Neuroendocrine-Immune Function Ntawm Cistanche Deserticola Thiab Nws Cov Nplej Wine Steaming Khoom hauv Glucocorticoid-Induced Rat Model-Ⅱ

3.2.4. Immunohistochemistry

Neuronal apoptosis yog tswj los ntawm ntau yam apoptotic thiab antiapoptotic genes, feem ntau yog cov apoptosis-txhawb cov noob Bax thiab cov noob tshuaj antiapoptotic ntawm Bcl -2 gene tsev neeg. Bcl -2 yog nyob rau hauv reactive oxygen-ua organelles xws li mitochondria, endoplasmic reticulum, thiab nuclear membranes. Bcl-2 muaj kev cuam tshuam me ntsis rau cov pa oxygen dawb radicals tab sis tuaj yeem ua tautiv thaiv oxidative puas hauv hlwb. Bax thauj los ntawm cytoplasm mus rau mitochondrial membrane, hloov cov mitochondrial membrane qauv, thiab txhawb kev tso tawm ntawm cytochrome c thiab induction ntawm apoptosis.

NATURAL CISTANCHE TUBULOSA rau kev txhim kho kev sib deev PHGS75% ECH 30% ACT 12%

Muaj ib qho kev qhia hauv adrenal cytoplasm ntawm Bax thiab Bcl -2 protein. Cov staining pom tag nrho cov khoom daj. Piv nrog rau cov pab pawg BC, qhov nruab nrab optical ntom ntom ntawm Bax protein nyob rau hauv pawg MC tau nce ntau (P< 0.01). Compared to that of the MC group, the average optical density in the CD-LD and WCD-LD groups decreased significantly (P < 0.01) (Figure 5(a)).

Daim duab 5(b) qhia tau hais tias piv rau cov pab pawg BC, qhov nruab nrab optical ntom ntawm Bcl-2 protein nyob rau hauv pawg MC poob qis heev (P< 0.05). Compared to that of the MC group, the average optical density in the WCD-HD and CD-HD groups increased significantly (P < 0.01), while those of the WCD-MD and CD-MD groups increased (P < 0.05).

Lub caspase-3 protein yog nyob rau hauv lub cytoplasm nyob rau hauv nws cov thawj daim ntawv nyob rau hauv ib txwm cov ntaub so ntswg thiab muaj tsawg qhia. Cov cim zoo yog daj ntseg daj lossis xim av daj. Piv nrog rau cov pab pawg BC, qhov kev qhia ntawm caspase-3 hauv pawg MC tau nce ntau heev (P< 0.05). Compared to that of the MC group, the expression of caspase-3 in the WCD-HD and WCD-MD groups increased (P < 0.05) (Figure 5(c)).

Hauv cov ntaub so ntswg ib txwm, qhov kev qhia ntawm Fas gene raug txo, thiab cov teeb liab zoo rau hauv situ hybridization yog xim daj daj. Raws li pom nyob rau hauv daim duab 5(d), piv rau cov pab pawg BC, qhov nruab nrab optical ceev ntawm Fas protein nyob rau hauv pawg MC nce (P< 0.05). Compared to that of the MC group, the average optical density in the WCD-HD and WCD-LD groups improved (P < 0.05) (Figure 5(d)).

FasL protein nyob rau hauv lub cell membrane los yog cytoplasm, thiab nws cov cim zoo yog xim av. Piv nrog rau cov pab pawg BC, qhov nruab nrab qhov muag pom qhov ntom ntom ntawm FasL noob hauv pawg MC tau nce ntau (P< 0.05). Compared to that of the MC group, the average optical density of the FasL gene in the WCD-HD, WCD-MD, CDHD, and CD-MD groups increased significantly (P < 0.01) (Figure 5(e)).

