Application and Effectiveness of Chinese Medicine in Regulating Immune Checkpoint Pathways.

Immunotherapy targeting immune checkpoint molecules has emerged as a key approach in cancer treatment, representing the forefront of antitumor research. However, studies on immune checkpoint molecules have mainly focused on targeted therapies. Chinese medicine (CM) research as a complementary medicine has revealed that immune checkpoint molecules also undergo disease-specific changes in the context of autoimmune diseases. This review article presents a comprehensive analysis of CM studies on immune checkpoint molecules in the last 5 years, with a focus on their role in different diseases and treatment modalities. CM research predominantly utilizes oral administration of herbal plant extracts or acupuncture techniques, which stimulate the immune system by activating specific acupoints through temperature and needling. In this study, we analyzed the modulation and mechanisms of immune checkpoint molecules associated with different coinhibitory and costimulatory molecules, and reviewed the immune functions of related molecules and CM studies in treating autoimmune diseases and tumors. By summarizing the characteristics and research value of CM in regulating immune checkpoint molecules, this review aims to provide a useful reference for future studies in this field.

[Effects of herbal cake separated moxibustion on Tim-4 and ubiquitination of PD-1 in rabbits with immunosuppression].

OBJECTIVE: To observe the effects of herbal cake separated moxibustion on macrophage effector molecule T-cell immunoglobulin and mucin-domain containing-4 (Tim-4) and ubiquitination of programmed cell death protein 1 (PD-1) in rabbits with immunosuppression, and to explore the possible mechanism on herbal cake separated moxibustion in improving immunosuppression. METHODS: Thirty-two big-ear white rabbits were randomly divided into a normal group, a model group, a moxa stick moxibustion group and a herbal cake separated moxibustion group, 8 rabbits in each group. Except the normal group, the immunosuppression model was established by intraperitoneal injection of cyclophosphamide of60 mg/kg in the other 3 groups. "Shenque" (CV 8), "Shenshu" (BL 23), "Zusanli" (ST 36), etc. were selected in both the moxa stick moxibustion group and the herbal cake separated moxibustion group. Moxa stick moxibustion was applied in the moxa stick moxibustion group, one cone at each acupoint; herbal cake separated moxibustion was applied in the herbal cake separated moxibustion group, 5 cones at each acupoint. The intervention was given once every other day for 10 times in both groups. Leukocyte content in peripheral blood was detected by blood cell analyzer; the positive expression of PD-1 in CD+4 T lymphocytes, CD+8T lymphocytes and CD+68 macrophages in peripheral blood was measured by flow cytometry, the serum levels of interleukin 2 (IL-2), CD8, CD68 and Tim-4 were detected by ELISA, and the expression of Tim-4 and F-box only protein 38 (FBXO38) in the liver and spleen tissues was measured by immunohistochemistry. RESULTS: Compared with the normal group, in the model group, white blood cell count (WBC) and percentage of neutrophils (NEU%) were decreased while percentage of lymphocyte (LYM%) was increased (P<0.01) in peripheral blood; the positive expression rates of PD-1 in CD+4 T lymphocytes, CD+8T lymphocytes and CD+68 macrophages in peripheral blood were increased (P<0.01); the serum levels of IL-2, CD68 and Tim-4 were increased (P<0.01), the serum level of CD8 was decreased (P<0.01); the average optical density (AOD) of Tim-4 in the liver tissue and FBXO38 in the liver and spleen tissues was increased (P<0.01). Compared with the model group, in the moxa stick moxibustion group and the herbal cake separated moxibustion group, WBC and NEU% were increased (P<0.01); the positive expression rates of PD-1 in CD+4 T lymphocytes, CD+8T lymphocytes and CD+68 macrophages in peripheral blood were decreased (P<0.01); the serum levels of IL-2, CD68 and Tim-4 were decreased (P<0.01), the serum levels of CD8 were increased (P<0.01); the AOD of Tim-4 and FBXO38 in the liver tissue and FBXO38 in the spleen tissue was decreased (P<0.01, P<0.05). Compared with the moxa stick moxibustion group, in the herbal cake separated moxibustion group, the positive expression rate of PD-1 in CD+68 macrophages in peripheral blood was increased (P<0.05); serum level of Tim-4 was increased (P<0.01); AOD of Tim-4 in the liver tissue was decreased (P<0.05). CONCLUSION: Herbal cake separated moxibustion can improve immunosuppression by regulating the expression of macrophage effector molecule Tim-4 and the FBXO38 mediated ubiquitination of PD-1, Tim-4 may be one of the specific indexes of immunomodulation involving with herbal cake separated moxibustion.

