Qinghua decoction(清化饮)improves chronic nonbacterial prostatitis possibly via regulating the chromogranin A/nerve growth factor/tyrosine kinase A signaling pathway mediated by inflammatory factors

OBJECTIVE:To explore the mechanism by which Qinghua decoction(清化饮)regulates neuroendocrine inflammation in chronic nonbacterial prostatitis(CNP)model rats and provide an experimental basis for clinical treatment.METHODS:The rats were randomly divided into six groups:normal control,model,Qianlie Tongyu capsule,low-dose Qinghua decoction,medium-dose Qinghua decoction,and high-dose Qinghua decoction group with six rats in each group.Rats in each group were sacrificed on the 29th day of treatment,and blood and prostate tissues were collected.Serum levels of tumor necrosis factor-alpha and interleukins 1-beta,6,8,and 10(TNF-αand IL-1β,-6,-8,and-10,respectively)were measured using enzyme-linked immunosorbent assay.The pathological changes in the rat prostate tissue in each group were observed under a light microscope.The expression levels of chromogranin A(CgA),nerve growth factor(NGF),and tyrosine kinase A(TrkA)were detected using reverse transcription quantitative polymerase chain reaction.Western blotting was used to detect protein expression of CgA,NGF,and TrkA.RESULTS:In the model group,the prostate capsule membrane and stroma were significantly dilated with more inflammatory cells infiltrating the stroma and perivessels.TNF-α,IL-1β,-6,and-8,CgA,NGF,and TrkA levels increased,whereas the content of IL-10 decreased,which was statistically significant compared to that in the normal control group(P<0.05).Prostate tissue cells in the high-dose group were neatly arranged with no obvious inflammatory cell infiltration.When compared with the model group,the high-dose Qinghua decoction group showed a significant improvement in these indices(P<0.05).CONCLUSION:Qinghua decoction led to inhibition of pathological changes in the prostate tissue of rats with CNP,regulation of inflammatory cytokine expression,and inhibition in the expression of CgA,NGF,and TrkA.This mechanism may be primarily related to regulation of the CgA/NGF/TrkA signaling pathway mediated by various inflammatory factors.

补肾调经方在种植窗期经STAT3信号通路改善超促排卵小鼠胚胎着床的作用及机制

目的观察补肾调经方对控制性超促排卵(COH)小鼠种植窗期子宫内膜信号转导和转录活化因子3(STAT3)信号通路的影响,探讨其改善胚胎着床的作用及机制。方法昆明小鼠60只,随机分为模型组、补肾调经方组和正常组,每组20只。妊娠第4天每组处死10只小鼠,放射免疫法检测血清雌二醇(E_2)、孕酮(P)水平,Western blot法检测子宫内膜白血病抑制因子(LIF)、磷酸化信号转导和转录活化因子3(P-STAT3)蛋白的含量,免疫组化法检测子宫内膜瘦素及瘦素受体(OB-R)的表达。妊娠第8天处死各组剩余小鼠,观察平均胚胎着床数。结果与正常组比较,模型组小鼠血清E_2降低(P<0.05),P升高(P<0.01);子宫内膜LIF、P-STAT3的含量下降(P<0.01),内膜上皮细胞瘦素及OB-R蛋白的表达降低(P<0.01);平均胚胎着床数减少(P<0.01)。与模型组比较,补肾调经方组小鼠血清P降低(P<0.05);子宫内膜LIF、P-STAT3蛋白的含量升高(P<0.01),内膜上皮细胞瘦素表达增加(P<0.05);平均胚胎着床数增多(P<0.01)。结论补肾调经方可改善COH小鼠胚胎着床,其机制可能与调控种植窗期子宫内膜STAT3信号通路相关蛋白有关。

大黄?虫丸对ox-LDL诱导血管内皮细胞泛素蛋白酶体系统的影响

目的探讨氧化低密度脂蛋白(ox-LDL)损伤人脐静脉血管内皮细胞(HUVEC)泛素蛋白酶体系统(UPS)的变化以及大黄[庶虫]虫丸对泛素(Ub)、泛素激活酶(UbE1)、核转录因子(NF-κB)的影响。方法体外培养HUVEC,加入ox-LDL共同孵育制成细胞损伤模型。将药物血清作用于细胞,实验分为正常对照组、ox-LDL组、大黄[庶虫]虫丸(1 400 mg·kg^-1、700 mg·kg^-1)干预组、辛伐他汀阳性对照组。观察各组细胞形态,ELISA法检测ICAM-1含量,免疫组化法检测Ub表达,RT-PCR法测定UbE1、NF-κB、IκB mRNA表达,Western Blot法检测UbE1、NF-κB、I-κB的蛋白表达。结果大黄[庶虫]虫丸干预后细胞形态随浓度变化趋于正常;ICAM-1含量降低(P<0.05);Ub呈弱阳性;UbE1、NF-κB mRNA表达减少(P<0.05),蛋白表达量下降;IκB mRNA上调(P<0.01),蛋白表达量增加。结论大黄[庶虫]虫丸可通过阻抑泛素蛋白酶体系统活化NF-κB干预基因的表达,从而减轻动脉粥样硬化(AS)的炎症反应。

