E_(2)(P<0.05),and decrease in the number of early follicles and luteum in the ovary(P<0.01).Besides,the apoptosis rate of granulosa cells increased significantly(P<0.01);the expression of p-PI3K,p-Akt,p-mTOR and Bcl-2 in ovarian tissue decreased significantly,while the expression of Bax increased significantly(P<0.01).Compared with the model group,the number of early follicles in the ovary increased and the apoptosis rate of granulosa cells decreased after intervention in each administration group.In addition,the protein expressions of p-PI3K,p-Akt,p-mTOR and Bcl-2 increased,while the expression of Bax decreased,especially in Progynova+Zuogui Pill high dose group,the differences were statistically significant(P<0.05,P<0.01).[Conclusions]Zuogui Pill may protect the radiation-injured ovary through activating the expression of PI3K/Akt/mTOR protein in ovarian tissue,increasing the amount of Bcl-2 protein and inhibiting the expression of Bax protein.

Network pharmacological study of Zuogui pill and Yougui pill for the treatment of postmenopausal osteoporosis

Background:Postmenopausal osteoporosis(PMO)is the most common primary osteoporosis in older women.This condition imposes a huge economic and medical burden on society.Aside from western medicine,traditional Chinese medicine is also widely used for the treatment of PMO,especially in Asian countries.Zuogui pill(ZGP)and Yougui pill(YGP)are classical formulas for the treatment of PMO with potent clinical effects.This study aimed to explore the potential active compounds,potential targets,and potential mechanisms of ZGP and YGP for the treatment of PMO.Methods:Compounds from ZGP and YGP were collected from three online databases,and these compounds were screened by oral bioavailability and drug-likeness.Their corresponding targets were determined from the Medicine Systems Pharmacology Database and Analysis platform.The corresponding targets for PMO were obtained from two disease databases.The target protein-protein interaction was identified through the STRING platforms and the core targets were ascertained by analyzing the protein-protein interaction network by using Cytoscape software.PMO-related genes were identified via Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses to identify specific biological processes,cellular components,molecular functions and key signalling pathways of ZGP and YGP for PMO treatment.Results:A total of 68 compounds were screened from ZGP with 168 putative potential target genes,whereas 99 compounds were screened from YGP with 214 potential target genes associated with PMO.Most of the core components included quercetin and kaempferol,and most of the core targets were TP53,AKT1,MAPK1,JUN,TNF,HSP90AA1,APP,IL6,VEGFA,RELA,MAPK8,NR3C1,EGFR,CXCL8,ESR1,FOS and MAPK14.Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed that ZGP and YGP treat PMO primarily via regulation of bone formation and resorption,anti-inflammatory immunity and hormonal regulation.Conclusion:Our data provided insights into the mechanisms,by which ZGP and YGP treat PMO.This study points out a direction for further research into ZGP and YGP monomers and pathways and offers a new clinical treatment option for non-traditional Chinese medicine clinicians in the treatment of PMO.

Effects of Zuogui Pill (左归丸) on Wnt Singal Transduction in Rats with Glucocorticoid-induced Osteoporosis

Objective: To reveal the mechanism of Zuogui Pill (左归丸) in treatment of glucocorticoid-induced osteoporosis from the angle of the Wnt signal transduction pathway and to provide further experimental evidence for expounding the scientific connotation of "the kidney dominating the bones" in TCM. Methods: Forty-two male Wistar rats were selected and randomly divided into three groups, control group (n=12), model group (n=15) and Zuogui Pill group (n=15). Form the beginning, The rats were injected dexamethasone for eight weeks to make the model of osteoporosis, and the Zuogui Pill were administered intragastrically to the rats of Zuogui Pill group for eight weeks. The relative morphological parameters were measured in the undecalcified tibial slices. And the protein expression levels of Wnt1, LRP-5 and β-catenin in rat tibial osteoblasts (OB) and bone marrow stromal cells (BMC) were detected by immunohistochemistry. Results: Compared with the control group, TBV% and TFS% decreased significantly, while TRS% increased significantly, and the protein expression of Wnt1, LRP-5 and β-catenin in OB and BMC decreased significantly in the model group. And compared with the model group, TBV% and TFS% increased significantly, and expression levels of Wnt1, LRP-5 and β-catenin proteins increased significantly in the Zuogui pill group. Conclusion: Zuogui Pill can prevent and treat glucocorticoid-induced osteoporosis in rats by up-regulating the expression of the key signal molecules Wnt1, LRP-5 and β-catenin in Wnt signal transduction pathway.

