Hemopoiesis is a pluripotent hematopoietic stem cell（hematopoietic stem cells，HSCs）Under the network regulation of specific hematopoietic microenvironment, the dynamic balance process of proliferation and differentiation into various mature blood cells. humanHSCsIt mainly exists in bone marrow and a small amount ofHSCs。HSCsIt has a high self-renewal and proliferation ability, and can differentiate into various blood cell precursor cells (such as hematopoietic progenitor cells), and then redefine under the action of hematopoietic microenvironmentTo proliferate and differentiate into various primitive and mature blood cells. Finally, these mature blood cells enter the blood from the bone marrow to complete hematopoiesis^[1]. due to bone marrowHSCsIt has low differentiation, high proliferative activity, and is sensitive to chemotherapy drugs, radiation and other factors. Therefore, chemotherapy drugs and radiation are vulnerableinjuryHSCs, hematopoietic progenitor cells and bone marrow hematopoietic microenvironment, causing bone marrow depression^[2-3]And the suppression of bone marrow hematopoietic function caused by various reasonsHSCsClosely related,HSCsThe function inhibition caused by aging is the main cause of hematopoietic dysfunction. Therefore, in-depth studyHSCsThe mechanism of aging has important scientific significance for the prevention and treatment of diseases related to low hematopoietic functionHSCsAging is an important strategy for the prevention and treatment of hematopoietic dysfunction.

The most famous compatible application of Astragalus and angelica is the Astragalus and Angelica ratio created by lidongyuan5∶1Danggui Buxue Tang, made byAstragalus membranaceusOneortwoangelicaIt is mainly used to treat the syndrome of qi deficiency and fever. Danggui Buxue Decoction has a good effect in the treatment of various anemia and low hematopoietic function. Previous studies have shown that the compatibility of Astragalus and Angelica can increase the number of nucleated cells in bone marrow and hematopoietic growth factor in blood（HGF）Horizontal, suppressedHSCsAging and promoting its proliferation^[4-5], increase peripheral blood picture^[6-7]Astragalus membranaceus and Angelica sinensis1∶1The hemopoietic effect of compatibility was the strongest. Further withHSCsAging model study found that Astragalus membranaceus-angelica1∶1Compatibility can promote agingHSCsProliferation, promote cell cycle transition^[8]. Tangrong et al^[9]Research shows that Astragalus membranaceus-angelica1∶1The main components absorbed by the small intestine during compatibility are ferulic acid, pistil isoflavones, pistil isoflavone glycosides, astragaloside A and formononetin5Active ingredients. Since most traditional Chinese medicines enter the body after oral administration and absorption, it is speculated that5The three active ingredients may be the main pharmacodynamic substances for its hematopoietic promotion. But this5Whether the compatibility of three ingredients can play the role of Astragalus membranaceus and Angelica sinensis, and what is its mechanism of promoting hematopoiesis have not yet been clarified. This experiment adoptsHSCsAging model to study the effects of the compatibility of the main active components of Astragalus and Angelica on AgingHSCsTo determine the main effective substances of its hematopoietic promoting effect and provide scientific basis for its rational application.
(experiment process is omitted)
 

discuss

HSCsIt is the seed cell of the body's hematopoietic system, with strong multi-directional differentiation and self-renewal potential^[1]。HSCsAnd progenitor cell proliferation and differentiation are important for normal development and maturation of blood cells and maintenance of normal hematopoietic functionThe basis of energy. Aging of the body or under the conditions of radiotherapy and chemotherapy will causeHSCsAging,HSCsThe remarkable feature of aging is the decrease of self-renewal and proliferation ability, which is manifested as agingHSCsDecreased homing ability to bone marrow, weakened self-renewal ability and abnormal differentiation function, fineThe cell proliferation cycle is blocked, and the proliferation reaction is obviously delayed, which shows the characteristic manifestations of hematopoietic system aging such as anemia and immune function decline, the increase of tumor incidence, and the difficulty in repairing tissue and organ damage^[10-11]Chemotherapy, radiotherapy and reactive oxygen species（ROS）And other factorsHSCsAging and cell cycle arrest.At present, it is believed that,HSCsThe generation of aging is related to a variety of mechanisms, among whichp53/p21The signaling path plays an important role. Under stress conditions (e.gDNAInjury, hypoxia, infection, etc.) activationp53, resulting inp21High expression,p21Can inhibit cyclin dependent kinase（CDK）Activation, causing cell cycle arrest; up-regulationp14 ARFornutlin-3causep53Activation andp21High expression can lead to cell aging^[12]。SA-β-GalStaining is an important method to identify cell aging, and this method can only stain aging cells, but pre-aging cells, quiescent cells and tumor cells will not stain. The experimental results are reliable and not easy to be disturbed by other heterogeneic cells^[13]。

In the past, the effects of Astragalus membranaceus and Angelica sinensis on aging were studied by plasma pharmacologyHSCsThe results show that,10%Astragalus membranaceus-Angelica combined with medicated plasma can significantly promoteHSCsProliferation, and with Astragalus membranaceus-angelica1∶1The mechanism of compatibility is related to the regulation of cell cycle^[8]The experimental study of intestinal absorption in rats showed that^[9], Astragalus membranaceus-angelica1∶1The main components of intestinal absorption were ferulic acid, pistil isoflavones, pistil isoflavone glycosides, astragaloside A and formononetin5Active ingredients. Therefore, in this experiment, in order to further compare whether the compatibility of effective ingredients has Astragalus membranaceus-angelica1∶1Compatible with similar promotersHSCsProliferative effect of Astragalus membranaceus-angelica1∶1The compatibility of drug containing plasma and blank plasma was compared to explore the similarities and differences of compatibility between drug containing plasma and active components.

