Cistanche Tubulosa Tiv Thaiv Dopaminergic Neurons Los Ntawm Kev Cai Ntawm Apoptosis Thiab Glial Cell-Drived Neurotrophic Factor: Hauv Vivo Thiab hauv Vitro-Ⅱ

Kev Xeem Cwj Pwm

Kev xeem ua luam dej (Zhu li al., 2014)

Kev sib koom tes ntawm lub cev txav hauv cov nas tau ntsuas los ntawm Kev Ntsuas Ua Si. Cov nas tau muab tso rau hauv ib lub tank dej (25 cm nyob rau hauv qhov siab thiab 10 cm nyob rau hauv txoj kab uas hla) uas muaj 10 cm dej thiab sim nyob rau hauv ib tug nyob ntsiag to ib puag ncig los sau lawv nyob twj ywm ntev tshaj 5 min.

Qhib Field Test (Kawai li al., 1998)

Locomotor kev ua haujlwm tau ntsuas los ntawm kev siv Open field test. Cov nas tau sim nyob rau hauv ib qho chaw ntsiag to thiab tsis pom kev thiab muab tso rau hauv ib tus zuj zus hauv 30 cm × 30 cm × 15 cm pob tshab acrylic thawv nrog 6 cm × 6 cm sib cais daim phiaj hauv qab. Cov nas tau muab 10 min kom hloov mus rau ib puag ncig thiab tom qab ntawd qhov kev hloov pauv ntawm tus lej kab sib chaws thiab kev txhawb nqa ntawm cov nas ib leeg tau ntsuas tsib zaug sib law liag kom tau txais txiaj ntsig zoo.

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

Brain Tissue Sampling

Ua ntej cov ntaub so ntswg sampling, nas tau pub ad libitum nrog kev nkag mus rau dej dawb thiab tau txais kev cuam tshuam tshuaj rau 14 hnub sib law liag. Plaub nas nyob rau hauv txhua pab pawg raug xaiv thiab decapitated sai. Lub SN (Bregma: −2.75 mm −2.92 mm) los ntawm txhua tus tsiaj raug cais thiab muab tso rau hauv dej khov. Cov ntaub so ntswg hauv hlwb tau muab yaug nrog 0.9% dej khov-txias sodium chloride tov kom tshem tawm cov ntshav thiab ziab ntawm daim ntawv lim ua ntej khaws cia ntawm −80◦C. Plaub nas los ntawm txhua pab pawg tau txhaj tshuaj intraperitoneally thiab lawv lub hauv siab qhib. Tom qab ntawd ib rab koob infusion tau muab tso rau hauv sab laug ventricle ntawm txhua tus tsiaj. Txhawm rau tshem tawm cov ntshav hauv cov hlab ntsha, txoj cai atrial appendage raug txiav thiab tus tsiaj tau ntim nrog 4◦C ib txwm saline kom txog thaum lub siab tig daj ntseg kom ntseeg tau tias muaj kev sib txuas ntxiv. Thaum cov effluent ntawm txoj cai atrium tau pom tseeb, txhua tus tsiaj tau perfused nrog 4% paraformaldehyde fixative. Tom qab perfusion, lub paj hlwb ntawm txhua tus tsiaj tau raug txiav kom zoo thiab tom qab kho hauv 4% paraformaldehyde rau 24 teev. Tom qab ntawd cov ntaub so ntswg tsau tau muab yaug hauv dej ntws, lub cev qhuav dej hauv cov tshuaj ethanol, thiab tshem tawm hauv cov tshuaj xylene. Qhov no yog ua raws li los ntawm paraffin immersion thiab embedding.

Hloov ntawm DA Quantity Measured by HPLC

Cov nanopowdered SN los ntawm txhua pab pawg tau muab tso rau hauv ib da dej dej khov uas muaj 0.9% sodium chloride tov (1: 9 ratio). Lub hlwb cov ntaub so ntswg tau homogenized siv lub ultrasonic cell disruptor thiab centrifuged ntawm 1200 rpm rau 20 min ntawm 4◦C kom tau lub supernatant. Rau HPLC, Hypersil AA-ODS kem (2.1 mm × 200 mm, 5 µm) ntawm 30◦C kem kub tau siv. Kev kuaj pom fluorescence tau ua tiav ntawm 280 nm λex thiab 340 nm λem. Kev txhaj tshuaj yog 10 µL.