Effects of Zuogui Pill(左归丸) and Yougui Pill(右归丸) on the Expression of Brain-Derived Neurotrophic Factor and Cyclic Adenosine Monophosphate/Protein Kinase A Signaling Transduction Pathways of Axonal Regeneration in Model Rats with Experimental Autoimm

Objective：To study the effects of Zuogui Pill（左归丸,ZGP）and Yougui Pill（右归丸,YGP）on the expressions of brain-derived neurotrophic factor（BDNF）and cyclic adenosine monophosphate（cAMP）/protein kinase A（PKA）signaling of axonal regeneration in the Lewis rats with experimental autoimmune encephalomyelitis（EAE）,in order to explore the possible mechanism of ZGP and YGP on promoting axonal regeneration.Methods：The rats were randomly divided into normal control（NC）,model（MO）,prednisone acetate（PA）,ZGP and YGP groups.The EAE model of rat was established by injecting antigen containing myelin basic protein（MBP）68-86.The brain and spinal cord were harvested on the 14th and 28th day postimmunization（PI）,the protein and mRNA expression of BDNF and PKA in the brain and spinal cord of rats were detected by Western blot analysis and real-time quantitative polymerase chain reaction（PCR）,and the cAMP levels were detected by using enzyme-immunoassay method.Results：（1）On the 28th day PI,the mRNA expression of BDNF in brain white matter and spinal cord of rats in ZGP and YGP groups were up-regulated,especially in YGP group（P〈0.05 or P〈0.01）.（2）On the 14th day PI,the cAMP levels in brain white matters significantly increased in PA and YGP groups compared with MO group（P〈0.05 or P〈0.01）,and the cAMP level in YGP group was higher than that in ZGP group（P〈0.05）.The cAMP level in spinal cord also significantly increased in YGP group compared with MO,PA and ZGP groups,respectively（P〈0.01）.（3）On the 14th day PI,the PKA expression in spinal cord of rats in ZGP group was significantly decreased compared with MO and YGP groups,respectively（P〈0.05）.（4）On the 28th day PI,there was a positive correlation between cAMP and PKA expression in the brain white matter of YGP rats.Conclusions：The results suggest that ZGP and YGP may promote axonal regeneration by modulating cAMP/PKA signal transduction pathway,but the targets of molecular mechanism of ZGP may be different from those of YGP.

Research on the Mechanism of Zuogui Pill(左归丸)and Yougui Pill(右归丸)in Promoting Axonal Regeneration in Model Rats of Autoimmune Encephalomyelitis

Objective：To study the molecular mechanism of Zuogui Pill（左归丸,ZGP） and Yougui Pill（右归丸,YGP） on axonal regeneration in rats with experimental autoimmune encephalomyelitis（EAE）.Methods： EAE rat model was established by bilateral rear pedes subcutaneous injection of antigen made by mixing myelin basic protein（MBP） and complete Freud＇s adjuvant（CFA） in the volume ratio of 1：1.The pathological changes of axonal injury and regeneration in the brain and the spinal cord were observed on the 14th（the acute stage） and the 28th day（the remission stage） after modeling,with hematoxylin-eosin（HE） staining,silver stain, and immunohistochemical staining.The rats treated with prednisone acetate were taken as controls.Results： Observation under the light microscope with HE staining showed a sleeve-like change in rats＇ cerebrospinal parenchyma with inflammatory cell infiltration around the small vessels and neuronic denaturation,while silver staining showed excessive tumefaction and abscission of axon,and immunohistochemical analysis showed decreasing of nerve growth factor（NGF） expression at the acute stage of EAE,which was even more remarkable at the remission stage,showing significant difference as compared with the normal control（P〈0.05）. And the expressions of Nogo A,an axon growth inhibitor,and its receptor（Nogo-66 receptor,Ng R） were significantly higher than those in the normal control at the acute stage（P〈0.01）.However,after the intervention of ZGP and YGP,the pathological changes and axon damage in rats＇ brain and spinal cord were much more alleviated,and the NGF expression was significantly higher than that in the model group at the acute stage （P〈0.05）.The expression of NGF was even stronger during the remission stage,and a better effect was shown by YGP.As for Nogo A and Ng R expressions,they were significantly lower than those in the model group at the acute stage（P〈0.05）,but a better effect was shown by ZGP.Conclusions：ZGP and YGP can prevent axonal injury and promote the axonal regeneration in rats of EAE,and the possible mechanism is to increase the expression of NGF and reduce the expression of Nogo A and its receptor.However,some differences are observed between the two Chinese preparations in their acting times and points,which provides a certain basis for revealing the modern connotation of the Chinese medicine theory on tonifying Shen（肾）-yin and Shen-yang.

