| Aging refers to the process of the decline of the normal physiological functions of biological individuals,which can be manifested as skin folds,gray hair,slow movement,related hormone secretion disorders,memory loss,and degenerative changes in various tissues and organs.Aging is an important risk factor for the development of geriatric diseases.Premature aging is accompanied by various functional declines throughout the body and the premature appearance of geriatric diseases.Brain aging is the earliest and most important manifestation of the body’s decline.Brain aging includes Alzheimer’s disease,Parkinson’s disease and other progressive diseases of the central nervous system,with cognitive dysfunction as the main manifestation in the early stage,and there is currently no effective treatment.Objective:This study systematically sorted out the theoretical literature on aging,premature aging and brain aging in traditional Chinese medicine,discussed the pathogenesis and treatment of premature aging and brain aging from the essence-qi-spirit theory of qi-collateral doctrine,and also focused on the relationship between the functions of qi transformation of qi-collateral,qi-collateral warming and nourishing,information transmission,and homeostasis regulation,and premature aging as well as brain aging,revealing the guiding value of qi-collateral theory in preventing and treating premature aging and brain aging.In the experimental part,the role and mechanism of Bazi Bushen capsule(BZBS)in preventing and treating premature aging and brain aging were explored from different aspects such as overall functional state and brain function by establishing a D-gal-induced premature aging mouse model.This subject provided a theoretical and experimental basis for the prevention and treatment of premature aging and brain aging guided by the essence-qi-spirit theory of qi-collateral doctrine.Method:1.Theoretical discussion:The theoretical part of this research systematically sorts out and summarizes the understanding of the disease names,etiology and pathogenesis,treatment principles and methods,and prescriptions of aging,premature aging,and brain aging in traditional Chinese medicine;To explore the relationship between qi transformation of qi-collateral and abnormal effects of warming and nourishing,information conduction,and homeostasis regulation under the qi transformation and premature aging and brain aging.2.Experimental research:Experiment 1:8-week-old male C57BL/6J mice were randomly divided into normal group(100mg/kg/d sterile injection water+0.5%CMC-Na),model group(aging group,100mg/kg/d D-gal+0.5%CMC-Na),NMN control group(NMN group,100mg/kg/d D-gal+100mg/kg/d NMN),BZBS low-dose group(LD-BZ group,100mg/kg/d D-gal+lg/kg/d BZBS)and BZBS high-dose group(HD-BZ group,100mg/kg/d D-gal+2g/kg/d BZBS)after 1 week of adaptive feeding according to body weight.At the 12th week,modeling and treatment were started.Each mouse was subcutaneously injected(s.c.)into the skin on the back of the neck at a volume of 0.1mL/10g body weight,and each mouse was treated by intragastric injection(i.g.)at a volume of 0.2mL/10g body weight,once a day for 60 days.During the experiment,the initial body weight of the mice in each group was recorded,and the mice were weighed and recorded every week before the modeling and treatment.The mental state,activity level,and coat state of the mice were observed every day,and pictures were recorded on the first day after modeling and treatment.Suspension,grip,rotarod,Barnes maze and other behavioral tests were used to evaluate the muscle strength,endurance,motor coordination,learning and memory functions of mice.The blood pressure and cardiac function of mice were evaluated by non-invasive blood pressure test and cardiac ultrasonography.At the end of the experiment,the mice were fasted for 12h,and then sacrificed after anesthesia,and blood samples were collected to detect the content of aging-associated secretory phenotype(SASP)factors and anti-inflammatory factors in the blood of mice.The abdominal cavity and skull of the mice were opened,and the thymus,brain,kidney,spleen,and whole heart were weighed and saved for subsequent hematoxylin-eosin staining(H&E staining),Nissl staining,SA-β-Gal staining,immunohistochemistry staining(IHC),tissue immunofluorescence staining(IHF),western blotting(WB),real-time fluorescent quantitative PCR(qRT-PCR),magnetic Luminex assay.Experiment 2:The morphology of neurons and Nissl bodies in the brain tissues of each group mice was detected by H&E staining and Nissl staining.The protein expression of postsynaptic dense protein-95(PSD95)and ionic calcium-binding adaptor 1(Iba1)in brain tissue were determined by WB and 1HF assays.QRT-PCR was used to detect PSD95,Iba1,glial fibrillary acidic protein(GFAP)mRNA and different phenotypes of microglia CD 11b,Arg-1,Trem-2,CD206,Fizz1,Ym1 mRNA expression,and different phenotypes of astrocytes H2-D1,CD109,Ptx3,Sphk1,Tm4fs1 mRNA expression.To evaluate the effect of BZBS on brain tissue function.The protein expressions of cyclin-dependent protein kinase inhibitor CDKN2A/p16INK4a(p16INK4a)and proliferating cell nuclear antigen(PCNA)in brain tissue were detected by WB and IHC.Magnetic Luminex Assay was used to detect SASP factors,anti-inflammatory factors,and pro-inflammatory factors in blood and brain tissue,including GM-CSF,IGFBP-3,Endoglin(CD105),PAI-1,CCL3,CCL2,Expression of TNF-α,IL-1β,IL-1α,IL-2,IL-6,ICAM-1,MMP-3,MMP-9,CRP,IL-10.To evaluate whether cellular senescence is involved in the chronic inflammatory process of D-gal-induced brain aging and the effect of BZBS on cellular senescence.Result:1.Theoretical discussion:Traditional Chinese medicine believes that the