There is certainly evidence that DN T cells participate in the aggravation of certain syndromes and kidney injury in MRL/lpr mice [23]. of DN T and plasma cells in MRL/lpr mice. Moreover, Acetanilide LCHJ significantly suppressed the accumulation of CD138+ T cells in MRL/lpr mice, which led to the decreased production of the anti-dsDNA antibody in vivo. LCHJ significantly decreased CD4+CD138+ T cells originated from CD4+CD138- T cells, which subsequently prevented the accumulation of CD138+ T cells in MRL/lpr mice. Our results MET indicated that LCHJ alleviated renal injuries and prevented the enlargement of the spleen and lymph nodes by suppressing DN T cell accumulation, and reduced anti-dsDNA antibody secretion by preventing the accumulation of CD138+ T cells. values of less than 0.05 were considered statistically significant. Results Identification of chemical components in LCHJ using UPLC-MS/MS analysis UPLC-MS/MS analysis was employed to identify the chemical components of LCHJ. The total positive (Physique 1A) and unfavorable (Physique 1B) ion chromatograms showed 34 major components in the LCHJ decoction. The 34 major components in LCHJ including salvianolic acid A, astragaloside I, rutinum, icariine, salidroside, atractylenolide I, gentiopicroside, tanshinone I, and formononetin were shown in Table 2. Open in a separate window Physique 1 Identification of some components in LCHJ decoction. Base peak ion chromatogram of LCHJ in positive (A) and unfavorable (B) ion modes are shown as indicated. Table 2 Characterization of chemical components in LCHJ decoction using UPLC-MS/MS analysis thead th align=”left” rowspan=”1″ colspan=”1″ Peak no /th Acetanilide th align=”center” rowspan=”1″ colspan=”1″ Identification /th th align=”center” rowspan=”1″ colspan=”1″ tR (min) /th th align=”center” rowspan=”1″ colspan=”1″ Ion mode /th th align=”center” rowspan=”1″ colspan=”1″ Measured [M-H]- (m/z) /th th align=”center” Acetanilide rowspan=”1″ colspan=”1″ Predicted [M-H]- (m/z) /th th align=”center” rowspan=”1″ colspan=”1″ ppm /th th align=”center” rowspan=”1″ colspan=”1″ Formula /th th align=”center” rowspan=”1″ colspan=”1″ Drive from /th /thead 1Phenylalanine4.69ESI+166.086166.0862-1.20C9H11NO2A, D2Tryptophan6.07ESI+205.0968205.0971-1.46C11H12N2O2A3Plantamajoside6.15ESI+641.2071641.2076-0.78C29H36O16H4Salvianic acid A6.29ESI+199.0599199.0601-1.00C9H10O5J5Salidroside6.46ESI+301.1275301.1281-1.99C14H20O7E6Gentiopicroside6.83ESI-355.1039355.10234.51C16H20O9L7Loganic acid7.23ESI-375.1292375.12851.87C16H24O10L8L-Epicatechin7.33ESI+291.0856291.0863-2.41C15H14O6K9tyrosine7.34ESI+182.0808182.0811-1.65C9H11NO3A10Ferulic acid7.47ESI+195.0647195.0651-2.05C10H10O4I, L11Hyperoside8.01ESI+465.1022465.1027-1.08C21H20O12F12Astragalin8.2ESI+449.1069449.1078-2.00C21H20O11F13Calycosin-7-glucoside8.26ESI+447.1275447.1285-2.23C22H22O10C14Rutinum8.35ESI+611.1595611.1606-1.80C27H30O16E15Isoacteoside8.36ESI+625.2121625.2126-0.80C29H36O15H16Specnuezhenide8.76ESI+685.2355685.23382.48C31H42O17E17Acteoside8.77ESI-625.2122625.2126-0.64C29H36O15H18Isoquercitrin8.78ESI+465.1017465.1027-2.15C21H20O12F19Quercetin8.79ESI+303.0494303.0505-3.63C15H10O7F, N20Quercitrin9.28ESI+449.1069449.1078-2.00C21H20O11F21Kaempferol9.28ESI+287.0544287.0555-3.83C15H10O6F22Isorhamnetin9.33ESI+317.0651317.0655-1.26C16H12O7F23Daidzein9.73ESI+255.0645255.0651-2.35C15H10O4K24Icariine10.68ESI+677.2419677.2439-2.95C33H40O15G25Formononetin10.97ESI+269.08269.0808-2.97C16H12O4K26Atractylenolide