[Effect of herbal cake-separated moxibustion on behavioral stress reactions and blood lactic acid level and muscular AMPK/PGC-1α signaling in rats with chronic fatigue syndrome].

OBJECTIVE: To observe the effect of herbal cake-separated moxibustion (HCSM) on serum lactic acid (BLA) level and AMPK/PGC-1α signaling pathway in the quadriceps femoris in chronic fatigue syndrome (CFS) rats, so as to explore its mechanisms underlying improvement of CFS. METHODS: According to the random number table, 50 SD rats were divided into blank control, model, HCSM, sham HCSM and medication (herbal medicine gavage) groups, with 10 rats in each group. The CFS model was established by using chronic restraint and exhaustive swimming, alternately, once daily for 21 days. The herbal cake was made of Xiaoyao Powder (Mental Ease Powder, composed of [Danggui (Radix Angelicae Sinensis), Baishao (Radix Paeoniae Alba), Chaihu (Radix Bupleuri), Fuling (Poria), Baizhu (Rhizoma Atractylodis, Macrocephalae), etc.]. The HCSM was applied to "Shenque" (CV8), "Guanyuan "(CV4), bilateral "Zusanli" (ST36) and "Qimen" (LR14), 5 moxa-cones for each acupoint, once daily for 10 days. For sham HCSM, the excipient was instead of herbal cake, and the same 5 moxa-cones was given as the HCSM group. Rats of the medication group received gavage of Xiaoyao Powder suspension (60 mg·kg-1), once daily for 10 days. The open field test and tail suspension test were conducted for determining the animals' locomotor activity. The blood sample was taken from the abdominal aorta under anesthesia for assaying the levels of serum BLA, chemokine ligand CXCL9 and ß-endorphin (EP) by ELISA. Bilateral quadriceps femoris were sampled for observing histopathological changes after staining with conventional H.E. technique, and for detecting the expression levels of phosphorylated AMP-activated protein kinase (p-AMPK) and peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) by using immunohistochemistry. RESULTS: Compared with the blank control group, the number of rearing and horizontal grid-crossing times, struggling times of tail suspension test were significantly decreased (P<0.05), and the immobility time was obviously prolonged (P<0.05) in the model group. Compared with the model group, both HCSM and medication groups had a significant increase of rearing, horizontal grid-crossing times and struggling times (P<0.05), and the immobility time had a significant decrease (P<0.05). But there were no significant differences in the total movement distance among the 5 groups (P>0.05), and in the 5 indexes of behavioral measurements between the HCSM and medication groups (P>0.05). The sham HCSM could also evidently increase the struggling times and reduce the immobility time (P<0.05). The contents of serum BLA, CXCL9 and ß-EP were obviously higher in the model group than in the blank control group (P<0.05), as well as remarkably lower in the HCSM and medication groups than in the model group (P<0.05). Whereas the expression levels of muscular p-AMPK and PGC-1α were considerably lower in the model group than in the blank control group (P<0.05), and significantly increased in both HCSM and medication groups relevant to the model group (P<0.05). Compared with the sham HCSM group, the contents of BLA, CXCL9 and ß-EP in serum of the HCSM group and contents of CXCL9, ß-EP in medication group were significantly decreased (P<0.05), and the protein expressions of p-AMPK and PGC-1α in quadriceps femoris in both HCSM and medication groups were significantly increased (P<0.05). H.E. staining showed smaller intercellular space, uneven cytoplasmic staining in some muscle fibers, nucleus pyknosis and condensation, and inflammatory cell infiltration in the model group, which was milder in both HCSM and medication groups. CONCLUSION: HCSM can mitigate the stress behavioral state in CFS rats, which may be related with its functions in lowering the levels of serum BLA, CXCL9 and ß-EP, and activating AMPK/PGC-1α signaling pathway (balancing energy metabolism) in the quadriceps femoris.