Effect of Zuogui Pill(左归丸) on Monoamine Neurotransmitters and Sex Hormones in Climacteric Rats with Panic Attack

Objective： To explore the effects of Chinese medicine prescription Zuogui Pill（左归丸, ZGP） on monoamine neurotransmitters and sex hormones in climacteric rats with induced panic attacks. Methods： Forty-eight climacteric female rats were randomized into 6 groups with 8 rats in each group： the control group, the model group, the low-, medium- and high-dose ZGP groups and the alprazolam group. Rats in the low-, medium- and high-dose ZGP groups were administered 4.725, 9.45, or 18.9 g/kg ZGP by gastric perfusion, respectively. The alprazolam group was treated by gastric perfusion with 0.036 mg/kg alprazolam. The control and model groups were treated with distilled water. The animals were pretreated once daily for 8 consecutive weeks. The behaviors of rats in the open field test and the elevated T-maze（ETM） were observed after induced panic attack, and the levels of brain monoamine neurotransmitters and the plasma levels of sex hormones were measured. Results： Compared with the control group, the mean ETM escape time and the levels of 5-hydroxytryptamine（5-HT） and noradrenalin（NE） of the model group were significantly reduced（P〈0.05）, Compared with the model group, the mean ETM escape time and the 5-HT and NE levels of all the ZGP groups increased significantly（P〈0.05 or P〈0.01）. However, no significant difference was observed in the levels of sex hormones between the groups. Conclusion： Pretreatment with ZGP in climacteric rats may improve the behavior of panic attack, which may be related to increased 5-HT and NE in the brain.

左归丸联合丁苯酞佐治帕金森病轻度认知障碍疗效评价

目的探讨左归丸联合丁苯酞佐治帕金森病轻度认知障碍的临床疗效。方法选取医院2023年1月至3月收治的帕金森病轻度认知障碍患者80例,按随机数字表法分为观察组和对照组,各40例。两组患者均予常规抗帕金森病西药治疗,并予丁苯酞软胶囊口服,观察组患者加服左归丸。两组均连续治疗8周。结果观察组总有效率为90.00%,显著高于对照组的70.00%(P<0.05)。治疗后,观察组患者的腰膝酸软、胁肋疼痛、头晕头痛中医证候积分均显著低于对照组(P<0.05);观察组患者的沉默信息调节因子1(SIRT1)蛋白表达水平显著高于对照组,核因子κB(NF-κB)蛋白表达水平显著低于对照组(P<0.05);观察组患者的蒙特利尔认知评估量表(MoCA)、简易精神状态量表(MMSE)及日常生活活动能力(ADL)量表评分均显著高于对照组(P<0.05);观察组患者的统一帕金森病评定量表(UPDRS)评分显著低于对照组(P<0.05)。观察组与对照组不良反应发生率相当(5.00%比2.50%,P>0.05)。结论常规抗帕金森病治疗基础上佐以左归丸联合丁苯酞,可降低帕金森病轻度认知障碍患者的中医证候积分,改善SIRT1及NF-κB蛋白表达水平,提升认知功能及日常生活能力。

