香京julia种子在线播放

Glioblastoma (GBM) is the most common and aggressive primary intracranial tumor of the central nervous system (Barthel et al., 2019; Miller et al., 2019). Most of them are induced by genetic mutations of high penetrance genes related to rare syndromes, mainly manifested as increased intracranial pressure, neurocognitive dysfunction, and seizures, resulting in central nervous system damage and endangering the lives of patients (Zanders et al., 2019). The standard treatment for GBM is surgery, drug therapy, and radiation therapy, and the median survival time of patients is only 15 months (Kumar et al., 2019). With the changes in eating habits, living environment, and work pressure, the incidence of GBM is increasing and getting younger. Surgical resection combined with postoperative radiotherapy, chemotherapy, and immunotherapy will inevitably damage the body’s normal function and cause adverse reactions. Multi-drug resistance, especially temozolomide (TMZ), leads to frequent GBM recurrences, which is a challenge in treating GBM, and its underlying molecular mechanism is still unclear (Yin et al., 2019). Since the blood-brain barrier (BBB) can prevent the accumulation of charged or macromolecules in the tumor microenvironment at a physically relevant concentration, thereby exerting an oncolytic effect, the content of TMZ in the brain is only 40% percent of the content in the blood, and new component pharmacological methods must be developed to enhance the curative effect of the current treatment, prolong the median survival time of the patient to exceed the median survival time of 15 months (Kumar et al., 2019).

The plants of traditional Chinese medicine (TCM) were used for the treatment of various cancers (Dai et al., 2016), such as GBM (Wang et al., 2019). The use of TCM to promote health and adjuvant therapy is becoming increasingly popular worldwide (Khan and Tania, 2020). The active components of Salvia miltiorrhiza can inhibit the proliferation of U87 cells, induce apoptosis, and enhance the efficacy of TMZ (Wang et al., 2019). Lycium chinense can up-regulate CD3+T, CD8+T, and TNF-α, inhibit the proliferation of mouse C6 cells, and up-regulate CD4⁺CD5+T cells to prolong survival and regulate the BBB (Wang et al., 2019). Magnolol inhibits the migration and proliferation of GBM cells through the JAK-STAT3 signal pathway, mainly by inhibiting the production of GBM stem cell-like cells (Fan et al., 2019). However, the clinical application value of TCM in the treatment of GBM has not been promoted, and more molecular mechanism studies are needed to verify it. Therefore, our research aims to provide new potential for the treatment of GBM with a medicinal plant.

The TCM Siraitia grosvenori (SG) is a perennial herbaceous plant of the Cucurbitaceae family with huge resource reserves and native to southern China, also known as monk fruit and Luo Han Guo, which is a medicinal food homologous species granted by the China Food and Drug Administration with significant clinical effects (Xia et al., 2018). Mogroside has an excellent biological development, which can inhibit the excessive activation of Signal Transducer and Activator of Transcription 3 (STAT3) and promote tumor cell apoptosis (Liu et al., 2018), and targeting STAT3 can improve tumor progression and anticancer immunity response (Lee et al., 2019); reversing emergency medical technician (EMT) and destroying the cytoskeleton to inhibit hyperglycemia-induced lung cancer cell metastasis (Guan et al., 2019). Mogroside IIV and IIIV activate AMP-activated protein kinase (AMPK) and produce anti-hyperglycemic and anti-lipid properties in the body (Abdel-Hamid et al., 2020); mogroside V can cross the BBB and affect schizophrenia-like behavior (Ju et al., 2020) and can also exert neuroprotective activity (Xia et al., 2013); mogroside IVe may be potentially used as a bioactive phytochemical supplement for the treatment of colorectal cancer and laryngeal cancer (Liu et al., 2016). Monk fruit also has other pharmacological effects, such as up-regulating Sirtuin 1 (SIRT1) to reduce oxidative stress and alleviate the decline in oocyte quality during in vitro aging (Nie et al., 2019).

Network pharmacology is based on the high-throughput multi-omics data analysis to clarify the mechanism of multi-component/multi-target/multiple action pathways in medicinal plants (Hopkins, 2008). The newly network pharmacology analysis was employed to integrate active compounds, targets and pathways prediction, and network analysis which may provide novel insights into the therapeutic effects and molecular mechanisms of SG in the treatment for GBM (Abdel-Hamid et al., 2020). Then, we offered a new flowchart to explain the potential target of Siraitia grosvenorii (Luo Han Guo) for GBM (Figure 1).