Cov txiaj ntsig tau pom tias CD thiab WCD tuaj yeem ua tauinduce apoptosis nyob rau hauv inflammatory cellsthiab txo qhov kev puas tsuaj rau adrenal los ntawm kev txhawb nqa kev qhia ntawm FasL protein thiab khi rau Fas protein, ua kom cov caspase-3 protein thiab ua rau caspase cascade. Cov nyhuv ntawm WCD-HD pawg yog qhov zoo tshaj plaws.

3.3. Kev txiav txim siab ntawm kev tiv thaiv kab mob

3.3.1. Immune Organ Index

Piv nrog rau cov pab pawg BC, thymus coefficient thiab cov poov xab index ntawm pawg MC poob qis heev (P< 0.01). Compared to the MC group, the WCD-MD group had the greatest difference (P < 0.05). The thymus coefficient was almost back to the normal level. For the spleen index, the WCDMD, WCD-LD, and CD-MD groups showed significant differences compared with that of the MC group (P < 0.01) (see Figure 6). Therefore, WCD and CD treatment could protect the immune organs, and the effects of WCD were better.

NATURAL CISTANCHE TUBULOSA rau kev txhim kho kev sib deev PHGS75% ECH 30% ACT 12%

3.3.2. Kev kuaj pom T Lymphocyte Subsets hauv cov ntshav Peripheral

Qhov feem pua ntawm CD4+ cov hlwb nyob rau sab laug sab laug thiab CD8+ cov hlwb nyob rau sab xis qis qis hauv BC thiab txhua pawg tshuaj tau siab dua li hauv pawg MC. Piv nrog rau pawg BC, qhov piv ntawm CD4+ThCD8+ poob qis hauv pawg MC (P< 0.01). Compared to that of the MC group, the ratio of CD4+ThCD8+ increased significantly in the PC group, the WCD group, and the CD-HD group (P < 0.01) (Figure 7).

3.3.3. Kev txiav txim siab ntawm Qib IL-10, IL-6, IL-2, TNF-Α, Thiab IFN-C hauv cov ntshav

Daim duab 8 qhia tau hais tias piv rau cov pab pawg BC, qib ntawm IL-6, TNF- , thiab IFN-c tau nce ntau (P< 0.01). The level of IL-10 and IL-2 decreased in the MC group. Compared to that of the MC group, the level of IL-6, TNF-α, and IFN-c decreased and IL-10 and IL-2 increased in serum in each of the drug groups. The effect of WCD was better than the effect of CD, especially in the WCD-MD group, in which the effect was close to that of the PC group.

3.3.4. Cov txiaj ntsig ntawm Lymus Tissue Microscope Observation Experiment

Daim duab 9 qhia tau hais tias nyob rau hauv pawg BC, muaj coob tus lymphocytes nyob. Lymphocytes thiab reticulocytes tau xoob nyob rau hauv qhov chaw nrog tawg pleb los yog oval thymocytes. Tsis muaj necrosis, tsis muaj inflammatory cell infiltration, thiab tsis muaj qhov txawv txav.

Pab pawg MC tau pom ntau cov ntaub so ntswg lymphoid necrosis, nrog rau kev tsis sib haum xeeb ntawm lymphocytes thiab reticulocytes hauv qhov chaw. Round los yog oval thymocytes ploj, loj heev cell degeneration los yog necrosis tshwm sim, thiab muaj inflammatory cell infiltration nrog txawv pathological hloov.

Hauv CD pawg, cov yam ntxwv morphological ntawm lymphocytes tau rov qab mus rau qhov qub, thiab kev npaj ntawm central lymphocytes thiab reticulocytes yuav luag zoo li qub. Round lossis oval thymocytes tau zoo lawm, tab sis ib tug me me ntawm degenerative los yog necrotic lymphocytes thiab infiltrating inflammatory cells tseem nyob.

Hauv pawg WCD thiab pawg PC, cov yam ntxwv morphological ntawm lymphocytes tau kho zoo dua. Cov lymphocytes thiab reticulocytes nyob rau hauv qhov chaw tau zoo sib xws, lub puag ncig los yog oval thymocytes yuav luag zoo li qub, tab sis tseem muaj tsawg tsawg ntawm infiltrating thiab degenerative lymphocytes.