Kev nthuav qhia ntawm TH, GDNF, GFR 1 thiab Ret Pom los ntawm Immunohistochemistry

Paraffin seem (5 µm tuab) ntawm ib tus neeg lub hlwb raug cais tawm ntawm txhua tus tsiaj thiab muab tso rau hauv 40◦C dej sov da dej rau flattening thiab adhering rau iav slides. Tag nrho cov ntaub so ntswg tau muab tso rau hauv qhov cub 60 ◦C rau 3-6 h, ua raws li xylene dewaxing, gradient ethanol dehydration, thiab antigen retrieval los ntawm incubating nyob rau hauv ib tug citric acid tsis thiab cua sov nyob rau hauv lub microwave rau 20 feeb. Cov ntaub so ntswg tau muab tso rau hauv 3% H2O2 tov ntawm chav tsev kub rau 10 min. Tom qab tau ntxuav peb zaug hauv PBS, cov ntaub so ntswg tau muab tso rau hauv cov kua dej ib txwm nyob hauv chav kaw ntawm chav sov li 20 min. Immunohistochemical staining tau ua raws li cov chaw tsim khoom cov lus qhia. Motic Med 6.0 duab analyzer tau siv los xam tus nqi ntawm kev sib xyaw ua ke ntawm qhov muag pom hauv cov hlwb zoo.

Western Blot Analysis nyob rau hauv lub hlwb cov ntaub so ntswg ntawm nas

Txoj kev tshawb no tau soj ntsuam cov protein qhia ntawm tyrosine hydroxylase (TH), GDNF, GFR 1, Ret, Bcl2, thiab Bax. Lub hlwb lysate los ntawm txhua pab pawg tau homogenized rau 3{{10}}} min rau dej khov, ua raws li qhov kub thiab txias centrifugation, 20,000 rpm, ntawm 4 ◦C rau 5 feeb los sau cov supernatant. Cov qauv protein tau muab cais raws li qhov siab tas li siv 10% SDS-PAGE gel raws li tau piav qhia saum toj no. Thawj cov tshuaj tiv thaiv kab mob: TH 0.15 mg / ml, GDNF 0.5 mg / ml, GFR 1 0.8 mg / ml, Ret 0.63 mg / ml, Bcl -2 0.34 mg / ml, thiab Bax 0.11 mg/ml. Cov txheej txheem yog tib yam li saum toj no.

NATURAL CISTANCHE TUBULOSA FOR ANTI AGING ANTI-ALZHEIMER PHGS75% ECH 30% ACT 12%

Kev txheeb cais

Txoj kev tshawb no tau siv SPSS 20.0 statistical software rau kev ua cov ntaub ntawv thiab tshuaj xyuas. Parameter qhov tseem ceeb tau qhia tias txhais tau tias ± tus qauv sib txawv (¯x ± S). ANOVA tau siv rau kev txheeb xyuas cov ntaub ntawv ib leeg. LSD lossis Games-Howell xeem tau siv los sib piv cov pab pawg. P< 0.05 (or P < 0.01) was considered as a statistically significant difference.

TSEEM CEEB

Cov khoom xyaw nquag ntawm C. tubulosa Nanopowder

Hauv qhov ntau ntawm 200-400 nm scanning, ECH hauv C. tubulosa thiab VER hauv 330 nm muaj qhov siab tshaj plaws nqus, uas tshwm sim hauv 20 min. ICA muaj qhov nqus siab tshaj plaws ntawm 270 nm thiab tshwm sim tom qab 20 min (Daim duab 1A). Cov txiaj ntsig tau pom tias cov qauv tsis zoo tsis cuam tshuam nrog kev kuaj pom (Daim duab 1B). Cov qauv thiab kev tswj tau zoo tib yam chromatographic peaks thiab cov qauv tsis zoo tsis muaj. Qhov no qhia tau tias lwm cov khoom xyaw hauv cov qauv tsis cuam tshuam nrog cov khoom ntsuas. Ntxiv mus, peb lub Cheebtsam thiab lub ncov uas nyob ib sab tuaj yeem ncav cuag lub hauv paus kev sib cais thiab qhov kev sib cais yog ntau dua 1.5.