maximum life expectancy of a person is 120 years old.Aging is not the same as old age.Older people can appear to be in a healthy state(elderly but not aging),impending sickness state(Gradual aging with age),disease state(aging state of the elderly).Middle-aged people show signs of aging(not yet old,but prematurely showing signs of aging),which is called premature aging.Brain aging is one of the most prominent manifestations in the process of degeneration of the body.The brain is the home of the primordial spirit,which dominates life activities,spiritual activities and sensory movement.In traditional Chinese medicine,the pathogenesis of premature aging and brain aging is attributed to deficiency,and deficiency of essence,qi and spirit is the main reason.The prescriptions of the past dynasties are based on invigorating kidney essence,nourishing vitality,nourishing kidney yin,and warming kidney yang.Qi collateral doctrine essence-qi-spirit theory believes that essence,qi and spirit are the internal factors of health and longevity.Essence is the origin of life,qi is the driving force of life,spirit is the manifestation of life.Essence is transformed into qi,qi is transformed into spirit,spirit can control the body.Qi collateral is the place where the essence,qi and spirit are transformed.Qi collateral transformation and its function of warming and nourishing,information conduction,and self-stabilizing regulation are the important guarantees for the transformation of essence,qi,spirit,and body.The Qi collaterals of the brain and marrow are closely related to the functions of the brain,spinal cord and nerves.The occurrence of premature aging and brain aging is based on deficiency of kidney essence.The main pathogenesis is that the essence does not transform into qi,the vitality is deficient,and the qi does not transform into the spirit.It is manifested by physical and spirit dystrophy and spirit apraxia.Established the treatment method of tonifying the kidney and filling the essence,regulating yin and yang,warming the vitality,and nourishing the body and spirit.The Bazi Bushen Capsule,created by extracting the essence of the traditional Chinese medicine kidney-tonifying theory,has the effect of nourishing the body and spirit,warming and nourishing the brain.2.Experimental research:Experiment 1:The results of this part of the experiment confirmed that subcutaneous injection of D-gal induced premature aging and brain aging in C57BL/6J model mice with hair loss,graying,decreased activity,and cognitive deficit.BZBS can improve the overall functional status of D-gal-induced premature aging mice,including coat status,muscle strength,endurance,flexibility,motor coordination,myocardial contractility,learning and memory,etc.According to the results of the overall status,BZBS has shown the effect of improving the overall functional status of the premature aging model mice in many aspects.Experiment 2:The results of this part of the experiment confirmed that BZBS can improve aging-related cognitive deficit by reducing neuronal damage,alleviating the decline in synaptic density and plasticity,inhibiting the activation of microglia and astrocytes,blocking and reversing the polarization of microglia and astrocytes from a neuroprotective phenotype to a neuroinflammatory phenotype,reducing neuroinflammatory factors,and increasing anti-neuroinflammatory factors,thereby protecting brain function.Cellular senescence is involved in the occurrence of a chronic inflammatory state of D-gal-induced brain aging.BZBS can alleviate cellular senescence,reduce the expression of pro-inflammatory factors,and increase the expression of anti-inflammatory factors.BZBS may improve aging-related cognitive deficit by reducing cellular senescence,promoting cell proliferation,and regulating SASP in brain tissue.Conclusions:1.Theoretical discussion:Qi collateral is the place where the essence,qi and spirit are transformed.The Qi collaterals of the brain and marrow are closely related to the functions of the brain,spinal cord and nerves.Qi collateral transformation and its function of warming and nourishing,information conduction,and self-stabilizing regulation are the important guarantees for the transformation of essence,qi,spirit,and body.The pathogenesis is that the deficiency of kidney essence,the essence does not transform into qi,the vitality is deficient,and the qi does not transform into the spirit.It is manifested by physical and spirit dystrophy and spirit apraxia.Among them,the abnormal of qi transformation of qi collateral and its function of warming and nourishing,information conduction,self-stabilization regulation induce abnormal transformation of essence,qi,spirit,and body are the key factors of premature aging and brain aging.Established the treatment method of tonifying the kidney and filling the essence,regulating yin and yang,warming the vitality,and nourishing the body and spirit,taking into account the transformation of qi collateral.The Bazi Bushen Capsule,created by extracting the essence of the traditional Chinese medicine kidney-tonifying theory,has the effect of nourishing the body and spirit,warming and nourishing the brain.2.Experimental study:BZBS can ameliorate the decline of overall functional status in D-gal-induced premature aging mice.BZBS can improve the abnormal brain tissue structure and function caused by aging.Its effect is to reduce cell senescence in brain tissue,inhibit the expression of pro-inflammatory factors released by senescent cells SASP,block and reverse the polarization of neuroprotective phenotype to neuroinflammatory phenotype,and promote cell proliferation,thereby protecting neurons and improving synaptic density and plasticity. |
