Differential Acupuncture Treatment of Hyperplasia of Mammary Glands

Hyperplasia of mammary glands is a common disease in the middle-aged women, and it is a precancerous lesion of mammary glands. For many years, the author has used Rugen (ST 18) of the Stomach Meridian of Foot-Yangming as the main point withcertain auxiliary points chosen on basis of the differentiation types to treat the disease and obtained satisfactory therapeutic effects. A report follows. ……

The Effect of Administration of Rutin on Plasma Levels of Estrogen, Prolactin, Growth Hormone and Gene Expression of Their Receptors in Mammary Glands in Ovariectomized Rats

The development of mammary glands, endocrine hormone concentrations and the gene expression of related receptors were measured in ovariectomized virgin rats after adminstration of an estrogen-like plant extract, rutin. Thirty-two ovariectomized virgin Wistar rats were randomly assigned to 4 treatments with 8 animals each： gastric infusion of 2 mL normal saline per unovariectomized rat per day （Sham）, gastric infusion of 2 mL normal saline per ovariectomized rat per day （Ova）, gastric infusion of 60 mg rutin kg-1 body weight （BW） per ovariectomized rat per day （Ova＋Rut）, or intramuscular injection of 60 ug estradiol kg-1 BW per ovariectomized rat weekly （Ova＋Est）. Samples of blood and mammary glands were harvested to determine the levels of estrogen （E2）, prolactin （PRL） and growth hormone （GH）, and the gene expression of estrogen receptors （ER）, prolactin receptors （PRLR） and growth hormone receptors （GHR） with radioimmunoassy （RIA） and RT-PCR technology, respectively. The E2 concentration in plasma and gland tissues from the rats of Ovx＋Rut or Ovx＋Est was higher than that of Ovx （P〈0.05）, but the plasma E2 concentration from the rats of Ovx＋Rut was lower than that of Sham （P〈0.05）. The order of the PRL concentration in plasma and gland tissues was Ovx〈Ovx＋Rut〈Ovx＋Est 〈Sham, and the difference in each treatment （P〈0.05）. The plasma GH concentration was lower in Ovx than in Ovx＋Rut or Ovx＋Est, and lower in Ovx＋Rut than in Sham （P〈0.05）. The GH concentration in gland tissues was lower in Ovx than in Ovx＋Rut or Ovx＋Est （P〈0.05）, and lower in Ovx＋Rut than in Sham （P〈0.05）. The gene expression of ER in gland tissues was increased in an order as Ovx〈Ovx＋Rut〈Ovx＋Est〈Sham （P〈0.05）, and PRLR, GHR showed the same trend. In conclusion, adminstration of rutin increased the E2 concentration in plasma and mammary glands, promoted pituitary PRL and GH release, up-regulated the gene expression of ER, PRLR and GHR, and stimulated mammary development in ovariectomized virgin rats.

AMPK-mTOR pathway is involved in glucose-modulated amino acid sensing and utilization in the mammary glands of lactating goats

Background:The local supply of energy-yielding nutrients such as glucose seems to affect the synthesis of milk components in the mammary gland(MG).Thus,our study was conducted to investigate the effects of locally available MG glucose supply(LMGS)on amino acid(AA)sensing and utilization in the MG of lactating dairy goats.Six dosages of glucose(0,20,40,60,80,and 100 g/d)were infused into the MG through the external pudendal artery to investigate the dose-dependent changes in mammary AA uptake and utilization(Exp.1)and the changes in mRNA and protein expression of the AMPK-mTOR pathway(Expt.2).Results:In Exp.1,total milk AA concentration was highest when goats were infused with 60 g/d glucose,but lower when goats were infused with 0 and 100 g/d glucose.Increasing LMGS quadratically changed the percentages ofαS2-casein andα-lactalbumin in milk protein,which increased with infusions from 0 to 60 g/d glucose and then decreased with infusions between 60 and 100 g/d glucose.The LMGS changed the AA availability and intramammary gland AA utilization,as reflected by the mammary AA flux indexes.In Exp.2,the mRNA expression of LALBA in the MG increased quadratically with increasing LMGS,with the highest expression at dose of 60 g/d glucose.A high glucose dosage(100 g/d)activated the general control nonderepressible 2 kinase,an intracellular sensor of AA status,resulting in a reduced total milk AA concentration.Conclusions:Our new findings suggest that the lactating MG in dairy goats may be affected by LMGS through regulation of the AA sensory pathway,AA utilization and protein synthesis,all being driven by the AMPK-mTOR pathway.