C. tubulosa Nanopowder Txo MPP+ -Induced Cytotoxicity hauv MES23.5 Cells

Kev ua tau zoo ntawm MES23.5 hlwb tau txo qis nrog kev nce ntxiv ntawm MPP +. Daim duab 2F qhia qhov tseem ceeb cytotoxicity ntawm qhov sib txawv ntawm MPP +.

C. tubulosa nanopowder txo MPP+-induced cytotoxicity thiab txhim khu kev vam meej ntawm MES23.5 hlwb. Daim duab 2G qhia tau hais tias C. tubulosa nanopowder ntau npaum li cas ntawm 10–250 µg/mL tau siv tshuaj tiv thaiv los ntawm MPP+- kho MES23.5 hlwb.

Cytomorphological nyhuv ntawm C. tubulosa Nanopowder

Ib txwm MES23.5 hlwb muaj cov cell adhesion zoo thiab yog spindle-zoo li tus nrog meej cell ciam teb thiab synapses. MPP+-MES23.5 hlwb ua rau pom cov cell adhesion tsis zoo thiab shrinkage, thiab ntau tau raug tshem tawm hauv xov xwm nrog kev cog lus synapses. Cov hlwb no tau sib sau ua ke, shrunken thiab puag ncig nrog vacuoles hauv, thiab cov nuclei tau tawg lossis tawg. C. tubulosa nanopowder nyob rau hauv ntau npaum li cas txhim kho lub cytomorphology ntawm MES23.5 hlwb nyob rau hauv txawv degrees los ntawm kev txhim kho cell adhesion thiab synaptic clearance ntawm lub tsheb pab pawg neeg. MES23.5 hlwb nyob rau hauv high-dose C. tubulosa pab pawg kho mob pom morphology uas zoo ib yam li pawg tswj ib txwm (Figures 2A–E).

Cov nyhuv ntawm C. tubulosa Nanopowder ntawm TH Expression thiab Apoptosis hauv Cells

Daim duab 3 qhia txog kev txo qis hauv TH protein qhia hauv pawg tsheb. Cov TH protein qhia tau nce txawv hauv cov pab pawg kho nrog ntau npaum li cas ntawm C. tubulosa. Txawm li cas los xij, kev kuaj LSD tau pom tias tsis muaj qhov sib txawv tseem ceeb ntawm peb pawg kho mob.

Daim duab 4 qhia cov txiaj ntsig ntawm kev ntsuam xyuas apoptosis siv cov cytometry ntws. Tus nqi ntawm apoptosis nyob rau hauv lub tsheb pab pawg neeg yog ho siab dua nyob rau hauv lwm pab pawg neeg. Cells kho nrog ntau npaum li cas ntawm C. tubulosa nanopowder tau pom qhov sib txawv ntawm kev poob qis hauv tus nqi apoptotic piv nrog cov pab pawg tsheb. Cells nyob rau hauv nruab nrab thiab high-dose C. tubulosa kev kho mob pab pawg muaj kev txhim kho tseem ceeb tshaj plaws nyob rau hauv apoptotic tus nqi piv nrog rau lwm pab pawg neeg kho C. tubulosa. LSD kuaj pom tias tsis muaj qhov sib txawv tseem ceeb ntawm ob pab pawg kho mob tab sis qhov sib txawv tseem ceeb ntawm pab pawg tsawg heev.

Cov nyhuv ntawm C. tubulosa Nanopowder ntawm Bcl2 / Bax Protein nthuav tawm hauv Cells

Daim duab 5 qhia tau hais tias qhov kev qhia ntawm Bcl2 protein nyob rau hauv lub hlwb ntawm lub tsheb pab pawg neeg tau qis dua piv nrog rau cov pab pawg tswj ib txwm. Nyob rau hauv sib piv, qhov kev qhia ntawm Bax protein nyob rau hauv lub hlwb ntawm lub tsheb pab pawg neeg yog ho siab tshaj nyob rau hauv ib txwm tswj pawg. C. tubulosa cov pab pawg kho mob tau pom tias muaj Bcl2 protein ntau ntxiv thiab txo qis Bax protein qhia hauv MPP+- kho MES23.5 hlwb. Nyob nruab nrab ntawm peb pawg kho mob, muaj qhov sib txawv tseem ceeb hauv LSD xeem. Cov teebmeem no yog nyob ntawm koob tshuaj.