加味桃核承气汤对糖尿病心肌病大鼠NLRP3炎症小体的影响

目的:探讨加味桃核承气汤对糖尿病心肌病大鼠NOD样受体蛋白3(NLRP3)炎症小体激活的影响。方法:选取3~4周龄SPF级雄性SD大鼠,随机分为正常组和造模组。造模组大鼠采用高脂饲料喂养4周后,给予链脲佐菌素(STZ)腹腔注射(35 mg·kg^(-1))制备糖尿病大鼠模型,再按照空腹血糖随机分为模型组、加味桃核承气汤低、高剂量组(11.7、23.4 g·kg^(-1))、盐酸二甲双胍组(67.5 mg·kg^(-1))。各组连续灌胃给药8周后经超声成像平台检测大鼠心脏功能及结构。股动脉取血检测大鼠空腹血糖(FBG)、甘油三酯(TC)、总胆固醇(TG);苏木素-伊红(HE)染色观察大鼠心肌病理形态学改变;酶联免疫吸附测定法(ELISA)检测大鼠血清白细胞介素^(-1)β(IL^(-1)β)及白细胞介素^(-1)8(IL^(-1)8)水平;蛋白免疫印迹法(Western blot)检测心肌组织NLRP3、ASC、Caspase^(-1)、p-NF-κB p65蛋白表达的影响。结果:与正常组比较,模型组大鼠FBG、TC、TG水平显著升高(P<0.01),左室射血分数(EF)、左室短轴缩短率(FS)明显降低(P<0.05),HE染色可见心肌细胞肥大、心肌纤维化,血清中IL^(-1)β、IL^(-1)8含量及心肌组织NLRP3、ASC、Caspase^(-1)、p-NF-κB p65蛋白表达均显著增高(P<0.01)。与模型组比较,加味桃核承气汤低、高剂量组及二甲双胍组可有效降低大鼠FBG、TC、TG水平(P<0.05),EF、FS均明显改善(P<0.05),大鼠心肌组织病理学损伤明显改善,同时血清IL^(-1)β、IL^(-1)8含量及心肌组织NLRP3、ASC、Caspase^(-1)、p-NF-κB p65蛋白表达均明显降低(P<0.05),其中加味桃核承气汤高剂量组改善更明显。结论:桃核承气汤加味可通过抑制NLRP3炎症小体激活从而发挥保护心肌作用。

资癸益冲方对卵巢储备功能下降模型大鼠卵巢氧化损伤的影响及其作用机制

目的探讨资癸益冲方对卵巢储备功能下降(DOR)模型大鼠卵巢氧化损伤的影响及其作用机制。方法采用环磷酰胺腹腔注射诱导DOR大鼠模型,将诱导造模成功的大鼠随机分为模型组、资癸益冲方高剂量组(28 g/kg)、资癸益冲方低剂量组(7 g/kg),另选10只动情周期规律的大鼠作为正常组。各组分别给予相应处理因素28 d后,选动情前期处死大鼠,检测血清雌二醇(E2)、抗苗勒管激素(AMH)、卵泡刺激素(FSH)、黄体生成素(LH)水平;HE染色观察卵巢组织病理形态学改变;检测卵巢组织超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)活力和丙二醛(MDA)含量;免疫组化和Western Blot法检测肿瘤抑制因子p53、周期蛋白依赖激酶抑制因子p21和核因子相关因子-2(Nrf2)的蛋白表达。结果与正常组比较,模型组大鼠血清E2、AMH水平降低,FSH、LH水平升高(均P<0.05);卵巢组织中各级卵泡数减少,闭锁卵泡增多,颗粒细胞层数减少;卵巢组织SOD、GSH-Px、CAT活力降低,MDA含量升高(均P<0.05);p53、p21、Nrf2蛋白表达增强(均P<0.05)。与模型组比较,资癸益冲方高、低剂量组大鼠血清E2、AMH水平升高,FSH、LH水平降低(均P<0.05);卵巢组织中闭锁卵泡减少,颗粒细胞层数增多;卵巢组织SOD、GSH-Px、CAT活力升高,MDA含量降低(均P<0.05);p53、p21和Nrf2蛋白表达下降(均P<0.05)。结论资癸益冲方能改善DOR大鼠的卵巢储备功能,减轻卵巢氧化损伤,降低卵巢组织中p53、p21和Nrf2的蛋白表达。