ELECTROACUPUNCTURE TREATMENT OF 30 CASES OF HYPERPLASIA OF MAMMARY GLANDS

In the present paper,30 cases of hyperplasia of mammary glands were treated withelectroacupuncture in different times following menstruation,showing satisfactory results.The totaleffective rate was up to 96.67％.In addition,the symptom of mastalgla abated abviously after treat-ment.

Comparative Expression Profiling of Lactogenic Hormone Receptor and It’s Signaling Molecules of Bovine Mammary Glands during lactation

Milk synthesis is known to be modulated by peptide hormones such as prolactin (PRL), growth hormone (GH), and insulin-like growth factor I (IGF-I). Previous studies suggested that PRL and IGF-I acted directly on mammary epithelial cells and were involved in lactation. Meanwhile, GH is thought to be indirectly involved in lactation by stimulating the secretion of IGF-I. It is controversial as growth hormone receptors (GHR) is expressed in the mammary epithelial cells. In order to clarify whether GH acted directly on mammary gland tissue, we investigated the prolactin receptors (PRLR), IGF-I receptors (IGF-IR), and GHR as well as the gene expression levels of the downstream signaling molecule for each receptor in the mammary gland tissue of Holstein cows during different stages of lactation. The results revealed that the mRNA expressions of PRLR and IGF-IR were highest during early lactation, and the mRNA expression of the GHR was highest during mid-lactation. We also found that the expression profiling of the signal transducer and activator of transcription 5 (STAT5) genes was similar to that of the GHR gene. On the other hand, the expression profiling of the PRLR gene was similar to that of the SHP2 gene. These results suggest that GH acts on the mammary glands directly, milk synthesis and secretion are chiefly stimulated in mid-lactation, and the timing of the action is different for PRL and IGF-I.

300 CASES OF HYPERPLASIA OF MAMMARY GLANDS TREATED WITH RED-HOT NEEDLE PLUS FILIFORM NEEDLE

Clinical observations on 300 cases of hyperplasia of mammary glands (group A) treated by red hot needle plus filiform needle are reported in the present study. "Ashi" points were used as the main acupoints. For comparison, other 300 cases (group B) were treated with routine acupuncture method, and Rugen (ST 18) was used as the main acupoint. In addition, other auxiliary acupoints were supplemented in the light of the concrete syndromes in these two groups. The treatment was given once daily, with 10 sessions being a therapeutic course. Of the 300 cases in group A, 240 were clinically cured, amounting to 80%; 58 were effective, amounting to 19.3%; 2 were failed, with a total effective rate of 99.3%. Of the 300 cases in group B, 113 were clinically cured, 165 effective, 22 were failed, with a total effective rate of 92.7%. The therapeutic effect of group A is significantly superior to that of group B (P<0.05).

CLINICAL OBSERVATION ON THE ACUPOINT INTRADERMAL EMBEDDING OF RINGHEADED THUMBTACK-NEEDLE THERAPY FOR HYPERPLASIA OF MAMMARY GLANDS

In the present paper, the author used acupoint intradermal embedding of ringheaded thumbtack-needle therapy combined with herbal medicine therapy to treat 110 cases of hyperplasia of mammary glands, and compared its therapeutic effect with that of herbal therapy alone. The observed indexes of the treatment were pain relief degree, the size of hyperplasic glands and the time of occurrence of marked effect. Following 3 courses of treatment,the results showed the therapeutic effect of the ringheaded thumbtack-needle needle plus herbal medicine therapy was superior to that of herbal medicine therapy alone in the above-mentioned 3 indexes (P ＜ 0.01 ).