Kev Xeem Cwj Pwm

Cov txiaj ntsig ntawm kev sim ua luam dej tau pom tias cov nas nyob hauv pab pawg lub tsheb muaj lub sijhawm nyob ruaj khov, uas tau nce ntxiv raws sijhawm. Hnub tim 14, cov nas nyob hauv pab pawg tsheb muaj lub sijhawm nyob ruaj khov ntev dua li cov nas hauv pawg tswj hwm ib txwm muaj. Lub sijhawm ruaj khov ntawm cov nas nyob hauv

koob tshuaj tsawgC. tubulosapab pawg kho mob tsis txawv ntawm cov nas nyob hauv pawg tsheb. Txawm li cas los xij, lub sijhawm nyob ruaj khov ntawm cov nas nyob rau hauv cov tshuaj tua kab mob siab C. tubulosa yog qhov tsawg dua li cov nas hauv pawg tsheb.

Cov txiaj ntsig ntawm Open field test qhia tias tom qab MPTP-vim kev puas tsuaj hauv cov nas, cov nas hauv pawg tsheb tau pom tias muaj kev poob qis hauv lawv lub peev xwm rau kev ua haujlwm raws li qhia los ntawm kev txhawb nqa ntau zaus. Tom qab ib hnub 14- kev tswj hwm ntawmC. tubulosa nanopowder, cov nas nyob rau hauv cov pab pawg kho mob nruab nrab- thiab cov koob tshuaj siab tau muaj kev txhawb nqa ntau dua piv nrog cov nas hauv pawg tsheb (Figures 6A, B).

Cov nyhuv ntawm C. tubulosa Nanopowder ntawm DA Cov ntsiab lus hauv nas

Kev hloov pauv hauv DA cov ntsiab lus ntawm SN raug txiav txim los ntawm HPLC. Nws tau pom tias DA cov ntsiab lus hauv lub hlwb ntawm pawg tsheb tau txo qis. DA cov ntsiab lus nyob rau hauv lub hlwb ntawm PD nas nyob rau hauv cov tshuaj uas tsis muaj C. tubulosa pab pawg neeg kho mob tsis txawv heev ntawm cov nas nyob rau hauv lub tsheb pab pawg neeg. Txawm li cas los xij,C. tubulosa khonce qib DA nyob rau hauv lub hlwb ntawm PD nas nyob rau hauv ib tug npaum li cas ntawm koob tshuaj. Lub hlwb ntawm PD nas kho nrog cov koob tshuaj siab C. tubulosa muaj cov ntsiab lus DA ntau dua li lub hlwb ntawm cov nas hauv pawg tsheb (Daim duab 6C).

Qhov cuam tshuam ntawm C. tubulosa Nanopowder ntawm TH Qhia hauv nas

Tus naj npawb ntawm TH-zoo hlwb thiab qib ntawm TH protein qhia hauv SN ntawm MPTP-induced PD nas tau qis dua piv nrog cov nas hauv pawg tswj hwm. Tom qab kev kho C. tubulosa, tus naj npawb ntawm TH-zoo hlwb thiab theem ntawm TH protein qhia nyob rau hauv SN ntawm MPTP-induced PD nas tau nce, nrog rau qhov sib txawv tseem ceeb ntawm cov kab mob siab C. tubulosa thiab pab pawg tsheb. los ntawm LSD test; thiab muaj qhov sib txawv tseem ceeb ntawm peb pawg kho mob (Daim duab 7).

Cov nyhuv ntawm C. tubulosa Nanopowder ntawm Protein Qhia ntawm GDNF thiab nws cov Receptors, GFR 1 thiab Ret hauv nas.

Cov protein qhia ntawm GDNF thiab nws cov receptors, GFR 1 thiab Ret, nyob rau hauv cov hlwb zoo, raug soj ntsuam siv immunohistochemistry. Western blot tsom xam tau siv los ntsuas cov protein qhia qib hauv SN ntawm ntau pawg nas. Cov kev tshawb pom rau cov pab pawg sib txawv tau zoo sib xws siv ob txoj kev tshawb nrhiav. Kev qhia ntawm GDNF thiab nws cov receptor proteins, GFR 1 thiab Ret, nyob rau hauv cov kab mob zoo stained hauv SN ntawm cov nas hauv pawg tsheb, tau qis dua cov nas hauv pawg tswj hwm ib txwm muaj. Kev siv ntau npaum li cas ntawm C. tubulosa kev kho mob tau nce tus naj npawb ntawm GDNF-, GFR 1-, thiab Ret-positive cells (Figures 8A–S).

