Makè Kalite Cistanches Herba ki baze sou relasyon spectre-efè antioksidan Ⅱ
Mar 19, 2024
3 rezilta
3.1 Etablisman kat anprent
Prepare 10 pakèt solisyon tès dapre metòd ki anba 2.1, sèvi ak kondisyon kwomatografik anba 2.2 pou deteksyon, jwenn anprent chak pakèt echantiyon (Figi 1), epi chwazi pik No 11 (C11) kòm pik referans nan. anprent. Enpòte done faz likid la. Fichye AIA nan anprent pwodwi tès Cistanche deserticola nan lojisyèl "Tradisyonèl Chinwa Medsin Chromatografik Anprent Resanblans Evalyasyon Sistèm" (vèsyon 2012). Yo te kalibre disèt pik chromatografik komen nan mòd komen an, epi konpare ak spectre estanda referans melanje (Figi 2), yo te idantifye yon total de 10 pik komen.
3.2 Rechèch sou efè antioksidan Cistanche deserticola
3.2.1 Efè Cistanche deserticola sou viabilite selil HEK 293 yo.
Konpare ak selil nan gwoup nòmal la, Cistanche deserticola pa t 'pwodwi sitotoksisite nan yon konsantrasyon mas 50 ~ 100 ug·mL-1, ak pousantaj siviv selil yo te pi gran pase 90%. Kòm konsantrasyon nan Cistanche deserticola ogmante, viabilite selil la diminye. Se poutèt sa, Cistanche deserticola ak yon konsantrasyon mas 100 µg·mL–1 te chwazi pou administrasyon ki vin apre.
3.2.2 Efè Cistanche deserticola sou ROS pwovoke pa H2O2 nan selil HEK 293
Yo te kalkile ROS IC50 Cistanche deserticola nan diferan konsantrasyon lè l sèvi avèk lojisyèl GraphPad Prism 8. Yo montre rezilta yo nan Figi 3 ak Tablo 2.
Pou plis konesans sou cistanche, an n pran yon gade nan ki jan yo fè distenksyon ant kalite cistanche. Bon jan kalite Cistanche depann sou kontni an nan engredyan yo aktif Echinacoside ak Acteoside, kontni an pi wo vle di fonksyon pi fò, jodi a nou pran 30% Echinacoside ak 12% Acteoside Sipleman Cistanche kòm yon egzanp pou ban nou yon eksplikasyon, jeneralman rele cistanche nan atik sa a.
KONBYEN TAN LI PRAN POU CISTANCHE TRAVAY?
3.2.3 Efè Cistanche deserticola sou SOD H2O2-pwovoke nan selil HEK 293
Jan yo montre nan Figi 4, konpare ak gwoup nòmal la, aktivite SOD nan selil yo nan gwoup modèl la te siyifikativman redwi, ak aktivite SOD nan gwoup administrasyon Cistanche deserticola ogmante nan diferan degre (P<0.001), indicating that Cistanche deserticola can increase the intracellular SOD enzyme activity.
3.2.4 Efè Cistanche deserticola sou H2O2-pwovoke CAT nan selil HEK 293
Jan yo montre nan Figi 5, H2O2 ka lakòz yon diminisyon enpòtan nan aktivite anzim CAT intraselilè (P<0.001), with the activity only reaching 30% of the normal group, indicating that H2O2 has seriously damaged the antioxidant system of cells. After Cistanche deserticola administration, intracellular CAT enzyme activity decreased significantly (P<0.001). The enzyme activity was significantly increased (P<0.001).
3.3 Cistanche deserticola tès depistaj Q-marker ki baze sou analiz korelasyon efikasite espèk
3.3.1 PCA sèvi ak valè pwòp > 1 ak pousantaj kontribisyon kimilatif divèjans > 85% kòm estanda pou ekstrè 4 eleman prensipal yo. Konplo scree la (Figi 6) nan twa premye eleman prensipal yo gen yon koub pi apik, ki vle di ke to kontribisyon li yo pi wo. gwo. Pousantaj kontribisyon kimilatif divèjans kat eleman prensipal yo se 87.640%, sa ki endike kat eleman prensipal sa yo ka reprezante 87.640% enfòmasyon varyab orijinal yo. Valè pwòp inisyal yo ak pousantaj kontribisyon divèjans yo montre nan Tablo 3. Matris eleman wotasyon an jwenn pa metòd nòmalizasyon maksimòm divèjans Kaiser, gade Tablo 4. Dapre valè chaj chak pik kwomatografik sou kat eleman prensipal yo, enfòmasyon yo reflete pa chak. eleman prensipal yo ka ekstrè. Sa vle di, pi gwo valè chaj la, se pi gwo enfliyans nan pik chromatografik la sou eleman prensipal la. Nan eleman prensipal 1, pik 1 a 3, 5, 7, 11, ak 13 a 15 gen pi gwo chaj absoli yo; nan eleman prensipal 2, pik 6 ak 16 gen pi gwo chaj absoli yo; pik 17 eksplike enfòmasyon prensipal eleman 3, pik 8 gen pi gwo enpak sou eleman prensipal 4, kidonk 13 eleman chimik nan pik 1 ~ 3, 5 ~ 8, 11, 13 ~ 17 yo te chwazi kòm baz materyèl prensipal Cistanche. deserticola pou analiz efikasite espèk ki vin apre.