ACUPUNCTURE FOR TRERTING 120 CASES OF HYPERPLASIA OF MAMMARY GLANDS

Objective: To observe the therapeutic effect of acupuncture therapy for 120 cases of hyperplasia of mammary glands. Methods: These patients were classified into liver-qi-stagnation type (n=53), phlegm-coagulation type (n=30) and Thoroughfare-Conception vessel maladjustment type (n=37). Acupoints used were Qimen (期门 LR 14), Wuyi (屋翳 ST 15), Weishu (胃俞 BL 21), etc.. Acupuncture treatment was conducted once daily and 30 treatments constituted one therapeutic course. Results: After 3 courses of treatment, of the 53, 30 and 37 cases of the liver-qi-stagnation, phlegm-coagulation and Thoroughfare-Conception vessel maladjustment types, 43 (81.1%), 24 (80.0%) and 29 (78.4%) were cured, 7 (13.2%), 5 (16.7%)and 5 (13.5%) markedly improved, 3 (5.7%), 1 (3.3%) and 2 (5.4%) improved respectively, and 1 (2.7%) of Thoroughfare-Conception vessel maladjustment type failed. A two-years’ follow-up showed that 3 of the cured 92 cases had a relapse, and after 2 more courses of treatment, they were cured again. Conclusion: Acupuncture therapy is effective for the treatment of hyperplasia of mammary glands.

A cell transcriptomic profile p ovides insights into adipocytes of porcine mammary gland across development

Background Studying the composition and developmental mechanisms in mammary gland is crucial for healthy growth of newborns. The mammary gland is inherently heterogeneous, and its physiological function dependents on the gene expression of multiple cell types. Most studies focused on epithelial cells, disregarding the role of neighboring adipocytes.Results Here, we constructed the largest transcriptomic dataset of porcine mammary gland cells thus far. The dataset captured 126,829 high-quality nuclei from physiological mammary glands across five developmental stages(d 90 of gestation, G90;d 0 after lactation, L0;d 20 after lactation, L20;2 d post natural involution, PI2;7 d post natural involution, PI7). Seven cell types were identified, including epithelial cells, adipocytes, endothelial cells, fibroblasts cells, immune cells, myoepithelial cells and precursor cells. Our data indicate that mammary glands at different developmental stages have distinct phenotypic and transcriptional signatures. During late gestation(G90), the differentiation and proliferation of adipocytes were inhibited. Meanwhile, partly epithelial cells were completely differentiated. Pseudo-time analysis showed that epithelial cells undergo three stages to achieve lactation, including cellular differentiation, hormone sensing, and metabolic activation. During lactation(L0 and L20), adipocytes area accounts for less than 0.5% of mammary glands. To maintain their own survival, the adipocyte exhibited a poorly differentiated state and a proliferative capacity. Epithelial cells initiate lactation upon hormonal stimulation. After fulfilling lactation mission, their undergo physiological death under high intensity lactation. Interestingly, the physiological dead cells seem to be actively cleared by immune cells via CCL21-ACKR4 pathway. This biological process may be an important mechanism for maintaining homeostasis of the mammary gland. During natural involution(PI2 and PI7), epithelial cell populations dedifferentiate into mesenchymal stem cells to maintain the lactation potential of mammary glands for the next lactation cycle.Conclusion The molecular mechanisms of dedifferentiation, proliferation and redifferentiation of adipocytes and epithelial cells were revealed from late pregnancy to natural involution. This cell transcriptomic profile constitutes an essential reference for future studies in the development and remodeling of the mammary gland at different stages.

Current status of outcome reporting in randomized controlled trials of traditional Chinese medicine for mammary gland hyperplasia: A systematic review

Objective:To assess outcome indicators in clinical trials and provide a reference for establishing a core outcome set to treat hyperplasia of mammary gland(HMG)with traditional Chinese medicine(TCM).Methods:Eight online databases were searched from their inception to December 31,2022,to assess outcomes reported in randomized controlled trials(RCTs)of HMG treated with TCM.The quality of the included studies was assessed according to the Cochrane Risk of Bias Assessment Tool.All outcomes were extracted,classified,and described.Results:A total of 8249 articles were initially retrieved.Of these,70 articles were eligible and involved 10618 participants with HMG.A total of 17 outcome indicators with a frequency of 271 times were involved and were collected according to six outcome domains.Conclusions:The core outcomes of RCTs of HMG treated with TCM are large and divergent.There are problems in evaluation standards,primary and secondary outcomes,TCM characteristic indicators,long-term prognosis,and standardization of reporting.It is recommended to strengthen the trial design and actively construct the core outcome sets with TCM characteristics for HMG.