Cov protein qhia ntawm GDNF, GFR 1, thiab Ret hauv SN ntawm cov nas hauv pawg tsheb tau qis dua cov nas hauv pawg tswj hwm. Nce kev kho mob concentration ntawmC. tubulosa nanopowerho txhim khu kev qha ntawm cov proteins no. Cov protein qhia ntawm GDNF, GFR 1, thiab Ret nyob rau hauv SN ntawm cov nas nyob rau hauv high-dose C. tubulosa pab pawg neeg kho mob yog ho siab dua cov nas nyob rau hauv lub tsheb pab pawg neeg (P< 0.01; Figures 8T, U). 

Cov nyhuv ntawm C. tubulosa Nanopowder ntawm Protein nthuav qhia ntawm Bcl2 / Bax hauv nas

Bcl2 protein qhia tau txo qis thiab Bax protein qhia tau nce ntxiv hauv SN ntawm cov nas hauv pawg tsheb (P< 0.01) compared with the mice in the normal control group. High-dose C. tubulosa treatment significantly increased Bcl2 protein expression and significantly reduced Bax protein expression in the brains of the vehicle mice (P < 0.01; Figure 9). LSD test showed there was no significant difference between the middle-dose and high-dose groups but a significant difference between the low-dose group. 

Kev sib tham

PD thiab Apoptosis

PD yog tus kab mob neurodegenerative. Raws li Zhang et al. (2005), qhov tshwm sim yog 10.7% nyob rau hauv Suav cov pej xeem hnub nyoog tshaj 55 thiab 1.67% nyob rau hauv cov neeg muaj hnub nyoog tshaj 65. Tus naj npawb ntawm cov neeg mob PD tau nce txhua xyoo nrog kev nrawm ntawm kev laus thoob ntiaj teb, muab lub nra hnyav rau cov tsev neeg ntawm cov neeg mob. thiab haiv neeg loj. Hauv lub hlwb, dopaminergic neurons feem ntau koom nrog hauv kev sib xyaw thiab tso tawm ntawm DA. Lawv tau dav faib rau hauv nruab nrab paj hlwb thiab nyob hauv feem ntau hauv SN (80%). TH yog qhov tseem ceeb ntawm tus nqi txwv enzyme rau DA synthesis. Yog li, qhov inhibition ntawm TH kev ua si txo DA synthesis (Huot thiab Niam Txiv, 2007). Lub ntsiab pathological thiab biochemical hloov pauv ntawm PD yog apoptosis ntawm dopaminergic neurons hauv SN, qhov txo qis ntawm nigrostriatal DA, thiab tsim cov Lewy lub cev hauv dopaminergic neurons (Dexter thiab Jenner, 2013). Lub etiology ntawm PD muaj feem cuam tshuam txog caj ces, ib puag ncig yam, thiab kev laus ntawm lub paj hlwb (Allam li al., 2005). Lub pathogenesis ntawm PD tseem tsis meej nyob rau hauv cov tshuaj niaj hnub. Txij li thaum xyoo 1960s, levodopa hloov kho tau siv tau zoo los kho PD thiab tau lees paub tias yog qhov hloov pauv loj hauv kev kho PD. Txawm li cas los xij, kev siv mus sij hawm ntev ntawm txoj kev kho no ua rau muaj kev mob tshwm sim thiab kev kho mob tsis kho qhov laj thawj ntawm PD (Del Sorbo thiab Albanese, 2008). Yog li ntawd, kev tshawb fawb nquag rau cov tshuaj tshiab lossis cov kev kho mob tsom rau kev tiv thaiv dopaminergic neurons yog qhov tseem ceeb rau kev kho PD.

Hauv cov kev tshawb fawb yav dhau los, daim ntawv thov ntawm lub davhlau ya nyob twg deoxynucleotidyl transferase-mediated dUTP nick end labeling qhia tias 0.6%–4.8% ntawm dopaminergic neurons hauv SN ntawm PD cov neeg mob pom apoptosis (Mochizuki li al., 1996). Electron microscopy pom apoptotic nta, suav nrog chromatic condensation thiab apoptotic lub cev hauv dopaminergic hlwb (Anglade li al., 1997). Tompkins et al. (1997). Lawv pom lub cev apoptotic nyob rau hauv txheej tuab ntawm SN hauv PD thiab DLBD cov neeg mob, muab pov thawj tseeb ntawm neuronal apoptosis hauv PD thiab cov kab mob ntsig txog. Yog li ntawd, kev txo lossis txo qis ntawm apoptosis hauv dopaminergic neurons yog qhov tseem ceeb rau kev kho PD.