3.3.2 PCC
ROS IC50 se yon endikatè negatif nan efikasite dwòg, kidonk yon koyefisyan korelasyon negatif vle di ke varyab depandan an pozitivman korelasyon ak efikasite dwòg. Koefisyan korelasyon nan 13 pik kwomatografik Cistanche deserticola yo tout negatif, sa ki endike ke yo tout pozitivman gen rapò ak retire ROS IC50, pami ki pik 14 ak 3, 7, 11, 1, ak 13 yo te trè fòtman korelasyon ak ROSIC50. (P<0.01), and peaks 2, 5, and 15 were strongly correlated (P<0.05); the 13 chromatographic peaks were all positively correlated with SOD, among which peaks 13 and 3 , 15, 11, 2 were extremely strongly correlated with SOD (P<0.01), and peaks 1, 14, 7, 5, and 6 were strongly correlated (P<0.05); 11 peaks were positively correlated with CAT, and peak 5 was positively correlated with CAT. There was a strong correlation (P<0.05). The results are shown in Table 5.
Konbine rezilta PCC ROS IC50, SOD, ak CAT, pik 1 ~ 3, 5, 7, 11, ak 13 ~ 15 yo te chwazi kòm gwoup engredyan aktif pou Cistanche deserticola pou fè egzèsis efè antioksidan.
3.3.3 PLS
In PLS, the ROS IC50 regression coefficients of 13 chromatographic peaks are all negative and positively correlated with the pharmacodynamic activity. Among them, the chromatographic peaks with VIP>1 are: 14, 3, 11, 1, 7, 13, 2, 5, 15 (Figure 7A); in the study of SOD enzyme activity, the coefficients of all peaks are positive, that is, all peaks are positively correlated with drug efficacy. Among them, the chromatographic peaks with VIP>1 are: 13, 3, 15, 11, 2, 1 , 14, 7 (Figure 7B); in the study of CAT enzyme activity, except for peaks 6 and 16, the other chromatographic peaks are positively correlated with drug efficacy, among which the chromatographic peaks with VIP>1 yo se: 5, 13, 14, 11, 7 (figi 7C). Konpoze yo reprezante pa pik 1 ~ 3, 5, 7, 11, ak 13 ~ 15 yo te depistaj kòm konpozan potansyèl antioksidan aktif nan modèl PLS la.
3.4 Verifikasyon aktivite Cistanche deserticola Q-marker
3.4.1 Efè engredyan aktif sou sitotoksisite HEK 293
Dapre rezilta analiz spectre-efè korelasyon, 6 engredyan ki gen ladan asid jenipinik, 8-asid epistrychnoic, echinaceoside, verbascoside, anthocynoside A, ak isorbascoside yo te chwazi pou verifikasyon aktivite. Sis engredyan aktif Cistanche deserticola yo pa montre okenn sitotoksisite nan konsantrasyon ki sòti nan 0.781 a 3.125 µmol·mL-1, ak pousantaj siviv selil yo te pi gran pase 90%. Kòm konsantrasyon an ogmante, viabilite selil yo inibe. Se poutèt sa, konsantrasyon dòz ki vin apre yo te chwazi kòm 0.781, 1.563, ak 3.125 µmol·mL-1.
3.4.2 Pè engredyan aktif
Efè antioksidan H2O2-pwovoke selil HEK 293 yo te verifye lè yo detekte efè scavenging chak engredyan aktif sou ROS intraselilè ak kapasite pou kontwole aktivite anzim SOD ak CAT pou verifye si li gen yon efè antioksidan. Nan eksperyans ROS scavenging, diferans ki genyen ant gwoup asid jenipinik ak dòz segondè yo ak gwoup modèl yo te estatistik enpòtan (P<0.01, P<0.001, Figure 8A). Compared with the model group, there were statistically significant differences between the low, medium and high dose groups of inulin, anthoside A, verbascoside and isorbascoside (P<0.001, Figure 8B, Figure 8C), indicating that the six monomers All have a certain degree of ROS scavenging effect; in the SOD and CAT enzyme activity measurement test, the enzyme activities of the low, medium and high dose groups of active ingredients were all higher than the model group, indicating that all 6 monomers can increase SOD and CAT Enzyme activity.