Efficacy of Lubeikangru formulation in mice with hyperplasia of the mammary glands induced by estrogen and progesterone

OBJECTIVE: To evaluate the protective effects of Lubeikangru formula(LF) on hyperplasia of the mammary glands(HMG) induced by estrogen and progesterone in mice.METHODS: Female mice were divided randomly into five groups: normal, model, tamoxifen(3 mg/kg),Rupixiao(900 mg/kg) and LF(900 mg/kg). All mice except those in the normal group were treated sequentially with estradiol and progesterone to induce HMG. From the tenth day of induction, mice in normal and model groups received distilled water and mice in the other groups were given the corresponding drugs by gavage, once a day, for 30 d.At the end of treatment, the mammary glands, ovaries, hypothalamus, and serum was collected for whole-mount and hematoxylin and eosin(HE)staining, enzyme-linked immunosorbent assays(ELISAs), or western blotting.RESULTS: Whole-mount and HE staining of mammary glands showed that LF rescued(at least in part) the hyperplasic morphology of the mammary glands, and the number of branch points decreased after LF treatment(P < 0.05). ELISAs revealed that levels of estrogen and progesterone were decreased following LF treatment, whereas levels of gonadotropin-releasing hormone, follicle-stimulating hormone, and luteinizing hormone were increased in serum and tissues. Western blotting confirmed that LF treatment led to a reduction in expression of phosphorylated(p)-Erk, p-p38 and p-c-Jun N-terminal kinase. LF was also confirmed to be safe by acute-toxicity tests.CONCLUSION: LF can protect the mammary glands of mice from estrogen-and progesterone-induced hyperplasia by adjusting hormone levels and regulating the mitogen-activated protein kinase pathway.

Thinking Path and Method for Establishing the Disease/ Chinese Medicine Syndrome Conjugated Pattern of a Mammary Hyperplasia Animal Model

Mammary hyperplasia （MHP） is the most commonly encountered mammary disease in women at the child-bearing stage. Especially, atypical hyperplasia which belongs to the precancerous category, is the disease for class Ⅰ prevention of breast cancer. Therefore, advancing the clinical efficacy of MHP treatment is of critical importance. Chinese medicine （CM） and drugs show a peculiar effect in this field; the clinical or experimental researches concerning MHP treatment by CM compounds or patent drugs have been increasing gradually in recent years, but the thinking paths and methods for establishing the MHP animal model are divergent. Particularly, the disease/CM syndrome conjugated model （D/S model） has rarely been studied. For this reason, the pathogenetic mechanism, the establishment of an animal disease model, as well as the thinking paths and methods for establishing the D/S model of MHP are discussed and summarized preliminarily in this paper by the authors. This could provide a new way of thinking and method for creating the MHP model in modern medicine.

Zfhx3 is essential for progesterone/progesterone receptor signaling to drive ductal side-branching and alveologenesis in mouse mammary glands

Progesterone(Pg)/progesterone receptor(PR) signaling drives mammary gland side-branching and alveologenesis, but the mechanisms through which Pg/PR signaling functions remain to be clarified.Using in vitro and in vivo models and histological and molecular analyses, we determined the role of Zfhx3 transcription factor in mammary gland development driven by Pg/PR signaling. Postnatal deletion of Zfhx3 in mouse mammary epithelial cells attenuated side-branching morphogenesis and alveologenesis. These effects were undetectable in the absence of Pg/PR signaling. During the estrus cycle, Zfhx3 expression corresponded to that of Pg, being at the highest level at the diestrus stage; Zfhx3 deletion inhibited mammary gland branching more potently at diestrus than estrus stage. Loss of Zfhx3 not only attenuated the expansion of stem/progenitor cells driven by Pg/PR signaling, but also impaired the function of Pg/PR signaling in the transcriptional activation of multiple genes. In addition, Pg/PR signaling significantly expanded PR-and Zfhx3-positive epithelial cells, and induced the physical association of ZFHX3 with PR. These findings establish Zfhx3 as an integral transcription factor of Pg/PR signaling in driving side-branching and alveologenesis during mammary gland development.