Cov kev tshawb fawb yav dhau los tau pom tias MPTP ua rau cov tsos mob zoo li PD. MPTP hla cov ntshav-hlwb teeb meem thiab yog metabolized los ntawm hom B monoamine oxidases hauv astrocytes. Tom qab ntawd nws tau hloov mus rau cov tshuaj lom MPP +, uas khaws cia hauv mitochondria ntawm dopaminergic neurons los ntawm kev noj cov protein ntau ntawm DA transporter. Nws yog li tsim cov pa oxygen dawb radicals ntau tshaj uas inhibit complex I kev ua ntawm mitochondrial respiratory saw thiab ATP synthesis. Cov xwm txheej no ntxiv txhawb kev tsim dawb radical thiab oxidative kev nyuaj siab, thiab nws thiaj li ua rau degeneration thiab tuag ntawm dopaminergic neurons. Yog li, txoj kev tshawb no tau siv MPP + los tsim ib lub tsheb hauv vitro hauv MES23.5 dopaminergic neurons thiab MPTP los txhawb lub tsheb nas rau kev sib tham. Raws li cov txiaj ntsig ntawm MTT kev soj ntsuam, MPP + txo qis qhov muaj peev xwm ntawm MES23.5 hlwb, qhia tias MPP + yog cytotoxic rau dopaminergic neurons. Cov txiaj ntsig kuj tau pom tias C. tubulosa tau txhim kho qhov kev qhia ntawm cov tshuaj tiv thaiv apoptotic thiab inhibited qhov nce ntawm MPP+-induced apoptosis.

PD thiab GDNF

GDNF yog ib qho neurotrophic yam, uas yog thawj zaug cais los ntawm Lin li al. (1993). Lin et al. (1993) kuj tau qhia tias GDNF muaj kev noj zaub mov tshwj xeeb rau dopaminergic neurons hauv nruab nrab ntawm cov nas. GDNF, neurturin (NTN), persephin (PSP), thiab artemin (ART) yog GDNF tsev neeg. Lawv yog cov qauv zoo sib xws thiab ua haujlwm muaj feem cuam tshuam txog cov protein ntau (Kotzbauer li al., 1996; Baloh li al., 1998; Milbrandt li al., 1998; Woodbury li al., 1998).

GDNF receptor muaj ob yam. Thawj cov khoom, GFR , yog tsau rau sab nrauv daim nyias nyias ntawm glycosylphosphatidylinositol (GPI) thiab khi rau saum npoo ntawm connexin. Qhov thib ob tivthaiv yog Ret protein. Kev tshawb fawb tau pom tias muaj plaub yam sib txawv ntawm GFR: GFR 1, GFR 2, GFR 3 thiab GFR 4. GFR 1 yog ib qho affinity receptor ntawm GDNF (Onochie li al., 2000; Chen li al., 2001; Lindahl. et al., 2001). Ret protein yog lub luag haujlwm receptor ntawm GDNF. Homodimer molecule ntawm GDNF ncaj qha khi rau GFR 1 los tsim cov complexes thiab cuam tshuam nrog Ret, uas ua rau lub dimerization thiab ua kom Ret. Vim yog autophosphorylation ntawm Ret, Ret qhib ntau txoj hauv kev TH teeb liab. Thaum tsis muaj Ret protein, GDNF ua rau cov protein phosphorylation ntawm MAPK, PI-3, thiab PLC-, ntxiv rau mRNA qhia thiab kev ua haujlwm ntawm C-fos los ntawm nws cov protein receptor, GFR 1 (Nws li al., 2008).