左归丸对帕金森病轻度认知障碍患者P38 MAPK信号通路及认知功能的影响

目的探究左归丸对帕金森病轻度认知障碍(PD-MCI)患者P38丝裂原活化蛋白激酶(MAPK)信号通路及认知功能的影响。方法80例肝肾阴虚型PD-MCI患者随机均分为观察和对照组,对照组采用丁苯酞软胶囊口服治疗,观察组在对照组治疗基础上加左归丸加味治疗,均治疗8周;观察两组患者的疗效、中医证侯积分、P38 MAPK蛋白表达、认知行为功能[蒙特利尔认知评估量表(MoCA)、日常生活行为能力量表(ADL)、简易智力状态检查量表(MMSE)]、炎症反应[尿酸(UA)、同型半胱氨酸(Hcy)]水平及治疗期间的不良反应。结果与对照组比较,观察组总有效率更高(P<0.05);治疗8周时,两组患者头或肢体震振、肢休拘痉、颈背僵直、腰膝酸软、胁肋疼痛、头晕头痛得分均较治疗前降低,且观察组低于对照组(P<0.05);两组患者P38 MAPK及p-JNK蛋白表达均较治疗前下降,且观察组低于对照组(P<0.05);与治疗前比较,治疗8周时两组肝肾阴虚型PD-MCI患者MMSE、MoCA得分均上升,ADL得分均下降;且观察组MMSE、MoCA得分高于对照组,而ADL得分低于对照组(P<0.05);与治疗前比较,治疗8周时两组患者肝肾阴虚型PD-MCI患者UA升高、Hcy下降,观察组Hcy低于对照组(P<0.05),治疗8周时两组UA水平比较,差异无统计学意义(P>0.05);两组患者治疗期间的不良反应发生率比较,差异无统计学意义(P>0.05)。结论左归丸联合丁苯酞治疗PD-MCI可改善患者的认知功能,其机制可能与调节P38 MAPK蛋白表达和炎症反应有关。

基于乙癸同源理论探讨左归丸对绝经后骨质疏松症患者骨代谢指标CTRP9、PINP、β-CTX与骨密度的影响

目的基于“乙癸同源”理论探讨左归丸对绝经后骨质疏松症患者骨代谢指标CTRP9、PINP、β-CTX与骨密度的影响。方法本次研究纳入2018年1月—2021年5月绝经后骨质疏松症女性90例作为观察对象,随机-编号分成2个组,对照组45例采用碳酸钙D3联合阿仑膦酸钠治疗;观察组采用左归丸治疗,持续治疗4周。治疗前后抽取血液标本测定血清β-骨胶原交联(β-CTX)、I型胶原羧基末端肽(PINP)、C1q/TNF相关蛋白9(CTRP9)和腰椎、股骨颈、股骨粗隆的骨密度,同时比较两组的治疗效果和不良反应状况。结果与治疗前比较,两组治疗后血清β-CTX、PINP水平降低,血清CTRP9水平升高;观察组治疗后血清β-CTX、PINP水平低于对照组,血清CTRP9水平高于对照组,相互比较有统计学意义(P<0.05)。与治疗前比较,两组治疗后腰椎、股骨颈、股骨粗隆的骨密度升高;观察组治疗后腰椎、股骨颈、股骨粗隆的骨密度高于对照组,相互比较有统计学意义(P<0.05)。对照组的有效率75.56%低于观察组91.11%,差异有统计学意义(P<0.05)。两组患者的不良反应率比较,无统计学意义(P>0.05)。结论基于“乙癸同源”理论的左归丸对绝经后骨质疏松症患者的治疗效果显著,可以改善骨代谢指标CTRP9、PINP、β-CTX,提高骨密度,而且安全性高,值得大力推广。

左归丸含药血清对Hedgehog-GLi通路关键因子mRNA表达及骨吸收功能的影响

目的探讨左归丸含药血清对成骨-破骨细胞共培养体系中Hedgehog-GLi通路关键因子mRNA表达及骨吸收功能的影响。方法建立成骨细胞、破骨细胞、骨磨片共培养体系,实验分为7组:空白对照组、假手术组、OVX组、阳性对照组、左归丸组、激动剂组和激动剂+左归丸组。选用50只雌性SD大鼠,分别制备相对应组含药血清,实验各组分别加入相对应组大鼠血清。通过实时荧光定量PCR检测成骨细胞Ihh、Ptch、Smo、GLi1、OPG及RANKL的mRNA表达;并采用甲苯胺蓝染色方法检测破骨细胞骨吸收功能。结果与假手术组相比,OVX组Ihh、Ptch、Smo、GLi1mRNA表达显著上调,同时OPGmRNA表达水平显著降低,RANKL mRNA表达水平显著增加,RANKL/OPG比值明显增高。与OVX组相比,阳性对照组与左归丸组Ihh、Ptch、Smo、GLi1mRNA表达明显下调,同时OPG mRNA表达水平显著升高,RANKL mRNA表达水平显著降低,RANKL/OPG比值明显下降。OVX组加入激动剂Shh重组蛋白后,Hedgehog-GLi信号通路关键因子的mRNA表达较OVX组明显增加。与激动剂组比较,激动剂+左归丸组Ihh、Ptch、Smo、GLi1mRNA表达以及RANKL/OPG比值均明显下降。与假手术组相比,OVX组的骨吸收陷窝面积显著增加;与OVX组相比,阳性对照组和左归丸组的骨吸收陷窝面积显著下降。结论左归丸含药血清可一定程度上抑制Hedgehog-GLi通路关键因子Ihh、Ptch、Smo、GLi1的活性,进而抑制OVX所致的骨吸收增强。