The LXRB-SREBP1 network regulates lipogenic homeostasis by controlling the synthesis of polyunsaturated fatty acids in goat mammary epithelial cells

Background:In rodents,research has revealed a role of liver X receptors(LXR) in controlling lipid homeostasis and regulating the synthesis of polyunsaturated fatty acids(PUFA).Recent data suggest that LXRB is the predominant LXR subtype in ruminant mammary cells,but its role in lipid metabolism is unknown.It was hypothesized that LXRB plays a role in lipid homeostasis via altering the synthesis of PUFA in the ruminant mammary gland.We used overexpression and knockdown of LXRB in goat primary mammary epithelial cells(GMEC) to evaluate abundance of lipogenic enzymes,fatty acid profiles,content of lipid stores and activity of the stearoyl-Co A desaturase(SCD1) promoter.Results:Overexpression of LXRB markedly upregulated the protein abundance of LXRB while incubation with si RNA targeting LXRB markedly decreased abundance of LXRB protein.Overexpression of LXRB plus T0901317(T09,a ligand for LXR) dramatically upregulated SCD1 and elongation of very long chain fatty acid-like fatty acid elongases 5–7(ELOVL 5–7),which are related to PUFA synthesis.Compared with the control,cells overexpressing LXRB and stimulated with T09 had greater concentrations of C16:0,16:1,18:1n7,18:1n9 and C18:2 as well as desaturation and elongation indices of C16:0.Furthermore,LXRB-overexpressing cells incubated with T09 had greater levels of triacylglycerol and cholesterol.Knockdown of LXRB in cells incubated with T09 led to downregulation of genes encoding elongases and desaturases.Knockdown of LXRB attenuated the increase in triacylglycerol and cholesterol that was induced by T09.In cells treated with dimethylsulfoxide,knockdown of LXRB increased the concentration of C16:0 at the expense of C18:0,while a significant decrease in C18:2 was observed in cells incubated with both si LXRB and T09.The abundance of sterol regulatory element binding transcription factor 1 precursor(p SREBP1) and its mature fragment(n SREBP1) was upregulated by T09,but not LXRB overexpression.In the cells cultured with T09,knockdown of LXRB downregulated the abundance for p SREBP1 and n SREBP1.Luciferase reporter assays revealed that the activities of wild type SCD1 promoter or fragment with SREBP1 response element(SRE) mutation were decreased markedly when LXRB was knocked down.Activity of the SCD1 promoter that was induced by T09 was blocked when the SRE mutation was introduced.Conclusion:The current study provides evidence of a physiological link between the LXRB and SREBP1 in the ruminant mammary cell.An important role was revealed for the LXRB-SREBP1 network in the synthesis of PUFA via the regulation of genes encoding elongases and desaturases.Thus,targeting this network might elicit broad effects on lipid homeostasis in ruminant mammary gland.

乳腺增生症的中西医认识及治疗研究进展

乳腺增生症(hyperplasia of mammary gland,HMG)中医病名为乳癖,是一种乳腺组织增生的疾病,其中医基本病机为情志失常,病机以肝内气机郁滞为主,兼有脾肾不足等,由此导致气滞、血瘀、痰凝。气、血、痰、瘀于乳络互结。西医病因病理尚未完全明确。西药目前尚无特异性疗法,中药治疗具有良好效果。文章将对乳癖的中西医病因病机进行论述,并结合临床案例及经验总结中医药治法治则及常用药物,以临床为角度基础,提出治疗乳癖的方法,并为乳癖病因病机的研究与探讨提供新思路。

蒲公英对乳腺疾病治疗作用机制的研究进展

乳腺疾病是我国常见的危害女性心理及健康的最主要因素,蒲公英在治疗乳腺疾病方面疗效确切,深入研究其药理作用及临床应用尤为必要。基于此,就近年来关于蒲公英对乳腺疾病的药理作用和临床应用的相关文献进行归纳分析,发现可通过含蒲公英的复方治疗乳腺增生病;其主要有效成分可通过调控核因子κB(nuclear factor-kappa B,NF-κB)信号通路和微生物相关分子模式(microbe-associated molecular patterns,MAMPs)信号通路,下调肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)和白细胞介素-6(interlukin-6,IL-6)等炎性因子的表达治疗哺乳期乳腺炎;通过中药复方合理配伍治疗非哺乳期乳腺炎;通过调控细胞周期以抑制增殖、促进凋亡、抗侵袭转移、调节自噬、影响雌激素、提高免疫力、抑制相关代谢途径等抗乳腺癌。总结梳理的关于蒲公英作用于乳腺疾病的药理作用及临床应用,可为进一步研究蒲公英在乳腺疾病的治疗提供新的思路和科学依据。