Cov kev tshawb fawb tau pom tias GDNF muaj kev tiv thaiv zoo tshaj plaws ntawm dopaminergic neurons (Rangasamy li al., 2010; Campos li al., 2012). Hauv tsheb siv MPTP thiab 6-hydroxydopamine (6-OHDA) los ua kom muaj kev puas tsuaj hauv dopaminergic neurons, GDNF tiv thaiv dopaminergic neurons los ntawm kev txo qis apoptosis thiab txhawb kev loj hlob ntawm axonal los txhawb cov qia cell sib txawv (Lucas li al., 2012; Littrell et al., 2013). Lin et al. (1993) tau qhia tias GDNF tshwj xeeb txhawb kev muaj peev xwm, kev sib txawv, thiab kev loj hlob ntawm axonal ntawm dopaminergic neurons los txhawb kev nce ntawm DA hauv cov neurons. Txoj kev tshawb no kuj tau qhia tias GDNF tsis tsuas yog tiv thaiv kev mob toxicity xwb tab sis kuj txo cov toxicity mus sij hawm ntev ntawm MPP+ los yog 6-OHDA hauv dopaminergic neurons, ntxiv rau tiv thaiv cell tuag nyob rau hauv kev nyuaj siab los yog puas hlwb (Yu li al., 2010). Tsis tas li ntawd, GDNF txhawb nqa neural qia cell proliferation thiab sib txawv rau dopaminergic neurons nyob rau hauv midbrain (Lindsay, 1995) los cawm dopaminergic neurons los ntawm retrograde degeneration (Hong-Juan li al., 2011).

Cov kev tshawb fawb tau pom tias GDNF qhia hauv SN tau txo qis hauv cov tsheb tsiaj (Yang li al., 2010). Qhov no qhia tias nws yuav yog ib qho ntawm cov txheej txheem ntawm pathogenesis hauv PD nas. Kev txhaj tshuaj ntawm 5–15 µg / d GDNF rau hauv lub ventricle lateral lossis striatum ntawm MPTP-vim lub tsheb tsiaj rau peb lub hlis sib law liag txhawb kev kho nigrostriatal ntawm dopaminergic system hauv lub tsheb tsiaj (Grondin li al., 2002). Kev tshawb fawb ntawm GDNF kev kho mob rau PD hauv cov tsiaj tsheb tau pom tias kev txhaj tshuaj intracerebral ntawm GDNF mus rau hauv cov cheeb tsam sib txawv ntawm lub paj hlwb, xws li SN, caudate nucleus, thiab lateral ventricle, txhim kho kev txav mus los cuam tshuam nrog PD tsiaj qauv, suav nrog kev ua haujlwm ntawm lub cev muaj zog, cov leeg rigidity thiab tremor (Grondin li al., 2002). Txawm li cas los xij, GDNF tsis tuaj yeem hla ncaj qha los ntawm cov ntshav-hlwb barrier. Yog li ntawd, kev txhaj tshuaj cerebral hauv zos ntawm GDNF cuam tshuam nrog kev pheej hmoo thiab teeb meem hauv kev siv tshuaj kho mob. Cov ntawv thov los qhia txog exogenous GDNF los ntawm kev tswj-tso microspheres, sustained-tso capsules, thiab viral noob tseem tab tom kawm (Liang li al., 2010; Yang et al., 2010; Qiao et al., 2012). Muab cov kev txwv ntawm ntau cov tswv yim los qhia exogenous GDNF rau hauv lub hlwb, cov tshuaj tiv thaiv neuroprotective uas txhawb kev tso tawm ntawm endogenous GDNF yog qhov tseem ceeb rau kev siv tshuaj kho mob.

PD thiabC. tubulosaNanopowder

PD feem ntau tshwm sim hauv cov neeg laus hnub nyoog nruab nrab thiab cov neeg laus. Txoj kev xav ntawm TCM suav hais tias PD feem ntau nyob hauv lub hlwb thiab feem ntau yog vim lub siab thiab lub raum tsis muaj zog, ntxiv rau qhov tseem ceeb ntawm lub zog thiab ntshav tsis txaus. Raws li qhov kev xav no, PD kev kho mob yuav tsum yog li ntawd tsom rau kev txhawb nqa lub raum thiab cov pob txha pob txha.Yang et al. (2010)siv randomized, ob-dig muag, thiab cov tshuaj placebo-tswj kev sim thiab pom tias kev sib xyaw ua ke siv Madopar thiab raum-tonifying zaub mov ua rau txo qis lub cev muaj zog ntawm cov neeg mob PD. Cov txiaj ntsig kev kho mob tau zoo dua li kev kho ib leeg siv Madopar. Kev kho mob ntawm TCM monotherapy los yog tshuaj sib xyaw ua ke hauv PD tau lees paub hauv PD tsiaj qauv thiab kev siv tshuaj kho mob. TCM daim ntawv thov rau tonifying raum thiab txhawb cov ntshav ncig txo cov koob tshuaj ntawm monotherapy rau PD siv Madopar. Qee cov kev tshawb fawb tau qhia tias TCM txhim kho cov tsos mob ntawm PD thiab tiv thaiv dopaminergic neurons, uas tej zaum yuav muaj feem cuam tshuam nrog kev txhawb nqa ntawm endogenous GDNF qhia (Hong-Juan et al., 2011; Qiao et al., 2012).