3.4. CaM Protein Gene Expression

RQ tus nqi (CaM mRNA -actin mRNA) tau txiav txim siab los tshuaj xyuas qhov qhia ntawm CaM mRNA. RQ ntawm pawg BC yog 1. Kev qhia ntawm CaM mRNA hauv hypothalamus hauv pawg MC tau nce ntau (P< 0.01), while the value decreased in the WCD-HD, WCD-MD, WCDLD, and CD-HD groups (P < 0.05) (Figure 10). The expression of CaM mRNA in the hypophysis in the MC group was lower than that in the BC group (P < 0.01). The expression in the WCD-HD, WCD-MD, WCD-LD, CD-HD, and CD-LD groups increased (P < 0.05), and the effect in the WCD groups was better than the effect in the CD groups.

4. Kev sib tham

Suav Pharmacopoeia [2] cov ntaub ntawv sau tseg tias CD thiab WCD tuaj yeem siv rau hauv tsev kho mob raws li hom tseem ceeb ntawm cov khoom decoction. Tom qab steamed nrog mov wine, lubantiaging thiab tonifying raum-yang teebmeemtau txhim kho. Los ntawm UPLC-QqQ-MS, peb tau kuaj pom qhov hloov pauv ntau ntawm cov khoom siv tshuaj lom neeg thaum lub sij hawm ua cov nplej wine steaming. Iridoid glycosides tsis ruaj khov thiab cov ntsiab lus ntawm cov glycosides txo qis, txawm tias ua tsis tau. Cov ntsiab lus ntawm phenylethanol glycosides kuj hloov pauv, uas tuaj yeem ua rau muaj kev hloov pauv hauv cov tshuaj pharmacodynamics thiab cov txiaj ntsig kho mob.

NATURAL CISTANCHE TUBULOSA rau kev txhim kho kev sib deev PHGS75% ECH 30% ACT 12%

Cov qauv ntawm lub raum-yang tsis muaj peev xwm tau rov ua dua los ntawm kev txhaj tshuaj subcutaneous ntawm corticosterone [19]. Kev noj tshuaj ntau dhau corticosterone tuaj yeem cuam tshuam qhov zais cia ntawm CRH hauv hypothalamus. Lub caij no, qhov zais cia ntawm ACTH kuj raug txwv. Qib ntawm adrenal cortex cov tshuaj hormones txo, thiab yog li, kev ua haujlwm ntawm HPA axis raug txo. Hypofunction ntawm HPA axis yog ib yam ntxwv ntawm lub raum-yang deficiency [20]. Tom qab ntawd cov nas pom qhov tshwm sim "depletion" thiab cov tsos mob zoo sib xws nrog cov mob raum-yang deficiency hauv tsev kho mob.

Hauv pab pawg MC, qib ntawm CRH, ACTH, T, thiab CORT hauv cov ntshav nas poob qis (P< 0.01). After being treated by CD and WCD, the content of CRH, ACTH, T, CORT, and cortisol all increased to some degree, and the effect of WCD-MD was the best, almost equal to that in the PC group. These results indicated that the relative hormone balance in the HPA axis had been destroyed in the MC group, CD and WCD could improve the functional recovery of abnormal adrenal axis hormone levels, but the effect of WCD was better.