色瓦绵羊GPIHBP1基因克隆及其组织表达特征分析

【目的】克隆色瓦绵羊乳腺组织糖基化磷脂酰肌醇锚定高密度脂蛋白结合蛋白1基因(GPIHBP1),明确其生物信息学特性及组织表达分布特征,为揭示GPIHBP1基因影响绵羊产奶性状的作用机制提供参考依据。【方法】克隆色瓦绵羊GPIHBP1基因编码区(CDS)序列,通过Prot Para、Prot Scale、SOPMA、SWISS-MODEL、TMHMM-2.0等在线软件进行生物信息学分析,并以实时荧光定量PCR检测该基因在色瓦绵羊乳腺、肌肉、心脏、肾脏、肝脏、小肠、瘤胃和卵巢等8个组织中的表达情况。【结果】色瓦绵羊GPIHBP1基因CDS序列长519 bp,共编码172个氨基酸残基,与山羊GPIHBP1氨基酸序列(XM_018058666.1)比对,有4处氨基酸发生突变(^(32)A→P、^(52)V→A、^(130)S→N和^(170)M→V);色瓦绵羊GPIHBP1蛋白分子量为18063.87 Da,理论等电点(p I)为4.29,不存在跨膜螺旋结构,但存在信号肽(位于第20~21位氨基酸处),是一种细胞外的酸性亲水性蛋白。色瓦绵羊GPIHBP1蛋白存在12个磷酸化位点(6个丝氨酸磷酸化位点,5个苏氨酸磷酸化位点,1个酪氨酸磷酸化位点),其二级结构以无规则卷曲状态(占55.07%)为主,其次是α-螺旋(占35.47%),延伸链、β-转角分别占5.81%和4.65%。GPIHBP1基因在色瓦绵羊乳腺、肌肉、心脏、肾脏、肝脏、小肠、瘤胃和卵巢等8个组织中均有表达,以乳腺中的相对表达量最高,其次是心脏和肝脏,均极显著高于肌肉中的相对表达量(P<0.01)。【结论】GPIHBP1基因在色瓦绵羊不同组织中均有表达,且以乳腺中的相对表达量最高,可能与乳脂相关基因表达的增减有关,其中,GPIHBP1基因与LPL基因共同作用而参与绵羊泌乳调控。

崂山奶山羊不同泌乳期乳腺组织可变剪接的鉴定与分析

本研究对崂山奶山羊泌乳初期、高峰期和末期乳腺组织RNA-Seq数据进行可变剪接分析,共鉴定到151503个可变剪接事件,对应17627个可变剪接基因,6323个差异剪接基因,显著富集到细胞器、蛋白质结合、mRNA加工和RNA剪接等相关GO条目,主要参与催产素信号通路、钙信号通路、内质网中的蛋白质加工等,与乳腺组织生理活性密切相关的KEGG通路,筛选出PTEN作为参与乳腺发育及泌乳相关生理过程中调控网络的核心基因。研究结果表明,可变剪接在奶山羊不同泌乳期的乳腺组织具有发育阶段特异性,特有可变剪接基因与乳腺组织生理活性特点密切相关,在调控奶山羊乳腺组织发育及泌乳生理中发挥重要作用。本研究可为探究可变剪接机制调控奶山羊乳腺发育与泌乳性状的分子机理提供理论依据。