III11.63ESI+249.1479249.1485-2.41C15H20O3B27Atractylenolide I11.64ESI+231.1374231.138-2.60C15H18O2B28Costunolide11.96ESI+233.1532233.1536-1.72C15H20O2B29Astragaloside A12.18ESI+785.4677785.4681-0.51C41H68O14C30Tanshinone I12.35ESI+277.0855277.0859-1.44C18H12O3J31Cryptotanshinone12.4ESI+297.1478297.1485-2.36C19H20O3J32Astragaloside I12.44ESI+869.4869869.4893-2.76C45H72O16C33Tanshinone II A12.71ESI+295.1323295.1329-2.03C19H18O3J34Pachymic acid13ESI+529.388529.3887-1.32C33H52O5D Open in a separate windows A: Pseudostellaria heterophylla (Miq.) Pax; B: Atractylodes macrocephala Koidz; C: Astragalus mongholicus Bunge; D: Poria cocos (Schw.) Wolf; E: Ligustrum lucidum W. T. Aiton; F: Cuscuta chinensis Lam; G: Epimedium brevicornu Maxim; H: Plantago asiatica L; I: Scleromitrion diffusum (Willd.) R. J. Wang; J: Salvia miltiorrhiza Bunge; K: Spatholobus suberectus Dunn; L: Gentiana macrophylla Pall; M: Cinnamomum cassia (L.) J. Presl; N: Bistorta officinalis Delarbre. LCHJ exhibited significant therapeutic effects on MRL/lpr mice After the administration of LCHJ, spleen and lymph node excess weight in MRL/lpr mice were significantly decreased, compared with vehicle-treated MRL/lpr mice (Physique 2B and ?and2C).2C). At the same time, spleen and lymph indices also showed a remarkable decline in MRL/lpr mice after LCHJ treatment (Physique 2A and ?and2C).2C). Treatment with prednisone alone and combined with LCHJ, also alleviated the enlargement of spleen and lymph node, decreasing spleen and lymph indices (Physique 2A-C). Our results indicated that LCHJ experienced significant effects on preventing the enlargement of spleen and lymph nodes Acetanilide in MRL/lpr mice. Open in a separate window Physique 2 Therapeutic effects of LCHJ on lupus in MRL/lpr mice. A. Body weight of MRL/lpr mice, n=10 per group. B. Image of the spleen (Top panel) and lymph nodes (bottom panel) in MRL/lpr mice. C. Spleen excess weight, spleen index, lymph node excess weight, and lymph node index in MRL/lpr mice; n=10 per group. D. Renal tissue pathological sections were stained with HE, PASM, PAS, and Masson, initial magnification: 400, level bar =60 m; n=4 per group. E. Representative frozen stained sections with Alexa Flour 488-conjugated goat anti-mouse IgG (H+L) to determine IgG deposits level, initial magnification: 200, level bar =100 m; Acetanilide n=5 per group. F. Urinary protein quantity; n=10 per group. G. ANA and anti-dsDNA antibody levels in the serum of MRL/lpr mice were measured using ELISA; n=6 per group. Data are offered as the mean SD; #P 0.05, ##P 0.01 by one-way analysis of variance, versus vehicle-treated mice. Next, we observed the effect of LCHJ on pathological injury in kidneys and measured renal histopathology and urinary protein levels in MRL/lpr mice. Hyaline deposits, interstitial and perivascular cellular inflammation, glomerular fibrosis, tubular cell necrosis, cellular crescent formation, and glomerulosclerosis were observed in kidneys of MRL/lpr mice (Physique 2D), whereas, all these renal injuries were alleviated after LCHJ.