The microenvironment cell population-counter method was chosen to evaluate the association between CCL5 and immune cell populations (Petitprez et al., 2020). We used immune cells markers and GBM transcriptome data to validate the strong correlation between CCL5 and 24 immune cells markers (Bindea et al., 2013). Gene mutations of GBM expression by “maftools” package (Mayakonda et al., 2018). CCL5 protein expression was detected by immunohistochemistry from the HPA database (https://www.proteinatlas.org/ENSG00000271503-CCL5/pathology/glioma#).

The microenvironment cell population-counter method evaluated the association between CCL5 and 10 immune cell populations from transcriptomic data. A strong correlation between CCL5 and CD8 T cells, T cells, B lineage, and fibroblasts were seen (Figure 5A). Then we further found the significant correlation (p-value < 0.05) between CCL5 and 9 of 24 immune cells markers (Figure 5B), such as the positive correlation in eosinophils (Figure 5C) and the negative correlation in Tcm (Figure 5D). In addition, exploring somatic mutations is helpful to understand the occurrence and development of GBM. The lollipop map shows the mutation distribution and protein domain of CCL5 with somatic mutation (Figure 5E). The distribution of the mutation spectrum of GBM samples can also be identified by a rainfall map (Figure 5F). The transition plot classified single nuclear variants into six categories (Figure 5G). Among them, the C > T mutation accounted for more than 50% of the total mutations. Furthermore, CCL5 protein expression can be detected by immunohistochemistry from the HPA database (Figure 5H).

GBM is the most frequent and the least treatable type of brain tumor, and the prognosis of GBM remains a challenge using the standard methods of treatment—TMZ, radiation, and surgical resection (Jiang et al., 2020b). TMZ is a novel methylating agent that demonstrated activity against recurrent GBM and is ineffective due to drug resistance (Wu et al., 2021). TCMs were considered anti-GBM auxiliary drugs, such as Solanum nigrum L. (Li et al., 2021), Panax ginseng, licorice, Lycium barbarum, Salvia miltiorrhiza bunge, Coptis rhizoma, and Sophora flavescens (Wang et al., 2019). TCM is a helpful complementary and alternative medicine, however, there are few studies on TCM for GBM (Wang et al., 2019). The anti-GBM effects of TCM extract provided the new medium for the treatment of GBM (Li et al., 2021).

We tried to find a new TCM complementary method to treat GBM and hope that through combining Chinese and Western medicine, TMZ resistance could be reversed and anti-tumor therapeutic effects could be achieved. Luo Han Guo is a TCM with the same medicine and food. The multiple compounds in Luo Han Guo not only act on the same target protein, but a single compound also acts on various target proteins and multiple pathways, which reflects the “multiple components, multiple targets, and multiple pathways” of Luo Han Guo’s synergistic effect. Luo Han Guo may work with ar-turmerone, methyl palmitate, lauric acid, beta-sitosterol, gamma-aminobutyric acid, coumarin, mogroside V, and squalene. GO functional enrichment analyses reflected that most of the active ingredients in SG might target nerve cells.

Through network pharmacology and bioinformatics analysis, we found that the CCL5 molecule is a potential target of SG, TMZ, and GBM, maybe the key to the clinical development of TMZ resistance (Figure 1). CCL5-CCR5 paracrine signaling could be an effective therapeutic strategy to improve chemotherapeutic efficacy against GBM (Zhang et al., 2021). CCL5 of glioma-associated microglia/macrophages regulates glioma migration and invasion via calcium-dependent matrix metalloproteinase 2 (Yu-Ju Wu et al., 2020). Knockdown or pharmacological inhibition of CCL5 increased the sensitivity of GBM cells treated with pericyte conditioned media to TMZ (Sprowls and Lathia, 2021). CCL5 was significantly highly expressed in GBM with poor prognostic. Uni-cox and randomForest were used to determine that CCL5 was a significantly important biomarker in GBM. CCL5 was also the target for SG and TMZ. The active ingredient of Luo Han Guo — squalene and CCL5 —show high binding efficiency. SG may be used as a new complementary therapy of the same medicine and food, acting on the target CCL5 and playing an anti-glioblastoma effect. Increasing the effective content of squalene in SG also needs further research. The radiation-related factors were the most critical in ROC c-index analysis. CCL5 plays a vital role in maintaining chemotherapy and radiation resistance.

Compared to genetically distinct syngeneic GBM models, the difference in mouse GBM models was eosinophils, reported in GBM (Khalsa and Shah, 2021). Eosinophils were associated with prognostic risk in the GBM microenvironment (Liang et al., 2020). We found that the SG-TMZ-GBM target, CCL5, a chemotactic ligand, is enriched and positively correlated in eosinophils. CCL5 is also the target of Luo Han Guo, and its effective active integrate compound – squalene—might act on CCL5, thereby affecting the immune microenvironment of GBM.