左归丸含药血清对成骨细胞中Ihh、Ptch、Smo、GLi1蛋白表达及破骨细胞数量的影响

目的 探讨左归丸含药血清对成骨细胞中Hedgehog-GLi通路关键蛋白表达及破骨细胞数量的影响。方法 选用50只雌性SD大鼠,随机分为空白对照组、假手术组、造模组。造模组大鼠切除双侧卵巢,12周后随机分为卵巢切除组(ovariectomy,OVX)、阳性对照组和左归丸组。阳性对照组大鼠灌服戊酸雌二醇混悬液,左归丸组大鼠灌服左归丸水煎液,连续干预7 d。末次给药1.5 h后,腹主动脉取血制备各组相应含药血清。建立成骨细胞、破骨细胞、骨磨片共培养体系,分为空白对照血清组、假手术血清组、OVX血清组、阳性对照血清组、左归丸含药血清组、激动剂组和激动剂+左归丸含药血清组,分别添加各组对应血清。免疫荧光法检测成骨细胞印度刺猬蛋白(Indian hedgehog,Ihh)、碎片蛋白(patched,Ptch)、润滑蛋白(smoothened,Smo)及GLi1蛋白表达;免疫组化法检测成骨细胞骨保护蛋白(osteoprotegerin,OPG)、核因子受体激活因子-κB配体(receptor activator for nuclear factor-κB ligand,RANKL)蛋白表达;对破骨细胞进行TRAP染色,计算TRAP+细胞数;ELISA检测共培养体系上清液中TRAP的含量。结果与假手术血清组比较,OVX血清组Ihh、Ptch、Smo、GLi1蛋白表达水平显著上调(P<0.01),RANKL/OPG比值增高;TRAP+细胞数和细胞上清液中TRAP含量亦明显增加(P<0.05)。与OVX血清组比较,左归丸含药血清组Ihh、Ptch、Smo、GLi1的蛋白表达明显下调,RANKL/OPG比值下降;TRAP+细胞数和细胞上清液中TRAP含量明显降低(P<0.05)。与激动剂组比较,加入左归丸含药血清后,Ihh、Ptch、Smo、GLi1的蛋白表达及RANKL/OPG比值均明显下降,TRAP+细胞个数及TRAP含量亦明显降低(P<0.05)。结论 左归丸含药血清可通过抑制异常活跃的Hedgehog-GLi信号通路,使Ihh、Ptch、Smo、GLi1的蛋白表达水平下调,降低RANKL/OPG比值,抑制破骨细胞的数量和活性,起到减少骨丢失作用。

