To determine the relationship of hemodialysis access with vital sign variability and hemodialysis-related headache (HRH). Adult outpatients receiving maintenance hemodialysis (MHD) were prospectively recruited, and 12 consecutive dialysis sessions were monitored. Intradialysis (hour-to-hour) and interdialysis (dialysis day-to-day) vital sign variabilities were assessed via three metrics: the difference between the maximum and minimum values, average real variability (ARV), and residuals. Multivariate logistic regression analysis was used to explore the factors triggering HRH. A total of 91 Chinese MHD patients (60.4% male) aged 58.5 ± 17.2 years were included, with 59 patients using radiocephalic arteriovenous fistulas (RCAVFs) and 32 patients using tunneled cuffed catheters (TCCs) for dialysis. The median dialysis vintage was 26.8 (12.0-44.7) months. Compared with the RCAVF group, the TCC group had significantly greater urea reduction (71.1 ± 9.3% vs. 61.7 ± 10.5%, p < 0.001) and clearance (1.5 (1.2-1.8) vs. 1.1 (1.0-1.4), p < 0.001) rates, higher intradialysis pulse variability and lower intradialysis diastolic blood pressure variability. Some of interdialysis variability indexes in pulse, systolic blood pressure (SBP), and SpO2 were significantly greater in the TCC group than that in the RCAVF group. Age (OR = 0.880, 95% CI = 0.785-0.986, p = 0.028), TCC use (OR = 22.257, 95% CI = 1.190-416.399, p = 0.038), intradialysis SBP-ARV (OR = 2.768, 95% CI = 1.069-7.171, p = 0.036), and blood sodium level (OR = 0.400, 95% CI = 0.192-0.832, p = 0.014) were shown to be independent risk factors for HRH. In conclusion, the use of TCCs has multifaceted effects on intradialysis and interdialysis vital sign variabilities and is independently associated with an increased risk of HRH.