[Clinical efficacy and safety evaluation of acupuncture combined with moxibustion in treatment of allergic rhinitis].

OBJECTIVE: To evaluate the clinical efficacy and safety of acupuncture combined with moxibustion on allergic rhinitis. METHODS: Using the random number table, 80 patients with allergic rhinitis were divided into a medication group and an acupuncture combined with moxibustion (acu-mox) group, 40 cases in each one. In the medication group, ioratadine tables were prescribed for oral administration, one tablet daily for 10 days as 1 session , 3 sessions of treatment were required. In the acupuncture combined with moxibustion group, bilateral Yingxiang (LI20), Yintang (EX-HN3), bilateral Hegu (LI4) and bilateral Shenshu (BL23) were selected as the main points and stimulated with acupuncture and moxibustion; and the acupoint prescription was modified according to symptoms. This combined treatment was given once every day, stimulating for 30 min each time, and 10 treatments made 1 course, for 3 courses of treatment totally. Before and after treatment, the scores for symptoms and physical signs, as well as the score of rhino-conjunctivitis related quality of life scale (R-QOL) were evaluated separately. The sample of the inferior turbinate mucosa tissue was collected and the distribution of eosinophil (EOS) was scored using HE staining and Sheldeny evaluation. Using enzyme-linked immunosorbent assay (ELISA), the contents of serum immunoglobulin E (IgE), retinoic-acid-receptor-related orphan nuclear receptor γt (RORγt), forkhead box protein P3 (Foxp3), interleukin-17 (IL-17), IL-27 and IL-33 were determined. The clinical efficacy was evaluated in the patients with allergic rhinitis of two groups and all the adverse reactions were recorded during treatment. RESULTS: The scores of symptoms and physical signs as well as the score of R-QOL, and EOS distribution score and the contents of serum IgE, RORγt, IL-17 and IL-33 were all reduced as compared with those before treatment in each group (P<0.05), and the contents of serum Foxp3 and IL-27 were increased as compared with those before treatment in each group (P<0.05). After treatment, the scores of symptoms and physical signs as well as the score of R-QOL, and the contents of serum IgE, RORγt and IL-33 in the acu-mox group were lower than those in the medication group (P<0.05), and the contents of serum Foxp3 and IL-27 were higher than those of the medication group (P<0.05). The total effective rate of the acu-mox group was 100.0% (40/40), significantly higher than 82.5% (33/40) in the medication group (P<0.05). No ob-vious adverse reaction was found in either group during and after treatment. CONCLUSION: Acupuncture combined with moxibustion is significantly effective and safe in treatment of allergic rhinitis. Its effect mechanism may be related to the balance modulation of Th1/Th2 and Th17/Treg cells mediated by naive CD4+T cells.

[Influence of electroacupuncture on myocardial NO and NOS and intracellular Ca2+ contents in myocardial ischemia-reperfusion injury rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on contents of myocardial nitric oxide (NO), nitric oxide synthase (NOS), and intracellular Ca2+ levels in experimental myocardial ischemia-reperfusion injury (MI/RI) rats, so as to provide some evidence for acupuncture treatment of cardiovascular disorders and for the correlation between the acupoints and the internal organs. METHODS: Fifty male Wistar rats were randomly divided into sham-operation (sham), model, Neiguan (PC 6), Lieque (LU 7), and Hegu (LI 4) groups (n = 10/group). MI/RI model was established by occlusion of the descending anterior branch of the left coronary artery for 40 min and reperfusion for 60 min. EA (30 Hz/100 Hz, 2-4 mA) was applied to the bilateral PC 6, LU 7 and LI 4 for 20 min. The myocardial tissue of the ischemic area was taken, homogenated and centrifuged, separated for assaying NO and NOS activity by using spectrophotometer and their contents with nitrate reductase colorimetric assay. Another part of the myocardium tissue was processed to be cell suspension (added with Fluo-3/AM dye) for assaying intracellular free [Ca2+]i concentration under the laser confocal microscope. RESULTS: Compared with sham group, myocardial NO and NOS contents of model group decreased mildly and significantly (P < 0.05), while myocardial fluorescence intensity (content) of intracellular Ca2+ increased evidently (P < 0.01). In comparison with model group, the contents of myocardial NO and NOS were upregulated considerably in PC 6 group (P < 0.05), and intracellular Ca2+ content in PC 6 group reduced significantly (P < 0.01). No significant differences were found between LU 7 and model groups and between LI 4 and model groups in the above mentioned 3 indexes (P > 0.05). CONCLUSION: EA of PC 6 can upregulate myocardial NO and NOS contents and downregulate myocardial intracellular Ca2+ level in MI/RI rats, which may contribute to its effect in relieving myocardial injury. Neiguan (PC 6) and the heart have a relatively specific correlation.