左归丸加减结合温针灸治疗早发性卵巢功能不全临床研究

目的观察左归丸加减结合温针灸治疗早发性卵巢功能不全临床疗效。方法研究纳入2020年1月—2022年7月收治的81例早发性卵巢功能不全患者进行分组研究,以随机数字表法分为观察组(41例)与对照组(40例),对照组患者采取雌孕激素序贯疗法治疗,观察组患者采取雌孕激素序贯疗法治疗+左归丸加减结合温针灸治疗,各组数据对比:临床疗效、治疗前后中医证候(五心烦热、腰膝酸软、闭经、头晕及潮热盗汗等)积分变化及卵泡刺激素(Follicle-stimulating hormone,FSH)、黄体生成素(Luteinizing hormone,LH)以及雌二醇(Estradial,E_(2))等水平变化、窦卵泡数以及卵巢面积、卵巢平均直径等指标变化、血清免疫球蛋白A/G/M(Immuno globulins A,Immuno globulins G,Immuno globulins M,IgA、IgG、IgM)指标变化、不良反应。结果对照组患者治疗总有效率(80.00%,32/40)比观察组患者(95.12%,39/41)低,P<0.05;各组患者治疗前中医证候(五心烦热、腰膝酸软、闭经、头晕及潮热盗汗等)积分、FSH、LH、E_(2)、窦卵泡数以及卵巢面积、卵巢平均直径、IgA、IgG、IgM等指标比较,P>0.05,各组患者治疗后中医证候(五心烦热、腰膝酸软、闭经、头晕及潮热盗汗等)积分、FSH、LH、E_(2)、窦卵泡数以及卵巢面积、卵巢平均直径、IgA、IgG、IgM等指标均明显改善,观察组患者治疗后中医证候(五心烦热、腰膝酸软、闭经、头晕及潮热盗汗等)积分、FSH、LH、E_(2)、窦卵泡数以及卵巢面积、卵巢平均直径、IgA、IgG、IgM等指标均显著优于对照组,P<0.05;对照组1例腹泻,1例恶心,1例呕吐,不良反应率为7.50%(3/40),观察组患者2例恶心(4.88%,2/41),两组不良反应均较为轻微,不良反应率较低,P>0.05。结论左归丸加减结合温针灸治疗早发性卵巢功能不全临床疗效显著,患者治疗后病症明显减轻,且患者卵巢功能恢复较好,免疫功能提升,不良反应轻微且较少,安全可靠,值得临床应用。

基于非稳态负荷评估雄性SD大鼠衰老进展及左归丸的干预作用

目的采用非稳态负荷(allostatic load,AL)的理论与方法评估衰老以及左归丸对自然衰老雄性SD大鼠模型的影响。方法自然衰老雄性SD大鼠,分别在2、5、8、14、18、21月龄剪尾取血,其中18月龄时随机分为老年对照组、左归丸低剂量组和左归丸高剂量组。左归丸干预3个月后,取用血样以及肌肉、肝等组织样本。血样用流式细胞术检测T淋巴细胞数量和凋亡率,并检测血清生化指标,包括低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-c)、高密度脂蛋白胆固醇(high-density lipoprotein cholesterol,HDL-c)、甘油三酯(triglycerides,TG)、总胆固醇(total cholesterol,TC)、游离脂肪酸(free fatty acids,FFA)、25羟维生素D(25 hydroxyvitamin D,25-OH-D)、皮质酮(corticosterone,CORT)、C反应蛋白(C-reactive protein,CRP)、白细胞介素-6(interleukin 6,IL-6)。将LDL-c、TC、HDL-c、FFA、TG、CORT、IL-6、CRP、25-OH-D、CD3^(+)T淋巴细胞计数、CD3^(+)T淋巴细胞凋亡率共11项指标用于AL评分。结果大鼠血清LDL-c、TG、TC、25-OH-D、CRP、IL-6随增龄发生显著改变,但变化模式有所不同。CD3^(+)T淋巴细胞计数随增龄显著降低(P<0.01),CD3^(+)、CD4^(+)、CD8^(+)T淋巴细胞凋亡率随增龄显著升高(P<0.01)。左归丸干预后,老年大鼠血清CORT水平显著提高(P<0.01)、CD8^(+)T淋巴细胞凋亡率显著降低(P<0.05)。AL评分自5月龄开始升高,至18月龄到达峰值。结论与单一指标比较,AL能更好地表征衰老过程。左归丸可以提高衰老大鼠的应激反应能力,延缓其免疫衰老。