Lub raum-tonifying compound siv nyob rau hauv txoj kev tshawb no,C. tubulosananopowder, muajCistanche, epimedium, thiabRhizoma polygonati tuaj. Kev tshawb fawb niaj hnub qhia tias cov tshuaj muaj pes tsawg leeg ntawmCistancheyog ECH, uas tiv thaiv dopaminergic neurons hauv SN hauv MPTP-induced PD nas thiab inhibits qhov txo qis ntawm DA thiab DA transporter (Zhao et al., 2010). Tsis tas li ntawd, nws tiv thaiv 6-OHDA-vim txo qis hauv DA thiab tiv thaiv striatal dopaminergic neurons (Chen et al., 2007). Epimedium inhibits kev ua haujlwm ntawm caspase-3 thiab ua haujlwm neuroprotective (Liu et al., 2011). Epimedium flavonoids zoo txhawb nqa neural qia cell proliferation thiab sib txawv (Yao et al., 2010).

Hauv qhov kev tshawb fawb no,C. tubulosananopowder antagonized qhov nce ntawm MPP+-induced apoptosis nyob rau hauv ib koob tshuaj-nyob ntawm tus yam ntxwv. Nws tau txhim kho TH qhia tau zoo hauv covhauv vitrotsheb thiab muaj cov teebmeem tseem ceeb los tiv thaiv apoptotic hauv dopaminergic neurons. MPTP-induced tsheb nas pom kev coj tus cwj pwm tsis zoo thiab txo qis TH qhia hauv cov ntaub so ntswg hauv nruab nrab thiab DA qib, uas yog cov yam ntxwv ntawm PD. Sib txawv dosages ntawmC. tubulosananopowder txo lub sijhawm nyob ruaj khov, txhim kho kev ua haujlwm ntawm tus kheej, txhim kho tus cwj pwm tsis zoo, nce qib DA hauv lub hlwb, thiab nce TH qhia hauv tsheb. Cov txiaj ntsig no qhia tiasC. tubulosananopowder tau txais kev tiv thaiv los ntawm dopaminergic neurons, yog li txhim kho kev coj tus cwj pwm tsis zoo ntawm lub tsheb. Sib txawv dosages ntawmC. tubulosananopowder nce qhov kev qhia ntawm GDNF protein thiab nws cov receptor proteins nyob rau hauv lub hlwb ntawm lub tsheb nas. Cov koob tshuaj ntauC. tubulosaKev kho mob tau nce qib Bcl2 qhia thiab txo Bax qhia, uas tau pom zoo tiasC. tubulosanano hmoovtej zaum yuav txhawb nqa GDNF kev qhia thiab tso tawm hauv MPTP-mob hlwb puas. Tsis tas li ntawd, nws yuav ua rau muaj kev tiv thaiv neuroprotective hauv dopaminergic neurons thiab txo qis neuronal apoptosis los ntawm kev txhawb nqa neurotrophic ntawm GDNF.

Txoj kev tshawb no qhia tau tiasC. tubulosananopowder tau txais kev tiv thaiv los ntawm dopaminergic neurons hauv ob qho tib sihauv vitrothiabhauv vivothiab nce TH qhia kom txhim kho DA cov ntsiab lus. Nws kuj tau txhim kho kev coj tus cwj pwm tsis zoo hauv MPTP-vim lub tsheb nas, tswj hwm cov protein qhia ntawm GDNF thiab nws cov receptor proteins hauv SN, thiab muaj kev tiv thaiv apoptotic hauv PD nas. Lub mechanism ntawm qhov chaw kho mob los ntawmC. tubulosananopowder hauv PD tuaj yeem koom nrog nce cov ntsiab lus ntawm endogenous GDNF hauv lub hlwb thiab yog li txo qhov kev puas tsuaj rau dopaminergic neurons.

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