Cov Cim Zoo Ntawm Cistanches Herba Raws Li Antioxidant Spectrum-Effect Relationship Ⅱ

3 ua

3.1 Tsim daim ntawv qhia ntiv tes

Npaj 10 batch ntawm cov kev daws teeb meem raws li txoj hauv kev hauv qab 2.1, siv cov xwm txheej chromatographic hauv qab 2.2 txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau txhawm rau ntsuas 10.10. ntiv tes. Ntshuam cov kua theem data.AIA cov ntaub ntawv ntawm cov ntiv tes ntawm Cistanche deserticola kuaj cov khoom rau hauv "Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System" software (2012 version). Kaum xya feem ntau chromatographic peaks tau calibrated nyob rau hauv cov hom, thiab muab piv nrog cov mix reference standard spectrum (Daim duab 2), tag nrho ntawm 10 ntau peaks tau txheeb xyuas.

3.2 Kev tshawb fawb txog cov nyhuv antioxidant ntawm Cistanche deserticola

3.2.1 Cov nyhuv ntawm Cistanche deserticola ntawm kev muaj peev xwm ntawm HEK 293 hlwb.

Piv nrog rau cov hlwb hauv cov pab pawg ib txwm muaj, Cistanche deserticola tsis tsim cytotoxicity ntawm qhov loj ntawm 50 ~ 100 ug·mL-1, thiab cov cell ciaj sia taus ntau dua 90%. Raws li qhov concentration ntawm Cistanche deserticola nce, cell viability txo. Yog li ntawd, Cistanche deserticola nrog qhov loj ntawm 100 µg·mL-1 raug xaiv rau kev tswj hwm tom ntej.

3.2.2 Cov nyhuv ntawm Cistanche deserticola ntawm ROS ntxias los ntawm H2O2 hauv HEK 293 hlwb

Lub ROS IC50 ntawm Cistanche deserticola ntawm ntau qhov sib txawv tau suav nrog siv GraphPad Prism 8 software. Cov txiaj ntsig tau pom hauv daim duab 3 thiab Table 2.

Yog xav paub ntxiv txog cistanche, cia peb saib yuav ua li cas paub qhov txawv cistanche zoo. Cistanche zoo nyob ntawm cov ntsiab lus ntawm cov khoom xyaw nquag Echinacoside thiab Acteoside, cov ntsiab lus siab dua txhais tau tias muaj zog dua, niaj hnub no peb noj 30% Echinacoside thiab 12% Acteoside Cistanche Supplements ua piv txwv los piav qhia rau peb, feem ntau hu rau cistanche hauv kab lus no.

Nws yuav siv sij hawm ntev npaum li cas rau CISTANCHE ua haujlwm?

3.2.3 Cov nyhuv ntawm Cistanche deserticola ntawm H2O2-induced SOD ntawm HEK 293 hlwb

Raws li pom nyob rau hauv daim duab 4, piv nrog rau ib txwm pab pawg neeg, SOD kev ua nyob rau hauv lub hlwb ntawm cov qauv pab pawg neeg tau txo qis, thiab cov kev ua ntawm SOD ntawm Cistanche deserticola pawg thawj coj tau nce mus rau qhov sib txawv (P<0.001), indicating that Cistanche deserticola can increase the intracellular SOD enzyme activity.

3.2.4 Cov nyhuv ntawm Cistanche deserticola ntawm H2O2-induced CAT hauv HEK 293 hlwb

Raws li pom hauv daim duab 5, H2O2 tuaj yeem ua rau muaj qhov txo qis hauv kev ua haujlwm ntawm CAT enzyme (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 Q-kos tshuaj ntsuam xyuas raws li kev txheeb xyuas kev sib raug zoo spectral