基于PI3K/AKT通路探讨左归丸改善卵巢衰老大鼠卵巢生殖干细胞干性的机制

目的基于磷脂酰肌醇3激酶/蛋白激酶B(PI3K/AKT)通路探讨左归丸通过缝隙连接蛋白43(Cx43)磷酸化改善卵巢衰老大鼠卵巢生殖干细胞(OSCs)干性的机制。方法将8周龄雌性SD大鼠随机分为正常组、模型组、脱氢表雄酮(DHEA)组、左归丸组,每组6只。采用环磷酰胺腹腔注射15 d复制卵巢衰老大鼠模型。模型复制结束后,DHEA组大鼠灌胃DHEA 8.1 mg·kg^(-1),左归丸组大鼠灌胃左归丸1.85 g·kg^(-1),每日1次,连续30 d。采用HE染色法观察大鼠卵巢组织病理变化及各级卵泡计数;ELISA法检测大鼠血清抗缪勒管激素(AMH)、卵泡刺激素(FSH)、雌二醇(E_(2))水平;Western Blot法检测卵巢组织中相关蛋白(PCNA、MVH、Oct4、Cx43、p-Cx43、EGFR、AKT、p-AKT)的表达水平。结果与正常组比较,模型组大鼠体质量明显下降(P<0.05),卵巢湿质量明显降低(P<0.05),始基卵泡数显著减少(P<0.01);血清FSH水平显著升高(P<0.01),E_(2)、AMH水平呈下降趋势(P>0.05);卵巢组织中Oct4、MVH、PCNA、EGFR、p-AKT/AKT、p-Cx43(Ser368)/Cx43、Cx43蛋白表达水平均明显降低(P<0.05,P<0.01)。与模型组比较,左归丸组大鼠体质量明显升高(P<0.05),卵巢湿质量、卵巢指数均显著升高(P<0.01),始基卵泡数显著增加(P<0.01);血清E2水平明显升高(P<0.05);卵巢组织中Oct4、MVH、PCNA、EGFR、p-AKT/AKT、p-Cx43(Ser368)/Cx43、Cx43蛋白水平表达均显著升高(P<0.05,P<0.01)。结论左归丸可能通过维持OSCs干性改善卵巢衰老,其作用机制与PI3K/AKT信号通路及缝隙连接蛋白磷酸化相关。

左归丸对孕期糖代谢异常大鼠肝损伤及其子代HPA轴功能的影响

目的通过左归丸孕期给药,观察其是否能缓解糖代谢异常孕鼠肝损伤及对子代/鼠下丘脑-垂体-肾上腺(hypothalamic-pituitary-adrenal,HPA)轴功能的影响。方法建立孕期糖代谢异常大鼠模型并予左归丸干预,孕20 d时,检测左归丸对模型大鼠体质量、进食量的影响,ELISA检测模型大鼠血糖变化,HE染色观察模型大鼠肝损伤情况。待其产子后,观察子鼠(0~10周)体质量变化、肝质量及肝质量/体质量百分比,Western blot和RT-qPCR检测子鼠10周时脑组织糖皮质激素受体(glucocorticoid receptor,GR)、促肾上腺皮质激素释放激素(corticotropin relesing hormone,CRH)、促肾上腺皮质激素(adrenocortico tropic hormone,ACTH)表达水平,ELISA检测子鼠10周时血清皮质酮(cortisol,CORT)含量。结果孕末期,与模型组比较,左归丸组孕鼠的体质量显著增加,且呈浓度依赖性(P<0.05)。左归丸组孕鼠的进食量、血糖水平明显低于模型组,且呈浓度依赖性(P<0.05)。经左归丸治疗的代谢异常孕鼠肝组织损伤程度显著低于模型组。左归丸组孕鼠子代的体质量明显高于模型组,且呈浓度依赖性(P<0.05),但左归丸对子代肝质量和肝质量/体质量百分比没有影响(P>0.05)。糖代谢异常孕鼠的子代GR、CRH、ACTH和CORT呈现高表达,左归丸干预能够降低子代GR、CRH、ACTH和CORT表达(P<0.05)。结论左归丸具有改善糖代谢异常孕鼠的肝损伤及调节其子代HPA轴功能的作用,为中医肾主生殖、发育,肝肾同源理论提供了实验依据。