[Effects of herbal cake-separated moxibustion on the expression of erythrocyte CD58 in different ages of healthy people].

OBJECTIVE: To observe the efficacy of herbal cake-separated moxibustion on the expression of erythrocyte CD58 in different ages of healthy people and explore the differences of the therapeutic effect in different ages and its mechanism. METHODS: A total of 82 health participants were divided into a young age group and a middle-old age group according to the ages. They were treated with herbal cake-separated moxibustion on Shenque (CV 8), Guanyuan (CV 4), Zusanli (ST 36), Pishu (BL 20), Shenshu (BL 23) with cake made by Shudihuang (Radiz Re hmanniae Preparata), Shanyao (Rhizoma Dioscoreae), Shanzhuyu (Fructus Corni ), etc. The treatment was given for 10 sessions once other day and each acupoint for 3 successive dosages. The mean fluorescence intensities of erythrocyte CD58 were measured by flow cytometry before and after moxibustion. RESULTS: After moxibustion, erythrocyte CD58 expression were significantly higher than that before moxibustion in two groups (both P < 0.01), particularly in young age group, which was significantly higher than that in middle-old age group (P < 0.01). CONCLUSION: The effect of moxibustion in youth is evidently superior to that in middle-old age. Its mechanism is connected with that moxibustion can enhance the expression of erythrocyte CD58.

[Survey of studies on mechanisms of acupuncture and moxibustion in decreasing blood pressure].

Mechanisms of acupuncture and moxibustion for treatment of hypertension have been studied extensively from poly-aspects, poly-levels and poly-links. The present paper reviews the studies on nervous regulation, humoral regulation and regulation of peripheral vessel resistance and other pathways and their relationships in recent ten years, and indicates further perfecting these studies from the following aspects: studies on the mechanism of section and combination of blood pressure-decreasing points, relationship of time-effect and dose-effect of acupuncture and moxibustion, and studies on mechanisms of functions of nerve-endocrine-immune system.

[Effects of acupuncture at the points of the Pericardium Meridian on the activity of myocardial Ca2+-ATPase and gene expression in the rat of myocardial ischemia and reperfusion injury].

OBJECTIVE: To investigate the mechanism of acupuncture at "Neiguan" (PC 6) and "Ximen" (PC 4) of the Pericardium Meridian in treatment of myocardial ischemia. METHODS: Rats were randomly divided into 5 groups, sham-operation group, ischemia-reperfusion model group, Neiguan acupuncture group, Ximen acupuncture group and Zhigou acupuncture group. Electroacupuncture (EA) was given at corresponding acupoints for 20 min in the later 3 groups, followed by ligation of the left anterior descending branch of the coronary artery and 40 min later, acupuncture was given at the points for another 20 min, reperfusion for 60 min, with ECG monitoring. Then the myocardial tissue sample (below the ligation site) of the left cardiac ventricle was taken for preparation of sarcoplasmic reticulum, and determination of Ca2+-ATPase activity according to quantitative analysis of phosphorum. The relative mRNA levels were determined by Northerm Bolt analysis. RESULTS: In the acupuncture groups, both the activities of Ca2+-ATPase and the gene expressions increased significantly as compared with the model group (all P < 0.01). CONCLUSION: The points of the Pericardium Meridian can obviously improve the Ca2+-ATPase activity and the gene expressions, reduce the degree of myocardial ischemia and reperfusion injury, and strength myocardial functions.