3.3.1 PCA siv eigenvalue> 1 thiab qhov sib txawv ntawm qhov sib txawv ntawm qhov nyiaj pab> 85% raws li tus qauv los rho tawm 4 lub ntsiab lus tseem ceeb. Cov phiaj xwm scree (Daim duab 6) ntawm thawj peb lub ntsiab lus tseem ceeb muaj qhov nkhaus steeper, uas txhais tau hais tias nws cov nyiaj pab ntau dua. loj. Qhov sib txawv ntawm qhov sib txawv ntawm plaub lub ntsiab lus tseem ceeb yog 87.640%, qhia tias plaub lub ntsiab lus tseem ceeb tuaj yeem sawv cev 87.640% ntawm cov ntaub ntawv ntawm thawj qhov sib txawv. Thawj qhov eigenvalues ​​thiab qhov sib txawv ntawm qhov sib txawv yog qhia nyob rau hauv Table 3. Lub rotated tivthaiv matrix yog tau los ntawm Kaiser normalization siab tshaj variance txoj kev, saib Table 4. Raws li tus nqi thauj khoom ntawm txhua chromatographic ncov ntawm plaub lub ntsiab lus tseem ceeb, cov ntaub ntawv cuam tshuam los ntawm txhua tus. Cov ntsiab lus tseem ceeb tuaj yeem muab rho tawm. Ntawd yog, qhov loj dua tus nqi thauj khoom, qhov ntau dua qhov cuam tshuam ntawm chromatographic ncov ntawm lub ntsiab lus tseem ceeb. Hauv cov ntsiab lus tseem ceeb 1, peaks 1 txog 3, 5, 7, 11, thiab 13 txog 15 muaj qhov loj tshaj plaws loadings; nyob rau hauv lub hauv paus ntsiab lus 2, peaks 6 thiab 16 muaj qhov loj tshaj plaws loadings; ncov 17 piav qhia cov ntaub ntawv ntawm lub hauv paus ntsiab lus 3, Peak 8 muaj qhov cuam tshuam loj tshaj plaws ntawm cov ntsiab lus tseem ceeb 4, yog li 13 cov tshuaj lom neeg ntawm peaks 1 ~ 3, 5 ~ 8, 11, 13 ~ 17 tau xaiv los ua cov khoom tseem ceeb ntawm Cistanche. deserticola rau tom ntej spectral efficiency tsom xam.

3.3.2 TSI

ROS IC50 yog qhov ntsuas tsis zoo ntawm kev ua tau zoo ntawm cov tshuaj, yog li qhov kev cuam tshuam tsis zoo ntawm kev sib raug zoo txhais tau hais tias qhov sib txawv ntawm qhov muaj feem cuam tshuam nrog kev siv tshuaj zoo. Qhov sib txheeb coefficients ntawm 13 chromatographic peaks ntawm Cistanche deserticola yog tag nrho qhov tsis zoo, qhia tias lawv txhua tus muaj feem cuam tshuam nrog kev tshem tawm ntawm ROS IC50, ntawm cov peaks 14 thiab 3, 7, 11, 1, thiab 13 tau muaj kev cuam tshuam nrog 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.

Kev sib xyaw ntawm PCC cov txiaj ntsig ntawm ROS IC50, SOD, thiab CAT, peaks 1 ~ 3, 5, 7, 11, thiab 13 ~ 15 tau raug xaiv los ua cov khoom xyaw nquag rau Cistanche deserticola kom muaj txiaj ntsig antioxidant.

3.3.3 TSI

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 yog: 5, 13, 14, 11, 7 (Fig. 7C). Cov tebchaw sawv cev los ntawm peaks 1 ~ 3, 5, 7, 11, thiab 13 ~ 15 tau raug tshuaj xyuas raws li cov khoom muaj peev xwm antioxidant hauv PLS qauv.

3.4 Cistanche deserticola Q-marker kev ua pov thawj

3.4.1 Cov nyhuv ntawm cov khoom xyaw nquag ntawm HEK 293 cytotoxicity

Raws li cov txiaj ntsig ntawm spectrum-effect correlation analysis, 6 cov khoom xyaw suav nrog genipinic acid, 8-epistrychnoic acid, echinaceoside, verbascoside, anthocynoside A, thiab isorbascoside raug xaiv rau kev ua pov thawj. Rau cov khoom xyaw nquag ntawm Cistanche deserticola tsis pom muaj cytotoxicity ntawm qhov ntau ntawm 0.781 txog 3.125 µmol·mL–1, thiab cov cell ciaj sia taus yog tag nrho ntau dua 90%. Raws li qhov concentration nce, cell viability yog inhibited. Yog li, cov koob tshuaj tom ntej tau raug xaiv raws li 0.781, 1.563, thiab 3.125 µmol·mL-1.

3.4.2 Cov khoom xyaw nquag ua khub

Cov nyhuv antioxidant ntawm H2O2-induced HEK 293 hlwb tau txheeb xyuas los ntawm kev kuaj xyuas cov nyhuv scavenging ntawm txhua yam khoom muaj nyob rau hauv intracellular ROS thiab muaj peev xwm tswj cov kev ua ntawm SOD thiab CAT enzymes los xyuas seb nws puas muaj cov nyhuv antioxidant. Hauv kev sim ROS scavenging, qhov sib txawv ntawm nruab nrab- thiab siab-dose genipinic acid pawg thiab pawg qauv yog qhov tseem ceeb (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.