左(右)归丸联合接骨续筋汤治疗脑卒中后偏瘫合骨质疏松临床研究

目的探讨左(右)归丸联合接骨续筋汤治疗脑卒中后偏瘫合骨质疏松的临床疗效,以及对患者运动功能、肢体功能和骨密度的影响。方法选取江苏省灌云县中医院2021年1月至2023年3月收治的脑卒中后偏瘫合骨质疏松患者120例,按随机数字表法分为对照组和观察组,各60例。两组患者均予常规康复训练,观察组患者加用左(右)归丸联合接骨续筋汤治疗,两组患者均连续治疗28 d。结果观察组总有效率为85.00%,显著高于对照组的66.67%(P<0.05)。治疗后,观察组患者骨密度显著高于对照组(P<0.05);骨代谢指标血清骨钙素水平显著高于对照组,β-交联胶原骨钙素、Ⅰ型前胶原氨基端肽水平均显著低于对照组(P<0.05);患侧Brunnstrom分期的高分期(Ⅴ-Ⅵ期)患者显著多于对照组(P<0.05);模拟视觉评分法(VAS)评分显著低于对照组(P<0.05),Fugl-Meyer评分量表(FMA)评分、Berg平衡量表(BBS)评分均显著高于对照组(P<0.05)。观察组和对照组不良反应发生率相当(6.67%比5.00%,P>0.05)。结论左(右)归丸联合接骨续筋汤治疗脑卒中后偏瘫合骨质疏松的临床疗效良好,可有效改善患者的运动功能和肢体功能,增强骨密度,且安全性良好。

中药内服、外敷联合西药治疗绝经后骨质疏松性胸腰椎压缩性骨折的疗效观察

目的:探讨中药内服、外敷联合西药治疗绝经后骨质疏松性胸腰椎压缩性骨折(OVCF)的疗效及安全性。方法:选择2020年2月至2022年10月于该院接受经皮椎体后凸成形术治疗的140例绝经后OVCF患者,依据随机数字表法分为对照组(70例,术后口服碳酸钙D3片、骨化三醇软胶囊和阿仑膦酸钠片)与研究组(70例,在对照组的基础上给予左归丸合桃红四物汤化裁汤剂口服联合海桐皮汤外敷),治疗6个月。比较两组患者的疗效和治疗期间内的安全性,观察治疗前后疼痛情况、Oswestry功能障碍指数(ODI)、骨密度、血清碱性磷酸酶(ALP)、血钙和血磷水平。结果:研究组患者的总有效率为94.29%(66/70),高于对照组的80.00%(56/70),差异有统计学意义(P<0.05)。治疗后,两组患者的视觉模拟评分法(VAS)评分、ODI评分、血清ALP和血磷水平低于治疗前,腰椎正位骨密度、股骨颈骨密度和血钙水平高于治疗前,差异均有统计学意义(P<0.05);且研究组患者的VAS评分、ODI评分、血清ALP和血磷水平低于对照组,腰椎正位骨密度、股骨颈骨密度和血钙水平高于对照组,差异均有统计学意义(P<0.05)。两组患者治疗期间均未出现血常规、尿常规、便常规、肝肾功能和心电图异常。结论:中药内服、外敷联合西药治疗绝经后OVCF安全、有效,能缓解患者疼痛,降低功能障碍程度,提高骨密度,并调节血清ALP、血钙和血磷水平。

三联疗法对膝骨关节炎疗效及对血清BMP-2、软骨寡聚体蛋白和炎症因子水平的影响

目的:探究左归丸、富血小板血浆(PRP)、间充质干细胞(MSCs)三联疗法对膝骨关节炎患者的治疗效果及对患者血清骨形态发生蛋白-2(BMP-2)、软骨寡聚基质蛋白(COMP)与血清炎症因子[白细胞介素-6(IL-6)、白细胞介素-17(IL-17)、C反应蛋白(CRP)水平]的影响。方法:选取2022年4月—2023年1月在新余市人民医院接受治疗的98例膝骨关节炎患者,按照随机抽签法将患者分入对照组和观察组,各49例。对照组接受膝骨关节炎的常规治疗,观察组在此基础上加用左归丸、PRP、MSCs三联疗法。对两组治疗前后中医证候积分、奎森功能演算指数(Lequesne)、西安大略和麦克马斯特大学骨关节炎指数(WOMAC)评分、BMP-2、COMP、IL-6、IL-17、CRP水平、疗效进行对比。结果:治疗后,与对照组相比,观察组中医证候各项积分、Lequesne评分、WOMAC评分、COMP、IL-6、IL-17、CRP水平均较低(P<0.05),BMP-2水平较高(P<0.05),疗效更优(P<0.05)。结论:左归丸、PRP、MSCs三联疗法有良好的治疗效果,可更有效地改善患者膝关节功能,缓解疾病带来的不适,提高患者生活水平。