Modulating qhov kev faib ua feem ntawm Bcl-2 cov neeg hauv tsev neeg yog ib qho ntawm cov txheej txheem tseem ceeb uas FasThFasL kho txoj kev tuag receptor, thiab Bcl-2 tsev neeg tswj txoj hauv kev mitochondrial [21]. Apoptosis-inhibiting factor Bcl-2 thiab apoptosis-inducing factor, xws li Bax, tag nrho cov koom nrog Bcl-2 tsev neeg [22]. Cell apoptosis tuaj yeem txiav txim siab los ntawm tus txheeb ze piv ntawm apoptosis-inhibiting yam thiab apoptosis-inducing yam [23]. Mitochondria muaj kev ua haujlwm ntawm lub hauv paus chaw kuaj xyuas rau ntau hom apoptosis [24]. Txoj kev tshawb fawb tam sim no tau pom tias CD-MD thiab WCDs tuaj yeem txiav tawm ntawm Bax thiab nce Bcl -2 rautxhim kho lub raum-yang deficiency. Fas yog qhov tseem ceeb tuag receptor uas induces apoptosis los ntawm ligation nrog Fas ligand (FasL) [25]. Kev nthuav qhia ntawm Fas thiab FasL hauv pawg MC tau nce ntxiv, thaum qhov kev qhia no tau nce ntau hauv WCD-MD pawg (P< 0.01) compared with the MC group. Both FasThFasL and BaxThBcl-2 systems transduce apoptotic signals to the caspase family [26]. Caspase-3 is one of the pivotal proteinases that initiate cell apoptosis. The activation of caspase-3 indicates that apoptosis enters an irreversible stage [27]. The expression of caspase-3 in the MC group increased and increased significantly in the WCD-HD and WCD-MD groups (P < 0.05).

Lub cev tsis muaj zog yog ib qho tseem ceeb tshwm sim ntawm lub raum-yang deficiency [28]. Hauv txoj kev tshawb no, peb pom tias qhov Performance index ntawm tus po thiab thymus ntawm WCD-MD pab pawg tau zoo heev (P< 0.01, P < 0.05). The immune response of the immune system is regulated mainly by the Th subgroups [29]. IL-2 is the growth factor for T cells, secreted by T cells and macrophages, participates in inflammatory regulation, regulates immunity, and increases infection resistance [30]. The level of IL-2 in the MC group decreased (P < 0.01), and after treatment with CD and WCD, the level of IL-2 recovered. IL-6 and TNF-α are proinflammatory cytokines. Increased levels of IL-6 and TNF-α cytokines cause cytokine imbalances and exacerbate inflammation. The levels of IL-6 and TNF-α increased in the MC group, while after treatment, the levels of IL-6 and TNF-α decreased. T cells are divided into CD4 cells and CD8 cells, and the CD4+ThCD8+ ratio can be used as an important index of immune regulation [31]. CD4+ThCD8+ in the MC group decreased (P < 0.01), while in the WCD and CDHD groups increased (P < 0.01).

HPA axis yog qhov kev tswj hwm tseem ceeb tshaj plaws vim tias nws tswj hwm glucocorticoid secretion thiab endocrine system los tswj cov kev ua haujlwm secretory ntawm adrenal cortex [32]. Kev nthuav tawm ntau dhau ntawm CaM mRNA hauv cov ntaub so ntswg hypothalamus yog cuam tshuam rau lub raum tsis muaj zog. CD thiab WCD tuaj yeem thaiv cov calcium channel, inhibit CaM mRNA qhia hauv hypothalamus, thiab txo cov calcium influx, yog li txo Ca2+·CaM complex kev ua haujlwm.

Kev tsim ntawm lub raum-yang deficiency yog ib txoj kev txhim kho kab mob ntev. Lub raum tsis yog lub tswv yim ntawm lub cev nkaus xwb, thiab nws yog ib lub ntsiab lus dav dav rau neuroendocrine-kev tiv thaiv kab mob thiab genitourinary systems. Neuroendocrine-immune (NEI) network yog ib qho tseem ceeb integrated tswj system [33]. CD thiab WCD tuaj yeem tswj hwm txoj haujlwm ntawm HPAT axis thiab txhim kho kev rov qab los ntawm lub raum-yang deficiency. Qhov txiaj ntsig no muab qhov kev sim hauv paus rau kev soj ntsuam kev tiv thaiv thiab kev kho mob raum-yang deficiency.