The results showed that ferulic acid, astragaloside A and anthocyanin from Radix Astragali and Angelica sinensis could promote aging in a dose-dependent mannerHSCsProliferation and retardationHSCsAging, for Astragalus, angelica antiHSCsThe main active ingredients of aging and promoting cell proliferation. And pistil isoflavones2.5～40.0 μg/mL, pistil isoflavone glycosides in1.25～20.00 μg/mLChronotropic agingHSCsThere was no significant effect on promoting proliferation and reducing cell aging rate. However, the orthogonal test showed that the combination of pistil isoflavones or pistil glycosides with ferulic acid, astragaloside A and formononetin could significantly promote the agingHSCsProliferation, reduce cell aging rate. The results showed that the application of pistil isoflavones and pistil glycosides alone did not promoteHSCsProliferate and reduce cell senescence, but3The compatibility of the main active ingredients can enhance the effect of the main active ingredients on promoting cell proliferation and reducing cell aging,5The compatibility of the two components has a synergistic effect on promoting cell proliferation and inhibiting cell senescence.

In the cell cycle, the cell cycleG₁Phase is the key period of cell aging, and also the key control point of its aging control. Therefore, throughG₁It can improve cell senescence.CDK4Is the core of cell cycle regulation^[14]，Cyclin D1It is a positive regulator, cell cycle dependent kinase inhibitor（CKI）Is a negative regulator^[15]。P16yesCKIRegulatory factors in the family, inG₁Key control points play an important role, and their mechanism is toCDK4Specific binding to preventCyclin D1/CDK4The formation of complexes, resulting inRbThe level of protein phosphorylation is reduced, which makes the cells irreversibly stagnate atG₁Period.CyclinIt acts on different stages of the cell cycle, causes cell division and proliferation, and accelerates the process of the cell cycle. stayCyclinIn the family,Cyclin DIn cellG₁/SThe transition point plays an important role, in whichCyclin D1yesG₁Phase specific expression proteins by acceleratingG₁To shorten the cell cycle^[16]。CDKIt plays a central role in the cell cycle regulatory network and is affected by cyclins,CDKIt is mainly through the binding with cyclin to positively regulate the cell cycle process. DifferentCyclinIdentify and combine differentCDK, composed of differentCyclin-CDKComplex, whereCDK4AndCyclin D1Combined byRbPhosphorylation of proteinsRbThe protein loses its blocking effect on the cell cycle, thus realizing the promotion and transformation of the cell cycle. The results showed that compared with the control group, the model group and blank plasma group had a static period（G₀/G₁Phase) cells increased significantly and proliferated（G₂/M＋SPhase) cells decreased significantly,CyclinD1andCDK4Protein expression decreased significantly. indicateHSCsThe expression of cyclin is down regulated during aging, which blocks the progress of cell cycle. Compared with the model group, ferulic acid, formononetin, astragaloside A and active ingredients couldG₀/G₁Phase I cells decreased significantly,G₂/M＋SPhase cells increased significantly,Cyclin D1andCDK4The protein expression was significantly enhanced. Compared with blank plasma group, Astragalus membranaceus-angelica1∶1Of the drug containing plasma groupG₀/G₁Phase cells also decreased significantly,G₂/M＋SPhase cells increased significantly,CyclinD1andCDK4Protein expression increased significantly. Compared with the active ingredient compatibility group, theG₀/G₁Phase II cell proliferation,G₂/M＋SPhase cell reduction,Cyclin D1andCDK4The protein expression decreased significantly. There was no significant difference between the drug containing plasma group and the active ingredient compatibility group. The results showed that ferulic acid, astragaloside A and formononetin were the compatible promoters of Astragalus and angelicaHSCsThe main pharmacodynamic substances of proliferation, and the secondary components pistil isoflavones and pistil isoflavones glycosides can enhance the promotion after the compatibility of the five componentsHSCsProliferative effect of Astragalus and Angelica1∶1Compatibility and5The compatibility of three active ingredients has the same promoting effectHSCsThe role of proliferation. These results showed that ferulic acid, astragaloside a, formononetin, and the compatibility of main components with pistil isoflavones, pistil isoflavone glycosides, and the compatibility of Astragalus and Angelica could increaseCyclin D1andCDK4Expression, thus promotingCyclinD1/CDK4The formation of the complex promotesHSCsfromG₁Phase entrySTo promote and transform the cell cycle.

The results showed that Astragalus membranaceus and Angelica sinensis1∶1Compatibility and its5The compatibility of ferulic acid, astragaloside IV, anthocyanin, pistil isoflavone and pistil Isoflavone Glycoside can improveHSCsAging, shortening cell cycle, promotingHSCsProliferation.5The compatibility of three active ingredients has synergistic effect. It may be that different active ingredients act onHSCsDifferent links of proliferation, so as to play a synergistic role.

References (omitted)  

To   Source: zhujiahuan, huangxiaoping, dengchangqing Study on the effect of the main active components of Astragalus and Angelica on the proliferation of aging hematopoietic stem cells [j]. Chinese herbal medicine, 2019, 50 (1): 111-119