采食高淀粉高油日粮奶牛的泌乳性能、乳成分变化及乳腺差异表达基因筛选

【目的】研究高淀粉高油日粮对奶牛泌乳性能和乳腺组织转录组的影响,探索乳脂下降综合征(Milk fat depression,MFD)的分子机制。【方法】选用8头荷斯坦泌乳奶牛,随机分为对照组和处理组(4头/组),采用2×2反转试验设计,分为2期,每期23 d,期间2组对调。每期的前16 d,对照组奶牛饲喂低淀粉低油的基础饲粮,对照组日粮的泌乳净能为6.78 MJ/kg;处理组奶牛在基础饲粮基础上添加266 g/kg(DM基础)细粉碎玉米和46 g/kg(DM基础)大豆油;后7 d 2组奶牛均喂基础饲粮,处理组日粮的泌乳净能为7.66 MJ/kg。分别于每期的第1、4、7、10、13、16、19和22天测定产奶量和乳成分,第16天采集乳腺组织用于检测转录本的变化。【结果】与对照组相比,处理组第13、16天奶牛的干物质采食量和产奶量显著降低(P<0.05);饲喂高淀粉高油饲粮7 d后乳脂率开始降低,其中,第10、13、16和19天的乳脂率和乳脂产量显著降低(P<0.05);第13、16和19天的乳蛋白、乳糖和非脂固形物含量显著提高(P<0.05)。在奶牛乳腺对照组和处理组中共检测到235个显著差异表达的基因,其中,64个上调、171个下调。GO分析表明,差异表达基因主要与炎症反应、α−氨基酸代谢、有机氮化合物代谢、细胞发育、脂质生物合成和脂肪细胞分化等相关;KEGG分析表明,差异表达基因主要与轴突导向、NF-κB信号通路、钙信号通路等相关。结合差异表达基因与泌乳性能的相关性分析,本研究筛选出了部分与脂质调控相关的基因作为研究奶牛MFD状态下脂质代谢的候选基因,包括在乳腺中下调表达的FGFR4、VDR、HTR2B、CCL21和TYRP1。【结论】高淀粉高油饲粮饲喂奶牛降低了干物质采食量、产奶量和乳脂率,提高了乳蛋白、乳糖和非脂固形物含量,下调了奶牛乳腺中脂肪酸合成相关基因的表达。本研究为科学配制日粮和研究MFD的分子机制研究提供了数据参考。

大肠埃希氏菌诱导小鼠乳腺纤维化模型中瘦素及其受体、MMPs和TIMPs表达的研究

为探究Leptin、Ob-R、MMP-2、MMP-9、TIMP-1和TIMP-2对小鼠乳腺组织纤维化的作用,用奶牛乳腺炎大肠埃希氏菌感染小鼠乳腺后,分别于感染后1、3、7、14、21 d采集小鼠乳腺组织,应用免疫组化染色检测Leptin、Ob-R、MMP-2、MMP-9、TIMP-1和TIMP-2在小鼠乳腺组织细胞中的表达与分布,应用qPCR、Western blot方法检测小鼠乳腺组织中Leptin、Ob-R、MMP-2、MMP-9、TIMP-1和TIMP-2 mRNA转录水平和蛋白表达水平。结果显示,小鼠感染E.coli前期(1~7 d),Leptin、Ob-R、MMP-2、MMP-9、TIMP-1和TIMP-2主要在上皮细胞,炎性细胞中表达,感染后期(14~21 d)主要在上皮细胞、成纤维细胞和炎性细胞中表达。qPCR检测mRNA转录水平,E.coli感染1~14 d,Leptin、Ob-R、MMP-2和MMP-9的转录水平均显著上升,TIMP-2的mRNA转录水平极显著下降,感染后期(14~21 d)Leptin、Ob-R、MMP-2、TIMP-1和TIMP-2的mRNA转录水平与空白组相比无显著性差异,MMP-9mRNA转录水平极显著上升。Western blot检测蛋白表达水平,E.coli感染后1~14 d,Leptin、MMP-2、TIMP-1整体蛋白表达水平上升,TIMP-2蛋白表达水平下降,Ob-R在感染后7~14 d显著上升,MMP-9在3 d和14 d显著上升,感染后14~21 d,Ob-R、MMP-2、MMP-9呈上升趋势,Leptin呈下降趋势,TIMP-1、TIMP-2与对照组的蛋白表达水平基本持平。研究表明,通过E.coli建立的小鼠乳腺纤维化模型,能够刺激Leptin、Ob-R、MMP-2、MMP-9和TIMP-1表达,抑制TIMP-2表达。初步探究了乳腺纤维化与Leptin及其受体、MMPs和TIMPs的关系,为进一步探索纤维化的分子作用机理奠定了基础。